Aquaculture 277 (2008) 287292

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Aquaculture
j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / a q u a - o n l i n e

Breeding and juvenile culture of the lined seahorse, Hippocampus erectus Perry, 1810
Qiang Lin a,b,, Junda Lin b, Dong Zhang b,c,
a b c

Key Laboratory of Tropical Marine Environmental Dynamics, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, PR China Vero Beach Marine Laboratory, Florida Institute of Technology, Vero Beach, Florida 32963, USA School of Environment and Public Health, Wenzhou Medical College, University Town, Wenzhou, Zhejiang 325035, PR China

a r t i c l e

i n f o

a b s t r a c t
All seahorse species worldwide have been placed under CITES Appendix II since 2004, because they have been over-exploited for traditional Chinese medicine and aquarium trades. Aquaculture has been recognized as a long-term solution for sustaining the seahorse trade while minimizing wild collection. In this study, we evaluated the breeding and juvenile culture of an important aquarium seahorse species, the lined seahorse Hippocampus erectus, Perry 1810. Pairing, mating and copulation behavior were observed. Gestation time and brood size were 17.33 2.94 days and 272.33 66.45 individuals/brood, respectively. Growth rates differed among juveniles from different broods. Effects of temperature on the growth rates and survivorship of the juveniles during the rst two weeks were compared. The highest growth rate and survivorship of the juveniles occurred at 2829 C among the temperatures tested (2433 C). Growth rate and survivorship of the juveniles during the rst 9 weeks at 28 C were investigated. The nal standard length and survivorship of the juveniles were 6.32 0.52 cm and 71.11 10.18%, respectively, and the relationship between the wet weight and the standard length of the juvenile seahorses can be expressed as: W = 0.0034 L2.5535 (r2 = 0.9903, n = 12, P b 0.01). These ndings suggest that H. erectus is a good candidate for commercial aquaculture. 2008 Elsevier B.V. All rights reserved.

Article history: Received 1 August 2007 Received in revised form 25 February 2008 Accepted 26 February 2008 Keywords: Hippocampus erectus Temperature Growth Survivorship Reproduction

1. Introduction All 33 recognized seahorse species (genus Hippocampus) in the world have been listed on Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES, 2004), due to overexploitation of the wild populations (Vincent, 1996; Lourie et al., 1999). The global trade in seahorses and their relatives as medicine, ornamentals and curios was estimated to be at least 20 million seahorses a year (Vincent, 1996), with at least 50 nations and territories around the world involved in buying and selling seahorses (Job et al., 2002). There has been growing interest in culturing and breeding seahorses in many countries to reduce the pressure on the wild stocks (Forteath, 1997; Jones et al., 1998; Hilomen-Garcia, 1999; Woods, 2000a; Adams et al., 2001; Job et al., 2002; Lu et al., 2002; Wong and Benzie, 2003). Research work has been conducted on H. kuda (Job et al., 2002; Lin et al., 2006), H. trimaculatus (Sheng et al., 2006), H. abdominalis (Woods, 2003a,b) and H. subelongatus (Payne et al., 1998). Commercial seahorse culture of mainly H. kuda and H.

Corresponding authors. Q. Lin is to be contacted at Key Laboratory of Tropical Marine Environmental Dynamics, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, PR China. Tel.: +86 772 231 6872. D. Zhang, School of Environment and Public Health, Wenzhou Medical College, University Town, Wenzhou, Zhejiang 325035, PR China. Tel.: +86 772 231 6872. E-mail addresses: linqiangzsu@163.com (Q. Lin), dzhang@t.edu (D. Zhang). 0044-8486/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.aquaculture.2008.02.030

trimaculatus has been carried out in China for several years for Chinese traditional medicinal products (Lin et al., 2006, 2007; Lu et al., 2001). However, seahorse aquaculture is a relatively new industry with many challenges such as low reproductive efciency and juvenile survivorship (Giwojna and Giwojna, 1999; Chang and Southgate, 2001; Woods 2003a,b; Foster and Vincent, 2004; Lin et al., 2007) and much research is still required to improve the efcacy and commercial viability of seahorse aquaculture. The age and associated size of parent H. kuda can inuence the embryonic size (Berglund et al., 1986; Ahnesjo, 1996; Masonjones and Lewus, 2000), fecundity, growth and survival of the offspring (Dzyuba et al., 2006). Temperature and diet can also greatly inuence seahorse gonad development, brood size and survivorship of juveniles (Wong and Benzie, 2003; Lin et al., 2007), and the growth and survival of the juveniles (Lockyear et al., 1997; Woods 2003a,b; Lin et al., 2006; Sheng et al., 2006). The lined seahorse, Hippocampus erectus Perry, 1810, is a highly valued species in both medicinal and aquarium trades (Correa et al., 1989; Vincent, 1990; Scarratt, 1995; Lourie et al., 1999; Foster et al., 2003). It is mainly found from Nova Scotia along the western Atlantic coast, through the Gulf of Mexico and Caribbean to Venezuela, in shallow inshore areas to depths of over 70 m (Lourie et al., 1999; Fritzche and Vincent 2002; Foster and Vincent, 2004). The species has been included on the IUCN (International Union for the Conservation of Nature) List of Threatened Species as Vulnerable (Foster et al., 2003). The present study aimed to investigate the captive reproductive process and tested the effect of temperature on the growth and

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Q. Lin et al. / Aquaculture 277 (2008) 287292

Table 1 The gestation time, brood size and newly hatched juveniles' standard length and wet weight of different broods of lined seahorse, Hippocampus erectus at 28 C Brood Brood 1 Brood 2 Brood 3 Brood 4 Brood 5 Brood 6 CF of father seahorses (100 BW g/ SL cm3) 0.191 (100 20.67/22.12 ) 0.223 (100 11.09/17.073) 0.246 (100 18.72/19.673) 0.204 (100 13.81/18.923) 0.193 (100 9.26/16.873) 0.263 (100 17.61/18.853)
3

Gestation time (day) 18 (3.214.06.2007) 14 (3.294.12.2007) 19 (4.024.21.2007) 22 (4.145.05.2007) 16 (5.035.19.2007) 15 (6.156.30.2007)

Brood size (individuals/brood) 189 298 314 362 267 204

SL of juveniles (cm) 1.13 0.062 1.08 0.090ab 1.03 0.091b 1.02 0.073 b 0.85 0.089c 0.91 0.088c
a

BW of juveniles (g) 0.0039 0.0003a 0.0028 0.0005bc 0.0033 0.0004b 0.0034 0.0005 b 0.0027 0.0005bc 0.0022 0.0004c

SL: body length; BW: body weight; CF: condition factor = 100 body weight (g) / body length (cm) 3. Signicant differences are indicated by different superscripts (P b 0.05).

survivorship of the juvenile H. erectus. These ndings can be used to develop the commercial culture of this important seahorse species.
2. Materials and methods 2.1. Seahorse broodstock Ten pairs of second generation (F2) seahorses, approximately 1.5 years old, ranging from 16.5 to 19.8 g in wet body weight and 17.1 to 20.7 cm in standard body length, were maintained in recirculating tanks (90 80 60 cm) at the Vero Beach Marine Laboratory, Vero Beach, Florida, USA. Seawater was pumped directly from the Atlantic Ocean and treated with double sand ltration. Seawater ow rate was kept at 0.50.8 L/min. Plastic plants and corallites were used as the substrate and holdfasts for the seahorses. The salinity, temperature, pH, dissolved oxygen (DO) and light intensity were maintained at (mean S.D.) 35 1.0, 28 0.5 C, 7.8 0.3, 6.5 0.5 mg/l and 3000 Lux, respectively. Photoperiod was maintained at 14 L (07302130) and 10 D (21300730). The seahorses were fed twice a day (0900 h and 1600 h) with frozen Mysis spp. shrimp (crude protein 69.5%, crude lipid 8.35%, and crude ber 2.75% in dry weight; 1.63 0.215 cm in body length) at a daily rate of approximately 15% wet body weight. When male and female seahorses completed their copulation, the male was isolated temporarily in a separate brooding/hatching tank (50 25 30 cm) lled with seawater with the same water quality and plastic plants and corallites as above until the juveniles were born. During the gestation period, the male was fed three times (0800 h, 1230 h and 1830 h) a day with frozen Mysis spp. and live adult Artemia. The light intensity was kept at 1000 Lux and faeces and uneaten food in the tanks were siphoned out two hours after each feeding. 2.2. Juvenile seahorse After being released from the male's brood pouch, 10 juveniles were collected, blotted dried on lter paper and weighed (Job et al., 2002). The standard length of each juvenile (the sum of the length from the tip of the tail to the mid-point of the cleithral ring and the length from the tip of the snout to the mid-point of the cleithral ring (Lourie et al., 1999)) was measured (Table 1) before being placed in the experimental tanks (50 25 30 cm). The juveniles were not fed the rst 6 to 10 hours after birth. Then they were fed with newly hatched Artemia nauplii (0.04 0.012 cm in body length) in excess. The salinity, dissolved oxygen (DO), light intensity and photoperiod in the tanks were 35 1.0, 6.5 0.5 mg/L and 1500 Lux, and 16 L (07002300 h): 8 D (2300 0700 h), respectively. The tanks were aerated gently so as not to form excessive air bubbles and turbulence. 2.3. Reproduction The reproductive behaviors of six pairs of parent seahorses were observed either directly or recorded with a video camera from March 21 to July 1, 2007. In the hatchery tank, the male and female seahorses were recorded using a video camera while they greeted each other, and when the female nished her egg-transfer, the male was transferred into a separate brooding/hatching tank (50 25 30 cm). When the male decreased his feeding rate and became inactive in the brooding/hatching tank, the video camera was used again to record the hatching behavior of the male until the juveniles were completely released from the male. Gestation time of each male seahorse was recorded. The number of juveniles released from each male was counted after the male was separated and placed in its own hatching tank. 2.4. Effects of seawater temperature In the rst experiment, 2-day old juveniles from two broods (released by two male seahorses) were used to test the effect of different temperatures on growth and survivorship. Four temperature treatments (24 C, 26 C, 28 C and 30 C) for the juveniles from brood 1 and ve temperature treatments (25 C, 27 C, 29 C, 31 C and 33 C) for the juveniles from brood 2 (Table 1), each with 3 replicates, were tested for 14 days. The wet weight and standard length of each seahorse at the beginning of the experiment were not signicantly different between the temperature treatments (P N 0.05). Transparent glass tanks (50 25 30 cm), which were the same to the brooding/hatching tanks, were used in this experiment and each experimental tank had 15 juveniles haphazardly sampled from the brooding/hatching tank. Individual

submersible heaters were used to maintain the tested temperatures and the variance of tested temperature was below 0.5 C. The salinity, DO and light intensity in the tanks were 35 1.0, 6.5 0.5 mg/l and 1500 Lux, respectively. The juveniles in different treatments were fed with newly hatched A. nauplii four times a day (0730 h, 1200 h, 1700 h and 2100 h) at an approximate Artemia density of 46/ ml. Faeces and uneaten A. nauplii were siphoned out at 2230 h daily. At least 3 juveniles were haphazardly sampled from each treatment to measure standard length and wet weight at the end of the 14-day study. The weight gain [WG = 100 (nal body weight initial body weight) / initial body weight (g)], specic growth rate [SGR = 100 (ln nal body weight ln initial body weight) / 7 days] and condition factor [CF = 100 body weight (g) / body length (cm)3] of the juveniles were calculated. Dead seahorses, if any, were removed upon daily inspection and recorded. 2.5. Growth in different broods Results from the above experiments showed that the juveniles from both broods (broods 1 and 2) had the largest growth rate at 2829 C (see the Results section). To assess the potential growth variations amongst the different broods, growth rates of juveniles (also 2-day old) from another two broods (broods 3 and 4) were similarly tested at 28 C, and then juveniles from the 4 broods (broods 1, 2, 3 and 4), each with three replicates, were cultured for 14 days. The wet weight and standard length of the juveniles were not signicantly different between the treatments at the beginning of the experiment (P N 0.05). During the experiment, the rearing conditions were the same as the experiment 2.4. At the end of the experiment, the nal wet weight and standard length of at least 3 juveniles in each treatment were measured. 2.6. Long-term juvenile culture The juveniles (15 juveniles at the beginning of the experiment) from brood 1 reared at 28 C were tested for an additional 7 weeks (total 9 weeks). The diets and feeding frequency were varied according to the age of the juveniles. The juveniles in 2 15 days post hatch (dph) were fed with newly hatched A. nauplii only at an approximate density of 46 /ml. From 16 to 30 dph, the juveniles were fed with Artemia 8 dph (0.68 0.216 cm in body length) enriched with microalgae Chlorella spp. and the sh oil emulsion SUPER SELCO (www.brineshrimpdirect.com). From 31 to 40 dph, the juveniles were fed with adult live Artemia cultured on Platymonus spp. and Chlorella spp. From 41 to 64 dph, the juveniles were fed with frozen Mysis spp. shrimp (7.21 1.42 mm in length, crude protein N 6.0%, crude lipid N 1.0%, crude ber b 0.5% and moisture b 92%) and adult live Artemia (0.93 0.275 cm in body length) cultured on Platymonus spp. and Chlorella spp. Low density of live Artemia (36 individuals/ juvenile/day) was also provided from 41 to 45 dph to wean the juveniles from live food to frozen food. During the experiment, 3 juveniles in each treatment were haphazardly sampled every 7 days and their standard length and wet weight were measured. Any dead seahorses were removed and recorded daily. 2.7. Data analysis Statistical analyses were conducted using the software SPSS 11.5 (Statistical Program for Social Sciences 11.5). One-way analysis of variance (ANOVA) was used to assess the differences of standard length, wet weight and survivorship of the juveniles among the treatments. All the variables were tested for normality and homogeneity. If ANOVA effects were signicant, comparisons between the different means were made using post hoc least signicant differences (LSD). Juvenile growth at different temperature was subjected to a regression analysis and growth performances (standard length, wet weight and age) were compared by one-way ANOVA between linear regressions. 3. Results 3.1. Reproduction 3.1.1. Mating Mating behavior of the six pairs of parent seahorses was observed. After reaching sexual maturity, the male became lighter in color and its brood pouch was inated, but the female did not change color. The male became active and constantly pursue a female seahorse of similar size. Prior to mating, the male and the female swam side by

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289

Fig. 1. Relationship between temperature and wet weight and standard length of juvenile Hippocampus erectus from broods 1 (a, b) and 2 (c, d).

side and the male often touched the female's belly with his head; the male and female exhibited head-pointing and their tails were often tangled together. After approximately 2 h, the male and female began to swim towards the water surface together. During this upward swim, the male opened his brood pouch through swaying his tail forward towards his belly and then the female deposited the eggs into the male's brood pouch. The egg-transfer process was usually repeated 3 times during each upward swim and egg-transfer lasted 13 days. Generally, copulation starts with daily greeting and ends with egg transfer, and the process is repeated a few times over a few days. When the male and female in the pair stopped greeting each other the following day, the copulation was considered completed. After mating, the female often settled at the bottom and became inactive, whereas the male kept actively swimming for approximately 20 min. After gestation and the release of his offspring, the male would generally seek out the same female for mating again unless the female was not available. 3.1.2. Birth Before releasing the juveniles, the male was generally inactive, hiding in the darker areas of the tank and attached to the articial plant tightly with his tail. During juvenile

releasing, the male swung his body up and down, widening the brood pouch opening. The juveniles were then released one by one for approximately 20 min. The male seahorses were found to prey on the newly released juveniles during hatching. The gestation time and brood size (number of juveniles produced per male) were different among the 6 pairs of the broodstock seahorses (Table 1). Gestation time varied from 14 to 22 days, with a mean S.D. of 17.33 2.94 days. Brood size varied from 189 to 362 juveniles per brood, with a mean S.D. of 272.33 66.45. The standard length (0.85 1.13 cm) and wet weight (0.00220.0039 g) of newly hatched juveniles differed among the broods (F4, 24 = 12.832, P b 0.05). Adult male size did not have an effect on the offspring size (Table 1). 3.2. Effect of temperature on juvenile growth rate and survivorship Growth rate of the juvenile H. erectus was signicantly affected by seawater temperature. The growth increments of wet weight and standard length of the juveniles during the rst 2 weeks differed signicantly among the different temperatures [F3, 11 = 61.711 (brood1), F4, 14 = 480.116 (brood2), P b 0.05]. The highest wet weight and standard length increases occurred at temperatures of 2829 C (Fig. 1).

Table 2 Effect of the temperature on the growth performance of the juvenile seahorses, Hippocampus erectus (broods 1 and 2) Temperature (C) Brood1 24 FBW (g) FSL (cm) WG (%) SGR CF SR (%) 0.052 0.003 3.02 0.12c 990 67.9b 17.1 0.44b 0.178 0.020a 64.44 7.70b
c

Brood2 26 0.073 0.001 3.43 0.06b 1432 19.3a 19.5 0.09a 0.182 0.009a 80.00 6.67ab
b

28 0.079 0.008 3.67 0.10a 1504 80.4a 19.8 0.36a 0.164 0.008a 86.67 6.67a
a

30 0.074 0.002 3.55 0.07ab 1427 52.0a 19.5 0.25a 0.168 0.007a 71.11 10.18ab
b

25 0.025 0.003 2.34 0.08c 561 60.2c 13.5 0.66c 0.193 0.038ab 42.22 10.18bc
b

27 0.035 0.007 2.59 0.05b 866 33.0b 16.2 0.24b 0.203 0.008a 53.33 6.67ab
ab

29 0.041 0.009 2.72 0.04a 1049 54.5a 17.5 0.34a 0.205 0.005a 57.79 13.88a
a

31 0.034 0.002 2.54 0.04b 827 38.2b 15.9 0.30b 0.208 0.010a 37.79 10.18bc
ab

33 0.017 0.001c 2.15 0.03d 344 15.2d 10.6 0.25d 0.167 0.013b 26.67 6.67c

Values were mean S.D. Initial body weight of the juveniles from brood 1 and brood 2 were 0.0048 0.0004 g and 0.0037 0.0003 g, respectively, and initial standard length of the juveniles from brood1 and brood 2 were 1.249 0.072 cm and 1.193 0.084 cm, respectively. The effect of temperature on the growth and survival of the juveniles from brood 1 and brood 2 was signicantly different (P b 0.05), and the values in the same row with different superscripts indicate signicant difference in brood 1 and brood 2, respectively (P b 0.05). FBW: nal body weight (g); FSL: nal standard length (cm); WG: weight gain = 100 (nal body weight initial body weight) / initial body weight (g); SGR, specic growth rate = 100 (ln nal body weight ln initial body weight) / 14 days; CF, condition factor = 100 body weight (g) / body length (cm)3; SR: survivorship (%). Signicant differences are indicated by different superscripts (P b 0.05).

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Q. Lin et al. / Aquaculture 277 (2008) 287292 3.3. Growth rate in different broods Incremental increases in the wet weight and standard length of the juveniles cultured at 28 C during the rst 2 weeks were signicantly different among the 4 broods (F3, 35 = 31.515 and F3, 35 = 4.983, P b 0.01) (Fig. 2). Juveniles in brood 1 had the highest weight increment at 0.050 0.005 g and juveniles in brood 4 had the highest length increment (2.002 0.261 cm). 3.4. Growth rate and survivorship over 9 weeks The wet weight and standard length of the juveniles increased steadily over time and reached 0.45 0.12 g and 6.32 0.52 cm, respectively, at the end of the 9-week culture (Fig. 3a, b). The relationship between the wet weight and the standard length of the juvenile seahorses (Fig. 3c) can be expressed as the following: W = 0.0034 L2.5535 (r2 = 0.9903, n = 12, P b 0.01) W: wet weight of the juveniles; L: standard length of the juveniles. The growth performances of the juveniles were signicantly different among the weeks (P b 0.01) (Table 3). The WG and the SGR of the juveniles in the rst 2 weeks were signicantly higher than the other weeks (F8, 80 = 76.411 and F8, 80 = 34.207, P b 0.01). The WG and SGR decreased sharply in the third week and the lowest values were 17.43 1.22% and 2.29 0.15, respectively, in the 6th week. The CF of the juveniles in the rst week was the highest at 0.22 0.016 g/cm3, and then became relatively stable from the second week on. The survivorship of the juveniles was 71.11 10.18% at the end of the 9-week study.

Fig. 2. Weight and length increments of juvenile Hippocampus erectus from different broods (1, 2, 3 and 4) cultured at 28 C.

The WG and SGR of the juveniles at 28 C were high in brood 1, but the CF did not differ signicantly among the temperature treatments (F3, 11 = 0.930, P = 0.433). In brood 2, the highest WG and SGR occurred at 29 C, and the highest value of CF was at 31 C (Table 2). Seawater temperature also signicantly affected the survivorship of the juveniles [F3, 11 = 5.588 (brood1), F4, 14 = 4.093 (brood2), P b 0.05] (Table 2). The highest survivorship was 86.67 6.67% at 28 C for brood 1 and 57.79 13.88% at 29 C for brood 2 (Table 2).

4. Discussion Studying reproductive behavior of seahorses could help improve reproductive efciency, which is one of the keys in seahorse aquaculture (Berglund et al., 1989; Masonjones, 2001; Lin et al., 2006). Generally, seahorse species attain sexual maturity at 68 months (Forteath, 1997; Wong and Benzie, 2003). Scarratt (1995) reported that the male lined seahorse H. erectus started to develop the brood pouch at 5 to 7 months of age, and the breeding season of this species mainly ranged from May to October in the wild (Teixeira and Musick, 2001; Foster and Vincent, 2004). In this study, the breeding season of H. erectus started in March in the laboratory. The reproductive behavior of H. erectus found in this study was similar to that found for H. zosterae (Masonjones and Lewis, 1996). However, during mating, two males were often found competing for one female. Therefore the species may not be monogamous in mating. In the present study, the gestation time of H. erectus was approximately 17 (1422) days, which is within the range generally found for other seahorse species (Lourie et al., 1999; Perante et al, 2002; Foster and Vincent, 2004), although shorter than that previously reported for H. erectus (2021 days) (Herald and Rakowicz, 1951; Lourie et al. 1999; Foster and Vincent, 2004). It is known that gestation time can be inuenced by species, environmental conditions (e.g. temperature), and food (Vincent and Sadler, 1995; Parente et al., 2002; Wong and Benzie, 2003; Lin et al., 2006, 2007). Newly hatched juveniles were measured at 0.851.13 cm in standard length, within the size range of other seahorse species (Foster and Vincent, 2004). Brood size of adult seahorses varies considerably amongst Hippocampus species, ranging from 35 in H. zosterae (Lourie et al., 1999, Lourie, 2004), 2001000 in H. reidi (Silveira, 2000), 400600 in H. kuda (Lin et al., 2006), to 1783 in H. subelongatus (Nguyen and Do, 1996). Environmental factors such as food and nutrition can also greatly inuence the brood size by affecting the gonad development (e.g. ovary development) and fertilization (Forteath, 1997; Chang and Southgate, 2001; Wong and Benzie, 2003; Foster and Vincent, 2004; Lin et al., 2006, 2007). In the present study, the brood size of the lined seahorse was 272 (189362), similar to what has been previously documented (250300) for this species (Lourie et al., 1999). Parent male seahorses were found to prey upon the newly hatched juveniles following their release, although only a few juveniles were eaten in each brood. This predation on progeny by the parent males appears to be unusual amongst seahorse species and should be considered by seahorse culturists in developing the husbandry protocols.

Fig 3. Relationship among the wet weight, standard length and age of juvenile seahorse, Hippocampus erectus cultured at 28 C over 9 weeks. Initial wet weight and standard length of the juveniles were 0.0039 0.0003 g and 1.13 0.062 cm, respectively.

Q. Lin et al. / Aquaculture 277 (2008) 287292 Table 3 The Weight Gain (WG), Specic Growth Rate (SGR) and Condition Factor (CF) of the juvenile seahorse, Hippocampus erectus, during the rst 9 weeks of culture at 28 C Time (week) WG (%) SGR CF 1 267.28 21.14 18.57 0.82a 0.22 0.016a
a

291

2 218.53 21.12 16.55 0.11b 0.16 0.019b

b

3 71.58 13.19 7.68 1.06c 0.15 0.009b

c

4 41.45 16.86 4.88 1.72d 0.15 0.016b

d

5 37.64 14.79 4.50 1.56d 0.15 0.007b

de

6 17.43 1.22 2.29 0.15e 0.16 0.006b

e

7 37.74 16.76 4.49 1.75d 0.18 0.020ab

de

8 19.05 14.14 2.42 1.69e 0.18 0.014ab

e

9 31.59 7.96de 3.90 0.87de 0.18 0.010ab

Values are mean S.D. Within the same row, signicant differences are indicated by different superscripts (P b 0.05). WG, weight gain = 100 (nal body weight initial body weight) / initial body weight (g); SGR, specic growth rate = 100 (ln nal body weight ln initial body weight) / 7 days; CF, condition factor = 100 body weight (g) / body length (cm)3.

Water temperature has a strong inuence on metabolic rate, therefore growth and development of sh (Il-Nam et al., 1995; Brodeur et al., 2001), including seahorses (Strawn, 1958; James and Woods, 2001). Wong and Benzie (2003) showed that oxygen consumption increased with increasing temperature from 17 to 26 C in juveniles of H. whitei. In addition, water temperature also affects the feeding behavior of juvenile H. trimaculatus and consequently their growth rates (Sheng et al., 2006). In the present study, juveniles of H. erectus grew the fastest at temperatures of 28 to 29 C, and this nding is consistent with the reports of adverse effects of low temperatures (under 25 C) on the growth of H. erectus (Correa et al., 1989). In the wild, the growth and development of juvenile of H. erectus are inuenced by the variable water temperature (Lourie et al., 1999; Foster and Vincent, 2004). In seahorse culture, the ability to provide constant temperature can therefore signicantly benet the juveniles. During this study, we found that the juveniles were inactive and rarely fed at the low temperature of 24 C. Therefore, 2829 C is a recommended temperature for the commercial culture of the juvenile of H. erectus. It is known that seahorse embryos possess a yolk sac and are nutritionally independent of the male seahorses (Azzarello, 1991), and that growth and survivorship of the juveniles are signicantly affected by environmental conditions (Vincent, 1990; Payne et al., 1998; Foster and Vincent, 2004). Broodstock size and brood pouch physiology can inuence the future growth and development of the offspring through the function of gaseous exchange, removal of waste products and osmoregulation (Linton and Soloff, 1964; Quast and Howe, 1980; Carcupino et al., 1997). In the present study, the growth rate of the newly hatched juveniles from 4 different male seahorses was compared and the results showed that the juveniles from the larger males had a higher growth rate (Table 1 and Fig. 2). This result is in agreement with that reported by Dzyuba et al. (2006) who showed that the large parent H. kuda produced offspring whose initial postnatal growth rates (weeks 1 to 3) were signicantly higher than those of the younger and smaller parents. In addition, the body condition of adult seahorses may also inuence the growth and survivorship of their offspring, and the relationships between brood size, parent male size and the juvenile quality should be further studied. Compared with other sh, seahorses grow relatively slowly in standard length and wet weight (Lourie et al., 1999). Generally, diet, together with the environment conditions, can greatly inuence the growth of juvenile seahorses (Lu et al., 2001; Woods, 2003a). For H. erectus, the average weekly WG (%) of the juveniles was 82.47%, which is much smaller than those of other sh (Wilson, 2002). The WG and SGR decreased sharply in the 6th and 8th weeks during this experiment (Table 3, Fig. 3). This was perhaps due to the fact that the juveniles were weaned from feeding on live Artemia to feeding on frozen Mysis spp. and Artemia. A major bottleneck of seahorse aquaculture is the low survivorship of early juveniles (Hilomen-Garcia, 1999; Woods, 2000b, 2003a,b; Adams et al., 2001; Chang and Southgate, 2001) and there is a signicant brood effect on survivorship of young juveniles (Dzyuba et al., 2006; present study). In this study, we found that mortality of juvenile H. erectus was very high during 47 days post hatch for brood

2 in the temperature experiment, which was consistent with ndings for juvenile H. kuda in commercial culture (Lin et al., 2006; Lu et al., 2001). We were able to achieve an average survivorship of 71.11 10.18% after the rst 9 weeks for H. erectus (2025% of the mortality occurred during 47 days post hatch). Improving reproductive efciency and survivorship of early juveniles are both needed to realize large-scale seahorse aquaculture. Acknowledgements The experiments comply with the current laws of the United States in which they were performed. We thank Sarah Jones for providing the seahorse broodstock and the staff of Maritech, Inc. for their help. This study was partially funded by the International Collaborative Research Project for Ph.D. students of Zhongshan University, China. References
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