sea cucumber

JOURNAL OF THEWORLD AQUACULTURE SOCIETY

Vol. 44, No. 5October, 2013

Spawning and Larval Development of the Four-Sided SeaCucumber, Isostichopus badionotus (Selenka 1867), under

Controlled Conditions

Magali Zacarías-Soto, Miguel A. Olvera-Novoa1,Saul Pensamiento-Villarauz and Itzel Sanchez-Tapia

Centro de Investigacion y de Estudios Avanzados del Instituto Politecnico Nacional (Cinvestav)Unidad Merida, 97310 Merida, Yucatan, Mexico

AbstractThe objective of this study was to generate information for the development of Isostichopus

badionotus culture protocols by determining optimal spawning methods, egg production, length andduration of larval stages, and larval habitat settlement preferences. Studies performed during thespawning seasons of 2010 and 2011 showed that this organism spawns between July and Novemberwithout mechanical or chemical induction. The larval development has five well-defined stages: early,mid and late auricularia, doliolaria, and pentactula. Juveniles (654.3 μm length) were obtained onaverage 25 days after fertilization (DAF), when larvae were incubated at 25 ± 1 C. No preference wasobserved for any of the materials used for settlement of the larvae. The easy adaptation of I. badionotusto the culture conditions and the competence of the larvae to complete metamorphosis while feedingsolely on microalgae concentrates make this species an excellent candidate for farming. However, itis necessary to continue designing techniques for larval culture and to establish appropriate captivityconditions that allow multiannual reproduction.

The high demand in Asian countries for seacucumbers and their products has led to theiroverexploitation worldwide, particularly in low-income countries (Purcell et al. 2011). This sit-uation is similar in America, as overfishing ofthe sea cucumber Isostichopus fuscus (Ludwig)in the East Pacific has brought the populationsdown to critical levels in both Ecuador andMexico (Toral-Granda 2008), while in recentyears, the four-sided sea cucumber, Isosticho-pus badionotus (Selenka), has been subject toan intense fishery in the Yucatan Peninsula,the Caribbean islands, and Central Americancountries. For this reason, in 2011, the Gov-ernment of Mexico established a closure periodfor this species from May 21 to December 31(DOF 2011). This situation has required thatprotective actions be taken, including precau-tionary fishery management and evaluation ofthe possible culture of I. badionotus . Further-more, there is an interest in understanding the

1 Corresponding author.

reproductive biology of these organisms as afirst step in the implementation of technologiesfor the establishment of sea farms in the area.

Isostichopus badionotus is an epibenthic,sediment-feeding holothurian belonging to thefamily Stichopodidae. It is localized primarilynot only in areas with sea grass but can alsobe found in coral reefs (Guzman and Guevara2002). This species can be found off theWestern Atlantic coast from South Carolina toBrazil, including the Antilles, and in Sao Tomeand the Gulf of Guinea in the Eastern Atlantic(Miller and Pawson 1984).

Guzman et al. (2003) described I. badiono-tus as a gonocoric organism without externalsexual dimorphism. The sex ratio of the pop-ulations in the fishing areas is usually 1:1,composed mainly of sexually mature organ-isms, indicating reproductive aggregations; con-sequently, the fisheries have a direct effect onreproductive sea cucumbers. Similar to othertropical holothurians, the reproductive cycle ofl. badionotus is characterized by spawning in

© Copyright by the World Aquaculture Society 2013

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LARVAL DEVELOPMENT OF ISOSTICHOPUS BADIONOTUS 695

the warmer months with peaks in July–August;this species has asynchronous maturation thatresults in multiple spawns throughout the warmmonths, with higher prevalence during the sum-mer. (Guzman et al. 2003; Foglietta et al. 2004).

This paper reports for the first time the con-trolled spawning of I. badionotus under cultureconditions. It describes preliminary studies onbroodstock management for obtaining sponta-neous reproduction in captivity, as well as thedevelopment and methods of larval culture toobtain juveniles under controlled conditions, asa contribution to the farming technology for oneof the most commercially important sea cucum-ber species in the tropical Western Atlantic.

Materials and Methods

Collection and Maintenance of Broodstock

The breeding culture was conducted duringtwo reproductive seasons at the CinvestavMarine Station in Telchac Puerto, Yucatan,Mexico. The first reproductive period tookplace from May to November 2010 with 78organisms (36 females and 42 males) and aninitial average wet weight of 533 ± 154 g. Thesecond period was carried out from April toOctober 2011 using 43 breeders (25 femalesand 18 males) with an initial mean wet weightof 870 ± 174 g, and mean length of 26 ± 3.5 cm.

Breeders were caught every year in Aprilby scuba diving at 24 m depth in front of theport of Sisal, Yucatan, Mexico, and transportedto the Marine Station facilities in coolersfilled with 24 C clean seawater, with aerationand oxygenation. In the laboratory, the seacucumbers were slowly acclimatized in thecoolers to the laboratory water temperature of25 ± 1 C and were then washed individuallywith fresh water to remove external parasitesbefore being placed into the reproduction tanks.

Culture Systems

The broodstock culture system consisted ofthree 700-L rectangular fiberglass tanks, eachcontaining 30-cm deep water in a tempera-ture and light-controlled laboratory (fluorescentlamps 75 W). Seawater for the system wasobtained from a 24-m deep beach well, passed

through a sand filter and treated with UV radi-ation prior to use. The tanks were part of aclosed recirculating system formed by a sandfilter, a biological filter, continuous UV radi-ation (2 × 40 W), and a water chiller fixedat 25 C. The water quality parameters in thetanks remained constant, with a temperatureof 25 ± 1 C, pH 8, salinity of 34 ± 1 g/L, anddissolved oxygen between 5 and 7 ± 0.1 mg/L.Feces were siphoned every third day and thewater lost during the process was restored. Thephotoperiod was maintained at 14:10 h (light :darkness).

The larval culture system consisted of twelve250-L square fiberglass hatchery tanks with30-cm deep water and was connected to theclosed broodstock system. Water entering thissystem was additionally treated using threecartridge filters (10, 5, and 1 μm). The waterquality parameters remained similar to thoseof the broodstock tanks. From the third dayafter hatching, approximately 30% of the totalvolume was replaced, a rate that was increasedto 100% replacement per day once the larvaereached the mid auricularia stage (7 days afterfertilization [DAF]).

Feeding Protocols

Two feeding protocols were tested: In thefirst year, the broodstock were fed with com-mercial tilapia and rabbit feed which wasground, supplemented with Spirulina powder,and blended with disinfected beach sand (30 gfood/kg sand). In the second year, the food waschanged to a mixture of ground Ulva sp., Sar-gassum sp., and Macrocystis sp. meals (BajaKelp, Ensenada, B.C., Mexico) blended withdisinfected beach sand (30 g algae mixture/kgsand). The food was changed every third dayto prevent fungus formation.

Larvae were fed twice a day with a mixtureof commercial concentrates of Tetraselmis sp.and Isochrysis sp. microalgae (Instant Algae,Reed Mariculture, Inc., Campbell, CA, USA)following the feeding protocol recommendedby Agudo (2006) of 20,000, 20,000–30,000,and 30,000–40,000 cells/mL for early, mid,and late auricularia, respectively. Once the

696 ZACARIAS-SOTO ET AL.

larvae reached the last auricularia stage, theanimals were cofed with live in situ-producedChaetoceros muelleri and/or concentrates ofThalassiosira weissflogii (Instant Algae, ReedMariculture, Inc.) to induce metamorphosisto doliolaria and the settlement of pentactulalarvae. This protocol was maintained until thejuveniles reached 3 cm in length, after whichthey were fed with powdered Macrocystissp. and Sargassum sp. sieved at 55 μm andenriched with Algamac-2000 (Bio-Marine, Inc.,Hawthorne, CA, USA).

Spawning

During May and June 2010, different pro-tocols to induce spawning were unsuccessfullytested using the methods recommended for theJapanese sea cucumber, Apostichopus japoni-cus (Selenka), including thermal stimulation,dry treatment, and water pressure (Yanagisawa1998; Renbo and Yuan 2004; Xilin 2004; Xiyinet al. 2004); however, the first spawn wasobtained spontaneously without any treatmentin July when the natural reproductive period ofI. badionotus began (Guzman et al. 2003). Dur-ing the first reproductive season, the broodstockreproductive behavior was monitored monthlyfrom the first full moon of May until Novemberand every half an hour from 2000 to 0100 h, 5d before and after the full or new moon. On thebasis of the information recorded, the number ofdays of observation for the 2011 reproductiveseason was adjusted, so the broodstock weremonitored only 2 or 3 d before and after thefull or new moon, maintaining the same sched-ule. No spawn was observed after November2010 and October 2011.

During 2011, two protocols were used forspawning. In the first one, females and maleswere maintained in the same tank, in whichthe reproductive behavior of females wasstimulated by the release of sperm. In thesecond protocol, sperm was collected directlyfrom the males’ gonopores and poured into thefemales’ tank. Water circulation was stopped,and approximately 1 h later, females began toshow spawning behavior. Once spawning wasimminent, the females were carefully moved to

250-L tanks for egg release, after which thefemales were returned to their respective tanks.

To estimate the total number of eggs pro-duced, some spawns were collected directlyfrom the gonopores in 1-L buckets after beingreleased by the females. Three random 1-mLsamples were taken per spawn, and the eggswere counted and measured using a stereo-scopic microscope (Leica Microsystems GmbH,Wetzlar, Germany) and a phase-contrast micro-scope (Motic Instruments, Inc., Richmond,British Columbia, Canada). A final concentra-tion of 1 × 106 sperm/mL was poured intoeach 250-L larval tanks for fertilization of thereleased eggs.

Larval Development

A mixture of eggs from multiple femaleswere maintained in the hatching tanks ata density of 1–2 eggs/mL without aerationor water replacement during the first 12 h;after that, a partial replacement of water wasperformed to remove the sperm, and a gentleflow of air was introduced to keep the ciliatedblastulas moving.

Embryonic development was followed con-tinuously during the first 24 h after spawning,and the larval development was followed everyday by taking at least 30 larvae per tank andrecording the total length and developmentalstage for each larva. Length was measuredusing the Leica Application Suite version 2.0.0(Build: 292; Leica Microsystems). Juvenileslonger than 3 mm were anesthetized with 2%ethanol–menthol solution in order to measureand manipulate them (Liang et al. 2010; Watan-abe et al. 2012). The data are expressed as themean ± standard deviation.

Settling Substrates

To determine whether the I. badionotus lar-vae have a preference for a type of substratefor settling, several artificial materials, includ-ing nylon mesh, polyethylene (PE) shade-meshwith 50 and 90% light retention, transpar-ent polycarbonate roof sheets, white polyvinylchloride (PVC) roof sheets, and plastic andlimestone plates, were tested.


The plastic plates (30 × 40 cm) were fixedequidistantly using grooved PVC pipes that hadbeen cut in half (Fig. 1A, B). The nylon or plas-tic nets were tied to PVC pipe frames of 25 ×25 cm, forming devices with three levels, eachone with a different material (Fig. 1C). Thelimestone plates were placed vertically by hang-ing in the middle or at the sides of the tanks.All of the substrates were preincubated forapproximately 15 d in tanks that had been inoc-ulated with the diatoms, Navicula incerta andNitzschia fonticola (Agudo 2006) before beingplaced haphazardly in the 12 hatching tanks; allthe substrates types were present in each tank.

Concurrently, it was determined whether thelarvae had a preference for the substrate colorused for settling. In a parallel experiment, blue,white, yellow, and green plastic plates wereused to test different substrate colors (Fig. 1A).The plates were coated with a mixture ofgelatin and Spirulina prior to immersion in thehatching tanks.

The different larval settlement devices wereintroduced into the incubation tanks once dolio-laria larvae had been detected. The number ofjuveniles that settled on each of the exper-imental materials was observed daily for aperiod of 60 d, and settlements were recordedmonthly.

Results

In this paper, spawning of I. badionotusin captivity and larval development undercontrolled conditions is reported. Both feedingprotocols used to feed the broodstock wereaccepted; the sea cucumbers maintained goodphysical condition and they spawned severaltimes without artificial stimulus during thereproductive season (July–November).

Both spawning protocols were successful instimulating females to spawn. Spawning ini-tiated late at night; male reproductive behav-ior was presented before that of the females,which started spawning approximately 1 h afterthe first male began releasing sperm. In eachspawning event, reproductive behavior was firstpresented by the males between 2100 and2300 h. During spawning, each female released

Figure 1. Devices with the different substrates used forthe settlement of Isostichopus badionotus (Selenka) lar-vae. (A) Devices made with different colored plastic platesfixed to polyvinyl chloride (PVC) tubes, (B) transpar-ent polycarbonate plates inserted on PVC tubes, and (C)devices for the settlement made with different meshes anda PVC frame.


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Figure 2. Number of spawns per week during the season 2010. The number above each bar represents the quantity ofeggs produced that week. n/a = data not available.

two or three pulses of eggs with a periodicity ofapproximately 30 min. The largest spawn num-bers were observed between 1 or 2 d before orafter the full or new moon, then the number ofspawning females were grouped per week afterthe first full moon of each reproductive period(Figs 2 and 3).

Eggs and Fecundity

During 2010, a total of 83 spawnings wererecorded, in contrast to 44 recorded in 2011. Itwas observed that during the reproductive sea-son the same female may spawn several times,with at least one spawn per month and an aver-age fecundity of 200,000 eggs per spawn (max-imum 1,066,000 eggs and minimum 13,500eggs). The number of eggs per female dimin-ished during the season, as shown by the differ-ences between July and September when therewere up to 1,066,000 eggs per female, andNovember when the records accounted for a

maximum of 25,000 eggs. The egg diametermeasured was 152 ± 6.19 μm on average, withvariations between 140.62 and 164.7 μm.

Embryonic Development

Table 1 shows the different phases of embry-onic development observed from fertilizationto gastrula. The first polar body was observedbetween 10 and 12 min after fertilization. Thefirst and second egg cleavages were observedat 45 and 60 min, respectively. The first ciliatedblastulas appeared between 3 and 4 h after fer-tilization (Fig. 4A), and the gastrula stage wasreached approximately 20–24 h after fertiliza-tion (Fig. 4B). This stage lasted almost 24 h, sothe first early auricularia larvae were observedapproximately 36–40 h after fertilization.

Larval Development

Under the established culture conditions,it was observed that I. badionotus has five


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Figure 3. Number of spawns per week during the season 2011. The number above each bar represents the quantity ofeggs produced that week. n/a = data not available.

Table 1. Embryonic development times of Isostichopusbadionotus (Selenka) (25 ± 1 C).

Embryonic stageTime after

fertilization (min)

Expulsion of the first polar body 10–12First division 45Second division 60Third division 90Fourth division 112Fifth division 129Sixth division 140Seventh division 171Blastula 180–240Gastrula 20–24 h

well-defined stages of larval developmentbefore reaching the juvenile stage: early, mid,and late auricularia, doliolaria, and pentactula.Table 2 includes the time in DAF in whicheach stage was reached and the mean larvalsize for each stage (n = 30 for each stage).

Table 2. Developmental stages of larval Isostichopusbadionotus. Time in days after fertilization (DAF) wherethe different larval stages and their average size (mean ±standard deviation) were observed ( n = 30).

Larval phase DAF Length (μm) Width (μm)

Early auricularia 2–3 584.9 ± 53.8 385.9 ± 38.7Mid auricularia 4–7 677.0 ± 61.3 464.7 ± 49.7Late auricularia 8–14 991.0 ± 83.9 714.7 ± 77.2Doliolaria 15–19 713.3 ± 57.6 529.6 ± 60.4Pentactula 19–22 447.8 ± 76.5 354.6 ± 74.2Juvenile 23–27 654.3 ± 93.4 395.7 ± 71.2

Early Auricularia

This stage began between 2 and 3 DAF, andlasted approximately 4 d. In this stage, lengthsranged from 440 to 697 μm and widths rangedfrom 325 to 89 μm. The larvae were transparentand had a complete digestive tract with mouth,esophagus, stomach, a very short intestine, andanal opening (Fig. 5A).


Figure 4. Embryonic development stages of Isostichopus badionotus. (A) Blastula (bar = 100 μm) and (B) gastrula (BP,blastopores) (bar = 200 μm).

Mid Auricularia

This stage was reached between 4 and 6DAF. In this stage, the larvae continued togrow, reaching lengths of 592–806 μm andwidths of 391–558 μm (Fig. 5B).

Late Auricularia

The first late auricularia larvae were observedaround the eighth DAF. At this stage, thelarvae reached their maximum length andwidth, with size ranges of 816–1131 and591–851 μm, respectively. Internally, it waspossible to distinguish folding of the axo-hydrocoel and lengthening of the intestine.As the larvae development progressed, it waspossible to observe five hyaline spheres oneach side of the peripheral folds of the cili-ated bands before metamorphosis to the dolio-laria stage (Fig. 5C). These internal and exter-nal changes make this stage the most sensi-tive, with the highest mortality rate of larvaldevelopment.

Doliolaria

During this stage, the larvae acquired thecharacteristic barrel shape surrounded by cil-iated bands. The mouth and anus had disap-peared, so the larvae had stopped feeding. Thelarvae were smaller compared with those inthe late auricularia stage, with lengths rangingfrom 609 to 910 μm and widths ranging from

432 to 704 μm. At the end of this stage, fivetentacles appeared in the medial-ventral regionand two podial pits appeared in the posteriorregion in preparation for metamorphosis to pen-tactula larvae (Fig. 5D).

Pentactula

The pentactula larvae appeared at approxi-mately 19 DAF. This stage is the final phase inthe metamorphosis process before the juvenilestage; these larvae migrate to the bottomwhere they settle and transform into benthicorganisms. Pentactula larvae have five anteriortentacles and two posterior ambulacral podiathat serve as anchors. Concurrently, the firstcross-shaped ossicles appeared over the larvalbody (Fig. 6A).

Juvenile

The organisms reached a mean length of654 ± 93 μm and width of 395 ± 71 μm. Thebody was covered by different shaped ossicles(Fig. 6B), and a calcareous ring surrounding themouth was clearly observed. As the juvenilesaged, the tentacle tips became ramified. Like-wise, they started to develop papillae and smallspots of red-orange pigment on the body wall.After 54 d, juveniles reached a mean size of3.1 ± 0.88 mm and were 10.1 ± 1.3 mm at 65 d,with an increase of 0.6 mm/day. At 90 DAF, thejuvenile population showed a broad size range,


Figure 5. Planktonic larval phases of Isostichopus badionotus. (A) Early auricularia, (B) mid auricularia, (C) lateauricularia, and (D) doliolaria. M, mouth; A, anus; E, esophagus; I, intestine; HS, hyaline spheres; AX, axohydrocoel;St, stomach; LS, left somatocoel; RS, right somotocoel; PT, primary tentacles; CB, ciliary bands; DT, digestive tract(bar = 200 μm).

with lengths between 5 and 40 mm (Fig. 7), aswell as a preference for dark places.

Effect of Material Type and Artificial SubstrateColor on Larval Settlement

Once doliolaria larvae began to appear,different devices for larval settlement wereplaced into the tanks and the number ofsettled juveniles was counted daily. Duringthe experimental period, settlement of juvenileswas not observed in the devices or substratestested, independent of material type or color,indicating that juveniles of this species preferto settle on the bottom of the tank in areasof reduced light. Occasionally, some settledjuveniles were observed on the 90% shadenet, but these juveniles generally migratedto the bottom of the tank 1 or 2 d later.

In contrast to juveniles of most species, I.badionotus formed aggregates underneath themesh devices, shelters, or PVC pipes that wereused as support.

Discussion

The reproductive behavior of I. badionotusunder culture conditions is similar to thatreported for the same species by Guzman et al.(2003) in the wild and also similar to thatreported for other members of the family. Thenumber of spawns as well as the number ofeggs obtained during both experimental periodsis consistent with the presence of matureor partially spawned gonads in accordancewith reports by these authors. Likewise, mostspawning occurred within 2 d before or after thefull or new moon; this reproductive behavior


Figure 6. Benthic developmental phases of Isostichopus badionotus: (A) Pentactula; ambulacral podia (AP), tentacles(Ten) (bar = 200 μm); (B) juvenile at 45 days after fertilization (DAF); tentacles (Ten), digestive tract (DT), ossicles (Os),ambulacral podia (AP), papillae (Pa), calcareous ring (Cr) (bar = 1000 μm).

Figure 7. Juveniles of Isostichopus badionotus 65 daysafter fertilization.

has been reported for other holothurians suchas the sand fish, Holothuria scabra (Jaeger)(Morgan 2000; Battaglene et al. 2002) andAustralostichopus mollis (Levin), which spawnpreferentially in the days following the fullmoon (Morgan 2009). In captivity, this speciesshows peaks of reproductive activity betweenJuly and November, which are the warmestmonths of the year, as was reported previouslyfor this organism in the wild by Guzman et al.(2003) in Panama and Foglietta et al. (2004) inVenezuela.

This study demonstrates that it is possible tokeep the same broodstock throughout the entire

spawning season if they are maintained undercontrolled conditions that permit gonad matu-rity (Xilin 2004), thus avoiding the need tocatch wild animals for each spawn, an aspectthat is especially important in areas where thenatural populations have been depleted by over-fishing. In the case of I. badionotus , brood-stock that are kept under culture conditions assuggested in this paper spawn spontaneouslywithout any type of induction. Similarly, theseorganisms retain a periodicity in their reproduc-tive behavior despite being in captivity, withfemales that spawned at least once a month dur-ing the reproductive season between July andNovember, which is a favorable feature in aspecies with culture potential.

Similar to other tropical species of seacucumbers, I. badionotus shows indirect lar-val development, with the presence of plank-tonic auricularia and doliolaria larvae prior tometamorphosis and settlement as pentactula,as has been reported for I. fuscus (Ludwig)(Mercier et al. 2004), H. scabra (James 2004),A. japonicus (Renbo and Yuan 2004; Xiyinet al. 2004), A. mollis (Morgan 2009), and theCurry fish, Stichopus sp. (Chaoqun et al. 2010),as well as other holothurian relatives. Isosti-chopus badionotus completes its larval periodin an average of 25 DAF at a temperature of25 ± 1 C. Its larval stages can be identified by


morphological changes and variations in size,so it is possible to easily distinguish the stagesof early, mid and late auricularia, doliolaria,pentactula, and finally the juveniles. It is impor-tant to emphasize that the larvae of I. badiono-tus accepted and assimilated the food suppliedin the form of commercial microalgae con-centrates, which makes it easier to supply thefood and therefore eliminates costs attributedto live food production, which are usuallyvery high.

The need to facilitate the collection of post-larval sea cucumbers and for increasing culturearea has motivated diverse studies to identifythe best substrates for larval settling. Althoughthe main materials that have been used arePE and polypropylene sheets impregnated withbenthic diatoms (Xilin 2004) or PE platesand monofilament nylon coated with naturalmicrobial films or N. closterium , the highestsettlement percentages have been obtained forPE (Li et al. 2010). Dong et al. (2010)tested with green, red, blue, white, yellow, andblack plastic plates and they observed that thejuveniles of A. japonicus preferred green, red,and blue substrates.

In this study, I. badionotus larvae did notsettle on any of the substrates tested, andinstead settled in the dark areas in the bottomof the tank, indicating a different behavior tothat reported for A. japonicus (Xilin 2004;Xiyin et al. 2004; Dong et al. 2010; Li et al.2010), which tends to settle on all of the arti-ficial substrates used. In nature, I. badionotusadults are characterized by their distributionmainly in deep protected areas far away fromthe effects of waves (Sloan and von Bodun-gen 1980). Similar behavior is observed injuveniles in culture. Yanagisawa (1998) notedthat settlement may not be related to the typeof substrate but to different factors, possiblya substance that also promotes the fixation ofdiatoms on plastic shelters. In this study, thereis a possibility that the periphyton growing inthe tank stimulated the settlement of the larvaeat the bottom, preferably in darker areas.

Despite low larval survival (2–5%), thetechniques used for culturing this species(maintenance of the broodstock, control of

the physical and chemical parameters, feed-ing protocols, and artificial fertilization tech-niques) are a technological advance. The spon-taneous spawning of this species in captiv-ity was obtained under controlled conditions,and juveniles reached lengths of more than3 cm over a 3-mo period, which is the recom-mended size for release or transfer to tanks inother species such as I. fuscus (Mercier et al.2004), A. japonicus (Xilin 2004), and H. scabra(Agudo 2006).

On the basis of these results, it is necessaryto continue working on the development ofprotocols and techniques for larval culturein order to establish the best conditions forthe management of incubators at a particulartemperature, culture density, feeding protocols,and the design of collectors for settlement,whereas studies on broodstock physiology andnutrition are needed to establish appropriatecaptivity conditions that permit multiannualreproduction.

Conclusions

Isostichopus badionotus is a species withhigh potential for aquaculture in the tropical andsubtropical Western Atlantic due to its excellentadaptation to the culture conditions and spon-taneous spawning in captivity. This organismspawns continuously during summer, with atleast one spawn per female per month fromJune–July to October. Its larval development issimilar to that of other holothurians, with plank-tonic auricularia (three stages), doliolaria lar-vae, and the benthonic pentactula larvae whichtransforms into the juvenile. Considering thehigh cost of the production of live feed, it wasproved that it is possible to raise the larvaeusing mixtures of at least two different speciesof commercial microalgae concentrates.

Although the technology for culture of thisspecies is still in the early stages of devel-opment, it is thought that advances in culturebiotechnology would satisfy the high demandfor the product and at the same time support theeconomy of marginal social sectors in severalLatin American countries that currently exploitthis valuable resource.


Acknowledgments

This research was supported by the Fun-dacion Produce Yucatan under the projects“Technology Development for Breeding andProduction of Sea Cucumber Offspring for itsCultivation in Yucatan” and “Development andTransfer of Technology for Sea Cucumber Fat-tening in Yucatan.” Special thanks to AliciaPoot and Enrique Poot for their enthusiasticsupport for the collection of breeders.

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