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By MATTHEW R. PALMTAG AND G.J.HOLT

Fire ( debelius) are a popular spe- cies in the marine aquarium trade. Wild collection of marine ornamental fishes and invertebrates, such as the fire shrimp, contribute to the pressures on natural popu- lations of coral reef . The development of cap- tive culture techniques would help safeguard coral reefs and is essentialfor the expansion of marine ornamental aquaculture technology. This paper presents a complete and descriptive protocol for the captive culture of fire shrimp. Larvae were fed rotifers (Brachionus plicatilis) and brine shrimp nauplii (Artemia sp.) both enriched with algae (Nannochloris oculata) for the first 12 days. From day 12 post hatch to metamorphosis, larvae re- ceived 0.5g of frozen, pureed shrimp and squid in addi- tion to brine shrimp nauplii enriched with algae. We raised 168 shrimp that metamorphosed into juveniles be- Fig. 1. Adult fire shrimp (Lysmata debelius) tween 75 and 158 days post hatching. All shrimp that metamorphosed into juveniles provided healthy individuals suit- shrimp that metamorphosed into juveniles between 75 and 158 able for the marine aquarium trade. days post hatching. All shrimp that metamorphosed into juve- niles were healthy and suitable for the marine aquarium trade. Introduction Techniques described in this paper lay the ground work for fu- Ornamental fish and invertebrate collections on coral reefs ture research and development on the production of fIfe shrimp, contribute to coral reef habitat destruction and degradation of possibly contributing to the preservation of marine ornamental wild populations. Cyanide fishing for food fish and ornamentals organisms and coral reefs. is reported to damage coral reefs, and is practiced in at least 15 countries (McManus et al 1997;UNEP/IUCN 1988). Captive Reproductive Biology cultivation of marine ornamental organisms is a possible "envi- Unlike the majority of decapod , fire shrimp are ronmentally friendly" solution to supplying the growing simultarieous hermaphrodites (Fletcher et al. 1995), meaning an aquarium hobby with coral reef organisms. While many fishes individual can simultaneously function as a male and female. and invertebrates sold in the freshwater aquarium industry are Similar findings were reported for a closely related , the cultured in captivity, more complex requirements of marine or- peppermint shrimp (Bauer and Holt 1998). namental organisms have impeded the development of marine ornamental aquaculture technology. Development of this tech- nology requires advancements in captive rearing protocols. We Broodstock Tank developed a protocol for the captive rearing of fIre shrimp (Fig- Broodstock are kept in 76- to l14-L (20- to 30-gallon) aquari- ure 1) becauseinformation is limited, it is a popular organism in ums with standard undergravel filtration, shell hash substrate, the aquarium hobby, and it demands a substantial price, which and small rocks arranged to imitate natural reef habitat. Mated could benefit the growth of marine ornamental aquaculture tech- 'pairs become territorial and need to be separatedfrom other fire nology. shrimp to prevent aggressionthat may result in maiming or death. Fletcher et at. (1995) published the only information pertain- In our experience, aggressionbetween the individuals of a mated ing to the captive rearing of this species. They provided infor- pair does not occur, however, aggression is common when three mation about rearing the fire shrimp in captivity, however they or more fire shrimp are together. Physical parameters for the did not provide a detailed protocol. The purpose of this work is successful breeding of fIre shrimp are as follows: temperature to provide the details of a protocol we used for rearing fIre shrimp 25° to 28°C (77° to 82°F), salinity 33 to 37 ppt, photoperiod 12 in captivity. The methods used in this paper are an enhancement hr light: 12 hr dark, and pH 8.0 to 8.2. Under these conditions, of culture techniques used to rear peppermint shrimp (Lysmata fire shrimp in our laboratory have produced 500 to 3,500 eggs wurdemanni, Riley 1994). With this protocol we raised 168 every 10 to 20 days. ... ~

Relocation of Broodstock with a 100-L (26-gallon) external biofilter (Figure 4). Six 18-L (5-gal- Spawning of fife shrimp can be Ion) PVC rearing chambers (Figure achieved in the brood stock tank, 5) with 48-um (0.DO2-in.)nitex mesh, however air stones, under gravel fil- are placed in the tank. Dimensions tration, and changing tank environ- of rearing chamber are: 25.4 cm (10 ments often result in high larval mor- in.) diameter and 46 cm (18 in.) tality. For successful captive cultiva- height. Rearing chambers concen- tion, brooding fife shrimp should be trate larvae and food, thus increas- removed from the tank and placed in ing food encounter rates. Rearing a hatching/rearing chamber in a lar- chambers also provide shelter for the val rearing system (described later) larvae and decreasethe possibility of 9 tol9 days after eggs appear on the physical damage from aeration, wa- swimmerets. Our protocol is to move ter exchange and heating elements. the brooding fire shrimp into the lar- The hatching chamber is a plastic val rearing system as close to hatch- cylinder used to contain the parent ing time as possible, and return the in the rearing chamber for hatching shrimp to the broodstock tank imme- and allows removal of the parent, diately following larval release. Fer- minimizing the chance of acciden- tilization of new eggs occurs after tally capturing and removing larvae. molting which can take place 0 to 24 The cylinder (Figure 3) is made of hours after larval release. rigid plastic mesh with 1 x 1 cm (3/8 Recognizing the eggs on the fifst in. x 3/8 in.) holes, 60 cm (24 in.) in day of adhesion to the swimmerets length, and 22 cm (8 3/4 in.) in di- is extremely difficult becauseof bold Fig. 2. Adult fire shrimp carrying eggs ameter, fits snug into the rearing pigmentation throughout the body of chamber. A raised level of 1 x 1 cm the adult shrimp. The swimmerets of (3/8 in. x 3/8 in.) rigid plastic mesh, a fire shrimp carrying eggs (Figure 22 cm (8 3/4 in.) in diameter, is fas- 2) will appear darker than the ab- tened in with cable ties 15 cm (6 in.) dominal segmentsabove, in contrast from the surface of the cylinder. to the lighter color of the swimmerets To provide water flow into the of a fife shrimp without eggs. Hours hatching and rearing chamber a 15 x before the eggs are ready to hatch the 3 cm (6 in. x 12 in.) L-shaped I-inch eyes of the larvae inside the egg can PVC standpipe equipped with a 3 cm be observed; this is an indication that (1 in.) air stone is used (Figure 6). A it is time to move the parent. hole, 3 cm (1 in.) in diameter, is cut Care must be taken when relocat- into the hatching chamber 15 cm (6 ing brooding shrimp to avoid dam- in.) above the raised level to provide aging the eggs. Instead of netting the access for the standpipe. The parent fire shrimp, a 500- to 1000- standpipe provides a steady flow of ml (15- to 30-fl. oz) beaker or jar is oxygenated water directly to the par- placed on the substrate of the tank ent fIfe shrimp, which will aid in the and the shrimp is chased into it by release of larvae as they hatch off the hand. Once the shrimp is completely swimmerets. in the beaker, the opening is covered A 5cm3 (2in3) rock is placed on by hand and slowly raised out of the the raised level of the rigid plastic tank. The beaker is then placed into mesh to mimic the parent fire shrimp the larval hatching chamber within tank living conditions (be sure the the rearing chamber (Figure 3) and rock cannot wobble or fall over onto the parent is acclimated over a pe- the broodstock). riod of at least 20 minutes in the bea- ker before it is released into the Removal of Parent Fig. 3. Hatching chamber hatching chamber. within the rearing chamber To remove the parent fire shrimp Larval Rearing System after the eggs have hatched, lift the hatching chamber until the raised The recirculating larval rearing level of the rigid plastic cylinder is system consists of a 400-L (lOS-gal- approximately 3 cm (1 in.) below the lon) round fiberglas tank equipped surface of the water. The parent is hatching brine shrimp nauplii, and Airline Rearing Rearing again chased into the beaker by hand rotifers and brine shrimp nauplii were \ Chamber Chamber and both the parent and mesh cylin- rinsed prior to release into the larval ~ l1~~~~~~tandPiPeTank Standpipe der removed. This method minimizes rearing chamber, using a 48 !lIn '"=;l the possibility of capturing larvae (0.002-in.) nitex mesh sieve). In ad- J with the adult. dition, every day beginning with day Return 1, 200 ml (6.8 fl oz) of algae Line Larval Rearing (Nannochloris oculata) at a concen- tration of -20,000,000 cells per ml Successfully raising the larvae to (0.034 fl oz), was added to each rear- J juveniles is still in the experimental ,. 7 ing chamber to provide nourishment stage. So far about 10 percent sur- for surviving rotifers and brine vival through the larval stage has shrimp nauplii. On days 6-11 algae been accomplished using the follow- .t-;t-Filter enriched brine shrimp nauplii were 1 ) I) "'Drain ing techniques. maintained at a concentration of "'- Pump Physical parameters of the larval -50,000 per rearing chamber. Dead rearing tank are as follows: tempera- "'- larvae, uneaten food and waste were Fig. 4. Recirculating larval rearing system ture 25° to 28°C (77° to 82°F), salin- removed from each rearing chamber ity 33 to 37 ppt, photoperiod 12 hr every other day by use of a vinyl si- light: 12 hr dark, and pH 8.0 to 8.2. phon tube with an inside diameter of The larvae were divided equally into 10 in. PVC 6.35 mm (5/32 in.). On day 10, lar- six 18 L (5 gallon) rearing chambers r- ~ vae in each rearing chamber were (approximately 250 larvae per cham- removed by beaker and transfeued ber). Larvae were moved by gradu- into clean rearing chambers.This was ally submerging a 250-ml (8 fl oz) repeated for all rearing chambers beaker into a mixture of water and 48 urn Nitex Mesh every 10 days. larvae, and transferring the larvae to On days 12-158, algae enriched ~Frame alternate rearing chambers. As the 46cm brine shrimp nauplii were fed at a density of larvae was reduced, the or concentration of -30,000 per rearing 18 in. volume of water in the rearing cham- chamber along with 0.5 g (0.0175 oz) ber was decreased by lifting it up, of frozen food. Frozen food com- keeping the water line above the bot- posed of 50 percent squid (Illex spp.) 48 urn Nitex Mesh tom of the rearing chamber.This con- and 50 percent shrimp (Penaeusspp.) centratesthe remaining larvae so they l was pureed in a blender and refro- can be collected. Larvae were re- zen. Portions of this mixture were Fig. 5. Rearing chamber leasedby submerging the beaker half thawed each morning and adminis- way through the surface of the water tered to the surface water of the rear- and slowly releasing the water and ing chamber. Beginning on day 14 larvae. The standpipe was added to and every fourth day thereafter 4 mls each rearing chamber after the lar- (0.13 fl oz) of Seachemreef calcium vae were divided. Air pressure of the (polygluconate calcium) was added. standpipe was set to a low level, en- abling larvae to slowly circulate around the rearing chamber. Slow Larval Development flow velocity, 100 to 400 ml per min. Fire shrimp pass through at least (3.3-13.3 fl oz per min.), allows the 10 larval stagesbefore molting to the larvae to grasp the sides of the en- postlarval form. Prior to stage 5 the closure; too much current will sweep fifth parapodia became flattened and them off. expanded into long paddle shaped On day 1, each of six rearing structures, which may assistin main- chambers were given -50,000 algae taining position and orientation in the (Nannochloris oculata) enriched ro- water column (Figure 7). Larvae also tifers (Brachionus plicatilis) and use these limbs for capturing food -20,000 algae enriched brine shrimp and holding onto solid structures in nauplii (Artemia sp.). This concen- the water column (Fletcher 1995). "- tration of rotifers and brine shrimp After the final molt to the juvenile Fig. 6. Stand pipe used in the hatching and nauplii was maintained through day stage,the compound eyes are located rearing chamber 5 (Note: cysts were removed after on the head rather than terminally placed on stalks, and the paddle-like tend for this information to enable appendagesbecoOle reduced to the scientistsand hobbyistsalike to re- typical adult appendages(Figure 8). fine this techniqueor developbetter At this tiOle the ShriOlpare no longer ones,expand research, and provide free swiInIning but instead settle onto an alternative to the capture of the bottoOl and sides of the rearing stressedand endangeredwild popu- chamber.This benthic transfonnation lations. Olay enhance their ability to prey on larger, Olore diverse foods and in- Acknowledgements creasesthe aniOlals feeding opportu- Cindy Faulk and Charlotte Kucera nities (Riley 1994). deserve a special thanks for assis- tance in every phase of the study and Fig. 7. Fire shrimp post-larvae (day 68) with ex- Metamorphosis Greg Dirnijian for photographing the panded fifth parapodia Therewere 400 larvaeon day 55, various life stages of the fire shrimp. whenthey werecounted for the fIrst We would also like to express grati- time. Larvae were not countedear- tude to the people and organizations lier for fearthat handling stress might that ultimately made this work pos- increasemortality. The first meta- sible with their financial assistance: morphosisfrom a larva into a juve- Hugh A. McAllister, Jr., Charitable nile occurred on day 75. Previous Foundation, The World Wildlife studiesindicate that the larval cycle Fund, and Texas Sea Grant College of fire shrimplasts for 11to 15weeks Program. (77 to 105 days) (Fletcher et al. L. C.t d 1995).Further research on larval nu- Iterature 1 e tritional quality may decreasethe Bauer, R.T.; and Holt, G.J. (1998). number of days necessary for Simultaneoushermaphroditism in the metamorphasisto thejuvenile stage. Fig. 8. Newlytransformed juvenile fire shrimp(day marineshrimp By day 158,42percent of the lar- 83) (: Hippolytidae): an vaecounted on day 55 hadmetamor- undescribedsexual system in the de- phosedinto juveniles, the remainderdied. We were .unableto capodCrustacea. Marine Biology, 132: 223-235. look at overall mortality and survival,however we estImate~at Fletcher,D.J.; Kotter, I., Wunsch,M. andYasir, I. (1995). Pre- approximately11 percentof the spawnsurvi~ed to thej~verule liminary observationson the reproductivebiology of orna- stage.Survival during the juvenile stagewas hlg.h (appro.Xlma~ly mentalcleaner prawns. Int. Zoo. Yb., 34, 73-77. 99 percent).Six monthsfrom the dateof hatchmg,~e J.uverules Haywood,Martyn; andWells, Sue.(1989). The manualof ma- rangedfrom 30 to 40 mm (1.2 to 1.6 in.) long. At thIStIme they rineinvertebrates.Salamander Books Ltd., Morris Plains,NJ. were suitablefor sale to marine ornamentalretail storesfor a McManus,J.W., Reyes, R. B. Jr., and Nanola,C.L. (1997).Ef- price of $10 each.Even late metamorphosingshrimp provided fects of SomeDestructive Fishing Methodson Coral Cover viable productfor the marineaquarium trade. andPotential Rate of Recovery.Environmental Management Vol. 21, No.1, pp. 69-78. Summary Riley, Cecilia M. (1994). Captive spawningand rearing of the The primary objective of this research was to provide a pro- peppermintshrimp (LysmataWurdemanni). Sea Scope Vol. tocol for the captive cultivation of fire shrimp. Protocols such as 11. this are essential for the development of marine ornamental Simoes,F.; Yasir I., Jones,D.A. (1998).Reproductive Biology aquaculture technology. This technology may allow aquaculture of Lysmatadebelius (Bruce 1983) and L. ambionensis(De to provide a form of coral reef habitat preservation and conser- Man 1888), (Crustacea,Caridea) Tropical Marine Cleaner vation in the near future. Using this protocol we were able to ShrimpsImportant In the SeaWater Aquarium Trade.Ab- raise 168 adult fire shrimp from a single hatch (estimated hatch stract,World AquacultureSociety, Las Vegas,1998,3 pp. 1,500 larvae, 11 percent successrate). Our results suggest that UNEP/IUCN. 1988. Coral reefs of the world, 3 vols. UNEP fire shrimp are appropriate subjects for future research and de- RegionalSeas Directories and Bibliographies.IUCN, Gland, velopment of marine ornamental aquaculture technology. We in- Switzerlandand Cambridge,UK/UNEP, Nairobi, Kenya.

A TEXAS SEA GRANT COLLEGE PROGRAM RESEARCH REPORT. The researchdescribed in this report was funded in part by Project R/M-61 of Institutional Grant NA86RGOO58to TexasA&M University from the National SeaGrant Office, National Oceanicand Atmospheric Administration, U.S. De- Texas partmentof Commerce.TAMU-02-201. September2001. 1M