Biochemical composition of wild and cultured seahorses (Hippocampus kuda Bleeker)

The seahorse has been used for thousands of years as an important traditional Chinese medicine (TCM) in China. Many species of wild seahorse have become endangered, but demand for them continues to grow rapidly. Therefore, it is urgent to cultivate seahorses to relieve the strain on natural populations and meet the market demand for this TCM. In this study, medicinal components of wild and cultured yellow seahorses (Hippocampus kuda Bleeker) were analysed and compared. Different parts of the body (cortex and bone), male and female specimens, and seahorses of different ages were analysed. Crude protein, crude fat, trace elements, calcium, amino acid, fatty acid, steroid and cholesterol content were measured. The results showed that moisture content accounted for 70% of wet weight and ash content increased with growth. Crude protein and fat contents were higher in cultured seahorses than in wild specimens, and the content of Zn in cultured seahorses (93.26 ± 10.44%) was significantly higher than that of same‐aged wild seahorses (74.20 ± 3.83%). Amino acid and fatty acid contents of cultured seahorses also were higher than those of wild seahorses. These findings show that cultured seahorses can also be used in TCMs and that 1‐year‐old cultured seahorses have the best properties.     

Effect of Temperature on Growth and Survival in Cultured Early Juvenile Pot‐bellied Seahorses,Hippocampus abdominalis

In Tasmania, Australia, commercial seahorse culture takes place in tank systems in which approximately 75% of the water is exchanged daily from the Tamar River estuary. As such, some water conditions such as temperature fluctuate on a daily and seasonal basis. The aim of this study was to examine the effect on growth, condition, survival of, and Artemia ingestion by, early juvenile seahorses, Hippocampus abdominalis, cultured for 6 wk at temperatures within the species' natural range (8–24 C) and above it (26 C). Seahorses cultured at 20 C were longer and heavier than those at 17 C, although not significantly different to 23 C. There were no differences in survival or Artemia ingestion of juveniles cultured at 17, 20, and 23 C. At 26 C, 100% mortality was reached on Day 15. This study demonstrates that H. abdominalis can be reared in captivity at a range of 17–23 C in early life stages without compromising growth and survival.     

A study on bacteria associated with the intestinal tract of farmed yellow seahorse,Hippocampus kuda (Bleeker, 1852): characterization and extracellular enzymes

The microenvironment of bacteria associated with the gastrointestinal tract of an animal influences the host in many ways, including the metabolism of several nutrients. Isolation, molecular characterization and enzymatic activities of culturable bacteria associated with the gastrointestinal tract of hatchery‐reared oceanic yellow seahorse, Hippocampus kuda, were investigated in the present study. 16S rRNA sequencing of bacterial isolates yielded 10 different bacterial genera belonging to two major groups: proteobacteria (62.9%) and firmicutes (37.03%). The predominant microbial flora belonged to the genera Vibrio spp., Enterovibrio sp. and Bacillus spp. Burkholderia cenocepacia, hitherto unreported from the gastrointestinal tract of marine fish, was retrieved from the seahorse gut, albeit in small numbers. Enzymatic studies indicated that the bacterial isolates possess the ability to degrade lipids, cellulose, xylan, starch and proteins. Lipase activity was exhibited by a majority of the isolates (70.4%), indicating that the bacterial flora associated with the intestinal tract of seahorses is capable of digesting foods rich in lipids. The possible implications for the role of these bacteria in the digestion and physiological processes are discussed. To our knowledge, this is the first time that the characterization and enzymatic activities of bacteria associated with the intestinal tract of farmed seahorses have been reported.     

Grow-out of juvenile seahorse Hippocampus kuda (Bleeker; Teleostei: Syngnathidae) in illuminated sea cages

This paper examines the feasibility of rearing 10–15-day- and 0.7–1.5-month-old seahorse Hippocampus kuda in illuminated sea cages to continue existing hatchery protocols to mass produce H. kuda for trade and enhance depleted wild stocks in their natural habitats. Thawed Acetes (a planktonic crustacean abundant in inshore seas) was fed to juvenile seahorses in lighted and unlighted sea cages while one group in lighted cages was not fed Acetes . After 10–12 weeks of rearing, both mean body weight and stretch height increased in all treatment groups, with lighted cage-reared seahorses fed Acetes being heavier (2 g) and longer (8 cm) than the other two treatment groups. Although instantaneous growth rates declined during the rearing period, these were generally higher among Acetes -fed seahorses in lighted cages (0.02–0.07) compared with those in the unlighted cages with Acetes and lighted cages without Acetes feeding. Mean survivorship in all groups ranged from 9% to 74% after the trials, but mean survivorship of juveniles in lighted cages with Acetes feeding (9–74%) was consistently lower than the two treatment groups as a likely result of crustacean and piscine predators being attracted by light and the odour of leftover Acetes in the lighted cages. These results demonstrate that light-attracted zooplankton prey supplemented by Acetes feeding may provide essential nutrients for the growth of H. kuda juveniles in illuminated sea cages. With further improvement in the grow-out protocol, it may provide a possible alternative livelihood to seahorse fishers and sufficient seed to re-populate depleted wild stocks of H. kuda .     

大海马(Hippocampus kuda Bleeker)幼体口吻部骨骼形态观察及其对摄食的影响

通过对10尾2-3日龄的幼体海马的固定、染色、消化的研究,用显微镜对其摄食器官的构成进行详细观察,明确了幼体海马管状鼻内骨骼的结构。结果显示,幼体海马在摄食时,其舌骨角、后翼骨、方骨、上颌骨、续骨起到支撑等重要的作用;海马幼鱼期续骨与后翼骨之间的夹缝结构,可使略大的饵料通过。因此,可以在养殖过程中通过适当地增加饵料的粒径,有效地提高幼体摄食效率,从而提高存活率。     

Biochemical composition of the wild and cultured seahorses, Hippocampus kuda Bleeker and Hippocampus trimaculatus Leach

The biochemical composition of wild and cultured seahorses Hippocampus kuda Bleeker and Hippocampus trimaculatus Leach was analysed in this study. The crude protein contents (% dry weight) of wild seahorses were high: 72.7±2.5% in H. kuda and 78.5±4.2% in H. trimaculatus . The crude lipid contents (% dry weight) of wild and cultured seahorses (1.1±0.1 vs. 1.4±0.2 for H. kuda and 1.2±0.1 vs. 1.2±0.2 for H. trimaculatus ) were low and did not differ significantly ( F _3, 12=1.461, P =0.12). The essential amino acids and flavour-enhancing amino acids in wild seahorses were higher than those in cultured seahorses, although the essential amino acid index in wild and cultured H. kuda and H. trimaculatus were generally low. The total amino acid content decreased with growth in cultured H. trimaculatus ( F _3, 12=14.927, P <0.05). The poly-unsaturated fatty acid (PUFA) contents in wild and cultured H. kuda and H. trimaculatus were 21.69%, 37.72%, 21.39% and 33.89% respectively. The fatty acid content in seahorses first decreased and then increased with growth both in wild and in cultured H. trimaculatus . The PUFA/saturated fatty acid ratio was 1.31 in cultured H. kuda and 0.87 in cultured H. trimaculatus . The trace metal concentrations in the wild seahorses were relatively low. The results presented in this study provide biochemical information necessary for understanding the medicinal value of H. kuda and H. trimaculatus and for elucidating the potential dietary requirements for their culture.     

Study on an integrated eco‐aquaculture system for rearing the yellow seahorse,Hippocampus kuda Bleeker

Seahorse aquaculture is challenged by insufficient feeding supply and lack of biological and nutritional knowledge, resulting in low survival rate and poor economical profitability. We report here an integrated eco‐aquaculture system in which the yellow seahorse and its natural prey were co‐cultured with fertilized water and seaweed in cement ponds. In the first stage, urea (10 g m^?3) and chicken manure (50 g m^?3) were used to fertilize the cultured water, 5–7 days later, rotifer and micro‐crustaceans were flourishing. Then, seahorse juveniles were stocked at 200 ind m^?3. After 2 weeks, seaweed Gracilaria lichevoides was transplanted into the ponds to regulate water quality, light and to provide holdfast attachment for seahorses. The optimal density of G. lichevoides was controlled between 0.5 and 2 kg m^?3. The introduction of seaweed provides the habitat for natural food of seahorse. Within the cluster of the seaweed, small crustacean density was over 450 individuals per 100 g of fresh weight. Initial size of seahorse juvenile was 1.03 ± 0.091 cm, After the 146‐day rearing period, seahorse survival rates were 70.8%, 57.7% and 42.5%, and body standard lengths were 11.33 cm, 10.84 cm and 10.04 cm in the integrated eco‐aquaculture system (GFA) and in monoculture systems of FA (fertilized + feeding) and BA (only feeding) respectively. This system incorporated traditional Chinese aquaculture technique in which feedings nature food organisms were cultivated by fertilization and ecological regulation. Results from these preliminary experiments suggest that the integrated system could be technically feasible, suitable and exemplary.     

Feeding selectivity of the seahorse,Hippocampus kuda (Bleeker), juveniles under laboratory conditions

This study examined the feeding selectivity of Hippocampus kuda juveniles under captive conditions and evaluates different food organisms that could be used to improve hatchery‐rearing of this species. Newly born H. kuda were reared for 10 days in 60‐L capacity tanks and fed rotifers (Brachionus rotundiformis), zooplankton (mostly Pseudodiaptomus annandalei and Acartia tsuensis) alone or both food sources. The size and amount of food ingested increased as seahorses grew. Selective feeding of seahorses appeared to change as they develop, preferring copepod adults over nauplii and rotifers. A. tsuensis was highly selected by juveniles over P. annandalei. Specific growth rate in terms of body weight (SGR‐BW, 15% day^–1) was the highest and mortality rate (9% at day 10) the lowest in seahorses fed a mixed food sources. Slowest growth rate (0.3% day^–1) and highest mortality rate (60% at day 7) were observed in seahorses fed rotifers alone. These results indicate that copepods are suitable food for seahorse juveniles, but a mixture of food organisms in the rearing tank environment enhances survivorship and growth of H. kuda, thus potentially providing a source of cultured rather than wild specimens for characterizing the life history of this threatened species.     

Substrate‐attachment Preferences of Cultured Newborn Pot‐bellied Seahorses,Hippocampus abdominalis (Lesson, 1827)

During a series of observations, newborn pot‐bellied seahorses, Hippocampus abdominalis, were offered three size choices of diameter and density of substrate. The seahorses exhibited a marked preference for the greatest diameter and the lowest density substrates.     

Growth of juvenile Trichogaster leerii (Bleeker, 1852) on diets with various protein levels

Abstract. This study concerns the effect of feeding artificial diets containing varying levels of dietary protein (13%—49%) on the growth of fingerling pearl gourami, Trichogaster leerii (Bleeker, 1852). The growth rate increased significantly with increasing dietary protein content, but it was observed that the increase in growth rate was faster within the 13–32% range than in the 32–49% range. Feed utilization efficiency increased, and protein efficiency decreased, with increasing dietary protein. On the basis of this study, the optimum dietary protein level for T. leerii is between 26 and 36%.     

温度对大海马摄食行为及生长速率的影响

选取体质良好、活力强的大海马(Hippocampus kuda Bleeker),体重(3.075 g±0.16 g)、体全长(11.55 cm±0.31cm)。设置18℃、23℃、28 3℃个实验组和1个对照组(CT组:自然变温),每组3个重复,实验周期28 d。结果显示,大海马在23℃时生长速率最快,摄食活跃;28℃大海马反应最迅速(5.8 s±0.74 s),摄食量最大(153.92 g±10.33 g),活动频繁,泳速大;而18℃和自然变温条件下对食物的反应不敏感,多缠绕在附着物上,摄食时呈聚集式分布;23℃和28℃时,相对活跃,会主动游向饵料摄食,摄食效率较高。综上所述,温度的改变对大海马的生长速率及摄食行为均有一定影响;在适宜温度下,大海马的活力大,摄食量多,生长速率相对较快。本研究旨在为大海马饵料开发和养殖环境优化提供科学数据。     

The Effects of Different Feed Enrichments on Survivorship and Growth of Early Juvenile Longsnout Seahorse,Hippocampus reidi

Culturing the longsnout seahorse, Hippocampus reidi, remains challenging because unlike most cultured seahorse species, Artemia is not an optimal first feed. Ideally, more nutritious live feeds such as wild plankton or copepods should be used, but are not always readily available. Therefore, facilities commonly utilize enriched rotifers. Numerous feed enrichment products exist, including Dan's Feed Artemia Enrichment^®, (Dan's Feed) which is a blend of commercially available enrichment products. The aim of this study was to assess the survivorship, growth, and condition factor (CF) of 0–28 DPR (days post release) H. reidi fed with rotifers and Artemia enriched with Dan's Feed. This enrichment was evaluated against (1) rotifers and Artemia enriched with live alga (Isochrysis galbana), and (2) wild plankton and Artemia enriched with Dan's Feed. At 28 DPR, in terms of survivorship and growth, seahorses fed with rotifers and Artemia enriched with Dan's Feed had a lower CF and outperformed those fed rotifers and Artemia enriched with I. galbana. Seahorses fed with rotifers and Artemia enriched with Dan's Feed performed on par with those fed with wild plankton and Artemia with no significant difference in CF. These findings demonstrated that the culture of H. reidi may be feasible with rotifers and Artemia enriched with Dan's Feed .     

Comparison of the Intestinal Bacterial Flora in Healthy and Intestinal‐diseased Seahorses Hippocampus trimaculatus,Hippocampus erectus,and Hippocampus spinosissimus

This investigation examined the intestinal microbial flora among healthy and intestinal‐diseased seahorses Hippocampus trimaculatus, Hippocampus erectus, and Hippocampus spinosissimus by utilizing polymerase chain reaction‐denaturing gradient gel electrophoresis (DGGE) and densitometric analysis. Results demonstrated that 16 disparate DGGE bands belong to six major bacterial groups, which were Vibrionace, Enterobacteriacea, Rhodobacteraceae, Sphingomonadaceae, Flavobacteriaceae, and Alcaligenaceae. It was found that Vibrionaceae was the dominant population among the healthy and intestinal‐diseased seahorses. Vibrio ponticus strain XJ3 and Vibrio neptunius strain WT82, especially V. ponticus strain XJ3 of high abundance, were identified for the first time in seahorses and concluded to be intestinal disease pathogens because of their co‐existence in three intestinal‐diseased seahorse species and other reported fish or oyster. In comparison, uncultured bacterium clone Alcaligenaceae, Vibrio sp., uncultured bacterium clone Rhodobacteraceae, Serratia nematodiphila strain, and Serratia marcescens strain comprised the basic intestinal bacterial flora of all healthy seahorses. This study is the first to report the presence of S. nematodiphila in seahorses.     

The effects of food and the sum of effective temperature on the embryonic development of the seahorse,Hippocampus kuda Bleeker

The effect of different food (live Acetes spp., live Mysis spp., frozen Mysis spp., and mixed food of 50% live Acetes spp. and 50% frozen Mysis spp.) on gonad development of seahorses, Hippocampus kuda Bleeker, was evaluated in this experiment. The developmental durations of testes and ovary of seahorses were significantly different among the four experimental treatments. The live Acetes spp. treatment presented the shortest developmental durations to stage V, which were 87.6 ± 3.84 days and 89.2 ± 3.71 days, respectively, for ovary and testes. The frozen Mysis spp. treatment had the longest developmental durations (F_3,15 = 13.284, P < 0.05). The relationship between developmental duration of the ovary and gonad developmental stages could be formulated: Y₁ = 12.04x + 24.36 (r² = 0.9722, n = 16, P < 0.05). The gonadosomatic index (GSI) of parent seahorses among the four treatments differed significantly (F_3,15 = 18.364, P < 0.05). The standard GSI of seahorses fed live Acetes spp. was 15.64 ± 1.65%, which was the highest. Feeding live food had a significant effect on the fecundity and spawning of seahorses. The fecundity and spawning number of the live Acetes spp. treatment were 598 ± 45.49 and 552 ± 49.19 individuals, respectively, which were dramatically higher than those of frozen treatment (F_3,19 = 34.152, P < 0.05). Live food also had a similar effect on the fertilization and hatching rate during the embryonic development of seahorses (F_3,19 = 11.386, P < 0.05). Food treatment could also induce an indirect effect on survival rate of juveniles through gonadal and embryonic development of the parents (F_3,15 = 14.519, P < 0.05). In this experiment, the mortality within parent seahorses in the frozen Mysis spp. treatment was the highest (15.1 ± 6.55%), and the survival of juveniles was the highest in the live Acetes spp. treatment (90.4 ± 2.26% at 10 days). In conclusion, feeding live Acetes spp. significantly benefited the gonadal and embryonic development of H. kuda.The effect of temperature (22 °C, 24 °C, 26 °C, 28 °C, 30 °C and 32 °C) on the hatching time of H. kuda was also studied. We demonstrated that the higher the temperature, the shorter the hatching time, as well as the higher the hatching speed. The relationship between hatching time and temperature could be expressed: T = − 39.337 t + 677.75 (r² = 0.9755, n = 30, P < 0.05). In this finding, we provided the sum of effective temperature (SET) and threshold temperature of embryonic development of H. kuda (14066.9 °C h^− 1 and 13.7 °C, respectively). This new information on the effect of feeding type and culture temperature is beneficial for the commercial rearing and breeding industry of this species.     

Diseases of captive yellow seahorse Hippocampus kuda Bleeker,pot‐bellied seahorse Hippocampus abdominalis Lesson and weedy seadragon Phyllopteryx taeniolatus (Lacépède)

Seahorses, pipefish and seadragons are fish of the Family Syngnathidae. From 1998 to 2010, 172 syngnathid cases from the Toronto Zoo were submitted for post‐mortem diagnostics and retrospectively examined. Among the submitted species were yellow seahorses Hippocampus kuda Bleeker (n = 133), pot‐bellied seahorses Hippocampus abdominalis Lesson (n = 35) and weedy seadragons Phyllopteryx taeniolatus (Lacépède; n = 4). The three most common causes of morbidity and mortality in this population were bacterial dermatitis, bilaterally symmetrical myopathy and mycobacteriosis, accounting for 24%, 17% and 15% of cases, respectively. Inflammatory processes were the most common diagnoses, present in 117 cases. Seven neoplasms were diagnosed, environmental aetiologies were identified in 46 cases, and two congenital defects were identified.     

Effect of Dietary n‐3 highly unsaturated fatty acids on Growth,Body Composition and Fatty Acid Profiles of Juvenile Black Seabream,Acanthopagrus schlegeli (Bleeker)

An 8‐wk feeding trial was conducted to investigate the effects of dietary n‐3 highly unsaturated fatty acids (n‐3 HUFA) on growth, body composition, and fatty acid profiles of juvenile Acanthopagrus schlegeli. Three replicate groups of fish (initial mean weight: 8.08 ± 0.09 g, mean ± SD) were fed diets with different levels of n‐3 HUFA (0.76%, HUFA_0.76; 0.83%, HUFA_0.83; 0.90%, HUFA_0.90; 0.97%, HUFA_0.97; 1.04%, HUFA_1.04; 1.12%, HUFA_1.12) at 12.9% of total lipid, with a constant eicosapentaenoic acid (EPA, 20:5n‐3) to docosahexaenoic acid (DHA, 22:6n‐3) ratio of about 2.1. Hepatosomatic index (HSI) and intraperitoneal fat (IPF) ratio were all linearly depressed by dietary n‐3 HUFA levels (P < 0.05), and condition factor (CF) was not affected. Adipocyte diameter in IPF decreased with the elevation of dietary n‐3 HUFA and significance occurred between group HUFA_0.90 and HUFA_1.12. Lipid content in dorsal muscle was significantly lowered by dietary n‐3 HUFA compared to fish fed diet HUFA_0.76. No significance was found in whole fish proximate composition. In liver, dorsal muscle and IPF, ∑SFA, 16:0 or ∑n‐3 HUFA were all positively correlated with dietary n‐3 HUFA, while DHA to EPA ratios remained constant in 2.68, 2.86, and 3.60, respectively. Fatty acid synthase (FAS, EC 2.3.1.85) activities of all treatments remained constant at first and then were significantly elevated by dietary n‐3 HUFA higher than 0.97% (P < 0.05). In contrast, hormone sensitive lipase (HSL, EC 3.1.1.3) changed following an opposite tendency. Quadratic analysis based on weight gain rate (WGR) indicated that dietary n‐3 HUFA requirement for juvenile black seabream was 0.94% of the diet in 12.9% lipid diets .     

Growth and survival of Procambarus acutus acutus (Girard, 1852) and P. clarkii (Girard, 1852) in competitive settings

Third‐instar young‐of‐the‐year (YOY) and juvenile Procambarus acutus acutus and P. clarkii stocked in aquaria, rice forage microcosms and a culture pond were evaluated in intraspecific‐ and interspecific‐competitive settings. Procambarus acutus acutus YOY, which were larger than P. clarkii YOY at stocking, grew and survived at significantly greater rates over fall‐, winter‐ and spring‐temperature courses in aquaria. Juvenile P. a. acutus starting an aquarium experiment with a mean total length (TL) size advantage survived at a significantly greater rate than P. clarkii juveniles. Growth rate of similar‐sized stocked P. a. acutus juvenile was significantly faster than that of P. clarkii juveniles in the same microcosms. Survival of juvenile P. clarkii in the microcosms with P. a. acutus was significantly less than when stocked alone. Estimated survival of 2200 P. a. acutus and 2200 P. clarkii YOY in a culture pond over a 174‐day grow‐out period was 84% and 57% respectively. Growth rate of these YOY P. a. acutus and P. clarkii was 0.48 and 0.40 mm TL day^?1. The body‐size advantage at hatching and through the faster growth of P. a. acutus was important in defining competitive interaction with P. clarkii.     

Comparison of Intestinal Microbiota in Intestinal Tracts of the Wild and Cultured Seahorses,Hippocampus trimaculatus (Leach)

Seahorse aquaculture has been a focus for meeting the demand of Chinese traditional medicine and aquarium, and one of the bottlenecks for aquaculture is disease. The present study focused on the intestinal microbiota between wild and cultured seahorses. Results showed that the Alcaligenaceae and Enterobacteriaceae were the dominant intestinal flora in intestinal tracts of seahorses, and Achromobacter sp., Serratia marcescens, and Serratia sp. were the dominant intestinal flora. It was the first time that two strains (TPL14 and yb97) of Achromobacter xylosoxidans were found in cultured seahorses and even in fishes. Interestingly, Vibrio, which accounted for 19.35% of the intestinal bacterial communities, only occurred in the cultured seahorses, and it was strongly involved in the intestinal diseases in seahorse H. trimaculatus.