Aquaculture Research, 2007, 38, 580^587

doi:10.1111/j.1365-2109.2007.01686.x

The effect of unilateral eyestalk ablation and diet on the reproductive performance of wild-caught

Farfantepenaeus aztecus (Ives, 1891) using a closed

recirculating maturation system
Ryan L Gandy,Tzachi M Samocha, Michael P Masser, Joe M Fox, Abdul-Mehdi S Ali, Delbert M Gatlin III & Michael Speed
Texas Agricultural Experiment Station ^ Shrimp Mariculture Research Facility, Corpus Christi,TX, USA
Correspondence: R L Gandy, 2085 Goldenrod St, Sarasota, FL 34239, USA. E-mail: ryanlgandy@hotmail.com

Abstract
Two studies were conducted to evaluate the eects of unilateral eyestalk ablation and diet on the reproductive performance of wild populations of Farfantepenaeus aztecus. In both studies, females in two treatments were unilaterally ablated while those in the control treatment were not. Shrimp in the nonablated treatment and one of the unilaterally ablated treatments received frozen bloodworms (8% BWday 1) and frozen squid (12% BWday 1). The bloodworm component of the diet of the third unilateral ablation treatment was replaced with frozen adult enriched Artemia sp. Ablated female population spawning per night, in both studies, was higher than non-ablated spawning (8.5 and 8.9 vs. 2.6%; 7.4 and 7.5 vs. 2.7% respectively; Po0.05). Replacement of bloodworms with adult enriched Artemia sp. had no negative eect on the number of eggs spawned per ablated female (124 000 vs. 115000 eggs spawn 1; 144 000 vs. 151000 eggs spawn 1 respectively; P40.05). The life span of ablated females fed adult enriched Artemia sp. was 8 and 40 days longer than ablated females fed bloodworms for the rst and second studies respectively. Replacement of bloodworms with adult enriched Artemia sp. resulted in higher hatch and larval survival rates (Nauplius 1 to Zoea 1) (55.0% vs. 46.9% and 44.8% vs. 37.2%), respectively, Po0.05.

Introduction Development of sustainable commercial bait shrimp aquaculture in the United States requires securing a supply of viral-pathogen-free seed-stock through domestication and the application of modern bio-secure maturation techniques. Seed stock used in the growout of Farfantepenaeus aztecus has historically been derived from wild-caught spawners, McKee (1986). Reliance on this source of larvae represents a risk to biosecurity and sustainability of future commercial ventures. Since the mid-1990s, a potentially catastrophic viral pathogen of shrimp, white spot syndrome virus (WSSV), has been consistently found in wild populations of Litopenaeus setiferus within the coastal waters of South Carolina, Texas and Mississippi (Browdy & Holland 1998). Therefore, interest has increased in the development of captive-breeding populations of F. aztecus and methods for commercial-scale maturation for further development of this industry. Eorts to induce maturation in captivity through unilateral eyestalk ablation of the three commercially important species native to the Gulf of Mexico has had limited success. Production of fertile eggs has been documented for the closed-thelycum shrimp, F. duorarum (Caillouet Jr 1972), and F. aztecus, AQUACOP (1975), as well as for the open-thelycum shrimp, L. setiferus (Brown Jr, McVey, Middleditch & Lawrence 1979). Studies by the AQUACOP group with F. aztecus were conducted in outdoor, shaded tanks (12 m2) with no environmental control and at

Keywords: Farfantepenaeus aztecus, maturation, closed recirculation, bait shrimp

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Aquaculture Research, 2007, 38, 580^587 Reproductive performance of F. aztecus under captive conditions R L Gandy et al.

stocking densities of 3.3^8.3 shrimp m 2 using a 1:1 male to female ratio. Broodstock were exposed to a natural photoperiod, ambient water temperature, salinity and pH with high rates of water exchange (200^300% day 1). As a result, waterquality factors showed a high level of variation on a daily basis. The authors mentioned daily temperature uctuations of 2^4 1C and short-term salinity variation due to heavy rain events. Although the AQUACOP (1975, 1977) studies reported that three generations of F. aztecus were produced in outdoor maturation systems, the reproductive performance was poor when compared with other species (e.g., L. vannamei, L. stylirostris, Fenneropenaeus merguiensis and Marsupeneaus japonicus). Ablation was the only method found to be eective in inducing ovarian development and spawning in these species (AQUACOP 1977). In the decades following these initial investigations, research eorts shifted away from species native to the Gulf of Mexico. An exotic species, L. vannamei, was eventually chosen as the principal species for development of commercial shrimp farming in the western hemisphere. Commercial culture of this species has currently beneted from two decades of intensive research into renement of maturation techniques. The use of closed recirculating systems for establishment of biosecure-controlled environments has allowed for the consistent production of viable eggs (Ogle 1992). Intensive research eorts to increase the reproductive performance of captive populations have also led to the development of a basal diet consisting of frozen bloodworms (Glycera dibranchiata) and squid (Loligo opalescens). This

nutritionally complete diet allowed for improved performance in the form of high-quality eggs (Middleditch, Missler, Ward, McVey, Brown & Lawrence 1979; Wyban, Cheng, Sweeney & Richards Jr 1987). In contrast, other studies have reported no adverse eect on reproduction when bloodworms were replaced by adult Artemia sp. (Browdy1985; Naessens, Lavens, Gomez, Browdy, McGovern-Hopkins, Spencer, Kawahigashi & Sogeloos 1997). Our study applies the advancements made in the development and renement of maturation techniques for open- and closed-thelycum species (Primavera 1985; Bray & Lawrence 1992; Browdy1992) as a basis for investigation of the reproductive performance of F. aztecus. Ultimately, the use of alternative nutrient sources for replacement of bloodworms could represent substantial savings to bait production facilities in the United States.

Materials and methods Maturation system The maturation system used in this study was adapted from a basic design described by Ogle (1992) and used by the Gulf Coast Research Laboratory (GCRL, Ocean Springs, MS, USA). Each of the three maturation tanks (9.6 m3; 4.0 m diameter, 0.8 m depth) was lined with a 0.25 mm PVC membrane and equipped with a centre drain (8 cm diameter). Water entering the centre drain of each tank owed by gravity into a common lter system housed in an adjacent room (Fig. 1). Euent from each maturation tank rst emptied into an inner 800 mm bag for

Figure 1 Schematic diagram of recirculating maturation system.

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capture of large particles (e.g., leftover feed, faeces and moults) and then an outer 100 mm bag, which collected small particles and any eggs spawned within the maturation tanks. These lter bags were located on a perforated high-density polyethylene (HDPE) plate used to deliver a homogenized water ow over the subsequent trickling biolter. Below this plate was a at sheet of vinyl acetate lter material (Ogle 1992) for further particle ltration. The trickling biolter (0.475 m3) was constructed of extruded cage netting lled with 2.5 cm (1in.) Bio Pac media (Aquaculture Supply, Dade City, FL, USA) positioned over a 4 m3 sump. A 1.5 hp centrifugal pump attached to the sump was used to discharge biologically treated water into a 5 mm Triton Nautilus FNS diatomaceous earth (DE) lter (Purex, Sanford, NC, USA). Approximately 5% of the water leaving the DE lter was diverted into a 7-L activated carbon lter (Ocean Clear J320 Chemical Filter; Aquatic Ecosystems, Apopka, FL, USA), which discharged directly to the sump. The remainder of the processed water, from the common sump, returned to the maturation tanks. A hydraulic ow meter (FL7403; Omega Engineering; Stamford, CT, USA) was installed on the inow line of each tank and used to maintain a ow rate of 32.2 L min 1. Excess water in the ltration system was diverted back into the sump through a relief valve. The DE lter and in-line-activated carbon lter components were used to improve water clarity and reduce daily system maintenance. Temperature in the system was maintained between 26.5 and 27.0 1C using two wall-mounted air conditioning units (total 1.9 107 J) and a 1.3 107 J submersible heater (Process Technology; Mentor, OH, USA) located in the sump. Lighting to maintain a photoperiod of 12L:12D over the maturation tanks was controlled using programmable timers (Intermatic, Spring Grove, IL, USA). Light banks were located 1.75 m above the water surface of each tank and consisted of 25, 50 and 75 W incandescent bulbs and two 40 W cool white uorescent bulbs. Sunrise simulation was accomplished by activation of each bulb in 15 min intervals from lowest to highest wattage. Once all orescent lights were activated, incandescent light sources were turned o completing the sunrise simulation process. Sunset was accomplished in reverse order to that of sunrise. The two uorescent 40 W cool white bulbs per tank yielded a maximum incident light intensity of 0.5 mE m 2 s 1. Biolter media was inoculated with Fritz^Zyme (Aquacenter, Leland, MS, USA) and pH of the maturation system was maintained at 7.75 0.5 during

both studies by addition of 500 g of sodium carbonate (light soda ash) every 5 days.

Spawning room In a separate adjoining room, 12 individual 200 L (76 cm diameter, 44 cm water depth) spawning tanks made of extrusion-welded black HDPE were used for spawning of individual female shrimp. Each spawning tank was equipped with one air stone (4 1.3 cm; Aquatic Ecosystems), stand pipe (2.54 cm diameter) and an HDPE lid.Water in spawning tanks was maintained at 29 1 1C with a 300 W submersible quartz heater (VT 301; Aquatic Ecosystems). Source water was ltered to o5 mm using a DE lter, foam fractionated and treated with 10 ppm ethylenedinitrotetracetic acid. After each spawning event, tanks were cleaned with a scrub brush and a solution of 10% sodium hypochlorite diluted to 10 ppm with tap water. Tanks were allowed to dry in an inverted position to allow for complete drying, without rinsing the bleach.

Water quality Temperature, salinity, pH and dissolved oxygen (DO) levels were monitored daily using a YSI model 55 DO meter (Yellow Springs, OH, USA), refractometer (model SR6; Aquatic Ecosystems) and a Cole Palmer Model 310 pH meter (Vernon, IL, USA) respectively. Nitrogen species (NH4^N, NO2^N and NO3^N) were measured weekly using HACH methods 10031, 8153 and 8039, Hach (1997), respectively, and a Perken^Elmer Lambda EZ201 spectrophotometer (Norwalk, CT, USA). Water in the experimental maturation system was maintained within the following parameters during the studies: temperature (26.5 1.1 1C), DO concentration (5.76 0.51mg L 1), salinity (35 1g L 1) and pH (7.75 0.5) during both studies. System water during Study 1 had mean NH4^N, NO2^N and accumulated NO3^N concentrations of 0.50 0.25, 1.65 0.95 and 48.5 mg L 1 respectively. Study 2 had mean NH4^N, NO2^N and accumulated NO3^N concentrations of 0.17 0.10, 1.05 0.75 and 52.6 mg L 1 respectively.

Daily system maintenance Excess feed, moults and dead shrimp were removed daily before rst feeding (08:00 hours), at which time the volume of excess feed, number and sex of moults,

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and tag number of dead shrimp were recorded. Dead females were replaced twice weekly with either ablated or non-ablated females. Female shrimp used for replacement were weighed and tagged upon introduction to the corresponding treatment tank. Activated charcoal and DE lters were recharged and changed, respectively, on a weekly basis.

the third tank (treatment 3) received a similar ration of squid; however, the bloodworm component was replaced with frozen adult enriched Artemia sp. To monitor the eect of diet and eyestalk ablation on reproductive performance, each female shrimp was tagged with a numbered latex ring mounted on one eyestalk, Browdy (1985).

Experimental populations of shrimp Two experiments were conducted to evaluate the effects of unilateral eyestalk ablation and diet on the reproductive performance of wild (F) populations of F. aztecus in the aforementioned experimental system. Wild populations of shrimp were collected on two separate collection cruises at a depth of 50 m in the Gulf of Mexico near Freeport, TX. Each population of broodstock was quarantined for 30 days at the Texas Agricultural Experiment Station-Shrimp Mariculture Research Facility (TAES-SMRF), Corpus Christi, TX. During the quarantine period, shrimps were screened for known infectious viruses, including infectious hypodermal and haematopoietic necrosis virus (IHHNV), Taura syndrome virus (TSV) and WSSV Upon conrmation of no viral presence, in the . samples tested by the Texas Veterinary Medical Diagnostic Laboratory (TVMDL, College Station,TX, USA), populations were subdivided into three treatment groups each in a separate tank as follows: the rst population of 180 shrimps was subdivided into three subgroups of 60 shrimps each and stocked into separate maturation tanks for Study 1. Each subgroup consisted of 40 males (20 3.6 g) and 20 females (36 3.3 g). The second population of 180 shrimp was also evenly distributed into three maturation tanks for Study 2 with 30 males (25 4.6 g) and 30 females (41.6 7.9 g) in each tank.

Data collection Shrimp were placed in the maturation system 2 weeks before initiation of each experiment. After acclimation, all shrimp in their respective treatment tanks were weighed. Because larger females are known to produce larger spawn sizes (Primavera 1985), the size consistency within a population at stocking was determined.Weight data were also used to achieve consistency among treatments to reduce the eect of weight dierences on the overall reproductive performance of a treatment group. Monitoring of reproductive performance commenced upon ablation and placement of identication tags on one eyestalk of females. Following ablation, female shrimp in maturation tanks were surveyed nightly at 16:00 hours for stage of ovarian development. Females with well-developed ovaries (stages 3^4) were removed from maturation tanks and placed individually in a spawning tank. The following morning (08:00 hours), previously mated females were removed from spawning tanks and their ovaries were visually examined. Extent of spawning was recorded as a proportion of residual egg mass remaining (e.g., full, partial, none) and females were then returned to their respective treatment tanks. Eggs numbers in spawning tanks were estimated by volumetric samples. Additionally, two sets of three 50 mL samples were removed from spawning tanks by a Hansen Stempel Pipette (1805-C42;Wildco, Saginaw, MI, USA) from the homogenized spawning tank and individually incubated in 100 Petri dishes at 29 1C. Triplicate Petri dishes were also used for nauplii (24 h incubation) and Zoea 1 (Z1) counts (48 h incubation). Stage of ovarian development at spawning, degree of spawn (e.g., partial or complete), total eggs spawned, hatch rate and survival from Nauplius to Z1 were obtained from these observations.

Maturation studies Duration of Study 1 and Study 2 was 142 and 132 days respectively. In both studies, females in two tanks were unilaterally ablated by cauterization. No eyestalk ablation was performed on the females in the third tank. In each study, shrimp in the non-ablated tank (treatment 1) received a diet of frozen bloodworms (8% of the total wet shrimp biomass per day) and frozen squid (12% of the total wet shrimp biomass per day). The same diet was fed to shrimp in a second tank (treatment 2) in which females were unilaterally ablated. Unilaterally ablated females in

Statistical analysis Data were evaluated using the Statistical Package For The Social Sciences (SPSS, version 11.0.1 2003, Chica-

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go, IL, USA). All data were evaluated to determine if requirements for normality and equality of error variances were met before analysis. In the event of inequality or presence of non-normal data, appropriate transformations were made to satisfy these requirements. Data represented as percentages (e.g., survival to Z1, hatch rate and per cent spawn per night) were subjected to arcsine transformation. Untransformed data were used to report descriptive statistics. Treatment dierences were evaluated using ANOVA (a 5 0.05). Signicant dierences in treatment means were identied using Tukeys highly signicant dierence (HSD).

Results Study 1 The reproductive performance of non-ablated females fed bloodworms was signicantly dierent when compared with ablated females fed bloodworms and those fed an adult enriched Artemia (Table 1). Non-ablated females exhibited longer life spans of 85 days vs. 45 days and 53 days, spawning was less frequent 2.61% vs. 8.53% and 8.88% of the female population spawning per night, attained a more advanced ovary developmental stage at spawning 3.7 vs. 3.37 and 3.40, and spawned more completely 2.85 vs. 2.56 and 2.56 than ablated females regardless of diet (Po0.05). Ablated females in both

diet treatments (bloodworm vs. Artemia) performed similarly with respect to the following: life spans 45 days vs.53 days, spawning frequency 8.53% vs. 8.88% of the female population spawning per night, ovary developmental stage at spawning 3.37 vs. 3.40 and completeness of spawn 2.56 vs. 2.56 (P40.05). Average egg production per female per spawn was similar for all three treatments (136 000, 124 000 and 115000 eggs spawn 1) (P40.05). Total egg production per treatment for Study 1 was 3.7 106, 18.0 106 and 18.0 106 respectively. Hatch rate for eggs from ablated females fed bloodworms (44.1%) was signicantly lower than ablated females fed enriched Artemia sp. (58.2%) (Po0.05), which performed similar to non-ablated females (57.2%) (P40.05). The three treatments showed similar results with respect to nauplii count per female (93000, 77 000 and 81000), survival to Z1 (43%, 46% and 55%) and production of Z1 larvae (74 000, 67 000 and 71000) (P40.05).

Study 2 Reproductive performance of non-ablated females fed bloodworms and ablated females fed enriched Artemia sp. was similar with regard to life span, ovary developmental stage at spawning, completeness of spawn and hatch rate (Table 2). Reproductive performance of ablated females fed bloodworms was inferior to non-ablated and ablated females fed enriched

Table 1 Maturation/reproduction performance, Study 1, population 2, (M):1 (F) sex-ratio

Treatment 1
Ablation Diet Study period (days) Female weight (g) Female life span (days) Female population spawning/night (%) Ovary stage at spawning (04) Spawning activity Egg production/female/spawn ( 103) Total egg production ( 106) Hatch rate (%) Nauplii count/female/spawn ( 103)w Survival (N1 to Z1)/female/spawn (%)w Zoea 1 count/female/spawn ( 103)w No Squid1BW 142 35.1 7.0a 85 45a 2.61 2.5a 3.7 0.56a 2.85 0.46a 136.0 69.0a 3.7 57.2 39.1ab 93.0 58.0a 43.3 37.7a 74.0 57.0a

Treatment 2
Yes Squid1BW 142 39.0 7.0b 45 39b 8.53 5.3b 3.37 0.55b 2.56 0.64b 124.0 53.0a 18.0 44.1 35.7a 77.0 50.0a 46.9 37.4a 67.0 49.0a

Treatment 3
Yes Squid1FEAA 142 36.4 5.2ab 53 49b 8.88 5.7b 3.40 0.55ab 2.56 0.05b 115.0 56.0a 18.0 58.2 33.3b 81.0 46.0a 55.0 32.7a 71.0 44.0a

Values are mean standard deviation. Means not sharing a common superscript letter within a row are signicantly dierent (Po0.05). Spawn score:0 5 none; 1 5partial; 2 5 full. wExcluding spawns with zero hatched eggs. BW, bloodworms; FEAA, frozen enriched adult Artemia sp.

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Table 2 Maturation/reproduction performance, Study 1, population 1, (M):1 (F) sex-ratio

Treatment 1
Ablation Diet Study period (days) Female weight (g) Female life span (d) Female population spawning/night (%) Ovary stage at spawning (04) Spawning activity Egg production/female/spawn ( 103) Total egg production ( 106) Hatch rate (%) Nauplii count/female/spawn ( 103)w Survival (N1 to Z1)/female/spawn (%)w Zoea 1 count/female/spawn ( 103)w No Squid1BW 132 34.7 4.4a 116 23a 2.67 0.63a 3.84 0.36a 1.96 0.19a 143.0 48.0a 4.0 62.9 26.3a 94.0 45.0a 51.6 35.5a 89.0 61.0a

Treatment 2
Yes Squid1BW 132 45.6 11.4b 82 39b 7.44 1.21b 3.59 0.49b 2.15 0.42b 144.0 48.0a 22.0 52.6 29.5b 76.0 53.0a 37.2 32.7a 63.0 46.0a

Treatment 3
Yes Squid1FEAA 132 45.7 10.3b 122 19a 7.50 1.92b 3.75 0.43a 1.95 0.23a 151.0 59.0a 23.0 55.7 31.7ab 83.0 59.0a 44.8 31.6a 73.0 55.0a

Values are mean standard deviation. Means not sharing a common superscript letter within a row are signicantly dierent (Po0.05). Spawn score: 0 5 None; 1 5Partial; 2 5 Full. wExcluding spawns with zero hatched eggs. BW, bloodworms; FEAA, frozen enriched adult Artemia sp.

Artemia sp. with respect to the aforementioned indicators of reproductive performance (82 days, 3.59%, 2.15% and 52.6%) (Po0.05). Spawning frequency of the ablated female population per night for those fed bloodworms was similar to that of those fed Artemia (7.44% vs. 7.50%) (P40.05), and both ablated treatments spawned more frequently than the non-ablated shrimp (2.67%) (Po0.05). Total egg production per treatment for Study 2 was 4.0 106, 22.0 106 and 23.0 106 respectively. Average egg production per female for all three treatments over the study period was similar (143000, 144 000 and 151000) (P40.05). No dierences in treatments were found for nauplii production per female (94 000, 76 000 and 83000), survival rate to Z1 (51.6%, 37.2% and 44.8%) and production rate of Z1 larvae (89 000, 63000 and 73000) (P40.05).

Discussion Farfantepenaeus aztecus was classied in 1985 by Primavera as adicult to spawnspecies. This classication cited the 1975 work of AQUACOP that found eyestalk ablation to be the only successful method of inducing egg ovarian development in female F. aztecus. Unilateral eyestalk ablation is a common practice to induce ovarian maturation in many of the species of dicult to spawn closed thelycum female broodstock (F. aztecus, F. duorarum, P. monodon and P. orien-

talis) (Primavera 1985). The ndings of this study corroborate those of AQUACOP (1975), which demonstrated that unilateral eyestalk ablation is a reliable method for producing consistent ovarian maturation in F. aztecus. In this study, no negative eect on spawn size was found between ablated and non-ablated F. aztecus females. The number of eggs spawned by ablated and non-ablated females was greater than that reported byAQUACOP (1977) for unilaterally ablated F. aztecus females. In contrast to the ndings of AQUACOP (1975), unablated females were able to mature and spawn viable eggs, albeit at a signicantly lower rate than ablated females. The contribution of the maturation system design and the stable physical environmental conditions created within the maturation system cannot be overlooked as a contributing factor to the maturation of non-ablated females. Maturation environment is known to inuence reproductive performance in other Penaeid shrimp (Chamberlain 1985; Bray & Lawrence 1992; Browdy, 1992; Browdy, McGovernHopkins, Stokes, Hopkins & Sandifer 1996). Improved environmental conditions within the maturation system and adequate diets are reasonable conclusions for the maturation and spawning of unablated females in this system. However, further research is necessary to determine the benecial eects of environment and nutrition on these unablated broodstock. Maturation diets for commercially cultured marine shrimp are under constant renement to increas-

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ing their eectiveness at enhancing reproductive performance and reducing production costs. The quality of the broodstock diet has a signicant aect on the reproductive performance of commercially cultured broodstock (Naessens et al. 1997). However, until this study was performed, the eects of advanced maturation diets on the reproductive performance of F. aztecus were unknown. When enriched Artemia sp. is substituted for bloodworms in F. aztecus maturation diets, no adverse eects on reproductive performance are found. Maturation diets of L. vannamei supplemented with enriched Artemia biomass show similar results of improved maturation, reproduction, ospring quality and higher broodstock survivals (Naessens et al. 1997; Wouters, Gomez, Lavens & Calderon 1999). These trends suggest that an enriched Artemia diet provides F. aztecus females with better nutrition to oset the stress of frequent handling and the energetic demand associated with ablation and the frequent spawning. Adequate maturation diet composition is essential in promoting higher broodstock survival, re-maturation, multiple successive spawning and better ospring quality (Wouters et al. 1999). In this study, the nutritional benet of feeding a frozen enriched adult Artemia sp. to the female broodstock of F. aztecus was not isolated to the broodstock performance. Larvae from ablated females fed a frozen enriched adult Artemia sp. achieved higher hatch rates and survival rates to Z1. The use of Artemia biomass as a carrier for nutrients and other dietary components to enhance the reproductive performance of L. vannamei broodstock and increase the quality of ospring was suggested in a 1997 study by Naessens et al. A follow-up study in 1999 by Wouters et al. found the best nutritional eects when the Artemia biomass is enriched with elevated concentrations of polyunsaturated fatty acids, cholesterol, vitamin C, vitamin E and astaxanthin. Therefore, further studies on F. aztecus should focus on specic nutritional proles to determine the adequate enrichment requirements for maximizing the reproductive performance of the species under controlled maturation conditions.

vances in maturation system technology and methods have allowed this study to produce signicant volumes of larvae under controlled maturation conditions. These ndings are signicant as they suggest that the development of commercial production of this species is feasible when these methods are followed. Further renement of commercial production techniques for native species will lead to the sustainable development of a farm-raised live-bait shrimp industry in the United States.

Acknowledgments This research was made possible through funding from the US Department of Agriculture, Small Business Innovation and Research (SBIR) Grant # 200103297. Additional resources and expertise were supplied by the Texas Agricultural Experiment Station ^ Shrimp Mariculture Research Facility, Corpus Christi,TX.

References
AQUACOP. (1975) Maturation and spawning in captivity of penaeid shrimp: P. merguiensis de Man, Penaeus japonicus Bate, Penaeus chinensis de Hann and Penaeus semisulcatus de Hann. Proceedings of the World Mariculture Society 6, pp.123^132. AQUACOP. (1977) Observation on the maturation and reproduction of penaeid shrimp in captivity in a tropical medium. Aquaculture Workshop ICES. 10^13 May 1977. Brest, France, 34pp. Bray W.A. & Lawrence A.L. (1992) Reproduction of Penaeus species in captivity. In: Marine Shrimp Culture: Principles and Practices (ed. by A.W. Fast & L.J. Lester), pp. 93^170. Elsevier, Amsterdam, the Netherlands. Browdy C.L. (1985) Aspects of the reproductive biology of Penaeus semisulcatus de Haan. Doctoral thesis. Tel Aviv University,Tel Aviv, Israel, 138pp. Browdy C.L. (1992) A review of the reproductive biology of Penaeus species. Perspectives on controlled shrimp maturation systems for high quality nauplii production. In: Proceedings of the Special Session on Shrimp Farming (ed. by J. Wyban), pp. 22^51. World Aquaculture Society, Baton Rouge, LA, USA. Browdy C.L. & Holland A.F. (1998) Shrimp virus risk management: a South Carolina case study. Aquatic Nuisance Species Digest 2, 25^35. Browdy C.L., McGovern-Hopkins K., Hopkins J.S., Sandifer P.A. & Stokes A.D. (1996) Factors aecting the reproductive performance of the Atlantic white shrimp, Penaeus setiferus, in conventional and unisex tank systems. Journal of Applied Aquaculture 6,11^25.

Conclusion This study was successful in demonstrating the hatchery potential of the Gulf Brown shrimp. Although previous studies have investigated the reproductive performance of this species, recent ad-

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Brown A. Jr., McVey J., Middleditch B.S. & Lawrence A.L. (1979) Maturation of the white shrimp Penaeus setiferus in captivity. Proceedings of the World Mariculture Society 10, 435^444. Caillouet A.C. Jr. (1972) Ovarian maturation induced by eyestalk ablation in the pink shrimp Penaeus duorarum Burkenroad. Proceedings of the World Mariculture Society 3, 205^225. Chamberlain G.W. (1985) Biology and control of shrimp reproduction. In: Texas Shrimp Farming Manual ^ An Update on Current Technology (ed. by G.W. Chamberlain, M.G. Haby & R.J. Miget), pp.1^41. Texas Agricultural Extension Service,Texas A&M University, Corpus Christi,TX, USA. Hach. (1997) WaterAnalysis Handbook (3rd. edn Hach, Loveland, CO, USA, 325pp. McKee D.A. (1986) An investigation of the live bait-shrimp industry of Texas and the culture and economic potentials for rearing two penaeid species as supplements to that industry. Doctoral Dissertation, Texas A&M University, College Station,TX, USA, 185pp. Middleditch B.S., Missler S.R., Ward D.G., McVey J.P., Brown A. & Lawrence A.L. (1979) Maturation of Penaeid shrimp: dietary fatty acids. Proceedings of the World Mariculture Society 10, 472^476.

Naessens E., Lavens P., Gomez L., Browdy C.L., McGovernHopkins K., Spencer A.W., Kawahigashi D. & Sogeloos P. (1997) Maturation performance of Penaeus vannamei co-fed Artemia biomass preparations. Aquaculture 155, 87^101. Ogle J.T. (1992) Design and operation of the Gulf Coast Research Laboratory penaeid shrimp maturation facility I. Litopenaeus vannamei. Gulf Coast Research Laboratory Technical Report 4, Ocean Springs, MS, USA, 42pp. Primavera J.H. (1985) A review of maturation and reproduction in closed thelycum penaeids. In: Proceedings of the First International Conference on the Culture Penaeid Prawns/Shrimps (ed. by Y.P. Taki), pp. 47^64. Aquaculture Department SEAFDEC, Iloilo, Philippines. Wouters R., Gomez L., Lavens P. & Calderon J. (1999) Feeding enriched Artemia biomass to Penaeus vannamei broodstock: its eect on reproductive performance and larval quality. Journal of Shellsh Research 18, 651^656. Wyban J.A., Cheng L.S., Sweeney J.N. & Richards W.K. Jr. (1987) Observations on the development of a maturation system for Penaeus vannamei. Journal of theWorld Aquaculture Society 18,198^200.

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