Ecology of aquaculture species and enhancement of stocks: proceedings of the thirtieth U.S. - Japan meeting on aquaculture, Sarasota, FL, Dec. 3-4, 2001

CONTENTS: I. U.S.-Japan Cooperation Open Ocean Aquaculture – A Venue for Cooperative Research Between the United States and Japan.............................................................................. 1 C. Helsley II. Growth, Nutrition and Genetic Diversity Daily Ration of Hatchery-Reared Japanese Flounder Paralichthys olivaceus as an Indicator of Release Place, Time and Fry Quality. In situ Direct Estimation and Possibility of New Methods by Stable Isotope............................ 7 O. Tominaga, T. Seikai, T. Tsusaki, Y. Hondo, N. Murakami, K. Nogami, Y. Tanaka and M. Tanaka Nucleic Acids and Protein Content as a Measure to Evaluate the Nutritional Condition of Japanese Flounder Paralichthys olivaceus Larvae and Juveniles........................................................................................................ 25 W. Gwak Genetic Diversity Within and Between Hatchery Strains of Flounder Paralichthys olivaceus Assessed by Means of Microsatellite and Mitochondrial DNA Sequencing Analysis...................................................................... 43 M. Sekino, M. Hara and N. Taniguchi Tracking Released Japanese Flounder Paralichthys olivaceus by Mitochondrial DNA Sequencing................................................................................ 51 T. Fujii Preliminary Aspects of Genetic Management for Pacific Threadfin Polydactylus sexfilis Stock Enhancement Research in Hawaii........................................ 55 M. Tringali, D. Ziemann and K. Stuck Enhancement of Pacific Threadfin Polydactylus sexfilis in Hawaii: Interactions Between Aquaculture and Fisheries............................................................. 75 D. Ziemann Aquaculture and Genetic Structure in the Japanese Eel Anguilla japonica..................... 87 M. Katoh and M. Kobayashi Comparative Diets and Growth of Two Scombrid Species, Chub Mackerel Scomber japonicus and Japanese Spanish Mackerel Scomberomorus niphonius, in the Central Seto Inland Sea, Japan.................................. 93 J. Shoji, M. Tanaka and Tsutomu Maehara iii Evaluating Stock Enhancement Strategies: A Multi-disciplinary Approach................... 105 T. M. Bert, R.H. McMichael, Jr., R.P. Cody, A. B. Forstchen, W. G. Halstead, K. M. Leber, J. O’Hop, C. L. Neidig, J. M. Ransier, M. D. Tringali, B. L. Winner and F. S. Kennedy III. Physiological and Ecological Applications Predation on Juvenile Chum Salmon Oncorhynchus keta by Fishes and Birds in Rivers and Coastal Oceanic Waters of Japan................................... 127 K. Nagasawa and H. Kawamura Interaction Between Cleaner and Host: The Black Porgy Cleaning Behavior of Juvenile Sharpnose Tigerfish Rhyncopelates Oxyrhynchus in the Seto Inland Sea, Western Japan............................................................................. 139 T. Shigeta, H. Usuki and K. Gushima IV. Case Studies Alaska Salmon Enhancement: A Successful Program for Hatchery and Wild Stocks............................................................................................... 149 W. Heard NMFS Involvement with Stock Enhancement as a Management Tool........................... 171 T. McIlwain Stock Enhancement Research with Anadromous and Marine Fishes in South Carolina...................................................................................... 175 T. I. J. Smith, W. E. Jenkins, M. R. Denson and M. R. Collins Comparison of Some Developmental, Nutritional, Behavioral and Health Factors Relevant to Stocking of Striped Mullet, (Mugilidae), Sheepshead (Sparidae), Common Snook (Centropomidae) and Nassau Groupers (Serranidae)........................... 191 J. W. Tucker Jr. and S. B. Kennedy Participants in the Thirtieth U.S.-Japan Meeting on Aquaculture................. Inside Back Cover iv (PDF has 204 pages.)

A comparison of seafloor habitats and associated benthic fauna in areas open and closed to bottom trawling along the central California Continental Shelf

Executive Summary: A number of studies have shown that mobile, bottom-contact fishing gear (such as otter trawls) can alter seafloor habitats and associated biota. Considerably less is known about the recovery of these resources following such disturbances, though this information is critical for successful management. In part, this paucity of information can be attributed to the lack of access to adequate control sites – areas of the seafloor that are closed to fishing activity. Recent closures along the coast of central California provide an excellent opportunity to track the recovery of historically trawled areas and to compare recovery rates to adjacent areas that continue to be trawled. In June 2006 we initiated a multi-year study of the recovery of seafloor microhabitats and associated benthic fauna inside and outside two new Essential Fish Habitat (EFH) closures within the Cordell Bank and Gulf of the Farallones National Marine Sanctuaries. Study sites inside the EFH closure at Cordell Bank were located in historically active areas of fishing effort, which had not been trawled since 2003. Sites outside the EFH closure in the Gulf of Farallones were located in an area that continues to be actively trawled. All sites were located in unconsolidated sands at equivalent water depths. Video and still photographic data collected via a remotely operated vehicle (ROV) were used to quantify the abundance, richness, and diversity of microhabitats and epifaunal macro-invertebrates at recovering and actively trawled sites, while bottom grabs and conductivity/temperature/depth (CTD) casts were used to quantify infaunal diversity and to characterize local environmental conditions. Analysis of still photos found differences in common seafloor microhabitats between the recovering and actively trawled areas, while analysis of videographic data indicated that biogenic mound and biogenic depression microhabitats were significantly less abundant at trawled sites. Each of these features provides structure with which demersal fishes, across a wide range of size classes, have been observed to associate. Epifaunal macro-invertebrates were sparsely distributed and occurred in low numbers in both treatments. However, their total abundance was significantly different between treatments, which was attributable to lower densities at trawled sites. In addition, the dominant taxa were different between the two sites. Patchily-distributed buried brittle stars dominated the recovering site, and sea whips (Halipteris cf. willemoesi) were most numerous at the trawled site though they occurred in only five of ten transects. Numerical classification (cluster analysis) of the infaunal samples also revealed a clear difference between benthic assemblages in the recovering vs. trawled areas due to differences in the relative abundances of component species. There were no major differences in infaunal species richness, H′ diversity, or J′ evenness between recovering vs. trawled site groups. However, total infaunal abundance showed a significant difference attributable to much lower densities at trawled sites. This pattern was driven largely by the small oweniid polychaete Myriochele gracilis, which was the most abundant species in the overall study region though significantly less abundant at trawled sites. Other taxa that were significantly less abundant at trawled sites included the polychaete M. olgae and the polychaete family Terebellidae. In contrast, the thyasirid bivalve Axinopsida serricata and the polychaetes Spiophanes spp. (mostly S. duplex), Prionospio spp., and Scoloplos armiger all had significantly to near significantly higher abundances at trawled sites. As a result of such contrasting species patterns, there also was a significant difference in the overall dominance structure of infaunal assemblages between the two treatments. It is suggested that the observed biological patterns were the result of trawling impacts and varying levels of recovery due to the difference in trawling status between the two areas. The EFH closure was established in June 2006, within a month of when sampling was conducted for the present study, however, the stations within this closure area are at sites that actually have experienced little trawling since 2003, based on National Marine Fishery Service trawl records. Thus, the three-year period would be sufficient time for some post-trawling changes to have occurred. Other results from this study (e.g., similarly moderate numbers of infaunal species in both areas that are lower than values recorded elsewhere in comparable habitats along the California continental shelf) also indicate that recovery within the closure area is not yet complete. Additional sampling is needed to evaluate subsequent recovery trends and persistence of effects. Furthermore, to date, the study has been limited to unconsolidated substrates. Ultimately, the goal of this project is to characterize the recovery trajectories of a wide spectrum of seafloor habitats and communities and to link that recovery to the dynamics of exploited marine fishes. (PDF has 48 pages.)

Gillraker analysis and speciation in the thread herring genus Opisthonema

ENGLISH: The thread herrings, genus Opisthonema, family Clupeidae, are marine fishes that occur only in tropical and subtropical waters off both coasts of the American continent. The genus is composed of one Atlantic species and four Pacific species. Opisthonema oglinum (LeSueur), the single species in the western Atlantic Ocean, occurs from Brazil to Massachusetts, at Bermuda, and throughout the West Indies. Opisthonema libertate (Günther), Opisthonema bulleri (Regan), and Opisthonema medirastre, new species, are Pacific coastal species which occur together from Peru to Mexico (with at least one occasionally in California). Opisthonema berlangai, new species, is confined to the vicinity of the Galapagos Islands. SPANISH: Los arenques de hebra, género Opisthonema, familia Clupeidae, son peces marinos que se encuentran sólo en aguas tropicales y subtropicales cerca a ambas costas del continente americano. El género está compuesto de una especie en el Atlántico y de cuatro en el Pacifico. Opisthonema oglinum (LeSueur), única especie que medra el Océano Atlántico occidental, se encuentra del Brasil a Massachusetts, en Bermuda, y las Indias Occidentales. Opisthonema libertate (Günther), Opisthonema bulleri (Regan), y Opisthonema medirastre, especie nueva, son especies costaneras del Pacífico que aparecen entremezcladas del Perú a México (ocasionalmente por lo menos una de ellas en California). Opisthonema berlangai, especie nueva, está confinada a la vecindad de las Islas Galápagos.

Ecology of Juvenile Walleye Pollock, Theragra chalcogramma: Papers from the workshop "The Importance of Prerecruit Walleye Pollock to the Bering Sea and North Pacific Ecosystems" Seattle, Washington, 28-30 October 1993

The Alaska Fisheries Science Center (AFSC), National Marine Fisheries Service (NMFS), hosted an international workshop, 'The Importance of Prerecruit Walleye Pollock to the Bering Sea and North Pacific Ecosystems," from 28 to 30 October 1993. This workshop was held in conjunction with the annual International North Pacific Marine Science Organization (PICES) meeting held in Seattle. Nearly 100 representatives from government agencies, universities, and the fishing industry in Canada, Japan, the People's Republic of China, Russia, and the United States took part in the workshop to review and discuss current knowledge on juvenile pollock from the postlarval period to the time they recruit to the fisheries. In addition to its importance to humans as a major commercial species, pollock also serves as a major forage species for many marine fishes, birds, and mammals in the North Pacific region. (PDF file contains 236 pages.)

Huevos y larvas de tres especies de peces marinos, Anchoa marinii, Brevoortia aurea y Prionotus nudigula

The description of the embryonic and early larval stages of three species of marine fishes: the anhovy, Anchoa marinii, the menhaden Brevoortia aurea and the gurnard, Prionotus nudigula is given. The time required from the fertilization to the hatching for each species was calculated. The eggs of these three species are found in the plankton collected in the zone situated in the vicinity of Mar del Plata. The eggs are only found in the plancton which was close to the shore. The anchoa marinii eggs are found in the sea from the middle of December at a water temperature of approximately 16,0°C to the end of April. Their greatest concentration takes place in January at 20,0-21,0°C. The eggs of Brevoortia aurea are found in the plakton from the beginning of October at a water temperature of approximately 10,0°C to the middle of December. Their greatest concentration takes place in November at 13,0-15,0°C. Only once were the menhaden's eggs can be found in the sea from the middle of November at the water temperature of aproximately 13,0° to the end of April. Their greatest concentration takes place in January and February at 20,0-21,0°C.

A review of IATTC research on the early life history and reproductive biology of scombrids conducted at the Achotines Laboratory from 1985 to 2005

English: For nearly a century, fisheries scientists have studied marine fish stocks in an effort to understand how the abundances of fish populations are determined. During the early lives of marine fishes, survival is variable, and the numbers of individuals surviving to transitional stages or recruitment are difficult to predict. The egg, larval, and juvenile stages of marine fishes are characterized by high rates of mortality and growth. Most marine fishes, particularly pelagic species, are highly fecund, produce small eggs and larvae, and feed and grow in complex aquatic ecosystems. The identification of environmental or biological factors that are most important in controlling survival during the early life stages of marine fishes is a potentially powerful tool in stock assessment. Because vital rates (mortality and growth) during the early life stages of marine fishes are high and variable, small changes in those rates can have profound effects on the properties of survivors and recruitment potential (Houde 1989). Understanding and predicting the factors that most strongly influence pre-recruit survival are key goals of fisheries research programs. Spanish: Desde hace casi un siglo, los científicos pesqueros han estudiado las poblaciones de peces marinos en un intento por entender cómo se determina la abundancia de las mismas. Durante la vida temprana de los peces marinos, la supervivencia es variable, y el número de individuos que sobrevive hasta las etapas transicionales o el reclutamiento es difícil de predecir. Las etapas de huevo, larval, y juvenil de los peces marinos son caracterizadas por tasas altas de mortalidad y crecimiento. La mayoría de los peces marinos, particularmente las especies pelágicas, son muy fecundos, producen huevos y larvas pequeños, y se alimentan y crecen en ecosistemas acuáticos complejos. La identificación los factores ambientales o biológicos más importantes en el control de la supervivencia durante las etapas tempranas de vida de los peces marinos es una herramienta potencialmente potente en la evaluación de las poblaciones. Ya que las tasas vitales (mortalidad y crecimiento) durante las etapas tempranas de vida de los peces marinos son altas y variables, cambios pequeños en esas tasas pueden ejercer efectos importantes sobre las propiedades de los supervivientes y el potencial de reclutamiento (Houde 1989). Comprender y predecir los factores que más afectan la supervivencia antes del reclutamiento son objetivos clave de los programas de investigación pesquera.

Evolutionary associations between sand seatrout (Cynoscion arenarius) and silver seatrout (C. nothus) inferred from morphological characters, mitochondrial DNA, and microsatellite markers

The evolutionary associations between closely related fish species, both contemporary and historical, are frequently assessed by using molecular markers, such as microsatellites. Here, the presence and variability of microsatellite loci in two closely related species of marine fishes, sand seatrout (Cynoscion arenarius) and silver seatrout (C. nothus), are explored by using heterologous primers from red drum (Sciaenops ocellatus). Data from these loci are used in conjunction with morphological characters and mitochondrial DNA haplotypes to explore the extent of genetic exchange between species offshore of Galveston Bay, TX. Despite seasonal overlap in distribution, low genetic divergence at microsatellite loci, and similar life history parameters of C. arenarius and C. nothus, all three data sets indicated that hybridization between these species does not occur or occurs only rarely and that historical admixture in Galveston Bay after divergence between these species was unlikely. These results shed light upon the evolutionary history of these fishes and highlight the genetic properties of each species that are influenced by their life history and ecology.

Reconstruction of original body size and estimation of allometric relationships for the longfin inshore squid (Loligo pealeii) and northern shortfin squid (Illex illecebrosus)

Quantification of predator-prey body size relationships is essential to understanding trophic dynamics in marine ecosystems. Prey lengths recovered from predator stomachs help determine the sizes of prey most influential in supporting predator growth and to ascertain size-specific effects of natural mortality on prey populations (Bax, 1998; Claessen et al., 2002). Estimating prey size from stomach content analyses is often hindered because of the degradation of tissue and bone by digestion. Furthermore, reconstruction of original prey size from digested remains requires species-specific reference materials and techniques. A number of diagnostic guides for freshwater (Hansel et al., 1988) and marine (Watt et al., 1997; Granadeiro and Silva, 2000) prey species exist; however they are limited to specific geographic regions (Smale et al., 1995; Gosztonyi et al., 2007). Predictive equations for reconstructing original prey size from diagnostic bones in marine fishes have been developed in several studies of piscivorous fishes of the Northwest Atlantic Ocean (Scharf et al., 1998; Wood, 2005). Conversely, morphometric relationships for cephalopods in this region are scarce despite their importance to a wide range of predators, such as finfish (Bowman et al., 2000 ; Staudinger, 2006), elasmobranchs (Kohler, 1987), and marine mammals (Gannon et al., 1997; Williams, 1999).

Population structure of king mackerel (Scomberomorus cavalla) around peninsular Florida, as revealed by microsatellite DNA

A total of 1006 king mackerel (Scomberomorus cavalla) representing 20 discrete samples collected between 1996 and 1998 along the east (Atlantic) and west (Gulf) coasts of Florida and the Florida Keys were assayed for allelic variation at seven nuclear-encoded microsatellites. No significant deviations from Hardy-Weinberg equilibrium expectations were found for six of the microsatellites, and genotypes at all microsatellites were independent. Allele distributions at each microsatellite were independent of sex and age of individuals. Homogeneity tests of spatial distributions of alleles at the microsatellites revealed two weakly divergent “genetic” subpopulations or stocks of king mackerel in Florida waters—one along the Atlantic coast and one along the Gulf coast. Homogeneity tests of allele distributions when samples were pooled along seasonal (temporal) boundaries, consistent with the temporal boundaries used currently for stock assessment and allocation of the king mackerel resource, were nonsignificant. The degree of genetic divergence between the two “genetic” stocks was small: on average, only 0.19% of the total genetic variance across all samples assayed occurred between the two regions. Cluster analysis, assignment tests, and spatial autocorrelation analysis did not generate patterns that were consistent with either geographic or spatial-temporal boundaries. King mackerel sampled from the Florida Keys could not be assigned unequivocally to either “genetic” stock. The genetic data were not consistent with current spatial-temporal boundaries employed in stock assessment and allocation of the king mackerel resource. The genetic differences between king mackerel in the Atlantic versus those in the Gulf most likely stem from reduced gene flow (migration) between the Atlantic and Gulf in relation to gene flow (migration) along the Atlantic and Gulf coasts of peninsular Florida. This difference is consistent with findings for other marine fishes where data indicate that the southern Florida peninsula serves (or has served) as a biogeographic boundary.

Bycatch of billfishes by the European tuna purse-seine fishery in the Atlantic Ocean

Billfishes are a component of offshore ecosystems; thus it is important to quantify the impact of the tuna fishery on these species in the world’s ocean. The aim of this study was to assess the bycatch of billfishes generated by the tropical tuna purse-seine fishery in the eastern Atlantic Ocean. Information on bycatch was collected by observers at sea during the European Union Bigeye Program. With a total of 62 observers’ trips, conducted on Spanish and French vessels between June 1997 and May 1999, this project is the biggest observer program ever carried out in the European tuna purse-seine fishery. This study showed that billfish bycatch by the purse seiners is very low (less than 0.021% of the total tuna catches and less than 10% of the total billfish catches currently reported). A Monte Carlo simulation was performed to account for some uncertainties in the fishing strategies of purse seiners operating in this ocean. One of the findings of this study indicated that the temporary moratorium on fishing with FADs (fish aggregating devices), adopted by the European purse-seine fishery in the eastern Atlantic Ocean, produced a decrease in incidental catches of marlins from 600–700 metric tons (t) to less than 300 t. In contrast, this trend was reversed for sailfishes, for which the bycatch increased from 25 t to 45 t. The difficulty of defining indices that express the conservation status in marine fishes and that gauge key ecosystem parameters and the need to promote an ecosystem approach for large-pelagic-resource management which takes into account biologic and socioeconomic criteria are briefly discussed.

Isolation of bile from fish and identification by thin layer chromatography

The paper deals with the collection of gall bladders, isolation of bile and identification of the constituents of the bile salts from different fishes. The yield of bile contents from fresh water fishes rohu, mrigal and catla was compared with that from marine fishes seer, tuna, shark and sardine. Considerable variation in yield was showed between marine and fresh water fish as well as between the species in both groups. It ranged from 0.04 to 0.06% of the body weight of fish in calla, mrigal and rohu. The bile constituents from rohu and mrigal were analysed by thin layer chromatography. The result showed that bile of rohu and mrigal contains mainly taurine derivative of lithocholic acid.

Influence of dietary proteins on cholesterol levels in albino rats

Six groups of albino rats were fed identical diets, differing in their protein sources for sixty days. The protein sources used were fat free casein, defatted groundnut cake, and defatted fish powders from three species of fishes, namely, the fresh water fish Labeo rohita, the marine fishes, Rastrelliger kanagurta and Otolithus argenteus and marine prawns, Parapenaeopsis stylifera. After sixty days, the levels of cholesterol in the serum, liver and heart of the rats were estimated. The casein group recorded the highest level of cholesterol. Compared to casein, the groundnut protein was distinctly hypocholesterolemic. The fish proteins had a still greater cholesterol lowering ability. Of the proteins used in this study, the proteins of prawns had the maximum hypocholesterolemic effect. An attempt is made to correlate the behaviour of the proteins in this respect, with their respective amino acid compositions.

Study of rainbow trout neoplasia in the region of Haraz

This study was carried out in order to determine neoplasia presence in rainbow trout in Haraz area. Neoplasia in various species of fishes including freshwater, brackish water and marine fishes was reported in other countries. But up to now there is no documented report of rainbow trout neoplasia in Iran. The study was performed in 20 farms of Haraz area during 2004-2005. All fishes of each farm firstly were observed in order to any abnormal mass which is suspicious to neoplasia. Besides in order to observation of external and internal organs, 20 fishes was sampled randomly from each farm and were examined clinically and necroptically. Any suspicious lesions were sent to pathology laboratory in 10% formalin followed by taking pictures of the lesions. Then histopathological evaluations were performed. From 400 fishes, 3 neoplastic cases including hepatocellular carcinoma along with bile duct papilloma, hepatocellular carcinoma and bile duct adenocarcinoma and intestine adenocarcinoma were found which all were in brooder fish.

Distribution and sighting frequency of reef fishes in the Florida Keys National Marine Sanctuary

This study analyzed species richness, distribution, and sighting frequency of selected reef fishes to describe species assemblage composition, abundance, and spatial distribution patterns among sites and regions (Upper Keys, Middle Keys, Lower Keys, and Dry Tortugas) within the Florida Keys National Marine Sanctuary (FKNMS) barrier reef ecosystem. Data were obtained from the Reef Environmental Education Foundation (REEF) Fish Survey Project, a volunteer fish-monitoring program. A total of 4,324 visual fish surveys conducted at 112 sites throughout the FKNMS were used in these analyses. The data set contained sighting information on 341 fish species comprising 68 families. Species richness was generally highest in the Upper Keys sites (maximum was 220 species at Molasses Reef) and lowest in the Dry Tortugas sites. Encounter rates differed among regions, with the Dry Tortugas having the highest rate, potentially a result of differences in the evenness in fishes and the lower diversity of habitat types in the Dry Tortugas region. Geographic coverage maps were developed for 29 frequently observed species. Fourteen of these species showed significant regional variation in mean sighting frequency (%SF). Six species had significantly lower mean %SF and eight species had significantly higher mean %SF in the Dry Tortugas compared with other regions. Hierarchical clustering based on species composition (presence-absence) and species % SF revealed interesting patterns of similarities among sites that varied across spatial scales. Results presented here indicate that phenomena affecting reef fish composition in the FKNMS operate at multiple spatial scales, including a biogeographic scale that defines the character of the region as a whole, a reef scale (~50-100 km) that include meso-scale physical oceanographic processes and regional variation in reef structure and associated reef habitats, and a local scale that includes level of protection, cross-shelf location and a suite of physical characteristics of a given reef. It is likely that at both regional and local scales, species habitat requirements strongly influence the patterns revealed in this study, and are particularly limiting for species that are less frequently observed in the Dry Tortugas. The results of this report serve as a benchmark for the current status of the reef fishes in the FKNMS. In addition, these data provide the basis for analyses on reserve effects and the biogeographic coupling of benthic habitats and fish assemblages that are currently underway. (PDF contains 61 pages.)

Deepwater reef fishes and multibeam bathymetry of the Tortugas South Ecological Reserve, Florida Keys National Marine Sanctuary, Florida

The Tortugas South Ecological Reserve, located along the margin of the southwest Florida carbonate platform, is part of the largest no-take marine reserve in the U.S. Established in July 2001, the reserve is approximately 206 km2 in area, and ranges in depths from 30 m at Riley’s Hump to over 600 m at the southern edge of the reserve. Geological and biological information for the Tortugas South Reserve is lacking, and critical for management of the area. Bathymetric surveys were conducted with a Simrad EM 3000 multibeam echosounder at Riley’s Hump and Miller’s Ledge, located in the northern and central part of the reserve. Resulting data were used to produce basemaps to obtain geological ground truth and visual surveys of biological communities, including reef fishes. Visual surveys were conducted using SCUBA and the Phantom S2 Remotely Operated Vehicle (ROV) at Riley’s Hump. Visual surveys were conducted using the ROV and the Deepworker 2000 research submersible along Miller’s Ledge, within and outside of the reserve. A total of 108 fishes were recorded during SCUBA, ROV, and submersible observations. Replicate survey transects resulted in over 50 fishes documented at Miller’s Ledge, and eight of the top ten most abundant species were planktivores. Many species of groupers, including scamp (Mycteroperca phenax), red grouper (Epinephelus morio), snowy grouper (E. niveatus), speckled hind (E. drummondhayi), and Warsaw grouper (E. nigritus), are present in the sanctuary. Numerous aggregations of scamp and a bicolor phase of the Warsaw grouper were observed, indicating the importance of Miller’s Ledge as a potential spawning location for both commercially important and rare deep reef species, and as a potential source of larval recruits for the Florida Keys and other deep reef ecosystems of Florida