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Notice: ©2005 American Fisheries Society. This article may be cited as: Tucker, J. W., Jr. (2005). culture. In A. M. Kelly and J. Silverstein (eds.), in the 21st century: Proceedings of an American Fisheries Society Symposium special symposium on aquaculture in the 21st century, 22 August 2001, Phoenix, Arizona. (pp. 307-338). Bethesda, MD: American Fisheries Society.

American Fisheries Society Symposium 46:307-338. 2005 © 2005 by the American Fisheries Society

Grouper Culture

JOHNW. TUCKER, JR. 1 Culture and Biology Department, Indian River Institute, Inc. 316 13th Avenue, Vern Beach , 32962, USA

Introduction and early to mid-stage larvae cannot swim very fast or far; therefore, both mostly drift with the are classified in 14 genera of the sub­ current. Larvae of most spend at least family Epinephelinae, which comprises at least their first few weeks drifting with the oceanic half the approximately 449 species in the family plankton. As they become juveniles, groupers . Throughout most warm and temper­ settle to the bottom, usually in shallow water, ate marine regions, serranids are highly valued where they can find hiding places. Then, until for food, and both small and large species are several centimeters long, they hide almost con­ kept in aquariums. Maximum size ranges from stantly. Their boldness increases with size, and about 12 em total length (TL) for the western At­ they move to deeper water.but mostspecies con­ lantic Setranus species and the Pacific creolefish tinue to stay near small caves for security. Wild Pantnthias countus to more than 4 m TL (~440 kg, grouper larvae at first eat copepods and other mature at 129 cm) for the king grouper, giant small zooplankton, then larger like grouper, groper, or brindlebass amphipods and mysid shrimp. Wild juveniles lanceolatus. and adults eat mainly fish, , shrimp, mantis Ages (months) at which some western At­ shrimp, , and mollusks. lantic serranids reach 454 g in nature are Goliath Most groupers that have been studied will grouper E.itajarn -12, black grouperMycteropcrcn mature within 2 to 6 years (Tucker 1998). Many bonaci 17, gag grouper M. microl-epis 18, red serranids are protogynous hermaphrodites (i.e., grouper E. moria 27, and black Cenirop­ most individuals mature first as females and ristis striata, subfamily 51. Nassau some of them become males later). Some of those groupers E. striatus reach 50 em (-2.5 kg) in 4--5 species, as a rule, change from female to male as years and Goliath groupers reach 50 cm (-2.7 kg) they grow older, while others might change only in -2 years (Bullock et aI. ] 992; Sadovy and if there is a shortage of males. In nature, Nassau Eklund 1999). Maximum reported sizes are 1'J'J groupers typically in large distinct aggre­ em TL and -27 kg for Nassau groupers (Sadovy gations (hundreds to thousands of fish) with a and Eklund 1999), 152 cm for black groupers sex ratio near 1:1. Gags spawn in harems, fre­ (Crabtree and Bullock 1998), 129 em for gags quently within aggregations, with a sex ratio of­ (Collins et al. 1998), -250 ern and 320 kg for ten near 1 male:10 females. For both species, in­ Goliath groupers (Sadovy and Eklund 1999). dividual spawning events usually involve small Nassau groupers live for at least 29 years, black numbers of fish (e.g., 2-5). Small serranids often groupers 33 years, gags 21 to 22 years, red grou­ spawn in pairs without aggregating. A few of the pers 25 years, and Goliath groupers 37 years small serranids are simultaneous hermaphro­ (Crabtree and Bullock]998). dites (male and female at the same time), but self­ Juveniles and adults of some grouper spe­ fertilization seems to be rare. cies live in coastal waters and estuaries, but oth­ ers prefer the cleaner waters of offshore reefs. Historical Overview Eggs and larvae are planktonic-the single non­ adhesive eggs are buoyant at normal salinities More than 20 grouper species have been raised commercially, mostly by growing out captured I E-mail: [email protected]

307 308 ruCKER

wild juveniles in the Southeast Asian region. eptesaltivelis (mature at 39 em, maximum 70 em) The Food and Agriculture Organization (2004) have become more important in the last few reported that 22,808 metric tons (mt) of grou­ years. Minor species are blaekspotted grouper pers and sea basses worth US$1l9,418,OOO were E. amblvcephnlu«, sixbar grouper E. eexfasciaius grown out in Taiwan (12,367), Indonesia (maximum 28 em), bluespotted grouper E. (7,057), Thailand (-1,442), Malaysia (1,399), trimaculatus (mature at 29 em, maximum49 em), Hong Kong (325), Philippines (115), Singapore E. quoymilis (mature at 24 em, (64), Korean Republic (~33), Brazil (3), and maximum 40 em), longtooth grouper E. bruneus Kuwait (-3) during 2002. Sadovy (2000, 2001) (mature at 54 em, maximum 100 em), leopard estimated that for the late 1990s, about 60 mil­ coraltrout Plectrop-omus leopardus (females ma­ lion juveniles grown out per year resulted in ture at 36 em, males at 42 em, maximum 120 23,000 mt of live table-size groupers sold in em), barred-cheek coraltrout P. ntaculnius (ma­ Southeast Asia; however, at least hundreds of ture at 54 ern, maximum 100 em), squaretail millions of wild grouper juveniles were caught eoraltrout P. areouiius (mature at 41 em, maxi­ and sold each year, so mortality from capture mum 73 ern), E. merra (ma­ and transportation had to be very high and ture at 19 em, maximum 31 em), camouflage wastefuL Despite improvements in recent grouper E. polyphekndioJ1 (mature at 58 em, years, until 2000/ only 20/000-80/000 juveniles maximum 109 em), and E. were produced annually by individual hatch­ unroina (mature at 61 em, maximum 107 em). eries in the region, not counting those in Tai­ Some reports on greasy groupers (actually rare wan or Japan (Sadovy 2000, 2001). About two­ in Southeast Asia) have been based on misi­ thirds of the groupers grown out in Taiwan dentification of orangespotted or Malabar were from hatcheries. During the 1990s, typi­ groupers; unless that has been confirmed for cal annual hatchery production of groupers (1­ references in this chapter, the name greasy 3 em) in Taiwan was about 20 million, mostly grouper is still used. For farming in the south­ orangespotted E. coioides and Malabar groupers eastern United States and Caribbean region, E. ntalabaricus. In 1998, about half of the grou­ Nassau groupers (Figure I), gags, black grou­ pers farmed there had been raised from wild pers, and Goliath groupers seem to have good juveniles (Cesar et al. 2000). In 2000, Taiwan­ potential. Dusky E. ntnrginatus and white grou­ ese hatcheries produced 300,000 brown­ pers E. aenaeus have been investigated in the marbled groupers and 2 million king groupers Med i terranean. (Chan 2001). King grouper culture has just Wild groupers tend to be very easy to been developed in the last several years; 5,000 catch, especially species that form large aggre­ 7-cm juveniles were produced in 1996 and gations for spawning like the 70,000 (-2% survival) in 1998 (Su 2002). (Dahlgren 1999). This grouper historically was The main farmed Asian species (Sadovy the most important through most of the Carib- 2001; total lengths mostly from Lau and Li 2000) have been orangespotted or estuary grouper (mature at 49 em, maximum ]20 cm) and (mature at 45 em, maxi­ mum 234 em). Other major species are dusky­ tail grouper E. bleekeri (mature at 42 em, maxi­ mum 76 em), redspotted or Hong Kong grou­ per E. akaara (mature at 30 em, maximum 5] em), E. atocara (mature at 34 em, maximum 60 em), squaretail grouper E. areolatus (mature at 23 em, maximum 47 em), and king grouper. Because of increased hatch­ ery production, brownmarbled grouper E. [uscoguiiaius (mature at 50 em, maximum 120 em), kelp grouper E. ntoara (maximum 60 em), Figure 1.-Nassau grouper Epinephclus striatus at and polka-dot or Croniil- Grand Cayman. (by John Tucker) GROUPER CULTURE 309 bean islands and coastal reefs, but has been of such catches will be. If heavy losses natu­ overfished to commercial extinction in many rally occur after the collected stage, then pro­ areas. Both commercial and recreational har­ tecting a number of the fish on farms actually vest now are prohibited in the United States. could increase the total numbers available for The gag and , because of their harvest. At least, if collection of maderate num­ abundance and availability to the fishery, are bers is small compared to those lost to natural economically the most important species in the mortality, significant losses of wild stocks United States. They and many other grouper would not result. Therefore, traditional grou­ species throughout the world have been over­ per farming (i.e., wild groupers grown in ponds fished, leading to reduced numbers and re­ or cages) in Asia has been encouraged by some duced average size (Beets and Friedlander environmental organizations. But at the same 1998; Huntsman et al, 1999; Chiappone et al. time, grouper hatchery tec1mology needs to be 2000; Coleman et al. 2000; Morris et al. 2000). perfected because the wild supply does flue­ One possible effect is an unbalanced sex ratio tua te and there is no guarantee thatit will con­ in protogynous species. If the males are larger, tinue indefinitely. Survival of collected juve­ they tend to be caught first. If males become niles also must be improved. too scarce, fewer offspring might be produced. In 1998, there were about 600 grouper This mechanism led to collapse of red porgy hatcheries and farms in Taiwan, producing Pagrus pagrlls populations, even though fe­ 5,000-7,000 mt/year, 90% of which were rnales were abundant. Also irnportant is the fact orangespott-ed and Malabar groupers (Cesar that most larger grouper species take several et al. 2000). The large number of participants, years to reach maturity and begin spawning. strong government support, intense research Usually, fishermen begin catching them before and specialization, availability of large areas thal point. For those species, can of coastal land, and mass production of eggs reduce the spawning population even though are factors contributing to the Taiwanese suc­ immature fish are abundant. For a number of cess. reasons, networks of marine reserves (no catch­ During 1999-2000, a commercial hatchery ing of groupers, especially in spawning areas) in Penang, Malaysia was producing about and habitat protection appear to be the best 900,000 orangespotted groupers and 360,000 ways to protect wild populations. Once grou­ brownmarbled groupers a year to about 6 em per production reaches a high enough level, (Sadovy 2000). The government hatchery at aquaculture also will help protect wild stocks Songkhla, Thailand was producing 100,000­ by reducing fishing pressure. 170,000 orangespotted and malabar groupers In some areas where wild juveniles are a year to 7.5 em. In 1999, a government hatch­ caught for grow out, supplies (catch per unit ery in Fujian Province, PR China, produced of effort) have decreased in recent years 30,000 redspotted groupers. In 2003, a govern­ (Johannes and Ogburn 1998), especially near ment hatchery in Japan raised more than Hong Kong, Taiwan, and China (Sadovy 2000, 400,000 juvenile kelp groupers. 2001). Possible factors include not only over­ There are more than 2,000 hatch­ fishing of juveniles, but also overfishing of eries in Indonesia, with about 10% near adults, habitat loss, toxic pollutants, and popu­ Gondol. In the last few years, many of those lation cycle variations. In other areas, supplies near Gondol have started raising groupers, of juveniles have been relatively constant, de­ too, based on results from the Gondol Research spite 20 or more years of collecting. As grou­ Institute for Mariculture (GRIM). The GRlM pers grow in nature, their numbers naturally produced 2,000 early juvenile polka-dot grou­ decrease. Harvesting small juveniles has less pers in 1998, then 54,000 (mean 25 mm TL) in impact than harvesting a similar number of n,',TO 10-m 3 tanks during 1999, with survival of larger fish and is less likely to be a determin­ 45% and 54% from hatching to 43 dah (Sial' et ing factor of population size. The more that a a1.2002; Sugama et al. 2002a, 2002b). Fertiliza­ given grouper population is limited by pre­ tion and ha tching rates each usually were 70'Yo. dation or lack of habitat for fish larger than Expected routine hatchery survival for polka­ those caught for grow out, the less the impact dot and brownmarbled groupers was 5% from 310 ruCKER

ha tching to 60 da h. After reaching 3 to 5 em in dally deep if horizontal swimming space is 50 to 60 d, juveniles are grown to about 10 cm sufficient, unless the broodfish are very large. in the nursery phase. In 2001, about 62 com­ Larvae have been reared mainly in fiberglass mercial hatcheries were able to produce 1.05 or concrete tanks (Figure 2) holding 1-60rrr' and million polka-dot grouper juveniles and 1.1 sometimes in concrete ponds of up to 500 m': million brownmarbled grouper juveniles. while a minimum of 3.5 m 3 is good, 7 m ' or larger is better, Tanks and cages (Figure 2 of Chapter 13, this volume) up to at least 75 m? Regulations and Permitting and larger ponds (Figure 3) have been used for grow out. Good environmental control is espe­ In regions where wild juveniles are caught for cially important for the broodstock and larvae, grow out, there is evidence and concern tha t but is not as critical for the other stages. wild stocks are being depleted (Sadovy 2001). In some areas where groupers are raised in cages, environmental pollution and self-pollu­ Broodstock tion have occurred. Caged groupers have been Broodstock can be captured or reared. Most known to spread diseases to wild fish and other groupers studied have quickly adapted to cap­ farmed fish. Shipping of infected wild or reared tivity. Adults usually are captured by traps or juveniles to distant grow-out areas has trans­ hook and line. Depending on species and cap­ ferred diseases, especially viral and parasitic ture depth, the gas bladder might expand too ones (Bondad-Reantaso et al. 2002). There are strong popular movements to have national and international regulations and agreements en­ acted to reduce and eventually prevent such problems. In the United States, existing regulations would not allow wild juveniles to be caught. Pollution and disease transmission would be strictly controlled. Harvest of Nassau groupers is prohibited in the United States and and restricted in much of the Caribbean. Har­ vest of Goliath groupers is prohibited in Florida and U.S. Exclusive Economic Zone waters and restricted in some other states. Capture and sale of other groupers are regulated to varying de­ Figure 2. -Concrete larval rearing tanks in Taiwan, grees. Raising and selling of protected species usually shaded when larvae are stocked. (by John would require special federal and/or state per­ Tucker) mitting in addition to the standard environmen­ tal and business permits.

Culture Methods

Facilities Salt water is needed for broodfish, eggs, and larvae. Juveniles and adults of many species can live in brackish but not freshwater. Groupers have spawned voluntarily in fiberglass, con­ crete, or plastic tanks with volumes of 1-21,200 m', concrete ponds of 1.8-860 m'', and cages of 26-75 m" (Tucker 1994, 1998). Even for reef spe­ Figure 3. -A pond used for growing out groupers cies, spawning tanks do not have to be espe- in Taiwan. (by John Tucker) GROUPERCULTIJRE 311

much for the fish to recover on its own (e.g., it conditions of ambient temperature and partial floats helplessly upside down). When defla­ or total natural light (Tucker 1994, 1998). Day tion is necessary, a hypodermic syringe with a length seems to be a less important stimulus 20-gauge needle attached and plunger removed for spawning than temperature. More than 30 can be used. While the fish is gently held up­ serranid species have spawned voluntarily in side down below the water surface, the needle captivity. Eggs are collected in automatic strain­ is gently pushed through the skin into the body ers or with soft, fine dipnets. It should be kept cavity, avoiding internal organs. The fish is de­ in mind that some species spawn near certain flated only enough for it to stay on the bottom moon phases and others spawn any day of the of the tank without struggling to stay down. If lunar month. With good timing and luck, grou­ . the gas bladder is expanded so much that the pers can be caught just before spawning and fish's insides (esophagus and maybe stomach) held in tanks or cages for up to a few days until protrude into the mouth, some fishermen punc­ they ovulate naturally. The eggs are stripped, ture the insides and push them back in; those or rarely, the fish are left in the tank for volun­ fish are less likely to survive, but a surprising tary or accidental fertilization to occur, if the number do. males are running ripe. Juvenile and adult Nassau groupers (and Forty female and nine male orangespotted many other groupers) are among the hardiest of groupers (protogynous) in a concrete tank fish. Their skin is so thick it has been made into spawned almost continuously for 50 d during leather clothing. In one case, a ripe female that April to June 1976 in Kuwait (Hussain and sat dry on a boat deck in the sun for 2 h recov­ Higuchi 1980). In the Philippines, one female ered and was induced to ovulate 2 d later. De­ orangespotted grouper with two males in a 48­ spite those examples, broodfish always should m" cage spawned 5-10 times a month for 4 be handled as gently (with soft nets, plastic bags, months, July-October; six females and four or hands, not towels) and infrequently as pos­ males in a 50-m J concrete tank spawned 5-17 sible, kept in well-oxygenated water, and fed times a monthfrom July to June {skipping May), well if kept for more than a few days. beginning near the last quarter moon (Toledo With protogynous hermaphrodites, sex con­ et al. 1990, 1992, 1993; Quinitio 1997). Maturity trol (Yamazaki 1983) sometimes is considered is reached at 25-30 em standard length (SL;Cesar necessary to ensure an optimal male:female ra­ et a1. 2000). tio for maximizing egg production. If males are During December 1989 to October 1990 in difficul t to collect or do not occur in reared Singapore, 10 female and 10 male brown­ broodstock when the females have become ma­ marbled groupers (protogynous) in a 75-rnJ cage ture, sex-reversal of females with dietary or in­ spawned 2-5 times during each of nine periods jected methyltestosterone could provide the of 2-6 d, usually starting between the last quar­ needed males (e.g., kelp grouper, Tsukashima ter moon and new moon (Lim et a1. 1990). and Yoshida 1984; blue-spotted grouper, Kuo et Eight female leopard coraltrout (proto­ al. 1988; greasy grouper, Chao and Lim 1991; gynous) in a pond produced eggs 110 times orangespotted grouper, Tan-Fermin et a1. 1994, during May to October 1989 in Taiwan (Chen et Quinitio 1997; dusky grouper, Clamuzina et a1. a1. 1991). Maturity is reached at -20 em FL 1998a). When too many females could become (Samoilys and Squire 1994). males, suppression of sex-reversal has been ac­ At 26-28°C, 24 female and 3 male polka­ complished by administering female hormones dot groupers (protogynous) in a 100-m J cylin­ like estradiol or by maintaining enough large drical concrete tank spawned 4-10 times a males in the tank (e.g., redspotted grouper, month, mostly near the new moon, during May­ Maruyama et al. 1994). With simultaneous her­ October in Indonesia, but they can spawn year­ maphrodites (e.g., , Tucker and round (Sugama et al. 2000a, 2000b). Females Woodward 1995),mature fish of similar size usu­ usually weigh 1-3 kg and males more than 2 kg ally can be paired. (Sugama et al. 2001). Voluntary spawning of captive groupers A pairof belted sandfish subligarius, has occurred mostly with well-fed uncrowded (subfamily Serraninae, simultaneous hermaph­ fish during the natural spawning season under rodites) in a 28-L aquarium spawned 79 times 312 nJCKER

in 81 d during February to May 1991 in Florida those fish were caught during thespawning sea­ (Tucker and Woodward 1995). Their offspring son and had adapted very quickly to captivity; were mature and began spawning at 16 months jf the interval between capture and attainment (70 mm TL). of spawning conditions was shorter, the fish In a very interesting project, hybrid grou­ probably would have spawned sooner than 15 pers were produced in a research hatchery on months). Nassau groupers reared from eggs the in Saudi Arabia by voluntary were mature by 5-6 years, but because tempera­ spawning between 12- to 14-kg female brown­ ture in their raceway could not be controlled and marbled groupers and 4- to 5-kg male camou­ fluctuated widely, they did not spawn immedi­ flage groupers (protogynous)(James et al. ately; in April 1998, temperature was more 1998a, 1999). Each species was held separately stable and the 8-year-old groupers spawned. at 1-2kg/m3 in 25-m 3 fiberglass tanks 2 m deep, Nassau groupers can be conditioned to spawn with 500%/d water exchange. They were fed during any month, mainly by temperature cy­ sardines injected with cod oil. Two days cling (i.e., raising or lowering temperature to the before the new moon, males were transferred spawning range 24-2.7°C). Males begin ap­ to the tank with females. Spawning began 3 d proaching females several days prior to spawn­ later and lasted for 3 d, with up to 700,000­ ing. On a spawning day, males turn bicolored 800,000 eggs produced per female per day, 71% (black above, white below) early in the after­ buoyancy, 96% fertilization, and an average noon, and the females become bicolored late in hatching rate of 80%. Maturity of camouflage the day at the time of ovulation, just before groupers is reached at 600 g (Tamaru et a1. 1996). spawning occurs. Nassau groupers (facultatively proto­ Average fertilization rate was only 19% gynous, but not obligatorily) have a spawning when 63 redspotted groupers (about 0.5-0.8 kg) season of 1-3 months during the winter in the voluntarily spawned for 2.5 months in an indoor tropics, spring nearJamaica, , and Florida, octagonal concrete tank 6 m wide by 2.7 m deep, and summer at Bermuda, over a wide range of with 1.7 m of water (Okumura et al. 2002). The day length (10.5-14.3 h) and with decreasing or shallow depth was blamed for reducing maneu­ increasing water temperature but mostly at 26 verability of the fish, but the narrow width could ± 1. °C (Tucker et al. 1. 993, 1996; Tucker 1998; have been more of a problem. The light level Sadovyand Eklund 1.999). Aggregations have also was very low, 2.0-800 lux during the day. been found at 6-50-m depths, with fernale:male Water depth also was thought to be the fac­ ratios ranging from 0.57:1 to 4.0;1. Both females tor preventing dusky groupers (1.9-21 kg, 40 and males can mature at 400-450 mm TL (54 fish in four tanks) from voluntarily spawning years), and most have by 500 mm (7 years). in outdoor 20-mJ tanks with l.5-m water depth A 6-kg female Nassau grouper can produce or tank spawning after hormone treatment in an about 900,000 eggs per day by na tural or hor­ indoor I5-m3 tank (Marino et al. 2001); however, mone-induced ovulation and 3.3 million eggs the tanks also were very narrow for voluntary in a 4-d period when spawning voluntarily. spawning of such large fish. Spawning by this species is among the most Greasy groupers (protogynous) in the Ara­ synchronized. bian Gulf spawned during March-June (El­ At Grand Cayman, one male and four fe­ Sayed and Abdel-Bary 1999). Less than 2% of male wild Nassau groupers spawned after 8 the fish caught in two studies were male. The months in a 26-m3 cage (Tucker et a1. 1993). In smallest mature females were about 250 mm TL, Florida, two males and three or four females and 50% were mature at 520mm (4-5 years). The spawned near the full moon in March and April smallest males reported were about 700 mill TL. after 15, 27, and 28 months in a 37-m 3 (11.2 x 3.4 Hormone-induced ovulation of ripe wild or x 0.9 m deep) concrete raceway; each female captive groupers and sea basses generally is spawned as many as nine times a day for 1-4 d reliable (Tucker 1994, 1998). More than 3] ser­ (Tucker et al, 1996). In both cases, spawning oc­ ranid species have been induced to ovulate. curred as soon as normal spawning tempera­ Typically, a female with fully yolked oocytes (de­ tures were reached (for the raceway fish, this veloping eggs) will ovulate (release mature eggs was 3-4 months later than in their home range; into the center of the ovaries) within 24-72 h (usu- GROUPER CULTIJRE 313 ally 36-50 h) after the first of 1-3 injections of 500­ spawns was 73-94%1. In later work, eggs with 1,000 IU human chorionic gonadotropin/kg close to 100% fertilization and survival to first body weight. Similar results have been ob­ feeding were routinely obtained. Hormone in­ tained for several species given 1-3 injections jection of wild fish was not always necessary of 10-50 ~g gonadotropin releasing hormone because newly captured wild female groupers analog (GnRHa)/kg body weight, and GnRHa that were about to ovulate could be identified implants also have worked. If newly caught during the early afternoon and held in 1-m3 broodfish are used, the hormone should be tanks until eggs were ready for release, usually administered as soon after capture as possible, at or just after sunset.Jf held with running-ripe to limit stress effects on oocyte development. males, those fish usually produced 100%-fertil­ For seven grouper species with egg diameters ized eggs in the holding tanks. of 800-1,000 urn, the minimum effective-oo­ Gags (protogynous) off northwestern Florida cyte diameter (seen in biopsy samples) before spawned mostly during February through April injection was in the range 41-61 % of the fer­ at depths of greater than 30.5 m (Collins et aL tilized egg diameter; ovarian biopsies are not 1998). Fewer than 1 in 20 fish were male. Mini­ necessary if external characteristics can be re­ mum size and age of spawning fish were: female lied upon as indicators of oocyte development. 577 mm rt, 3 years; male 981 mm, 8 years. Fe­ Females are handled as little as possible, but males can be mature at 450 mrn (Heppe1l1998). are monitored closely for swollen abdomen, Hood and" Schlieder (1992) reported that matu­ protruding genital papilla, stretching of the rity was reached by females at 2-6 years, -35 cm membrane holding eggs in, and spawning col­ TL and by males at 5 years, ..... 100 em. oration.They are checked more often (e.g., once McGovern et al. (1998) reported that gags an hour) just before the predicted time of ovu­ spawned off southern Florida from December lation. For Nassau groupers, the time from to mid-May, and off South Carolina from [anu­ ovulation to overripeness (deterioration of my through April, with a peak during March to eggs) is only 1-2 h at 26°C. mid-April. The percentage of male gags de­ When ovulation has occurred, milt is creased from 19.6% during 1976-1982 to 5.5°A.l stripped from one or more males and collected during 1994-1995. in 3-cc hypodermic syringes (without needles). Black groupers (protogynous) off western Then, eggs are stripped from the female into a Florida and the Florida Keys were in spawning beaker, milt is added, filtered water is added, condition mainly during December to May and the mixture is stirred. After 3-5 min, the (Crabtree and Bullock 1998). About 1 in 15 were eggs are transferred to a larger container and males. Minimum size and age at maturity were washed several times by water exchange or re­ 508 mm TL, 2 years for females and 947 mm, 6 peated transfer. Transparency, buoyancy, round­ years for males. Fully yolked oocytes had di­ ness, normal egg size, size uniformity, lack of ameters of 0.5-0.7 mm and hydrated ones 0.8­ stickiness, possession of a single oil globule, 1.2 mm. and normal oil globule size are initial signs of Goliath groupers (hermaphroditism notcon­ quality. High fertilization rate, normal cell di­ firmed) in the eastern spawned vision, high hatching rate, and successful first during June-December, wi th a peak in July-Sep­ feeding are subsequent signs. The eggs are in­ tember (Bullock et al, 1992). Aggregations were cubated under very clean, constant, optimal found at 30-46-m depths. Female.male ratio off conditions, often in conical tanks, which facili­ western Florida was 1.75:1 (Bullock et a1. 1992). tate removal of sunken dead eggs, if there are Females began reaching maturity at 120-135 cm any. Usually, eggs are transferred to rearing (6-7 years) and males at 110-115 cm (4 years). tanks just before hatching, or larvae are trans­ The season apparently is July-August in ferred just before first feeding, but it is best to and Puerto Rieo and September-October in Co­ avoid handling of grouper larvae. lombia (Sadovy and Eklund 1999). We first induced ovulation in Nassau grou­ InAsia, redspotted groupers (protogynous) pers during January 1987. With fresh milt and are mature at4-5 years, 0.5-1 kg (Doi et al. 1991), good water, fertilization rate was 85-86%. Sur­ and Malabar groupers (protogynous) at 45-50 vival from fertilization to first feeding for six cm SL (Cesar et al. 2000). In the rvTediterranean, 314 ll...'CEER

the smallest dusky grouper (protogynous) fe­ male caught was 367 mm SL and the smallest male was 685 mrn SL; sex ratio was 3.5 females:1 A~ male (Marino et al. 2001).ln the Gulfof Mexico, maturity of red groupers (protogynous) is reached at two or more years, by females at -30 em FL and males at -40 em (Brule and Deniel 1994). At Baja California, reared macuiatojasciatus were mature at 7.5 months, 19.5g, 66-116 mmSL (Martinez-Diaz et al. 2001).

Hatchery Larvae of most grouper species are small and fragile and have small mouths at first feeding (Figure 4). Known egg diameters are in the range 0.7-1.2 rom, and total length of hatchlings, 1.4-2.3 mm. Yolk and oil, which nourish the early larva until after feeding begins, tend to be exhausted quickly (range 2-5 d) (Kohno et al. 1994, 1997; Tucker 1998). Typically, the larval period is long (often -50 d, range 35-70 d), and groupers tend to require live food longer than most marine fish tha t have been reared. They have been stocked at 0.3-60 eggs or Iarvae/L, usually 5-20. Careful administration of thyroid hormones could be used to increase health and survival of grouper larvae and to accelerate Figure 4. -Development of Nassau grouper larvae. transformationiLam et al. 1994; Tay et al. 1994; (from Powell and Tucker 1992; A) 2.6 mrn TL 1 dah: de Jesus et al. 1998). B) 3.1mm TL 5 dah: C) 8.1 mm TL 25 dah; D) 18.0 Commercial-scale Asian hatcheries have rnm TL40 dah; with permission from the Bulletin of raised large batches of juveniles.with survival Marine Science and the U.S. Southern Regional Aquaculture Center) as high as 34% from hatchlings; smaller hatch­ eries have reported survival up to about 50%. The best survival has occurred in larger tanks by fright, strong current, toxins, pathogens, (60-500 m') under partial sunlight. Species malnourishment) larvae might appear ex­ with small mouths need small rotifers, tro­ hausted or stunned, swim erratically, drift with chophores ( or clam larvae), copepods, the current, and/or not feed well. Early grou­ or other zooplankton at first feeding. Larvae per larvae, especially when stressed, sometimes of some species (e.g., Nassau grouper) seem exude a large amount of mucus, whichcancause to be especially sensitive to noises, such as them to stick to each other, to the surface film, bumping of their tanks, which induces rapid, or to solid objects. Gorging onArte111 inis another frantic swimming. Providing the correct cause of mortality for mid-stage larvae, and can­ amount of turbulence in larval tanks is criti­ nibalism among early juveniles is a potential cal. With too little turbulence, the water strati­ problem. Gorging can be minimized by adding fies (maybe thermally), and zooplankton and the Artemia in small amounts and by feeding fish can aggregate dangerously (for example, rotifers and copepods for at least several days resulting in oxygen depletion, frequent colli­ after Anemia are started. Cannibalism can be sions, feeding difficulty) because they are at­ minimized by feeding the fish well, weaning tracted to brightareas of the tank. With too much them as soon as possible, and removing extra turbulence, the fish are battered. Stressed (e.g., large fish regularly (grading). GROUPER CULTURE 315

Nursery 2001; APEC 2002a, 2002b; Asia-Pacific Marine Finfish Aquaculture Network 2003). They In the Philippines and other areas, when fish sometimes are fed small and sometimes are too small to stockdirect] in grow-outponds y are polycultured with them. Wild fish often are or cages, the early juveniles are raised in con­ collected by attracting them with brush lures, crete or fiberglass tanks or small cages called rock mounds, or brush piles, then picking them hapas (Figure 5) inside ponds or larger cages, up with scoop or encircling nets (Baliao et a1. usually from 20 to 30 mm to 50-100 mm in 1998). Typical market size is 500-1,000 g, about 30-45 d (Baliao et al. 1998; APEC­ which can be reached in 6-8 months of grow SEAFDEC 2001). With high water exchange, out. TIle usual minimum size to begin grow stocking density in tanks (2-10 m") can be up to out/ 75-100 mm, -10 g (age 3-4 months), can 3 1/000/m or more. About 60 Iish z'm" or some­ be obtained in nursery tanks, cages, or ponds. times up to 700/m" are stocked in the cages (size Initially, they are stocked up to 60 fish Zm:' «1 range 1 x 2 x 1.5 m to 8 x 4 x 1.5 m, with 1- to 5­ J kg/m ) in cages. Density usually is reduced mm mesh). somewhat as they grow. In Taiwan, up to 2,000 juveniles are raised In the Philippines, 500- to 10,000-m 2 rectan­ per cage (1.2 x 0.8 x 0.8 m) in small ponds (-100 gular earthen or concrete ponds with depths of rn-) for 1-3 months until they reach -6 em TL 1-2 m and level bottoms are the preferred types (Rimmer 1998). Larger ponds are used mainly for grow out. Double water gates, aeration de­ for overwintering. Pond temperatures are -26°C vices, and emergency water pumps should be in summer and -20-24°C in winter. To limitcan­ present. Typically, juveniles are stocked at 5,000/ nibalism/ grading is performed at 5-7-d inter­ ha and grown from 50 to 100 mm to 400-1,000 vals. g. Foods include trash fish and sometimes tila­ pia living in the ponds. Cages (typically 5 x 5 x GrowOut 3 m with mesh size increasing from 8 to 38 mm) are stocked with 15-20 fish/rn''. In the Indo-Pacific and Middle Eastern regions, In Taiwan, undersized (25-mm) juveniles several species of grouper are farmed in cages, are raised in 100-m 2 nursery ponds (or tanks) to ponds, and tanks, butmostly they are raised from about 60 mm before stocking in 2,000-m2 grow­ wild juveniles and are fed trash fish (Anonymous out ponds. Sometimes cages (1.2 x 0.8 x 0.8 ~)/ 1986; Kohne et al. 1988; AQUACOP et al. 1990; with a maximum of 2,000 fish each, are used in Tookwinas 1990c; Chen and Liao 1991; Quinitio the nursery ponds. Intensive pond farms have and Toledo 1991; Aqua Farm News 1992, 1999; stocked juveniles at 60,OOO-B0,000/ha and har­ Tookwinas et al. 1994; Li 1995; Liao et al. 1995; vested 80% of them/ for a production density as 010U and Lee 1997; FFI 1998; Trai and Hambrey high as 30,000-40,000 kg/ha (usually 10,000­ 1998; Sadovy 2000, 2001; APEC/SEAFDEC 3D/DOD rnt), with aeration and 20%) water ex­ change/d; the groupers are fed mostly trash fish and can grow from 46 mm (2-3 months old) to 600 g in 12 months and 2 kg in 19 months (Anonymous 1992). Orangespotted grouperscan grow from 60 mm TL to 400-800 g after 8-10 months in cages or 10-14 months in ponds (Rimmer 1998). ICing groupers can grow from 75 nun to 2.4 kg in 1 year and 15 kg in 2 years. James et a1. (1998c) raised camouflage grou­ pers in 10-m3 circular fiberglass tanks at three densities (5, 15, 45 fish zm") on moist pellets for 9 months, then both moist pellets and trash fish for 3 months at 28-29°C and 42-43 parts per Figure 5. -A nursery cage (1 x 1 x 1 m) used for groupers and other fish at ~ingapor7' ~by John thousand (ppt) salinity. Except for lower sur­ Tucker; (Tom Tucker 1998; with perrrusSlOn from vival (84%), results were not significantly worse KluwerAcademic Publishers) at 45/m3; however, the harvest density of 17.33 316 llJCKER

kg/mJ was much better (59-513 g, 1.31 g/d, TL, 579-1,098 g, mean 909 g) were released and 0.60%/d, FCR 2.05). (FCR = feed conversion ra­ monitored by a diving tearn on an open-ocean tio = weight of dry feed necessary to produce a reef with depths of 1-15 mat St. Thomas, U.S. unit weight of wet fish; PCR = protein conver­ Virgin Islands (Roberts et al. 1995). They exhib­ sion ratio = weight of dry dietary protein neces­ ited behavior previously expected only from sary to produce a unit weight of wet fish.) wild groupers. Within 1 h after release from a Brownrnarbled groupers fed moist pellets holding cage, all the groupers had gobies and and trash fish in 10-m3 tanks for 7 months grew shrimp remove ectoparasites obtained while better at 45/m3 (127-618 g, 2.34 g/d, 0.75%/d, they were in the cage. Within 2 d, groupers were 3 FCR 1.24), than at 5 or 15/m , also with a better seen hunting alongside a moray eel or octopus. harvest density of 26.07 kg/m3 (James et a1. It was surprising that these behaviors known 1998b). for wild groupers were innate and did not have James etal. (1999)reported that hybrid grou­ to be learned by the hatchery fish. Although their pers (E. [uscoguttatns x E. polyphekadioll, from tags had been lost, two groupers seen at 297 d voluntary spawning) grew better than the par­ and one at 16 months probably were released ent species on moist pellets and trash fish dur­ fish. ing 7 months in 10-m3 tanks at 15/m3 (112-696 Those groupers were raised from eggs ob­ g, 3.02 gld, FCR 1.39). tained at Grand Cayman, which is about 1,800 Ahmad et al. (2000) grew out two sizes of krn west of St. Thomas. Because eggs and lar­ orangespotted groupers in I-m3 circular fiber­ vae can drift with ocean currents for at least 46­ glass tanks with different water flow rates for 75 d after hatching (dah), they can be distrib­ 14 weeks. When 40 small fish were stocked per uted great distances. Not surprisingly, prelimi­ tank, results were not significantly worse with nary studies indicate that Nassau groupers, at a loading rate of 6 kg/L/min (135-303 g, 1.71 least throughout this region, probably are ge­ gld, FeR 1.45) than with 0.5 kg/L/min (135­ netically homogeneous (Sadovy and Eklund 333 g; 2.02 gld, FCR 1.33). When 15 larger fish 1999). were stocked per tank, results were at least as Betweenweaning and 27 months (with very good with a loading rate of 3 kg/L/min (1,079­ little growth during 8 months at low tempera­ 1,502 g, 4.31 g/d, FeR 2.10) as with 1 or 6 kg/ ture), the tagged groupers had been fed dry L/min (1,046-1,400 g; 3.61 g/d, FCR 2.09). pellets. For 3 months before the release, they Orangespotted groupers weighed 3 g after were held in a 37-mJ concrete raceway and 2 months in the hatchery, 22 g after 3 more given live and frozen food in addition to pel­ months innursery ponds with trash fish for food, lets. The live foods, mainly goldfish Carassius and 194 g after 5 more months in grow-out auraius,with some minnows (Cyprinidae), pink ponds with a compound feed, at a low density shrimp Penaeus duorannn,and fiddler crabs (Uca of 0.5 fishv'm- (Bombeo-Tuburan et al. 2002). spp.), were dropped through a polyvinyl chlo­ Improvement of early growth rates (e.g., ride (PVC) pipe that opened at the tank bottom. better temperature control, earlier weaning) will Frozen food included a variety of fish, queen allow production of 600-g Nassau groupers conch Strombusgigas, pink shrimp, and . within 12 months, I-kg groupers within 18 The groupers showed no hesitation in eating months, and 2-kg groupers within 24 months. any of those foods, exceptfor live shrimp, which were not eaten immediately after being added Stock Enhancement the first time, but disappeared within a few hours. Later, the groupers consumed shrimp as In isolated locations where groupers have been fast as they were added to the tank. Live fish severely depleted, it might be possible to re­ and crabs were very easy for the groupers to plenish spawning stocks with reared fish. capture and usually everything was eaten Redspotted groupers from government hatch­ within 20 min. To prepare the groupers for en­ eries have been successfully released in Japan counters with predators, for several days before (Liao 1997; Okumura et al. 2002). they were shipped, we attempted to sharpen In an experimental study, 27 Nassau grou­ their reflexes by chasing them with nets and pers raised in tanks in Florida (309-367 mrn splashing the water surface. GROUPER CULrURE 317

TIle groupers were held in a 4 x 1.2 x 1.2 m for late larvae and early juveniles. Unless the chicken wire cage at the release site for 15-16 d water is very well mixed, the planktonic foods before being freed, to allow orientation and re­ are not distributed evenly in the tank, and av­ sumption of feeding. Because they were slow to erage prey density is only of theoretical value. begin eating foods offered by the diving team, What matters is that the fish can find and catch the fish were kept in the cage much longer than nutritious prey with a minimum of effort, maxi­ originally intended and several developed eye mize their intake of nutrients, and grow fast. injuries (Plastic coated wire would have been Depending on age and number of larvae, an safer but was not available). The caged groupers average density of 5-20 rotifers/mL seems ap­ also became accustomed to the divers and after propriate if larvae are feeding well enough to release let divers approach them. It would have prevent the rotifers from becoming so numer­ been easy to train the fish to fear people by ous that they reduce water quality too much and threatening and chasing them instead of giving crowd the fish. Arientia usually are stocked at them food. In areas where divers feed fish and about 1-2/mL when first given to the larvae, is uncommon, wild Nassau grou­ bu t as many as 6/mL can be used if the fish eat pers tend to be just as tame. Ifchased by people them within 12 h, before their nutritional qual­ under water, they become extremely wary. ity deteriorates. Microfeeds have been tried as a supple­ ment during the first week but probably are not Nutrition digested much until at least 2-4 weeks. Wean­ ing from live food to dry crumbles or small pel­ Foods forLetvee lets can be completed justbefore or during trans­ Grouper larvae usually are raised in green water, formation, which occurs at 35":'70 dah, depend­ mostly with the phytoplankton Nannochuiropsis, ing on species. Minced (e.g., muscle of Tetrasetmis, and/or Chlorella spp. stocked at 10­ fish, shrimp, ) often is used as an appe­ 500 cells /pL during 0--3 dah to 12-40 dah. At tizer or transitional food. first feeding, most species can easily eat small Redspotted groupers at 24DC hatch in about rotifers, but oyster (Su et £11. 1997), clam, or mus­ 24 hat 1.6 nun TL, first feed 2-4 dahl finish their sel eggs and larvae sometimes are used as a yolk (end of yolk sac = EYS) 4 dahl and trans­ supplement. Growth and survival tend to be form 45-50 dah (-30 mm). They have been suc­ better if copepods or mixed zooplankton are cessfully raised on a large scale in Japan in 60­ included in the diet, but care must be taken to m? tanks with green water (0-40 dah), rotifers avoid introduction of pathogens or predators. (2-37 dah), Arteinia (28-50 dah), and compound Beginning at 4 dah, brownmarbled grouper lar­ feed beginning at20 dah (Maruyama et £11. 1994). vae preferred copepod nauplii to rotifers (Ali In 1989, survival was 16% from eggs to 47 dah et £11. 1998). When large numbers of rotifers also and 34% from hatchlings to 47 dah. Mortality were added to rearing tanks at 7-10 dah, from nu tritional deficiencies sometimes oc­ orangespotted grouper larvae continued to pre­ curred at -20 dah. Mortality after 40 dah was fer copepcds, which they had been eating since mostly from cannibalism. They can be weaned 3 dah (Doi et a1. 1997; Toledo et al. 1997, 1999). by transformation at -50 dah. Certain ciliates (with widths of -20-80 urn) Kelp groupers at -25DC are 2.1 mm TL at could be added to improve survival during the hatching, first feed 3 dah, reach EYS 5 dah, and first few days of feeding, but some types are transform -40-45 dah (-37 mm). Larvae were not nutritious (Nagano et al. 2000). Cladocer­ raised in a 30-m 3 tank with small rotifers (3-6 ans sometimes are used for early to mid-stage dah), large rotifers (7-31 dah), formulated feed larvae. Arientia enriched with essential fatty ac­ (18-45 dah), Anemia nauplii (20-45 dah), and red ids can be a staple food beginning at 10-30 dah, sea bream Pagrus major eggs and larvae (32-45 but their abundance should be controlled to dah) (Sawada et a1. 1999). m..inimize gorging and incomplete digestion by Orangespotted groupers at 27°C hatch in the larvae. It is best to delay feeding of Artemin about 22 h at 1.7 mm TL, first feed 3 dah with as long as other foods are sufficient (until 20 mouth width of 268 urn (Ordonio-Aguilar et al. dah or later). Mysid shrimp sometimes are used 1995), reach EYS 4- dah, and transform 35-40 318 ruCKER dah (25-30 rom). At first feeding, the mouth mm) with a mouth width of 225 11m, all feed by gape was about 180 urn and small rotifers were -92 h, and reach EYS -1.5 h after all are feeding eaten (Duray 1994). By 20 dah (>6 mm TL), they (Kohno et al. 1994; Ordonio-Aguilar et al. 1995). preferred Artemui over rotifers. When Anemia Lim et al. (1990) reported that brownrnarbled were given to them at 15 dah, survival de­ grouper hatchlings were slightly larger than creased, but if Ariemia feeding was delayed those of greasy groupers, transformstion oc­ until 21 dahl survival was improved. curred at -40 dah (-31 mm TL), and the former Dry feeds were presented to orangespotted species survived better under similar hatchery groupers during 2-35 dah and minced fish start­ conditions. Brownmarbled grouper larvae were ing at 25 dah (Doi et al. 1991). Larvae were given raised in 900-L tanks with green water (2-12 only minced fish after 40 dah. In one 1990 trial, dah), rotifers (2-12 dah), and enriched Ariemia survival from fertilization to 10 dah was 38%, (10-40 dah). In 1990, the best survival from but by 43 dahl survival had decreased to 12.1% hatching to 40 dah was 13.2%. (still good for groupers). They were raised in Lim (1991) stated that the main advantage 5,OOO-L tanks with green water, oyster eggs and of brownmarbled groupers was that they could larvae (2-4 dah), rotifers (2-25 dah), copepods eat small rotifers at first feeding, while greasy and enriched Anemia (13-40 dah), compound groupers could not and had been fed feed (2-35 dah), and minced fish starting at 25 eggs and trochophores at first. Also the larval dah. period of greasy grouper was longer (40-50+ In 1995, survival of orangespotted groupers versus 35-40 d). Cannibalism, which became from hatching to 24 dah was better in 3,OOO-L serious at 30 dah in brownmarbled grouper and (20 versus 7%) than in 500-L tanks (Duray et al. at 35 dah in greasy grouper, and low stress re­ 1997). Growth (5.36 versus 4.23 mm TL) and sistance could have been related to nutritional survival (21 versus 11%) were better when roti­ deficiencies. fers fed during the first 2 weeks were sieved to Malabar groupers at 29°ehatch in about 19 h less than 90 11m (average lorica dimensions be­ at 2.3 nun TL, first feed and reach EYS -3 dah, fore sieving were 120 11m x 162 11m). Survival and transform 3~0 dah (-35 mm). Maneewong was better, but growth was worse when only et al. (1986) reported raising them in 26-m3 tanks rotifers were fed up to 35 dah (32%, 11.9 mm, with green water (2-20 dah), rotifers (2-25 dah), 4.6 mg dry.weight [DW]) than when Ariemia copepods (13-30 dah),Artemin nauplii (15-55 dah), alone (6%, 15.1 mrn, 8.8 mg) or rotifers plus cladocerans (30-? dah), Arteniia adults (35-? dah), Artemia (13.3%, 17.1 mrn, 10.7 mg) were pro­ and minced fish (45 dah onward). Tookwinas vided during 15-35 dah. Survival and growth (1990a) reported raising them in 26-m3 tanks with (starting with5 larvae/L) during 21-35 dah were green water (0-18 dah), rotifers (3-30 dah), Artemin better when Artemia nauplli were fed at 3/mL (12-15 to 50 dah), minced fish (45-? dah), and (30%, 12.0 mm TL, 4.9 mg DW) than at 1/mL sernimoist compound feed starting at about 50 (20%, IDA mm, 2.8 mg) or 2/mL (27(Yo, 11.9mrn, dah. Ruangpanit (1993) gave this schedule: 4.4 mg). Under similar conditions, survival was rnicroalgae (2-10 dah), rotifers (3-6 dah), enriched better with 12 Artemia/mL/d (50.3%) than with rotifers (7-30 dah), enriched Artemia (15-45 dah), 6/mL/d (39.7%) or 3/mL/d (18.0%). adult Anemia (30-60 dah), the cladoceran Moina Toledo (2002) reported improvement in sp. (30-60 dah), minced fish (40-60 dah), and com­ survival of orangespotted grouper larvae to 5.4­ pound feed (beginning 46 dah). 29.8% (hatching to 35 dah), when 2-36Iarvae/L Greasy groupers at 27°e hatch in about 24 were stocked in 5- or 10-m3 tanks, rotifers and hat 1.7 mm TL, first feed 3-4 dah, reach EYS-4 Artemia were enriched with highly unsaturated dahl and transform 36-50+ dah (~33 mm), Chen fatty acid (HUFA) and vitamin C, and thyroxin (1979) described feeding methods used for rais­ was administered in the water at 0.01 ppm or in ing larvae in 1,000-L tanks. Foods were boiled the Anemia at 0.5 ppm (to accelerate transfor­ chicken egg yolk 3-5 dah, rotifers 3-15 dahl cla­ mation) during 21-25 dah; however, survival at docerans (Dinpllal1osol1la sp.) from 10 dah until 55 dah (-50 mm TL) was 3%. transformation, rnysids (Mesopodopsis sp.) from Brownmarbled groupers at 28°e hatch in a few days before transformation until wean­ about 18 h at 1.3 rom TL, first feed -69 dah (2.8 ing to shredded fish or dry pellets at 55-60 mm GROUPER CUI.:n..iRE 319

TL. When weaning was attempted with smaller population). Survival during 1-10 dah was bet­ juveniles (33 mm), they had trouble digesting ter with a mixed diet of oyster trochophores and the pellets and did not survive well. The initial rotifers (T+R 15.6%) than with rotifers alone (R, weaning food was 80% ground trash fish mixed 9.7%) or a sequential diet of trochophores and with 20% pellets, becoming 100% pellets by rotifers (T~R, mixed during 5-7 dah, 2.6%). 1veaning day 6. Transformation began at 33 dah Growth was better with the rotifer and the tro- for large fish, but was not finished until 50-60 chophore + rotifer diets (R 4.1/ T+R 3.7, T~R 3.1 dah for small fish. mm final notochord length). Therefore, His likely Nassau groupers at 26°C hatch in about 26 that rotifers should be included from first feed­ h at 1.9 nun TL, first feed -2.5 dah, and finish ing even when trochophores are used. their yolk (EYS) and oil -5 dah (Figure 4). TI1e Polka-dot groupers at 27-28°C hatch in larval period is long wi th a wide time range for about 19 h at 1.5-1.6 mm TL, first feed at 2-3 transformation to the juvenile stage at 35-50 mID dah with a mouth width of about 140-150 urn, TL (Tucker and Woodward 1996). In 1990, lar­ and reach EYS at -4 dah (ACIAR 2000; Sugama vae reared in green water in 3,500-L tanks re­ et al. 2002a). They were raised on rotifers (su­ ceived rotifers enriched with Frippak Booster per-small/ 2-5 dah; small, 6-25 dah), Ariemin micro-capsules, Anemia nauplii, and Frippak (-]7-40 dah), and microdiets (15-50 dah), with Booster-enriched 2- to 5-d-old Artemia. Larvae a survival range from hatching to 50 dah (25 mm were planktonic until 42 dahl when they began TL) of 19-54%. orienting to the tank walls and bottoms. TI1en, at 20°C hatch in about 48 hat white plastic milk crates were added to provide 2.1 nun, first feed -3.5 dah, reach EYS 5.5 dahl cover. Transformation of all fish occurred dur­ and transform by 43-52 dah (~10 mm) (Tucker ing 46-70 dahl with individuals making the 1984, 1989). Berlinsky et al. (2000) raised larvae change in less than a week. Weaning to 1.6-mm in 40-L aquaria with green water, unenriched ro­ dry pellets was accomplished byfeeding minced tifers (3-12 dah), and enriched Ariemia (10-24 penaeid shrimp and crumbled pellets during dah), with weaning to dryfeed during 18-24 dah. 56-61 dah (4D--60 mm TL), but a small amount Taiwanese hatcheries use a range of sys­ of shrimp was given to the fish 63, 66, and 69 tems to raise grouper larvae, and survival of dah to enhance growth of smaller individuals. 7% to 4 weeks (25-30 mm) is considered good Mortality peaks were at 5 dah (yolk and oil ex­ (Rimmer 1998). Indoor systems include fiber­ hausted) and at about 20 dah, when consump­ glass or concrete tanks holding up to 100 m' tion of Ariemia nauplii was highest. In 1990, where larvae are reared in green (50,000-500/000 survival from fertilization to 98 dah was 5.0% cells/rnL) or clear water with oyster tro­ and would have been higher if fewer Ariemia chophores (-4-6 dah), small rotifers (beginning nauplii had been used (gorging, partial diges­ -4 dah), and Artemia and/or adult copepods tion, and death were observed). Transformation (beginning 16-22 dah). Outdoor systems in­ might have been accelerated by providing cover clude concrete or earthen ponds of 200-5/000 or feeding shrimp earlier. In 1994, trochophores, m? (sometimes 1. ha), which are filled through a rotifers, Ariemia, and copepods were used. in fine-mesh sock filter 1-2 d before stocking eggs one trial, survival from eggs to 15 dah in a 120­ to limit the size of zooplankters. Stocking rate 2 L aquarium was 25%. In another trial, survival is 600-1,500 eggs/rn • Larvae are held in float­ during 10-75 dah in a fiberglass tank was 59%. ing bags for the first 2 d of feeding (-4-6 dah) to In 1994, Watanabe et al. (1996) raised keep the trochophores concentrated. Rotifers, Nassau grouper larvae in 33.8-m J rectangular copepods, and other zooplankters raised in tanks on wild zooplankton (protozoans, copep­ separate ponds of 0.05-0.1 ha are pumped di­ ods), rotifers, and Ariemia in green water, with rectly into the fish ponds or collected in nets highest survival from hatching to 62 dah of and transferred. 1.4%. Preliminary research showed that survival was better (8.0 versus 2.1%) when rotifers sieved Feeds to less than or equal to 200 urn were fed during 2-10 dah than when rotifers were not sieved Wild juvenile and adult groupers eat mainly (lorica length range 50-264 urn in the culture fish, crabs, shrimp, mantis shrimp, lobsters, 320 'llJCKER and mollusks (Brule et al. 1994). Red groupers feeds likely would result in worse feed conver­ Epinepltelus mario seem to prefer crabs first, then sion and possibly slower growth. shrimp. Early juvenile Nassau groupers (50-75 Orangespotted grouper juveniles grew at mm TL) ate (by number) 37% gammaridean least as well when fish meal was replaced with arnphipods, 15% isopods, and 11% mysid processed solubles (ACIAR 2000). In the shrimp (Grover et al. 1998); from 200 to 299 mm range 6-4] g, orangespotted groupers grew as TL, they ate ~85% crustaceans and 11% fish; for well with 8% fish meal, 32% meat meal, 8% groupers larger than 299 mrn, foods were half blood meal, and 7% rice bran (growth 2.92%/d. crustaceans and half fish (Eggleston et al. 1998). FeR 1.04) as with 32% fish meal, 8% meat meal, Small juvenile leopard coraltrout «100 nun SL) 2% blood meal, and 13% rice bran (growth ate about the same amount of fish and crusta­ 3.13%/d, FCR 0.95); other ingredients were 10% ceans (mostly penaeid shrimp), but larger ones shrimp meal, 1% squid meal, 6% soybean meal, mostly ate all fish (St. John 1999). 15% wheat flour, and 6% cod liver oil (Milla­ In SoutheastAsia and other areas, groupers rnena 2002). ' have been fed mainly trash fish supplemented Malabar groupers (58-229 g) were raised with vitamins and minerals, secondarily moist on feeds containing still-dried muscle (23­ or semimoist pellets, and rarely high-protein 80%), dextrin (0-20%), alpha-cellulose (10-31%), dry pellets (Ruangpanit and Yashiro 1995). Bar­ carboxymethylcellulose (6%), cod liver oil (3%), rarnundi feeds work fairly well. Cesar et al. and com oil (0-14%), with protein contents of (2000)stated that8 mtof trash fish were required 20-70% and similar energy levels (Teng et al. to produce 1 mt of groupers and that compound 1978). The best growth (1.5-1.6%/d) and FCR feeds typically contained 50% fish. In the Phil­ (1.10) occurred with feeds containing 40% or ippines, nursery foods include minced fish, 50% protein, 13.5% or 9.1% fat, and 12.0% or mysid shrimp, and larger zooplankton (e.g., 11.1 DIr) digestible carbohydrate. adult Anemia, mysid shrimp, mosquito larvae) Chenand Chen (1986)raised Malabargrou­ given 4-6 times a day (Baliao et al. 1998;APEC/ pers on a series of feeds based on white fish SEAFDEC 2001). During grow out, a daily ra­ meal and dextrin. The best growth (2.12%/d) tion (DR) of 10%.chopped fish or 3% pellets is occurred with the feed containing the most pro­ fed in two meals until groupers weigh 200 g, tein (54%, moist) and fat (7.8(Yo, moist). then a DR of 5Wo fish or 2% pellets in one meal A preliminary feed formula used in Thai­ (APEC/SEAFDEC 2001). In Taiwan, nursery land for Malabar groupers consisted of 75% fish foods include live or chopped fish and shrimp, meal, rice bran, bananas, about 0.5% soybean adult Arlen/in, and eel feeds; initial feeding fre­ oil, vitamins, and minerals (Tookwinas 1990b). quency is 4-6 times a day dropping to twice a ln semipurified feeds for Malabar groupers, day by the time the fish are harvested at -6 cm 48% protein was optimal for growth, and 48 or TL (Rimmer 1998).When the groupers reach200 54% resulted in the best FCR (Chen and Tsai g, they are fed once a day in summer and every 1994). Shiau and Lan (1996) suggested that pro­ other day in winter. About 70% of Taiwanese tein can be reduced to 44% if gross energy is farmers were using some compound (mainly 3,400-3,750 callg because differences in growth moist) feeds in 1998. and conversion were small. Qin et al. (2000) found that 1,500-2,000 mg/ Greasy groupers (197-631 g) grew slightly kg ascorbic acid added to trash fish fed to yel­ but not significantly better (2.82 versus 2.34 g/d) low groupers improved growth rate and resis­ with experimental pellets (81.5%fish meal, 2.0% tance to Vibrio mllllificllS better than 1,000 mg/kg cod liver oil,12% tapioca (a-starch) thanwithtrash or less. Asuitablestarterfeed for groupers would fish (Chou and Wong 1985). The FeR was better contain 50-60% high-quality protein, 12-16% fat with trash fish (1.7 versus 2.4,by dry weight). (half or more from marine sources), 15% or less Greasy grouper (range 24-266 g) growth carbohydrate, less than 3% fiber, and less than and conversion apparently improved when fro­ 16% ash (Tucker 1998). By the time they reach zen fish was mixed 60:40 wi th a dry feed, but 500 g. Nassau groupers (and others) can be given the only significant difference was that final a feed with -45% protein, -9% fat, <')0% carbo­ weightwith the dry feed was lowest of the three hydrate, ~4% fiber, and

brown fish meal. 10% wheat flour, 8.6% wheat Nassau groupers (2.3-14.2 g) raised for 56 middlings, 4.2% fish oil, and 0.75% lecithin din 1993 grew better with an experimental dol­ (49.7% protein, 10.8% fat, 23.2(Yo carbohydrate). phin (62% protein/14.2% fat; 3.19%/d growth) The 60:40 pellet and 100% fish diets gave simi­ feed and a commercial Japanese carnivorous­ lar results. Results with the dry and moist pel­ fish (56/7.8; 3.05%/d) feed than with commer­ lets were: range 24-108,26-246 g; growth 0.84/ cial salmon (53/15.2; 1.71%/d) and (44/ 2.20 g/d; growth l.5L 2.25%/d; FCR 1.36/ 1.06; 5.9; 0.93%/d) growers (Ellis et a1. 1996).The FCR and PCR 0.68, 0.58. was 0.94 for the carnivorous-fish feed, 1.32 for Nassau groupers (40 mm TL/ 1 g) raised in the feed, 1.60 for the salmon grower, 1990were weaned from Ariemia to a starter feed and 5.55 for the trout grower. containing 60% herring meat 3.5% blood meal, Nassau groupers (1994) given a commer­ 5(Yo feather meal, 8% textured vegetable protein, cial starter feed (46% protein, 10% fat) grew fast­ 5% corn gluten meal, 5% high gluten wheat est (3.3-12.4 gin 63 d; 2.07%/d) at 31°C and had flour, 1% brewers yeast, and 7.5% menhaden oil slightly but not significantly better FeR (1.04) (60%protein, 16.2%fat, 13.4% total fish oil, 17.2% than at 22°C and 25°C (Ellis et al. 1997). carbohydrate, dry weight basis; feed JT9010 in Polka-dot groupers (4.9 g) grew well with Tucker and Woodward 1996 and Tucker 1998). 54.2% dietary protein in pellets with protein After thefish reached 6-7 gat -3 months, gro'wer mainly from fish meal plus small amounts of feeds developed during 1990-1992 with simi­ casein and squid liver meal (ACIAR, 2000). A lar ingredients, but less fish meal (40-45%) and preliminary report indicated that a feed with fish oil (6-8% total) were used. Then, under less 55.3% fish meal plus 10%"'soybean meal was than optimal conditions (including a 17-33°C better for 3-g fish than feeds with 63% FM + 0% temperature range), the groupers reached a SBM or 47.6% FM + 20°;;) SBM (ACIAR 2000). mean weight of 1.5 kg at 23 months (FCR rising Juveniles stocked in cages grew from 17.5 to 39.3 from 0.90 to 1.32) and 2.0 kg in 28 months (FCR gin 60 d (1.35%/d, DR 1.91, FCR 1.53) on a feed up to 1.80/ survival 97% during 3-28 months). containing: 66.9% local fish meal, 10% soybean On a 10% moisture basis, the starter feed con­ meal/4% fish oil, 3% squid oil, 6% wheat flour, tained 54% protein, 14.6% fat, and 8.6% carbo­ and 6% rice bran (Ahmad et al. 2002). Girl et a1. hydrate, and the grower feeds 53%/ 9.7%/ and (2003) concluded that 54% protein, 9% lipid, 15.8%. Growth rates ranged from 4.28%/d at 0­ 1.0% n3-HUFA, 30 mg/kg ascorbyl polyphos­ 6.1 g to 0.25%/d at 1.8 kg, and peR also was phate, and 7-28% dextrin, with about 118 mg good, 0.54-1.05. When the pelleted feed was crude protein/kcal and 4.77 kcal/g/ were suit­ supplemented with live and frozen food, able for polka-dot groupers in the size range 3­ growthwas slightly better. Reared Nassau grou­ 14 g. Research Institute for Coastal Fisheries pers almost always will eat immediately after (2003)recommended 45-48% protein, 12% lipid, being caught and moved, if handled gently. and 150 mg stable vitamin C (APNa)/kg for the Once they grow larger than the human hand, range -5-25 g. they become very tame. An hour after capture Dusky grouper Epinephelus marginatus wild in a trap, even 25-cm wild juveniles ate pellets juveniles grew faster on commercial pellets (7­ from my hand. 25 gin 69 d) than on octopus (7-21 g) (Castello­ Similar results were obtained by Johnston Orvay and Vilar 1993). Black sea bass grew from et a1. (2002). Feeds similar to those used in 1990 107 to 192 g in 14 weeks on a commercial feed by Tucker and Woodward (1996; Tucker 199B), with -57% protein and 20% fat (Berlinsky et al. except for addition of 5% squid liver powder 2000). and 0-9% cellulose, were used to test menha­ Belted sandfish, like larger serranids, den oil content at 0-9% (6/9,12, 15% total lipid) spawn readily if kept healthy (Tucker and and temperature (25, 30°C). Cannibalism and Woodward 1995). Giving a prepared feed to variable survival (treatment means 56-89%) broodfish is a good test of its quality. An ex­ were factors, but the highest biomass was perimental grouper feed OT9120in Tucker 1998) reached at 6% total lipid and 30°C (2.0-9.8 gin was the sale diet for a pair of belted sandfish 75 d, growth 2.09%/d, FCR 1.82/ peR0.72); 12% (-60 mm TL) in a 28-L aquarium during an 8­ or 15% lipid appeared to be too much. month period. The pair spawned 79 times in 3:22 TUCKER

81 d (February-May 1991), producing 65,054 Some natural light is good for all stages. eggs, with a mean combined fertilization and Continuous natural and artificial light at 1,000­ hatching rate of 92%. 3,000 lux should be suitable for larval rearing, but higher intensities have been used. Light should be as even as possible across most of Environmental Conditions the tank but can be reduced in danger areas, such as near surface drains. Tanks with diameters of 3-5 m or more are bet­ For most warmwater marine fish to be ter for grouper larvae than smaller ones because healthy; dissolved oxygen should be at least 5 surface film removal is safer, temperature and mg/L and hopefully nearsaturation. In the Phil­ water quality changes are moderated, and the ippines, 4.5-8.2 mg/L is considered acceptable fish contact the tank walls less often. Although for early juvenile groupers and 4.9-9.3 mg/L eggs or larvae have been stocked up to 40/L, for grow out (Baliao et al. 1998). 5-20/L is safer. Providing habitat (e.g., milk Safe limits for ammonia, nitrite, and nitrate crates) near the end of the larval stage can ac­ are only approximately known; more definitive celerate transformation to the juvenile stage. data will require testing. Ammonia should be Juveniles and adults of estuarine and coastal kept near zero (~1 l giL [ppb] unionized am­ species tend to be more tolerant of environmen­ monia nitrogen for larvae, ~5 i giL for adults); tal extremes, turbidity, and pollutants than those lOi giL unionized ammonia can be toxic to lar­ of offshore (especially reef) species. vae, and juveniles and adults are only slightly Most groupers that have been reared are more tolerant. Likely limits for nitrite nitrogen warmwater fish that spawn and grow best in are 0.1 mg/L (ppm) for larvae and 1 mg/L for the range 24-30°C; most can tolerate a range of older fish, but zero is best. It probably is best to 15°C to at least 35°C. The best temperature for keep nitrate nitrogen lower than 20 mg/L for Nassau grouper larvae is -24.....27°C; feed intake young stages and 50 mg/L for older fish. by juveniles and adults is minimal at less than Ammonia production data like that deter­ 18°C, but feeding and growth are good at 22­ mined by Leung et al. (1999a, 1999b, 1999c) for 30°C; best grow.th should occur near 30°C. In areolated groupers could be used in water qual­ the Philippines, 27-28.5°C is considered accept­ ity management of closed and flow-through able for earlyjuveniles and 24-31°C for grow rearing systems and to help estimate environ­ out (Baliao et al, 1998). Black sea bass larvae mental impacts of cage systems. grew faster at noc than at 18°C (Berlinsky et al. Juveniles and adults of estuarine and 200~. . coastal species seem to tolerate suspended sol­ Eggs of groupers that spawn at sea will re­ ids better than offshore (especially reef) species, quire a salinity of about 30 ppt or higher for which seem to suffer from gill irritation more them to float, but slightly lower salinity can be easily. tolerated even though the eggs sink. Salinity One of the greatest problems for grouper tolerance usually increases with age. Oceanic larvae is sticking to, suffocating at, or becoming species are healthiest at close to seawater salin­ intoxicated at the water surface. Film-producing ity (35 ppt), but some estuarine species can sur­ substances (e.g., polysaccharides, organic oils, vive at less than 10 ppt. Once they are fully de­ proteins, inorganic oils, soaps, plasticizers) need veloped juveniles, even Nassau groupers can to be controlled as much as possible, so that re­ tolera te 15 ppt for at least a few days. In the moval efforts are not necessary while larvae are Philippines, 19-24 ppt is considered acceptable too vulnerable. For the first several days of feed­ for early juveniles and 21-41 ppt for grow out ing/ skimming the surface wi th air jets and (Baliao et al. 1998). Toledo et a1. (2002) reported standpipes can be dangerous because early that 8 ppt was lethal, 16 and 24 ppt were suit­ grouper larvae tend to drift with the current able, and 32 ppt was too high (based on growth rather than swim against it and they cannot tol­ and survival) for orangespotted groupers at 1­ erate much turbulence, especially when they are 4 dah. Early juvenile black sea bass (3-8 g) grew near the surface. The larger the tank, the safer faster at20 ppt than at 32 ppt and could tolerate the larvae are from such localized disturbances and grow well at 10 ppt (Berlinsky et al. 2000). and from going down the drain. Standard skim- GROUPER CULTURE 323

ming devices can be used once larvae can swim Health Management well enough to avoid them. A possible approach to prevent larvae from Hatchery sticking to the surface is to lubricate the water surface with oil extracted from omega yeast for Good management of the microbial environ­ the first few days after hatching, but the oil has ment can protect larvae from pathogens and to be removed before larvae begin gas bladder eliminate the need for antibiotics and other inflation (Yamaoka et al. 2000). Sugama et a1. drugs. As a major management component, we (2001) recommended addition of squid or fish routinely inoculate larval rearing systems with feed oil on the water surface at 0.1 mL/m2 three probiotic (beneficial) bacteria isolated from times a day during 1-5 dah in concrete tanks. healthy fish raised in our hatchery (Kennedy et Sugama et al. (2004) obtained better survival of al. 1998). Without this seeding, the bacteria polka-dot grouper larvae when squid oil was populations can vary from bad to good. added during 1-9 dah than when it was added during ]-6 or ]-12 dah. Infections and Infestations Usually, a moderate growth of phytoplank­ ton is maintained in rearing water for grouper Viruses, gram-negative bacteria (Vibrio, larvae (greenwater culture). Besides being food Aeromonas, Pseudomonas, Cytoplwga, Ftavobacte­ for the zooplanktonic prey of the fish, the algae riutn, Flexibacter, Pasteurella spp.), gram-positive also can remove ammonia, generate oxygen and bacteria iStreptococcus, Mycobacterium), ectopara­ keep pH high, release anti-bacterial or growth­ sitic protozoans (Amyloodhlilil/1 ocellniutn, promoting substances, promote growth of ben­ Cntptocarfon itriians, Brooklynella spp., Ichlhy­ eficial bacteria, and stimulate behavior, feeding, ophihirius sp., Trichodina sp.), monogeneans or digestion. However, algae decomposition iBenedenia spp., Neobenedenia spp., Diplecttmutn products can contribute to formation of a sticky spp., Megnlocotyloides epinepheli, Pseudcrhob­ surface film. dosyl1oc!ws epinephelii, copepods, and leeches are Sugama et a1. (2001) recommended that among the most important pathogens of cul­ water transparency be reduced by maintaining tured groupers (Arthur and Ogawa 1996; Balian microalgae or dye in hatchery tanks during et a1. 1998; Chew-Lim et al. 1998; Lin and Ho most of the larval stage to limit aggregation 1998; Tanaka et a1. 1998; Tucker 1998; AI­ which can cause death by sticking to the sur­ Marzouk 1999; Chou et al. 1999; Koesharyani et face or each other (especially during 2-5 dah) al. 1999; Lee et al, 1999; Bondad-Reantaso et a1. or having spines tangle with those of other lar­ 2001, 2002; Koesharyani et al, 2001). Many in­ vae (10-25 dahl. ternal parasites (microsporans, myxosporans, Sugama etal. (2004) obtained bettersurvival trematodes, cestodes, nematodes, and acantho­ of polka-dot groupers up to 10 dah with zero cephalans) also have been reported. Nervous water exchange than with 100% or 200% ex­ suffering disease (signs could include gill, change per day. blood, gas bladder, liver, heart, brain, and nerve For a 9-m 3 tank (about 2.75 x 2.75 x 1.2 m damage) probably was caused by rancid di­ deep), Sugama et al. (2001) recommended use etary fats (Hua et al. 1994a, 1994b). of nine evenly spaced air stones during 0-10 Spinning grouper disease and viral nervous dah and 13 air stones (one centered, the rest necrosis could be the same. During the 1990s, against the wall) after 10 dah, with aeration high viral nervous necrosis (VNN) caused mass mor­ during 0-2 dah to keep larvae off the bottom, talities of larval and juvenile groupers, usually gentle during 3-10 dah to permit feeding and precededby whirling, in Taiwan (Chi et a1. 1998, limit surface sticking, increasing during 11-25 1999), Indonesia (Zafran et al. 2000), Philippines, dah to limit surface aggregation, then high af­ Singapore, Malaysia, Thailand, Japan, and Ko­ ter 25 dah to maintain dissolved oxygen. rea (Bondad-Reantaso et a1. 2002). Death rate Sugama et al. (2001) also suggested tha t tank was highest at 30-32°C and near zero at 23°C or bottom cleaning be started at 9-11 dah and sur­ lower. Infected adults in Korea exhibited anor­ face cleaning after compound feed is being exia, dark coloration, spiral swimming, and added (-17 dah), vertebral deformity (Sohn and Park 1998). Viral 324 'ruCKEI'.

nervous necrosis (paralytic syndrome) in Thai­ ten include darkening, hemorrhaging, ulcer­ land caused anorexia, dark coloration, equi­ ation, and erosion of skin (including the fins), librium loss, corkscrew swimming, and high cloudy eyes, anorexia, loss of coordination, mortality (Danayadol et al. 1993). Loss of coor­ overexpanded gas bladder, liver hemorrhage, dination, curled body, and upside-down swim­ and mortality. Vibrio algil1oIyticlls, Vibrio pamliae­ ming also are possible. molyticlls, other Vibrio spp., and Pseudomonas Grouper larvae and juveniles are very sus­ spp. have caused hemorrhagic septicemia of ceptible to VNN, the most serious disease in caged groupers (Baliao et al. 1998; Bondad­ SoutheastAsian grouper ha tcheries (Koesharyani Reantaso et aL 2002; Koesharyani et al. 2001). et a!' 2001; Yuasa and Koesharyani 2001). The Streptococcus spp. have caused systemic infec­ main signs of infection in polka-dot groupers tions with weakness, abnormal swimming, and are appetite loss and sudden mass mortality sometimes corneal hemorrhage (Baliao et al. during 10-20 dahl weak swimming and mass 1998). In Japan, pasteurellosis was a major dis­ mortality after a few days during 21-30 dah (the ease of young redspotted groupers (Sako 1996). most sensitive age), resting at the bottom with Parasitic ciliates and dinoflagellates mortality spread out during 31-120 dahl and tTrichodinn sPP" Cnjptocaruon irritans, and floating near the surface with some mortality Amyloodiniu11l sp.), have caused high mortality during 121+ dah (grow out). The PCR test re­ of early and older juveniles (Baliao et al. ]998). sults can be obtained in less than 6 h. Because Anorexia, scraping againstsolid objects, surface crowding promotes VNN, Sugama et al. (2001) swimming, tail rot, skin damage, mucus, and recommended that polka-dot grouper eggs be scale loss are common signs. Infestation by stocked at 4-5/L with the goal of 1.5 larvae/L Cryptocaryol1 irritanscombined with infection by at 10 dah. Vibrio hnrveyiled to mortality of brownmarbled Sleepy grouper disease in Singapore prob­ groupers (average 578 g) fed trash fish inearthen ably was caused by an iridovirus introduced ponds in Saudi Arabia (Afifi 2000). with wild juvenile groupers imported for cage The most common problem reported for farming (Chua et al. 1994) and also has occurred western North Atlantic species has been infes­ in Indonesia (Kaesharyani et al. 2001) and Tai­ tation of the gills, eyes, and skin by monoge­ wan (Bondad-Reantaso et al. 2002). Taiwanese nean or protozoan parasites, which feed on grouper iridovirus is suspected of causing high blood, skin, and mucus. They also frequently mortality of groupers in Taiwan since 1992; in­ infest caged Asian groupers, causing small hem­ fected fish were anemic and swam in circles orrhages and secondary bacterial infections (Chou et al. 1998). In southern Thailand, what (Baliao et aL 1998; Koesharyani et al. 2001). might have been an iridovirus caused lethargy, External bacterial infections have been dark coloration of tail and fins, and balance loss treated with freshwater, formalin, or iodine (100 in early juveniles (Baliao et al. 1998). In 2000, ppm for 1 h, Sugama et al. 2002b) baths. Proto­ this disease caused more than 80% mortality of zoans have been controlled by baths of fresh­ caged groupers in North Sumatra. Anorexia, water, chloroquine diphosphate, formalin, or liver hemorrhage, and external whitish blisters, copper salts (copper is dangerous in salt wa­ hemorrhage, and exudation are possible. ter). Monogeneans have been controlled .by Other viral pathogens and diseases include baths of freshwater, hydrogen peroxide, golden eye disease (astro-like virus), causing praziquantel, mebendazole, formalin, or trichlo­ anemia, suffocation, and mortality; red grouper rfon. Parasitic copepods have been treated with reovirus, causing anorexia, lethargy, and mortal­ hydrogen peroxide, freshwater, or formalin ity; herpes, causing dark patches, light and dark baths, and isopods with formalin baths or re­ nodules, paralysis, exhaustion, and mortality; moval with forceps. A ISO-ppm hydrogen per­ and Iyrnphocystis, causing numerous external oxide bath for 30 min was recommended for nodules but not much mortality. Preventing ex­ monogeneans and copepods by Sugama et al. posure is the best approach for viruses. (2001);however, Sugama et al. (2002b) also used Bacterial diseases usually result from me­ a 30-1ninfreshwater bath followed by a 24-h bath chanical injury (e.g.. from excessive handling) in 30 ppm formalin. A 50-ppm formalin bath for combined with low water quality, and signs of- 1 h was used to control leeches infesting GfmUPER CULTURE 3:25

orangespotted groupers (Cruz-Lacierda et a1. shipping groupers at 45-60 dah (15-28 mm 2000). Routine monitoring combined with the TL), 50-100/L, and 23°C for up to 8 h. Mid­ latest rapid detection methods will help prevent stage larvae (35 dah, 15 mm) were much more the spread of disease. The current published sensitive to handling. treatment methods and government regulations In the Philippines, groupers sometimes are always should be consulted before treating fish. harvested from ponds by trapping them in a tri­ Vaccines for certain grouper diseases might be angular stack of parallel bamboo or PVC pipes, available in the near future; (e.g., one for Vibrio which they enter during the night (Baliao et aI. vll1nificlIs was tested on yellow groupers by Qin 1998). Early in the morning, before the fish exit, et al. 2000;one for VNNwas tested onsevenband the pipes are raised with a liftnet, and the fish groupers by Tanaka et al. 2001; and another for are kept in a holding cage (4 x 8 x 1.5 m, mesh Vibrio algil101ytic1I5 was tested on yellow groupers 1-2 em) at 20/m3 for up to a week until being by Zhou et a1. 2002). Many drugs still used in transported. They are starved for at least 24 h, certain regions are dangerous to the environ­ then cooled at a rate of less than SoCper hour to ment, the fish, and/or the consumer. It is better ]8°C and shipped in double polyethylene bags to prevent disease by using vaccines if available inside styrofoam boxes. and practicing good sanitation methods such as stocking specific pathogen-free juveniles, not using raw trash fish for food, and raising fish in Processing and Marketing a clean environment. Spread of diseases from shipping of infected seedstock should be Nearly all species in this family large enough avoided. For maintenance of good health, fac­ to eat are preferred food fish. Groupers usually tors to be avoided are overstocking, underfeed­ are very easy to catch, and in many areas, stocks ing, low oxygen, low water quality, handling in­ have been highly depleted by overfishing. A juries, and lack of sanitation (Baliao et al. 1998). common pattern is for numbers of the primary targeted species to drop below a threshhold, then the nextlargestcommon species is targeted, Handling and Transporting and so on. In the Caribbean region, the shift in availability usually includes Nassau grouper Grouper eggs and larvae should be shipped in ~ E. guttatus ~ coney E. [uluus and 4-mil or thicker, food-grade, polyethylene bags E. cruenuttus, with weight decreasing about half full of oxygen and half full of water, from 14 to 23 kg to -100 g over the years. at the spawning temperature or a few degrees Typical market sizes are 400-700 g in the lower, inside an insulated rigid container. We Philippines, 600-900 g in Singapore, and more have shipped Nassau grouper eggs at 16,000/2 than 1 kg in Kuwait (Baliao et al. 1998). TIle L for 10 h at 23°C. Larvae of other species have usual minimum market size for groupers in the been shipped at 16,OOO/L for at least 2 h at 27°C. United States is 2 kg, but smaller fish can be Juvenile groupers have been shipped in bags sold in specialized markets, especially in large at 120 gil for 12 h, at normal or slightly lower cities. Recent (2002-2005) wholesale prices for than normal temperatures. One 900-g Nassau groupers in the eastern United States were in grouper can be shipped in 10 L of water at 23°C the range $6.10-10.50/kg depending on spe­ with oxygen in a box for 12 h. One 8.6-kg Nassau cies and season. Skinless, boneless yield grouper can be held in 100 L of water at 26°C for groupers is about 36-40%, at least as good with aeration for at least 8 h. On the way to the as channel , tilapia, and hybrid striped market, groupers sometimes are carried with bass. Fillets sold for $15.40-24.10/kg whole­ just enough water to keep them wet. sale and $24.20-37.40/kg retail. In east-cen­ Because orangespotted grouper eggs are tral Florida during July 2003-March 2005, gag most susceptible to stress from early cleavage to and black grouper fillets sold for $28.58-37.40/ gastrulation and least susceptible after eye for­ kg retail. In Nassau (Bahamas), live Nassau mation, handling should be minimized until groupers are sold for $11.00-22.00/kg by deal­ late development (Caberoy and Quinitio 1998). ers who buy directly from fishing boats. Estudillo and Duray (2003) recommended In Hong Kong, live groupers (0.5-2 kg) were 326 l1JCI\ER worth $22-44/kg wholesale, depending on spe­ high as 13.3% and 5.6/L. Recent survival of 20­ cies, but prices could drop to near $13/kg (Chan 40% has been reported (Rimmer et al. 2004). 2001) when supply was high. Cesar et al. (2000) Brownmarbled grouper is a good candidate reported that Hong Kong market prices were for farming in Southeast Asia (Lim et al. 1990; $15-65/kg in May 1999. In December 2002, Supriatna and KoOOo 1990; Chao and Lim 1991; Hong Kong prices were US$9-60 /kg wholesale Chao et a1. 1993; Kohne et a1. 1994; Tucker and and $21-92/kg retail; southern mainland China FitzGerald 1994). They spawn voluntarily in prices were $9-85/kg wholesale and $19-111/ tanks, cages, or ponds, potentially year-round, kg retail (McGilvray and Chan 2003). and can be induced to ovulate. They seem able to eat larger prey at first feeding than some other species, have a shorter larval period, and also Regional Status might grow faster. Larvae were raised at -28­ DC Indo-Pacific to Middle East 30 on rotifers and A rientia, in green water, with survival of 13% from hatching to 40 dah (Lim et Redspotted grouper is a major food fish fanned a1. 1990). When fed trash fish in cages, they can in Japan and nearby countries (Ukawa et a1. grow from 90 to 600-800 g in 6-8 months 1966; Tseng and Ho 1979, 1988; Maruyama et (Anonymous 1986) or 30 to 600 g within 10 a1. 1994; Furuya 1995). They spawn voluntarily months (Chao et al. 1993). in tanks or ponds and can be induced to ovu­ Malabar groupers are farmed in Thailand, late. Larvae can be raised on rotifers, Anemia, Taiwan, the Philippines, Singapore, and nearby and dry feed, in green water (Maruyama et a1. countries (Chua and Teng 1980; Chulavitay­ 1994). They grow to 25-30 mm TL by 40 dah. In anukool et a1. 1985; Huang et al. 1986; Kung­ 1989, 403,000 juveniles (29 mm TL) were pro­ vankij et al. 1986; Maneewong et al. 1986; duced at a Japanese hatchery. The highest sur­ Tookwinas 1990a, 1990b; Quinitio and Toledo vival reported was 34% during 1-47 dah (16% 1991; Ruangpanit 1993; Ogburn and Ogburn from eggs to 47 dah), one of the best for grou­ 1994; Liao et a1. 1995; Chou and Lee 1997). They pers. Malnutritionafter day20, cannibalism, and spawn voluntarily in tanks or ponds and can a viral disease were causes of mortality. be induced to ovulate. Larvae were raised on Orangespotted grouper is a major food fish rotifers, microencapsulated diets, and Artemia farmed in Malaysia, Thailand, Taiwan, nearby to 50 dah, with survival of -14% (Quinitio and countries, and the Middle East (Higuchi et a1. Toledo 1991). Wild Malabar groupers stocked 1980; Abdullah et a1. 1984; Doi et a1. 1991; To­ at 100/m) in cages and given compound feed ledo et a1. 1993; Liao et al. 1995; Duray et a1. grew from 27 to 587 g in 250 d. In the range 215­ 1996,1997; Almatar et a1. 1997;Ordonio-Aguilar 385 g. wild Malabar groupers stocked in a 0.1­ et a1. 1997; APEC 2002a). They spawn voluntar­ ha pond grew 2.43 g/d with FCR of 2.4 (dry) on ily in tanks or cages and can be induced to ovu­ a moist locally made feed (Ogburn and Ogburn late. Larvae were raised at 28-32°C on oyster 1994). A pond harvest density of 1.4 kg/m2 for eggs and larvae, rotifers, copepods, Ariemia,dry 1.25-kg fish was reported (Ruangpanit and feed, and minced fish, in green water, with sur­ Yashiro 1995). Polyculture with tilapia (1 grou­ vival of 12% from eggs to 43 dah (Doi et al. per/15 tilapia) has been successful. 1991); better nutrition probably was the reason Camouflage groupers spawn voluntarily in for the increase in larval survival compared with tanks or ponds and can be induced to ovulate. earlier trials. Later, larvae were raised on only Wild adults were induced to ovulate, and lar­ rotifers and Artetnia, then minced fish was given vae were raised to 20 dah (AQUACOP et a1. to juveniles; survival was 3.2% from hatching 1988). Larvae from voluntary spawning were to 60 dah (Duray et al. 1997). Overdependence raised at 28-29°C in 2.8-m 3 tanks on rotifers, on Artetniawas a cause of mortality. Use of larger Artentia, and compound feed, with survival as tanks (3 m"), starting with small rotifers, pro­ high as 4.7% (Al-Thobaiti and James 1997). longing the use of rotifers (up to 35 dah), and When given a formulated feed in tanks for 12 splitting Arieinia into four meals a day contrib­ months, juveniles stocked at 5/L grew from -57 J uted to better survival. Almatar et al, (1997) re­ to -572 g (-2.8 kg/m ) and those stocked at45/ 3 ported larval survival and harvest densities as L grew to -478 g (-18.2 kg/m ). GROUPER CULTURE 327

Greasy groupers are raised in Singapore, Mediterranean Taiwan, and nearby countries (Chen 1979; Anonymous 1986; Lim et al. 1990; Chao and Lim E. aeneus maturation and growth 199]; Lim 1991; Liao et al. 1995; Chou and Lee in captivity were studied (Popper et al. 1994). A 1997). They will spawn voluntarily in tanks, but growth trial was conducted with dusky grou­ mostly are induced to ovulate. Because wild per Epineplielus nuirginaius juveniles (Caste1l6 males rarely are caught, it is not easy to set up Orvay and Gracia L6pez 1997). Dusky groupers captive broodstock of this species. Hormone­ were induced to ovulate and early larvae were induced sex-reversal of females has been prac­ reared (Glamuzina et a1. 1998b, 1998c). Dusky ticed. Greasy grouper larvae need smaller first x white grouperhybrid eggs were obtained and food (e.g., trochophores) than brownmarbled early larvae reared (Glamuzina et al. 1999). groupers. When fed trash fish in cages, they can grow from 90 g to 600-800 g in 6-8 months Western Atlantic (Anonymous] 986). In the Philippines, greasy groupers stocked at 45 perm3-cage and fed trash Nassau grouper induced-spawning and larvi­ fish grew from 102 to 609 gin 5 months (Aqua culture methods were developed during 1987­ Farm News 1992). 1990 in the and Florida (Tucker Blackspotted groupers were induced to et a1. 1991; Powell and Tucker 1992; Tucker and ovulate, and larvae were raised to transforma­ Woodward' 1996). In the first complete rearing tion (-40 dah, -35 mm TL), with survival of study, 129 juveniles were produced from one -10% during 1-40 dah (Tseng and Chan 1985). spawn in 1990. From fertilization to 98 dah, the Sevenband groupers E. septemjascintus were in­ best survival was 5.0%; during 98-243 dah, over­ duced to ovulate, and larvae were raised to all survival was 97%. Sorrte of those fish were transforrna tion (-60 dah, -30 mrn TL) and be­ tagged and released (Roberts et al. 1995), and yond (Kitajirna et al. 1991). some were raised for more than 14 years. The Leopard coraltrout have been induced to minimum market size of 1.8 kg can be reached ovulate and have spawned voluntarily; they in less than 24 months, with FCR rising from 0.9 were raised to at least 152 dah (Chen et al. 1991; to 1.8. Masuma et al. 1993); culture in saltwater cages In 1994, Watanabe et al. (1996) raised 5,651 is being developed in Taiwan (Liao et al. 1995) Nassau groupers to 62 dah in two 33.8-mJ tanks and Japan (Liao 1997). Induced ovulation ofripe in a 70%-shade greenhouse at 24-26°C. The best squaretail coraltrout is routine (Tucker and survival from hatching to 62 dah was 1.4%. FitzGerald 1994). Gags were induced to ovulate and eggs and King grouper production in commercial early larvae were described (Roberts and numbers by Taiwanese hatcheries has been Schlieder 1983). Red groupers were induced to reached during the last few years (Chan 2001). ovulate, larvae were raised, and six juveniles Broodfish are large and difficult to handle, but fed pinfish Lngodon rhomboides in aquaria dur­ this species grows very fast (Rimmer 1998) and ing 66-557 dah reached -168 rum SL (Colin et is in much demand. al. 1996). The Gondol Research Institute for Maricul­ Goliath groupers produce many eggs and ture at Bali reported raising 54,000 early juve­ grow fast, but the broodfish are large and would nile polka-dotgroupers (mean 25 rnm TL) in two be difficult to handle and expensive to main­ 10-m3 tanks during 1999, with survival of 45% tain. Wild juvenile Goliath groupers fed fresh and 54% from hatching to 43 dah (Siar et al. 2002; fish in floating cages grew from 787 to 1,788 g Sugama et a1. 2002a, 2002b). Fertilization and in 90 d (Cervigon 1983). In circular tanks, 3- to hatching rates each usually were 70%. Expected 6-kg Goliath groupers reached 12-13 kg in 480 routine hatchery survival for polka-dot and d. Similar studies have been done with black brownmarbled groupers was 5% from hatching groupers (Alfonso et a1. 1983). to 60 dah, Average size of larval rearing tanks at Black sea bass, a very high quality serra­ Bali is about 10 m", and eggs are stocked at about nine tha t reaches abou t 3.5 kg, has been lOlL. After reaching 3-5 em in 50-60 d, juveniles spawned and raised experimentally (Hoff]970; are grown to about 10 em in the nursery phase. Roberts et al. 1976; Tucker 1984,1980; Berlinsky nCKER

et a1. 2000; Howell et al. 2003). However, it does weight gain 25-84 g, harvest size S50-1AOD g, not grow very fast, in nature to only 44 g the and time 16-18 months (age -19-21 months). first year, 126 g the second year, and 220 g (198 Greasy groupers performed best, reaching 1,400 mm SL) the third year (Cupka et al. 1973; Mer­ gin 16 months (age -19 months). In 1995, costs cer 1989). Possibly, growth rate can be doubled and revenues per 1 m' of cage volume were or tripled in culture (Petrocci 1995). Berlinsky $27.16 and $56.46 for greasy grouper and $22.06 et a1. (2000) reported that the largest 10% of and $58.01 for redspotted grouper. reared black sea bass weighed 156 g at 380 dah. In 1994 in Thailand, a l,300-g live grouper Sand Diplecirum !OrTILOSIl111 were in­ could sell for as much as $20.78, about 2.5 times duced to ovulate with HCG, and larval devel­ the price of a 500- to 800-g dead grouper opmentwas described (Manrique 1987a, 1987b). (Ruangpanit and Yashiro 1995). Ten-centimeter juveniles (age 3-4 months) stocked at 10-16/ 3 Eastern Pacific m" in 28- to 50-m cages reached 600-800 g in 7­ 8 months and 1,200-1,400 gin12-]4 months (age Spotted sand bass were raised from voluntar­ -15-18 months). A cage could produce at least ily spawned eggs and grown out in cages to 300 kg of l,200-g fish with a value of $3,431 per adults, which were selectively bred (Aviles­ cycle. Costs were: cage $61.76; juveniles $294.12, Quevedo et al. 1995; Martinez-Diaz et al. 2001). feed $423.53, and labor $141.18, leaving Paralnbmx nebuiifer spawned $2,510.78 net income. voluntarily (Gadomski and Caddell 1996). Liao et al, (1995) reported that the price of wild 25-mm groupers in Taiwan was as low as $0.20-0.40 in fall and winter, but 60- to 90-mrn Economics juveniles could be sold for $2-3 in the spring when demand was high. To grow groupers from Although studies have been done on spawning 60 mm (age 3 months) to 400-800 g in ponds, and larval rearing of dozens of serranids, grou­ stocking density was 20,000-70,000 fish/ha, FCR per farming fluctuates because of the lack of 4-5 for trash fish and 1.2-1.4 for moist pellets, sustained hatchery production for most species time 10-14 months (age -13-17 months), and and variability in the (mostly decreasing) sup­ harvest density 10,000-30,000 kg/ha. ply of wild ivveniles. Inconsistent quantity and During 1998 in Taiwan, orangespotted quality of trash fish combined with a lack of grouper eggs sold for $0.0001-0.0007 and 7-cm. economical compound feeds also has been a juveniles for $1.10; however, king grouper eggs handicap in some areas. The extended larval were worth $0.396, 2.5-em juveniles $2.30, and period and variable survival make the effortand 7-cm juveniles $7.00 (Su 2002). cost of producing grouper juveniles in hatcher­ During 2001-2002 in Bali, prices for polka­ ies higher than for most other types of fish. Once dot and brownmarbled groupers were about the juvenile stage is reached, survival should $0.00025 and $0.0001-0.00015 for eggs, $0.60 and be near 100%. Good growth and feed conver­ $0.30 at 4 em, $1.30 and $0.90 at 11 ern, and $2.75 sion will make grow out economical; however, and $1.75 at 17 cm (Siar et a1. 2002). water quality must be maintained at a higher In a bioeeonomic analysis of Malabar grou­ level than for purely estuarine or . per grow out in Taiwan, Miao and Tang (2002) In many areas, cages have been preferred for recommended the use of concrete rather than grow out, but there has to be good water ex­ earthen ponds, salinity of 29-30 ppt, and change and a minimum of 3-5 m of water be­ ground shrimp rather than fish in moist pellets. low the cages to limit diseases and self-pollu­ tion. Ponds are not necessarily better. If the value of the fish is high enough, recirculating Future Prospects tank systems can be justified. Li (1995) reported that when four grouper With one larval rearing tank, six nursery tanks, species (typically 1,350 juveniles stocked in a 3 six phase-I grow-out tanks, and 24 phase-If x 3 x 3 m cage) were grown out in China, FCR grow-out tanks, it would be possible to raise with trashfish was 7-8, survival 60-70%, monthly 54,545 kg of Nassau groupers worth $382,000- G]{OUPER CULTIiRE 329

$545,000 in 2 years (Tucker 1999). Hatchery Juveniles and adults are among the hardiest of costs would be less than $1.00 per weaned, 70­ fish and their feed conversion is good. d-old, 50-mm juvenile. In nursery tanks, those Year-round egg production in environmen­ early juveniles would be raised to robust 125­ tally controlled spawning tanks will permit a mrn juveniles (pellet size increasing from 1.6 to year-round supply of market-size groupers. 2.4 mm). During grow out, those juveniles Good management of broodfish and the larval would be raised to 230 mm (pellets up to 4.0 rearing environment (including the bacteria) mm) in the first phase and 450 rnm, 1.8 kg (pel­ will allow production of specific-pathogen-free lets up to 9.5 mm) in the second. The culture juveniles, which greatly reduces the risk for period would be about ]15 d shorter for 1.36- grow-out operations. Vaccination of groupers .kg fish, but to take advantage of their fast growth for major diseases (e.g., vibriosis) is likely in in this size range (~4.0 g/d), the groupers the future. should be raised to at least 1.8 kg. Overall feed cost would be $0.70-1.40/kg of fish (feed $0.55­ References 1.] O/kg, overall FCR 1.275, overall PCR 0.757). The loB-kg groupers would be worth at least Abdullah, M. A. S., S. Akatsu, K. Al-Abdul-Elah, and $7-10/kg whole and at least $17-22/kg as fil­ S. K. Teng. 1984. Refinement of spawning and lets (wholesale). larval rearing techniques in hamoor (Epinephelus A $36.3 million recirculating intensive tank tauvinav. Kuwait Institute [or Scientific Research, fann (including hatchery and processing plant) Annual Research Report (1983):55-57. Sufat. in the southern United States that can produce ACIAR (Australian Centre for International Agricultural 7.8 M kg of live marine warmwater fish per year Research). 2000. Improved hatchery and grow-out technology [or grouper aqnaculrure in the Asia­ with gross sales of $50.3 M has been estimated Pacific region. Australian Centre for International to have $18.7 M annual operating costs and $2.9 Agricultural Research, Project AS/97173, Canberra. M depreciation (Tucker 1999). Itemized operat­ Afifi, S. H. 2000. Association of Vibrio harveyi with ing costs would be feed 33% (assuming $0.55/ mortal ities in the brown-, kg), utilities 12%, labor 40%, marketing, sales, Epinephelus [uscoguttatus (Forsskal) raised un­ freight 10%, and miscellaneous 5%. der grow-out culture conditions. Assiut Veterinary For fish farming in the southeastern United Medical Journal 42:175-182. Ahmad, 1'., Rachrnansyah, and N. N. Palinggi, 2002. States and Caribbean region, Nassau groupers, The availability and use oflocal ingredients in fish gags, black groupers, and Goliath groupers are feed for humpback grouper grow out. Pages 95­ good candidates. In the United States, grow out 101 in Report of the Regional Workshop on Sus­ most likely will occur in recirculating tanks or tainable Seafarming and Grouper Aquaculture, offshore cages. Medan, Indonesia, 17-20 April 2000. Asia-Pacific Traditional Asian-style grouper farming is Economic Cooperation Publication No. 202-FS­ 04.1, Singapore. not appropriate for the United States. Inshore Ahmad, 1'.A., S. M. El-Dakour, and C. R. El-Zahar. 2000. areas suitable for cage culture (e.g., well-flushed Growth and survival of the grouper Epinephelus deep bays) are very rare in the southern United coioides(Hamilton) at different loading rates in tanks. States. Collection of wild juveniles is seasonal, Aquaculture Research 31 :603--608. unreliable, and in most cases, unethical and il­ Alfonso, A., C. Fermin, and G. Robaina. 1983. legal. The use of trash fish for food is wasteful Aspectos biologicos del genero Mvcteroperca y and could result in disease transmission. It su potencial de cultivo in Mernorias V Simposio, Asociacion Latiuournerica de Acuicultura, would be unwise to initia te suchnonsustainable Septiernbre 5-9, 1983. Valdivia, Chile, 1:2. methods. Ali, A., M. T. Mohn Sallch, and A. Z. Siti Noraziah. Sophisticated hatcheries with good environ­ 1998. Food preference of early larvae of brown­ mental control and very dedicated staffs are nec­ marbled grouper. Aquaculture Asia 3(4):39-43. essary for raising groupers from eggs. Obtain­ Al-Marzouk, A. E. 1999. Association of Pseudomonas ing eggs from most species is relatively easy. anguilliseptica with mortalities in cultured marine orange-spotted grouper Epineplielus coioides in However, larvae of most are fragile, and re­ Kuwait. Fish Pathology 34: 167-168. ported survival from eggs to juveniles often has Alrnatar, S., T. Al-Ahrned, S.-K. Teng, and K. Al­ been only 0-1 %. With a good hatchery and staff, Abdul-Elah. 1997. Advances in research on routine survival of at least 10% is attainable. aquaculture of the blue-fin porgy (Sparidcntex 330 TUCKER

liasta Valenciennes) and orange-spotted grouper la cabrilla arenera. Centro Regional Investiga­ (Epillephelus coioides Hamilton 1822) in Kuwait. ciones Pesquera, La Paz, Mexico. Pages 15-17 in Martinique 97: island aquaculture Baliao, D. D., M. A. de los Santos, E. M. Rodriguez, and tropical aquaculture. European Aquaculture and R. B. Ticar. 1998. Grouper culture in Society, Oostende, Belgium. brackishwater ponds. Southeast Asian Fisheries Al-Thobaiti, S. A., and C. M. James. 1997. Breeding Development Center, Aquaculture Extension and grow-out production of the grouper Epin­ Manual No. 24, Tigbauan, Iloilo, Philippines. ephelus polyphekadion (Bleeker) in Saudi Arabia. Beets, J., and A. Friedlander. 1998. Evaluation of a Pages 20-21 in Martinique 97: island aquaculture conservation strategy: a spawning aggregation and tropical aquaculture. European Aquaculture closure for red hind, Epinephelus guttatus, in the Society, Oostende, Belgium. U.S. Virgin Islands. 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