Safety Evaluation of Transgenic Tilapia with Accelerated Growth

Recent advances in modern marine biotechnology have permitted the generation of new strains of economically important fish species through the transfer of growth hormone genes. These transgenic fish strains show improved growth performance and therefore constitute a better alternative for aquaculture programs. Recently, we have obtained a transgenic tilapia line with accelerated growth. However, before introducing this line into Cuban aquaculture, environmental and food safety assessment was required by national authorities. Experiments were performed to evaluate the behavior of transgenic tilapia in comparison to wild tilapia as a way to assess the environmental impact of introducing transgenic tilapia into Cuban aquaculture. Studies were also conducted to evaluate, according to the principle of substantial equivalence, the safety of consuming transgenic tilapia as food. Behavior studies showed that transgenic tilapia had a lower feeding motivation and dominance status than controls. Food safety assessment indicated that tilapia growth hormone has no biological activity when administered to nonhuman primates. Furthermore, no effects were detected in human healthy volunteers after the consumption of transgenic tilapia. These results showed, at least under the conditions found in Cuba, no environmental implications for the introduction of this transgenic tilapia line and the safety in the consumption of tiGH-transgenic tilapia as an alternative feeding source for humans. These results support the culture and consumption of these transgenic tilapia. Received: March 9, 1998; accepted June 25, 1998.     

The status of marine fish larval-rearing technology in Australia

Over 20 marine fish species have been studied forfarming or stock enhancement in Australia. However,commercial production has been dominated by the cageculture of Salmo salar in Tasmania, Thunnusmaccoyiiin South Australia, and to a lesser extent Latescalcarifer in Queensland. A major impediment to thecommercial production of new species has been thelarge-scale production of juvenile fish. Thedevelopment of marine fish larval rearing technologyin Australia has had four main influences over thelast decade: culture system technology from France,live food culture and nutritional enhancement fromBelgium, artificial diets from Japan and extensivepond culture from the USA. Microalgae and live foodculture is based on traditional aquaculture speciesand methods. Recent Australian research has focusedon induced spawning, the role of stress in inhibitingovulation, factors influencing initial swim bladderinflation in larvae, larval nutrition, extensiveculture and diagnosis of disease. Over the next 5years, Australian aquaculturists should be able toproduce industrial quantities of a range of nativemarine fish, either in intensive fish hatcheries, orin combination with extensive pond culture.     

Gene transfer technology in aquaculture

The gene transfer technique, transgenesis, has permitted the transfer of genes from one organism to another to create new lineages of organisms with improvement in traits important to aquaculture. Genetically modified organisms (GMOs), therefore, hold promise for producing genetic improvements, such as enhanced growth rate, increased production and efficiency, disease resistance and expanded ecological ranges. The basic procedure to generate transgenic fish for aquaculture includes: (1) design and construction of transgenic DNA; (2) transfer of the gene construct into fish germ cells; (3) screening for transgenic fish; (4) determination of transgene expression and phenotype; (5) study of inheritance; and (6) selection of stable lines of transgenics.GMOs offer economic benefits, but also pose environmental threats. Optimising the mix of benefits and risks is of fundamental importance. The potential economic benefits of transgenic technology to aquaculture are obvious. Transgenic fish production has the goal of producing food for human consumption; thus the design of genetic constructs must take into consideration the potential risks to consumer health, as well as marketing strategies and product acceptance in the market.     

Growth Performance and Gonadal Development of Growth Enhanced Transgenic Tilapia Oreochromis niloticus (L.) Following Heat-Shock-Induced Triploidy

Triploid induction offers a way of considerably reducing fertility in fish, and could therefore be employed to help ensure that any adverse environmental impact of transgenic fish was markedly less. In order to produce sterile growth-enhanced transgenic fish, we have induced triploidy in two lines of transgenic tilapia. Growth performance and gonadal development were analyzed following triploidization by heat shock. Ploidy status was confirmed by nuclear size measurement of erythrocytes. Erythrocytes of triploids were found to be 1.5 times larger than diploids. Observations of growth enhancement and gonadal development were made on diploids and triploids from both transgenic and nontransgenic full sibling batches. In both lines, transgenic diploids were superior in growth performance, followed by transgenic triploids, nontransgenic diploids, and nontransgenic triploids. Although the testes of transgenic triploids were significantly smaller than those of nontransgenic triploids and nontransgenic diploids, histologically they did not show signs of gross deformation. There were also some spermatozoa present in the testes of some triploids, which could be indicative of reproductive functionality. However, the ovaries were devoid of oocytes, underdeveloped, and deformed in all triploids and were completely nonfunctional. Although the best growth performance was shown by the fertile diploid transgenics, the triploid transgenic females could offer a good option for aquaculture purposes because they showed superior growth performance over the normal wild-type tilapias with the advantage of sterility to ensure nonhybridization and noncontamination with the local gene pool. However, careful monitoring of potential gene flow would be required prior to commercial use. Received December 1, 1998; accepted May 18, 1999.     

Biofouling in marine aquaculture: a review of recent research and developments

Biofouling in marine aquaculture is one of the main barriers to efficient and sustainable production. Owing to the growth of aquaculture globally, it is pertinent to update previous reviews to inform management and guide future research. Here, the authors highlight recent research and developments on the impacts, prevention and control of biofouling in shellfish, finfish and seaweed aquaculture, and the significant gaps that still exist in aquaculturalists’ capacity to manage it. Antifouling methods are being explored and developed; these are centred on harnessing naturally occurring antifouling properties, culturing fouling-resistant genotypes, and improving farming strategies by adopting more sensitive and informative monitoring and modelling capabilities together with novel cleaning equipment. While no simple, quick-fix solutions to biofouling management in existing aquaculture industry situations have been developed, the expectation is that effective methods are likely to evolve as aquaculture develops into emerging culture scenarios, which will undoubtedly influence the path for future solutions.     

转基因鱼的研究进展与商业化前景

转基因技术为鱼类育种开辟了新的途径。目前已培育出转生长激素基因鲤、鲑和罗非鱼,转荧光蛋白基因斑马鱼与唐鱼等可稳定遗传的转基因鱼品系,其中快长转生长激素基因鱼的获得对于提高水产养殖的产量与养殖效益具有十分重要的意义。文章简要综述了转基因鱼应用研究的成就、相关技术及生态安全方面的研究进展。显微注射仍是目前基因转植的常用方法,应用转座酶或巨核酸酶介导的转基因新技术可提高基因转植效率与整合率。转基因元件的选择应尽量考虑"全鱼"基因或"自源"基因,以减少转基因鱼食用安全方面的顾虑同时也有利于转植基因的表达与生理功效的发挥。生态安全是转基因鱼商用化面临的最大问题。虽然有研究显示转基因鱼与传统的选育鱼类相比适合度较差,但由于环境与基因型间的相互作用,根据实验室获得的转基因鱼对生态影响的结果,难以预测转基因鱼一旦逃逸会对自然水生态环境产生怎样的影响。因此应建立高度自然化的环境以获得可靠的数据客观评价生态风险,有效的物理拦截、不育化处理等生物学控制策略仍是保证转基因鱼安全应用的关键措施。     

Progress in the evaluation of transgenic fish for possible ecological risk and its containment strategies

Genetically improved transgenic fish possess many beneficial economic traits; however, the commercial aquaculture of transgenic fish has not been performed till date. One of the major reasons for this is the possible ecological risk associated with the escape or release of the transgenic fish. Using a growth hormone transgenic fish with rapid growth characteristics as a subject, this paper analyzes the following: the essence of the potential ecological risks posed by transgenic fish; ecological risk in the current situation due to transgenic fish via one-factor phenotypic and fitness analysis, and mathematical model deduction. Then, it expounds new ideas and the latest findings using an artificially simulated ecosystem for the evaluation of the ecological risks posed by transgenic fish. Further, the study comments on the strategies and principles of controlling these ecological risks by using a triploid approach. Based on these results, we propose that ecological risk evaluation and prevention strategies are indispensable important components and should be accompanied with breeding research in order to provide enlightments for transgenic fish breeding, evaluation of the ecological risks posed by transgenic fish, and development of containment strategies against the risks.     

Present status of fisheries in Turkey

Turkey’s natural and ecological situations are very suitable for aquaculture. Turkey also has a wide variety of freshwater and marine species comprising trout, carp, sea bass, sea bream, turbot, mussel, crayfish, etc. The total production of fish and shellfish was 646,310 tons in 2008. The contribution of freshwater catch to total fishery production is relatively small. Capture fisheries production amounted to 494,124 tons whilst aquaculture production was 152,186 tons in the same year. In Turkey, Engraulis encrasicholus (anchovy) is the main caught sea fish species. Fisheries in the Black Sea are the most important fishery by far and show the greatest variations in total catch. Alburnus tarichii (a local species belonging to Cyprinidae) and Cyprinus carpio (the common carp) are the most important species caught from freshwaters. Aquaculture is going to play an increasingly important role in the Turkish economy, as fishery products are the only products of animal origin that can be exported to the EU. There has been a fast increase in the aquaculture production in Turkey with the implementation of scientific and technological modernization. For example, total aquaculture production for 1986 and 2008 was 3,075 and 152,186 tons, respectively. The percentage of aquaculture in total fish production has been rising every year. The ratio of cultured fish production to total fish production was 1.5% in 1990 s, 13.57% in 2000 and more than 20% in 2005. It was 23.55% in 2008. Trouts are the main cultured freshwater fish species. Raceways and floating cages are employed in culture of trout. Carps are also important cultured freshwater fish species. Sea bass and gilthead sea bream are grown marine fish species. Floating cages, off-shore and earthen ponds are used for marine fish species culture. There has been an increase in fishery exports and imports in recent years. It was more than US500 million in 2008, but that of 2004 was just over US 500 million in 2008, but that of 2004 was just over US 233 million. However, aquaculture production is still far away from the production targets and fisheries sector is not an important part of the economy at present.     

Bacterial Interactions in Early Life Stages of Marine Cold Water Fish

Abstract The intensive rearing of various fish species in aquaculture has revealed intimate relationships between fish and bacteria that eventually may affect establishment of a ``normal' mucosal microflora or result in disease epizootics. Interactions between bacteria and mucosal surfaces play important roles both at the egg and larval stages of marine fish. Bacterial adhesion and colonization of the egg surface occur within hours after fertilization. The diverse flora which eventually develops on the egg appears to reflect the bacterial composition and load of the ambient water, but species-specific adhesion at the egg surface may also play a role in development of the egg epiflora. Proteolytic enzymes produced by members of the adherent epiflora may cause serious damage to the developing egg and may also affect further adhesion of the epiflora. Ingestion of bacteria at the yolk sac stage results in establishment of a primary intestinal microflora which seems to persist beyond first feeding. Establishment of a gut microflora is likely to undergo several stages, resulting in an ``adult' microflora weeks to months after first feeding. Ingested bacteria may serve as an exogenous supply of nutrients or essential factors at an early life stage. Early exposure to high bacterial densities is probably important for immune tolerance, and thus for the establishment of a protective intestinal microflora. Successful rearing of early life stages of several marine fish species depends on knowledge of the complex interactions among the cultured organisms and the bacterial communities which develop at the mucosal surfaces and in the ambient water and rearing systems. The routine use of antibiotics during rearing of fish larvae is not advisable, since it may increase the risk of promoting antibiotic resistance and adversely affect the indigenous microflora of the larvae. The use of probiotics has proven advantageous in domestic animal production, and the search for effective probiotics may have a great potential in aquaculture of marine organisms. Bacteria with antagonistic effects against fish pathogens have been successfully administered to several fish species, resulting in decreased mortality or increased growth rate. Received: 14 December 1998; Accepted: 7 April 1999     

A review of the use of copepods in marine fish larviculture

In spite of the growing interest and success obtained using cultured-copepods, their use in marine aquaculture remains sporadic. Besides, mass culture of several marine copepods has been well established by several authors. However, the upscale of copepod cultures to commercial levels is still a challenge. The practice of using wild copepods from natural ponds which thus increases the risk of parasitic infections of most species has limited their application in aquaculture. The present paper thus emphasizes on recent research efforts focused on the use of chemical treatments and freeze-thawing methods to eradicate procercoids from copepods. Research efforts focused on copepod culture systems which subsequently improved and refined their culture in marine fish larviculture are also well discussed. Advances in the use of copepod eggs as potential source of nauplii for marine fish larvae with special emphasis on the viability, storage conditions and biochemical compositions of the copepod eggs are underscored. Additionally, recent advances in the biochemical compositions (protein, amino acids, pigments, and vitamins) of copepods, which has received relatively little attention compared to researches on the lipid and fatty acid compositions are well emphasized. Specific recommended areas for further research are also proffered.     

稻渔综合种养的科学范式

21世纪是渔业的世纪。中国和世界水产业历经数十年的发展为人类应对食品危机做出了巨大贡献。然而,我国传统的水产业对产量的片面追求导致养殖环境日趋恶化,养殖生态系统不断退化,养殖业的可持续发展受到限制。传统稻田其氮素的流失亦是导致农业面源污染的主要原因之一。我国当前的环境问题源于复合生态系统演化进程的缺陷,解决当前的环境问题,必须从优化复合生态系统演化进程着眼。采用优化的生态循环水产养殖模式,如综合水产养殖则可以大大提高氮、磷等养分物质的利用率。稻渔综合种养是一种科学的复合生态模式,可以概括为三种模式,一种是在我国传统稻田养鱼的基础上,逐步发展起来的一种稻渔共生模式,可采取稻鱼、稻蟹、稻虾等多种共作形式;二是稻田作为湿地用于处理水产养殖尾水的模式,属于异位处理形式;三是将稻渔共生和水产养殖相耦合的模式,尤其是与多种水产养殖形式结合或与复合水产养殖系统相结合,甚至是与农牧系统相结合。这第三种稻渔共作模式又称为陆基生态渔场,具有高产、高品质、高生态可持续性等特点,应加强对该创新养殖模式中有机碳、氮、磷等营养收支平衡和循环利用的相关机制以及复合生态系统对外源营养输入的整体响应机制开展研究。概括而言,尾水排放是传统池塘养殖中氮源的主要流失途径,颗粒物吸附沉降是池塘养殖中磷源的主要流失途径,而系统中的碳源则主要是通过鱼类等生物的呼吸作用进行消耗。基于生态循环的"稻渔共生-池塘复合生态系统"则恰好可以解决这三大类营养物质在生态系统中的高效保持和利用问题,实践业已证明该复合系统拥有较高的产量、品质和生态效益,是一种可持续的农业发展模式。稻渔复合生态系统的创新模式因其特有的生态循环机制及系统的高弹性、高缓冲性、高可持续性,将成为我国乃至世界应对农田、渔业生态系统的退化,复合高效解决渔业、农业或农牧业生态环境问题的典型范式。     

Progress in the evaluation of transgenic fish for possi-ble ecological risk and its containment strategies

Genetically improved transgenic fish possess many beneficial economic traits; however, the commercial aquaculture of transgenic fish has not been performed till date. One of the major reasons for this is the possible ecological risk associated with the escape or release of the transgenic fish. Using a growth hormone transgenic fish with rapid growth characteristics as a subject, this paper analyzes the following: the essence of the potential ecological risks posed by transgenic fish; ecological risk in the current situation due to transgenic fish via one-factor phenotypic and fitness analysis, and mathematical model deduction. Then, it expounds new ideas and the latest findings using an artificially simulated ecosystem for the evaluation of the ecological risks posed by transgenic fish. Further, the study comments on the strategies and principles of controlling these ecological risks by using a triploid approach. Based on these results, we propose that ecological risk evaluation and prevention strategies are indispensable important components and should be accompanied with breeding research in order to provide enlightments for transgenic fish breeding, evaluation of the ecological risks posed by transgenic fish, and development of containment strategies against the risks. Supported by the Development Plan of the State Key Fundamental Research of China (Grant Nos. 2007CB109205 and 2007CB109206), the National Natural Science Foundation of China (Grant No. 30430540), and the ‘863’ High Technology Project (Grant No. 2006AA10Z141)     

Environmental and Socioeconomic Constraints to the Development of Freshwater Fish Aquaculture in China

In this article, we provide an overview, on freshwater fish aquaculture in P.R. China, with special emphasis on pond fish culture. We describe the history, ecology (trophic structure and species reared), and technological aspects (including inputs/outputs, yields, labor productivity, and fossil energy use) of freshwater fish production and analyze its role in relation to the socioeconomic context. We discuss the prospects for intensification of production. In China, freshwater fish aquaculture has always been closely linked to cultivation of crops and animal husbandry, that is, feed inputs are in the form of agricultural wastes. The close integration with the farming system at large results in an efficient use of nutrients, low environmental loading, and little dependence on fossil energy inputs. About 7 to 9 different fish species, mainly herbivores, are kept in the same pond and efforts are made to maintain as much as possible the natural mechanisms of matter regulation and energy flows within the pond ecosystem. However, ecological compatibility is paid for by relatively low productivity, both per hectare of waterbody and per hour of labor input. If the throughput of freshwater fish production per unit of area and labor are to be dramatically increased, the equilibrium of the traditional integrated system will be difficult to maintain.     

Transgenic tilapia and the tilapia genome

The tilapia fish (Oreochromis niloticus) has an important place in the aquaculture of the developing world. It is also a very useful laboratory animal, and readily lends itself to the transgenic technology. Through the use of reporter genes, a range of potential gene promoters have been tested in tilapia, both through transient and stable expression of the reporter construct. Using the transgenic technology, growth enhanced lines of tilapia have been produced. These fish have no abnormalities and offer a considerable growth advantage for future exploitation. It is however crucial that transgenic fish, to be exploited in aquaculture, be sterile, and various methods of achieving sterility are considered. These include triploidy, gene knock out of crucial hormone encoding genes via homologous recombination, and knock down of the function of the same genes via ribozyme or antisense technologies.Transgenic tilapia also offer the potential for exploitation as biofactories in the production of valuable pharmaceutical products, and this is also discussed.     

The Protozooplankton–Ichthyoplankton Trophic Link: An Overlooked Aspect of Aquatic Food Webs1

ABSTRACT. Since the introduction of the microbial loop concept, awareness of the role played by protozooplankton in marine food webs has grown. By consuming bacteria, and then being consumed by metazooplankton, protozoa form a trophic link that channels dissolved organic material into the “classic” marine food chain. Beyond enhancing energy transfer to higher trophic levels, protozoa play a key role in improving the food quality of metazooplankton. Here, we consider a third role played by protozoa, but one that has received comparatively little attention: that as prey items for ichthyoplankton. For >100 years it has been known that fish larvae consume protozoa. Despite this, fisheries scientists and biological oceanographers still largely ignore protozoa when assessing the foodweb dynamics that regulate the growth and survival of larval fish. We review evidence supporting the importance of the protozooplankton–ichthyoplankton link, including examples from the amateur aquarium trade, the commercial aquaculture industry, and contemporary studies of larval fish. We then consider why this potentially important link continues to receive very little attention. We conclude by offering suggestions for quantifying the importance of the protozooplankton–ichthyoplankton trophic link, using both existing methods and new technologies.     

Parasitic diseases in marine cage culture--an example of experimental evolution of parasites?

Rapid development of fish culture in marine cages has been associated with an emergence of parasitic diseases. There is a general trend to an increase in infections with ectoparasites with direct life cycles and a reduced diversity of parasites in aquaculture. Some mariculture creates conditions that are similar to serial passage experiments, which are used to study adaptation during experimental evolution of pathogens. In particular, increased density of fish, repeated introduction of naive hosts, homogenous host populations, fast growth and a potential decrease in genetic diversity are attributes of both aquaculture and serial passage experiments. Some free-living organisms, for example Neoparamoeba spp. and Uronema spp. parasitise fish in culture, but have not been reported from wild populations. Farming fish in marine cages can increase the risk of outbreaks of parasitic diseases, including those caused by opportunistic parasites. However, aquaculture has the potential to control parasitic diseases through selective breeding, vaccination and general fish health management.     

Applications and needs of fish and shellfish cell culture for disease control in aquaculture

This review focuses on the application of cellculture and in vitro methods to addresssome of the key elements identified asstrategies for integrated health management inaquaculture, including the standardization andvalidation of diagnostic methods, thedevelopment of new safe therapeutants, and theimplementation of effective disease controlmethodologies. It is expected that anylong-term rise in seafood production willdepend on the future progress of aquaculture.However, the development of aquaculture faces anumber of problems, of which diseases –particularly the emergence of new pathogens –represent serious risks for the production ofaquatic animals, but also for the health offish farmers and of the consumers ofaquaculture products. The complex interactionsunderlying disease outbreak and progression maybe better studied using in vitro modelsthat use cell/tissue culture methods andexperimental systems, in which the interactionsbetween aggressors and the host can bedissected. Researchers have developed invitro assays for fish toxicological,pathological, and immunological studies, as thein vitro assays allow higher control ofthe conditions of the experiments. Thecombination of cell/tissue cultures and invitro assays reduces also the variability ofthe in vivo responses, which are due tothe unavoidable effects of stress and ofenvironmental influences, and to the disparategenetic backgrounds of farmed fish andshellfish species.     

The use of inter-specific hybrids in aquaculture and fisheries

Inter-specific hybrid fishes have been produced for aquaculture and stocking programmes to increase growth rate, transfer desirable traits between species, combine desirable traits of two species into a single group of fishes, reduce unwanted reproduction through production of sterile fish or mono-sex offspring, take advantage of sexual dimorphism, increase harvestability, increase environmental tolerances, and to increase overall hardiness in culture conditions. Hybrids constitute a significant proportion of some countries' production for certain taxa; for example, hybrid striped bass in the USA, hybrid clarid catfish in Thailand, hybrid characids in Venezuela, and hybrid tilapia in Israel. Despite its widespread use, there is a general impression that inter-specific hybridization is not a very useful tool for aquaculture. We believe this impression stems from inaccurate reporting of some useful hybrids, limited testing of strains used for hybrids, and from early work on salmonids that did not result in hybrids of commercial advantage.Experimentation with new hybrid fishes is ongoing, especially in marine culture systems where sterile fish may be preferred because of the concern that fish may escape into the marine and coastal environment.Hybridization has been used in tandem with polyploidization to improve developmental stability in hybrid progeny. The results of inter-specific hybridization can be variable and depend on the genetic structure (including the sex) of the parent fish. Inadvertent hybridization and backcrossing can lead to unexpected and undesirable results in hybrid progeny, such as failure to produce sterile fish, loss of color pattern, and reduced viability.Hybridization is only one tool to improve aquaculture production and will require knowledge of the genetic structure of the broodstock, good broodstock management and monitoring of the viability and fertility of the progeny. Hybridization does represent a genetic modification wherein genes are moved between different species; implications for biodiversity conservation and regulation of this type of modification are discussed.     

The impact and control of biofouling in marine aquaculture: a review

Biofouling in marine aquaculture is a specific problem where both the target culture species and/or infrastructure are exposed to a diverse array of fouling organisms, with significant production impacts. In shellfish aquaculture the key impact is the direct fouling of stock causing physical damage, mechanical interference, biological competition and environmental modification, while infrastructure is also impacted. In contrast, the key impact in finfish aquaculture is the fouling of infrastructure which restricts water exchange, increases disease risk and causes deformation of cages and structures. Consequently, the economic costs associated with biofouling control are substantial. Conservative estimates are consistently between 5–10% of production costs (equivalent to US$ 1.5 to 3 billion yr^?1), illustrating the need for effective mitigation methods and technologies. The control of biofouling in aquaculture is achieved through the avoidance of natural recruitment, physical removal and the use of antifoulants. However, the continued rise and expansion of the aquaculture industry and the increasingly stringent legislation for biocides in food production necessitates the development of innovative antifouling strategies. These must meet environmental, societal, and economic benchmarks while effectively preventing the settlement and growth of resilient multi-species consortia of biofouling organisms.