Development of a Standardized Modular System for Experimental Coral Culture

The increasing interest in coral culture for reef restoration, biotechnological applications, and to supply the marine aquarium trade has prompted researchers to optimize coral culture protocols, with emphasis on ex situ production. However, the diversity of experimental systems employed to investigate ex situ coral production may be a bottleneck to the advance of the state of the art, as it impairs reliable comparisons between experiments, as well as the replication and optimization of culture protocols. This study presents a versatile modular culture system for experimental coral production ex situ, assembled using materials and equipments available from suppliers all over the world and thus allowing researchers worldwide to truly replicate experimental setups. The validation of the modular culture system was performed using the soft coral, Sarcophyton cf. glaucum, as a model organism. The validation experiment tested the effect of different light spectra on the photosynthetic performance, symbiont density, chlorophyll and carotenoid pigments concentration, survival, and growth of coral fragments. The validation experiment confirmed the potential of this modular culture system, which ultimately enables researchers to perform direct comparisons among experiments, and more efficiently contribute to advance ex situ coral aquaculture.     

Ex situ Culture of Colonial Marine Ornamental Invertebrates: Concepts for Domestication

The worldwide market for ornamental saltwater invertebrates supplies the needs of millions of aquarium hobbyists, as well as for public exhibition (zoos, aquaria), universities, and research institutions. The large-scale continuous collection of marine organisms is responsible, in many places, for the destruction of habitats, including coral reefs. The perceived expansion of the animal trade further threatens these fragile habitats. In the present paper, several concepts for the domestication of marine ornamental invertebrates (mainly colonial species) are discussed, offering an alternative commercial approach. The major rationale is based on future ex situ propagation, not field collections; a strategy aimed to circumvent the need for wild-harvested animals. This strategy is based on: (1) collection, settlement and metamorphosis of large numbers of larvae from marine organisms or of naturally shed germ cells under aquarium conditions, where survivorship exceeds several orders of magnitude than that in nature; (2) fragmentation of very small pieces (such as the size of a single polyp in colonial corals or blood vessel ampullae in tunicates) for the production of new colonies; (3) the development of replicates and inbred-lines from chosen ornamental species; (4) the use of cryopreservation of larvae and germ cells which will support the supply of material year-round; (5) several concepts for husbandry methods. Some benefits and deficiencies associated with the strategy for ex situ cultures are discussed, revealing its importance to the future of the trade.     

Investigating fragment size for culturing reef-building corals (Porites lobata and P. compressa) in ex situ nurseries

Culturing small propagules of coral has the potential for high yield with low environmental impact, provided that mortality is low. This study investigated the size-specific mortality and growth of minute fragments (as small as 5 polyps) of two of the most abundant reef-building corals in Hawaii (Porites lobata and P. compressa). Two ex situ nursery systems differing in cost, design complexity, and labor intensity were compared. The first nursery experiment lasted for four months in a large tank with high water motion/surge and extensive manual cleaning to remove competitive algae. The corals were then transferred to a simple low-flow tank containing sea urchins (Tripneustes gratilla) and reduced cleaning, where they were grown for six more months (Nursery II). ‘Nursery I’ resulted in 92% of P. lobata and 73% of P. compressa fragments surviving and nearly doubling in area, in spite of a brief infestation by a nudibranch (Phestilla sibogae) that primarily fed on larger P. compressa fragments. There was a significant positive relationship between fragment size and growth rate, and survivorship was significantly different between donor colonies (genets), but there was no evidence of size-specific mortality. ‘Nursery II’ on the other hand had clear size-specific mortality and higher urchin damage for smaller fragments, resulting in moderate survivorship (78% for P. lobata and 76% for P. compressa), and only a slight increase in the total area covered by coral tissue. Fragments larger than 3 cm² were undamaged and had the highest survival and growth rates. This study illustrates how size-specific mortality can be reduced by ex situ nursery conditions.     

Feeding of scleractinian coral, Galaxea fascicularis, on Artemia salina nauplii in captivity

Satisfying nutrient requirement of corals is still a major constraint for maintaining corals in marine aquariums. Corals are polytrophic in nature. Heterotrophic feeding on zooplankton is one of the corals’ strategies to overcome nutrient deficiency. Artemia salina nauplii are commonly used as biocarriers for many fish larvae in aquaculture and can also serve as a biocarrier for coral in aquariums, provided coral acceptability, optimal feeding rate, and digestibility of the nauplii are well understood. Feeding rate and digestibility of coral fed on A. salina nauplii at 100, 2,000, 4,000, 6,000, and 10,000 ind. l⁻¹ under light and dark conditions was assessed in this study. The maximum feeding rates of Galaxea fascicularis under light and dark conditions was 113.6 ind. polyp⁻¹ h⁻¹ and 76.9 ind. polyp⁻¹ h⁻¹, respectively. The daily feeding rates of G. fascicularis varies and depends on nauplii density. Light plays an important role in coral feeding. Nevertheless, the quantity of A. salina nauplii consumed by the coral under light and dark conditions was not significantly different (P > 0.05). A. salina nauplii are well accepted by G. fascicularis. Complete nauplii digestion was observed after 180 min. Digestibility of A. salina nauplii by G. fascicularis was positively correlated with digestion time.     

Performance and cost‐effectiveness of sexually produced Acropora granulosa juveniles compared with asexually generated coral fragments in restoring degraded reef areas

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海月水母螅状体无性繁殖方式观察及一种新发现报道

无性繁殖是水母螅状体有效补充种群数量及扩展栖息地的重要方式,为正确掌握海月水母的无性繁殖方式进而深入了解其生活史,在显微镜下常年观察并详细描述了海月水母螅状体进行匍匐茎生殖、芽体生殖、足囊生殖、繁殖体生殖、纵向分裂生殖和破裂生殖等复杂无性繁殖方式的过程,发现繁殖体生殖可产生浮游态繁殖体芽,在水中漂浮并生长发育成螅状体,新发现一种破裂生殖方式,仅发生在螅状体极度饱食的状态下。对6种无性繁殖方式的行为特征和功能作用进行了对比分析,认为匍匐茎生殖和破裂生殖有利于螅状体种群快速增加个体数量,芽体生殖和纵向分裂生殖有利于子代幼体生长,足囊生殖有利于种群抵御不良的环境条件,繁殖体生殖有利于种群随水流进行长距离迁移,以扩展或转移栖息地。研究表明,不同的繁殖方式是海月水母应对不同环境条件的适应性繁殖对策。     

Survival and Growth of Isolated Polyps of Galaxea fascicularis (Linnaeus 1767) on Six Kinds of Culture Substrates: Implications for Mariculture,Aquarium Culture,and Conservation

Coral aquaculture is viewed as a sustainable method of providing colonies for use in the aquarium trade, pharmacological studies, and coral reef rehabilitation. The production of large, healthy specimens depends on providing conditions that mimic or surpass natural reef environments. Materials with a high degree of variability in physical and chemical properties are utilized as substrates in coral aquaculture, but until fragment–substrate interactions are isolated and independently studied, the possible inhibitory and facilitative effects of each material will remain unclear. To understand potential key roles that substrates can play in coral culture, a comparison was made about the role of various materials on fragment survival and growth. The experiment examined the interactions between isolates of a common scleractinian coral, Galaxea fascicularis, and six commonly used substrates (coral rock, terracotta, polyvinyl chloride, steel, rubber tire, and concrete) and how these affected survival and growth. No differences were observed in the survival, colony size, and polyp production of the resulting microcolonies. The results show that, for G. fascicularis, growth is not affected by substrate type. The results demonstrate the use of a wide variety of materials for the successful aquaculture of scleractinian corals.     

长江鲟类资源现状及保护

归纳了白鲟、长江鲟、中华鲟近40年自然资源量变化情况,统计了长江鲟和中华鲟人工增殖放流数据,评估了增殖放流成效。因物种特性和分布的差异,放流无效的原因迥异。过度捕捞使人工放流长江鲟在放流后6个月之内难逃被“误捕”,而中华鲟放流数量不足及长江和近海过度捕捞导致中华鲟人工增殖放流的贡献甚微,其结果是长江鲟和中华鲟增殖放流均无法达到自然繁殖群体的补充水平。本文探究了近40年来白鲟、中华鲟和长江鲟在保护、管理和决策上存在的误区和不足,提出在生态大保护的背景下,(1)应编制和实施《长江鲟拯救行动计划》优先项目;(2)应以《中华鲟拯救行动计划》为指引,在就地保护、迁地保护和遗传多样性保护方面提出更具有可操作性的优先计划;(3)应设立中华鲟和长江鲟物种拯救行动计划专项,对现有涉栖息地或保护区生态补偿项目进行优化整合。唯有以恢复长江鲟和中华鲟自然繁殖为核心,才有望延续和恢复其自然种群,实现人与自然的和谐共处,推动长江流域社会经济绿色可持续发展。     

Influence of light intensity on feeding, growth, and early survival of leopard coral grouper (Plectropomus leopardus) larvae under mass-scale rearing conditions

This study investigated the effect of different light intensities on feeding, growth and survival of early stage leopard coral grouper Plectropomus leopardus larvae. Four different light intensities (0, 500, 1000 and 3000 lx) were used and larvae were kept under constant light conditions from 0 day after hatching (DAH) to 5 DAH. The larvae were fed a small S-type of Thai strain rotifers at a density of 20 individuals/mL from 2 DAH. The number of rotifers in larval digestive organ and total length of larvae were examined at 3 h intervals between 04:00 and 22:00 h on 3 DAH, and thereafter at 6 h intervals until the end of the experiment (5 DAH). Four experimental trials of the larval rearing were repeated using by 60 kL mass-scale rearing tanks. The results indicate that coral grouper larvae are visual feeders and their food intake increases with increasing light intensity. Food intake of larvae reared at 3000 lx was significantly higher than those reared at 0–1000 lx on 3 DAH despite being the first-feeding day (P < 0.01). On 4 DAH, total length of larvae reared at 3000 lx was significantly larger than those reared at the lower light intensities (0, 500 and 1000 lx), and thereafter light intensity significantly influenced larval feeding and growth until the end of the experiment. Survival on 5 DAH did not show a significant difference between light intensities, but survival rate at 3000 lx and 1000 lx had a tendency to be higher than those reared at the lower light intensities (0 and 500 lx). In contrast, larvae reared at 0 lx exhibited stagnant and/or negative growth. These results indicate that light intensity is significantly the factor affecting larval feeding, growth, and survival in coral grouper larvae under the rearing conditions.     

Growth of wild pearl oysters <Emphasis Type="Italic">Pinctada fucata,Pinctada margaritifera</Emphasis> and <Emphasis Type="Italic">Pinctada sugillata</Emphasis> (Bivalvia: Pteriidae) in Taiwan

In order to understand growth features of pearl oysters in the genus Pinctada, i.e. Pinctada fucata, Pinctada margaritifera, and Pinctada sugillata in Taiwan, a total of 3062 wild individuals of these species from juvenile to adult were collected monthyly from March 2001 to April 2002 in Jukeng, Pingtung County, south-west Taiwan. Quantitative measurements of live oysters were conducted for shell height (SH), shell length (SL), shell width (SW), hinge length (HL), and wet weight (WW). Different cohorts were identified through multiple length frequency analysis on SH of P. fucata and P. margaritifera, and growth curves with seasonal variation were estimated for these species. Pinctada fucata in Taiwan had a different seasonal growth pattern from the Japanese population, but had similar growth rates during the high growth period. The growth rate of P. margaritifera in Taiwan was slower than in French Polynesia, the Solomon Islands, and the Red Sea. Comparisons of morphological growth features among the three species show large differences in the SW-related features. Pinctada fucata in Taiwan had larger SW than in Japan and Korea. The differences in growth rates and morphological features suggested that the wild Taiwanese oysters may retain genetically pristine characteristics, thus genetic conservation might be urgently needed.     

Comparison of three types of adhesives in attaching coral transplants to clam shell substrates

• 1. Nubbins from 12 coral species were transplanted onto dead giant clam shells at three sites in a lagoon near Bolinao, north‐western Philippines. Transplants were attached using three types of adhesives: cyanoacrylate glue (SG), epoxy putty (EP) and marine epoxy (ME) and were monitored over five months for detachment, in situ mortality and natural self‐attachment by tissue growth.
• 2. Corals attached with SG showed a significantly higher rate of detachment (logrank test) than those attached with either EP or ME (P<0.001 in each case). Also, those attached with EP showed a higher rate of detachment than corals attached with ME (logrank=6.46, P<0.05).
• 3. ANOVA (P>0.05) and survival analysis (logrank=2.85, P>0.05) showed no among‐adhesives differences in in situ mortality. Among the species used, Porites cylindrica, Pavona frondifera, Heliopora coerulea and Porites rus had the highest survivorship with 82%, 80%, 76% and 73%, respectively, of nubbins alive at the end of the experiment and also displayed the highest cumulative rates of self‐attachment. Species with the lowest survival rates included Acropora muricata and four pocilloporids.
• 4. Although there was no significant difference in the number of self‐attaching transplants among the adhesives, corals transplanted using ME and EP self‐attached earlier than those transplanted using SG (logrank=11.19, P<0.01).
• 5. Results from this study highlight the importance of selecting the appropriate means of attaching corals to substrates and carefully considering species‐specific traits of candidate corals for transplantation.

Copyright © 2008 John Wiley & Sons, Ltd.     

Use of Microbacterium sp. and Exiguobacterium mexicanum to improve the survival and development of Artemia under xenic conditions

The effect of Microbacterium sp. strain 8L and Exiguobacterium mexicanum strain 8N was evaluated in the diet of Artemia under xenic conditions. Viable cultures of bacteria were provided to xenic cultures of Artemia in combination with Sacharomyces cerevisiae, cornflour or Spirulina, and the effect on the survival and growth was recorded. The use of these bacterial strains improves significantly the survival of Artemia independently of the used food (P < 0.05), and variable results were observed in the growth.     

单环刺螠Vasa基因的早期发育表达图式

Vasa基因编码DEAD-box家族成员中一种ATP依赖的RNA解旋酶,是决定生殖系发育的重要调控因子之一。采用同源克隆策略及SMART-RACE技术,克隆了海洋经济螠虫动物单环刺螠Vasa基因的全长cDNA,该序列长4 080 bp,开放阅读框2 322 bp,编码733个氨基酸,具有DEAD-box蛋白家族共有的全部9个保守域。整体原位杂交和半定量RT-PCR结果显示,Vasa基因在未受精卵、受精卵、2～8细胞的早期胚胎中均有明显的表达,显示其母源性提供的特点;从囊胚开始,表达量明显降低,在原肠胚表达信号主要集中在内中胚层细胞中;至担轮幼虫,表达信号进一步集中在消化道处;当发育至体节幼虫时,阳性细胞分布于消化道和体节的隔膜处,进入蠕虫状幼虫,信号仅在头部的腹刚毛附近以及后肠周围的细胞中表达。实验结果为探知刺螠动物生殖系的起源和分化以及低等型生殖腺的发生提供重要的数据。     

Applying forest restoration principles to coral reef rehabilitation

• 1. Forest restoration through silviculture (gardening) programs revives productivity, biodiversity, and stability. As in silviculture approaches, the coral ‘gardening’ strategy is based on a two‐step protocol.
• 2. The first step deals with the establishment of in situ and/or ex situ coral nurseries in which corals are farmed (originating from two types of source material: asexual [ramets, nubbins], and sexual [planula larvae, spat] recruits).
• 3. The second is the reef rehabilitation step, where maricultured colonies are transplanted into degraded sites.
• 4. We compare here the rationale of forest restoration to coral reef ecosystem restoration by evaluating major key criteria. As in silviculture programs, a sustainable mariculture operation that focuses on the prime structural component of the reef (‘gardening’ with corals) may promote the persistence of threatened coral populations, as well as that of other reef taxa, thus maintaining genetic diversity. In chronically degrading reef sites this may facilitate a halt in biodiversity depletion.
• 5. Within the current theoretical framework of ecosystem restoration, the recovery of biodiversity indices is considered a core element since a rich species diversity provides higher ecosystem resilience to disturbances.
• 6. The gardening measure may also be implemented worldwide, eliminating the need to extract existing colonies for transplantation operations. At degraded reef sites, the coral gardening strategy can assist in managing human and non‐human stakeholders' requirements as is done in forest management.

Copyright © 2003 John Wiley & Sons, Ltd.     

A comparison of the growth and survival of two freshwater crayfish species, Astacus leptodactylus Eschscholtz and Pacifastacus leniusculus (Dana), under different temperature and density regimes

Growth experiments carried out with two juvenile crayfish species, Astacus leptodactylus Eschscholtz and Pacifastacus leniusculus (Dana), at different temperatures and densities highlighted the problem of cannibalism under conditions aimed at intensifying crayfish production. Cannibalism proved to be much lower in A. leptodactylus than P. leniusculus, suggesting that the former might be the better candidate for astaciculture. In the first of two population survival trials, A. leptodactylus showed a higher survival rate than P. leniusculus, with 26% of P. leniusculus surviving in the first replicate, compared to 42.3% of P. leniusculus, and 40.76% surviving in the second replicate, compared to 47.6% of P. leniusculus. The same trend was observed in the second experiment, with P. leniusculus showing survival rates of 48.5 and 55.3% (first and second replicates, respectively) in comparison to 65.2 and 68.2%, respectively, for A. leptodactylus. It is therefore highly likely that if P. leniusculus were to become established in Turkey, it would outcompete the native A. leptodactylus even if it were not to be devastated by the crayfish plague that has decimated the native species. The experiments also highlighted the problem of differential growth, with some juveniles hardly growing at all while others reached a relatively large size. The range in carapace length (CL) was 9–18 mm for A. leptodactylus and 8.5–18.5 mm CL for P. leniusculus at the end of the first experiment. While both species grew quickly, P. leniusculus hatched earlier, giving it an advantage over A. leptodactylus; consequently, by the end of the summer, the juveniles of the former were larger than those of A. leptodactylus. Specific growth rate values showed that the juveniles of the two species had similar growth rates. The results also revealed that growth was not significantly affected by density in both species at 15°C, but at 25°C, growth was significantly better at a density of 234 juveniles m⁻² than at 468 juveniles m⁻², and better at 468 juveniles m⁻² than at 937 juveniles m⁻² (P < 0.01 and P < 0.001, respectively). We conclude that P. leniusculus is a good candidate for aquaculture as it has a rapid growth rate and early hatching and maturity; however, its aggressive behavior may make it a less attractive proposition than A. leptodactylus, which is also fast growing but less aggressive. This study is a part of PhD study of M.M. Harlıoğlu, who is supported by Fırat University Elazığ, Turkey.     

核糖体DNA在厦门白姑鱼和大黄鱼染色体上的比较定位

为了探究石首鱼核型微观结构上的变化,实验利用荧光原位杂交(fluorescence in situ hybridization,FISH)比较定位了厦门白姑鱼和大黄鱼18S rDNA和5S rDNA的分布特征。结果表明,厦门白姑鱼与大黄鱼在宏观核型以及18S rDNA和5S rDNA染色体分布等3个方面均存在较大差异。厦门白姑鱼的核型公式为2n=48t,臂数FN=48;单对18S rDNA信号分布于1号染色体臂间;单对5S rDNA信号分布于3号染色体近着丝粒区域。大黄鱼的核型公式为2n=2sm+4st+42t,臂数FN=50;单对18S rDNA信号分布于18号染色体短臂端部;5S rDNA信号9~11对,除一对分布于臂间外,其余全部分布于着丝粒端或短臂端部。综合其他石首鱼核型数据可以推断:厦门白姑鱼呈现原始核型特征,而大黄鱼核型是原始核型经染色体重排和/或转座衍生的特化核型;石首鱼宏观核型和18S rDNA分布模式总体保守,仅少数物种存在变化,而5S rDNA位点的分布模式存在高度的种间变化。本研究首次揭示了石首鱼物种间核型微观结构的变化,为进一步开展石首鱼分子细胞遗传学研究奠定了基础。     

Effect of density on survival and growth of cyprinid fish fry

One of the bottlenecks in Israel's cold water ornamental fish industry is the large loss of fish during the post-larval stage. As a first step towards increasing survival rates, the optimal stocking density in earthen ponds for fry of each species should be determined. The results of five consecutive experiments with fry of goldfish (Carassius auratus), common carp and koi (respectively, edible and ornamental morphotypes of Cyprinus carpio) are presented here. The experiments were run in 18 cages of 1 m³ and 0.8 mm mesh size placed in a 0.02 ha earthen pond, stocked at 50,100 and 200 fry per cage. The variables analyzed were growth rate, harvesting weight, biomass and survival. Equations to estimate growth rates under different conditions are given. The three fish types present different growth rates and survival patterns in relation to stocking density and season, which lead to different management implications. Koi can be stocked up to 2 million fry per ha without showing negative density effects. Goldfish should be stocked at low density (500,000–1 million fry per ha) in spring and at higher rate (2 million fry per ha) in summer. Common carp stocking density should be adjusted to obtain the required fish size at harvest.     

致急性肝胰腺坏死病副溶血弧菌(VpAHPND)自然感染三疣梭子蟹

近年来包括急性肝胰腺坏死病(AHPND)在内的多种新发疫病的流行,使我国甲壳类养殖业遭受了严重的经济损失。为了筛查导致山东潍坊某养殖场中一虾蟹混养池塘内患病三疣梭子蟹感染的可能病原,本研究采用分子生物学检测方法,对三疣梭子蟹样品进行了白斑综合征病毒(WSSV)、传染性皮下及造血组织坏死病毒(IHHNV)、虾血细胞虹彩病毒(SHIV)、致急性肝胰腺坏死病副溶血孤菌(Vp_(AHPND))、虾肝肠胞虫(EHP)、偷死野田村病毒(CMNV)、黄头病毒(YHV)和肝胰腺细小病毒(HPV)等8种病原的检测,并对样品进行了组织病理和原位杂交分析。分子生物学检测结果显示,患病三疣梭子蟹样品呈Vp_(AHPND)阳性,而呈现WSSV、IHHNV、SHIV、EHP、CMNV、YHV和HPV阴性。对样品进行Vp_(AHPND)套式PCR第二轮扩增产物的序列测定、比对和进化树分析,结果显示,扩增产物序列与致病副溶血弧菌质粒上pirA~(vp)毒力基因片段具有99%的同源性,该序列与已报道的多个致病副溶血弧菌PirA聚在进化树的同一主分支上。组织病理学分析显示,患病三疣梭子蟹的肝胰腺小管上皮细胞坏死,心肌纤维呈溶解样病变,鳃丝上皮柱突细胞明显坏死,胸神经节的神经细胞损伤严重,并且这些组织中还可见大量的细胞核固缩现象;原位杂交结果显示,肝胰腺、心肌、鳃组织及胸神经节中的病变部位均存在Vp_(AHPND)探针的蓝紫色杂交信号。以上表明,虾蟹混养池塘中三疣梭子蟹在自然状态下感染了Vp_(AHPND),并导致肝胰腺、心肌、鳃和胸神经节发生了严重病理损伤。本研究首次在养殖三疣梭子蟹中检测到Vp_(AHPND)感染并揭示了感染所致的病理变化,相关结果为揭示Vp_(AHPND)自然宿主种类和养殖三疣梭子蟹病害防控提供了基础信息。     

Decapsulated Artemia cysts as dietary supplement for juvenile crayfish (Pacifastacus leniusculus, Astacidae) at different food supply frequencies from the onset of exogenous feeding under controlled conditions

Considering that the use of decapsulated Artemia cysts as direct food for juvenile crayfish could be an alternative to live nauplii, a 100-day experiment was carried out under controlled conditions to evaluate the effects of cysts, comparing with nauplii, as supplement to a dry diet for salmonids on the survival and growth of juvenile signal crayfish (Pacifastacus leniusculus) from the onset of exogenous feeding (stage 2). The reduction of feeding frequency was also tested. According to a bifactorial design, six treatments, differing in the supplement and feeding frequency, were tested: the dry diet supplemented with Artemia nauplii or decapsulated cysts was supplied once a day, once every two days and once every three days. Survival rates ranged from 56.7% to 81.7%, rising significantly with increasing the feeding frequency. The highest growth (12.94 mm carapace length and 593.08 mg weight) was reached by the crayfish that received the dry diet supplemented with cysts once a day, with significant differences from the rest of the treatments. Considering the supplement, the cysts supported significantly higher growth than the nauplii. Regarding the feeding frequency, growth was higher when the food was supplied once a day, showing significant differences from the other two frequencies (once every two days and once every three days). This study shows that decapsulated cysts are better dietary supplement than live nauplii. In crustacean culture, this is the first report of successful use of Artemia cysts from the onset of exogenous feeding.