Technical and economic feasibility of integrating seahorse culture in shrimp/oyster farms

Integrated multi‐trophic aquaculture systems (IMTA) are building ecosystems designed to produce aquatic organisms with less environmental impact. We have developed a simple system and a management strategy for introducing seahorse (Hippocampus spp.) culture in shrimp/oyster farms to increase economic sustainability. The system includes broodstock, nursery and grow‐out phases of Hippocampus reidi G. culture. We performed a test in a 42.4‐ha marine shrimp/oyster farm, which showed that this system is technically feasible. Then, we performed a partial budget analysis including cost‐return, cash flow and financial feasibility analysis. More than 12 000 marketable size seahorses may be produced in 402 m² annually. Initial investment was only US$ 21 103.00. The benefit–cost ratio was ~US$ 20.00 for each US$ 1.00 spent, internal rate of return was 131% and payback period <2 years. In conclusion, the introduction of seahorse in shrimp/oyster IMTA system is technically feasible, profitable, resilient and shows high liquidity. This system may be easily established in developing countries and has potential to provide seahorses to supply different markets (live specimens for the marine aquarium trade and dried specimens for traditional Chinese medicine and the curio trade). In addition, it can indirectly contribute to preserve natural populations.     

Effects of Anesthetic Disposal on the Physiological and Behavioral Responses of the Lined Seahorses,Hippocampus erectus

Anesthetic disposal for fish often results in physiological and behavioral responses. There is limited information involved on the metabolic and behavioral responses of the seahorses. The lined seahorse, Hippocampus erectus, is of great interest in the sciences because of its unusual S‐shaped morphology and male pregnancy behavior. This study found that the lined seahorse could be effectively anesthetized into Stage II at the concentrations of 10 mg/L clove oil or 20 mg/L MS‐222 based on the comparison of six different treatments. The ventilation frequency, oxygen consumption, and ammonia‐N excretion of the seahorses were decreased significantly as soon as they were exposed to the anesthetic agents. A higher O : N ratio was found in seahorses anesthetized by clove oil as compared to those exposed to MS‐222. After the anesthetic treatment, the feeding frequencies of recovered seahorses were low at the beginning and then increased during the 7‐d culture. Moreover, there was no incremental difference of wet weights among the tested seahorses (P = 0.534). The findings indicate that the anesthetic process has few effects on the feeding and short‐term growth of seahorses in spite of the occurrence of physiological and behavioral responses.     

Rearing the spotted seahorse Hippocampus kuda by feeding live and frozen copepods collected from shrimp ponds

This study evaluated the use of live and frozen copepods collected from shrimp ponds for rearing juveniles of the spotted seahorse Hippocampus kuda. Protein and HUFA contents in frozen copepods were all higher than in Artemia nauplii, the conventional live food for seahorse juveniles. The results of this study showed that copepods can be used as feed for rearing seahorse fry and juvenile. The spotted seahorse showed obvious preference for live copepods and rarely fed on dead copepods on the tank bottom. Furthermore, the combination of frozen copepods and live Artemia nauplii resulted in highest growth and highest survival of the experimental seahorses. Further research on possible effects of DHA:EPA ratio on survival and growth of young H. kuda is recommended.     

The administration of Lactobacillus plantarum directly to food: An effective way to improve the survival and growth of juvenile lined seahorses (Hippocampus erectus)

Previously, we found Lactobacillus plantarum had probiotic effects on lined seahorse (Hippocampus erectus) juveniles by the addition Lactobacillus directly into rearing seawater containing juveniles and their food (i.e. copepods). However, how the juveniles consumed Lactobacillus, directly from water or from copepods in whose guts Lactobacillus may have accumulated by ingestion, is still unclear. To clarify this, three experiments were successively carried out in this study. In the first experiment, the manner by which juveniles consumed Lactobacillus, namely, directly from food, was discovered via a 2 × 2 factorial design with two factors of water and food, and two levels with and without L. plantarum. In the second experiment, a 5 × 4 factorial design with five levels of L. plantarum doses (D: 5, 10, 20, 40 and 80 × 10⁸ cfu/L) and four levels of enrichment time (T: 0.5, 1.0, 1.5 and 2.0 hr) was conducted to analyse lactic acid bacteria (LAB) contents in the enriched copepods under different treatments. In the third experiment, six kinds of Lactobacillus‐enriched copepods chosen from the second experiment that carried significantly different LAB contents were fed to juveniles, and it was found that the copepods containing LAB greater than 5 log₁₀colonies/g had the most improved effects for survival and growth. In conclusion, the effective manner of L. plantarum administration in seahorses is by the addition to their food. Additionally, D40 * T1.0, whose LAB content was more than 5 log₁₀colonies/g, is generally the most economic copepod treatment for improving survival and growth in H. erectus rearing.     

Nutrient removal efficiency of Hydropuntia cornea in an integrated closed recirculation system with pink shrimp Farfantepenaeus brasiliensis

In this study, we have tested the effect of seaweed stocking density in an experimental seaweed biofilter using the economically important red seaweed Hydropuntia cornea integrated with the cultivation of the pink shrimp Farfantepenaeus brasiliensis. Nutrient removal efficiency was evaluated in relation to seaweed stocking density (2.5, 4, 6 and 8 g fw L^?1). Total ammonia nitrogen (TAN) was the main nitrogen source excreted by F. brasiliensis, with concentrations ranging from 41.6 to 65 μM of NH₄⁺‐N. H. cornea specific growth rates ranged from 0.8 ± 0.2 to 1.4 ± 0.5% day^?1 with lowest growth rates at higher seaweed stocking density (8 g fw L^?1). Nutrient removal was positively correlated with the cultivation densities in the system. TAN removal efficiency increased from 61 to 88.5% with increasing seaweed stocking density. Changes in the chemical composition of the seaweed were analysed and correlated with nutrient enrichment from shrimp effluent. The red seaweed H. cornea can be cultured and used to remove nutrients from shrimp effluents in an integrated multi‐trophic aquaculture system applied to a closed recirculation system. Recirculation through seaweed biofilters in land‐based intensive aquaculture farms can also be a tool to increase recirculation practices and establish full recirculation aquaculture systems (RAS) with all their known associated benefits.     

Colonization of Aeromonas salmonicida subsp. masoucida strains in Atlantic salmon (Salmo salar L.) during infection

Aeromonas salmonicida subsp. masoucida (ASM) is classified as atypical A. salmonicida and brought huge economic damages to the local salmonid aquaculture in China. An ASM strain named AS‐C4 was used to investigate the colonization of ASM in Atlantic salmon (Salmo salar L.) by an immersion challenge with the control group (T0, no AS‐C4), group T1 (2.67 × 10⁴ CFU/ml AS‐C4) and group T2 (2.67 × 10⁷ CFU/ml AS‐C4). The numbers of AS‐C4 copies in different fish tissues (gill, intestine, skin, blood, muscle, spleen, liver and kidney) were determined at different time points post challenge using the quantitative real‐time PCR (qRT‐PCR). AS‐C4 were detected in the gill and intestine as early as 0 hr after the challenge both in T1 and T2 groups, suggesting that the gill and intestine were probably the portals of entry of AS‐C4 into salmon. Although AS‐C4 could not be detected in the skin until 24 hr after the challenge in T1 group, it could be detected in the skin as early as 0 hr after the challenge in T2 group, indicating that the skin may also be a portal of entry of AS‐C4 into salmon. AS‐C4 was immediately detected in the blood within 3 hr after it entered the host, suggesting that AS‐C4 successfully invaded the bloodstream of fish. After AS‐C4 colonized the host, it colonized the internal tissues, such as the spleen, liver, kidney and muscle. The results of this study will contribute to the understanding of the pathogenesis of the ASM strains and give a broader understanding of the infection route of ASM in it's host, providing more information for the development of new therapeutic strategies to protect against this pathogen in aquaculture.     

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.     

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.     

Identification of RAPD‐SCAR marker linked to white spot syndrome virus resistance in populations of giant black tiger shrimp,Penaeus monodon Fabricius

White spot disease (WSD) caused by white spot syndrome virus (WSSV) creates severe epizootics in shrimp aquaculture industry worldwide. Despite several efforts, no such permanent remedy was yet developed. Selective breeding using DNA markers would be a cost‐effective strategy for long‐term solution of this problem. In the present investigation, out of 30 random primers, only one primer produced a statistically significant (P < 0.01) randomly amplified polymorphic DNA (RAPD) marker of 502 bp, which provided a good discrimination between disease resistant and disease susceptible populations of Penaeus monodon from three geographical locations along the East coast of India. Because RAPD markers are dominant, a sequence characterized amplified region (SCAR) marker was developed by cloning and sequencing of 502 bp RAPD fragment, which generates a single 457 bp DNA fragment after PCR amplification only in the disease resistant shrimps. Challenge experiment was also conducted to validate this 457 bp SCAR marker, and the results suggested that the WSSV loads were 2.25 × 10³ fold higher in disease susceptible than that in disease resistant shrimps using real‐time PCR. Therefore, this 457 bp DNA SCAR marker will be very valuable towards the development of disease‐free shrimp aquaculture industry.     

灰海马染色体制备及核型分析

为了解灰海马的细胞遗传学特征,便于今后开展灰海马的种质评价与鉴定、规模化人工繁育、亲缘关系研究及人工选育等工作,实验以雌雄灰海马的背鳍为材料,采用秋水仙素浸泡和常规热滴片法制备染色体标本。借助Photoshop图像软件,将同源染色体配对、拼贴,做出染色体核型图。采用Image J软件的自定义曲线测量功能,以着丝粒的中心位置为起点,顺着染色体弯曲的形态,到染色体臂末端为终点,测量线段长度,得出图片中染色体的臂长,根据相同放大倍数下标尺的测量值,换算出染色体的实际臂长。根据臂比值,将染色体进行配对、分类后,得出灰海马的染色体核型公式。结果显示,灰海马的背鳍组织可作为其染色体制备的理想材料,实验选用的染色体制备方法能获得图像清晰、形态良好的细胞分裂相。此法简单、效果好,解决了海龙科鱼类染色体制备的难题;灰海马具有22对染色体,二倍体染色体数目2n=44,雄鱼的染色体核型公式为2n=2sm+20st+22t,雌鱼的染色体核型公式为2n=1m+2sm+20st+21t,雌鱼存在性染色体异型的现象,因此灰海马的性染色体为ZW/ZZ型。