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)     

城市化区域生态风险驱动力及管控策略——以北京市为例

快速城市化导致城市及周边区域生态风险不断增加,认知城市化区域生态风险,探究其变化的驱动因素,提出行之有效的风险管控策略,对城市化区域生态环境与社会经济协调发展具有重要意义。以北京市为例,基于生态系统服务对生态风险进行了定量表征,分析了城市化因素对生态风险的驱动作用,探究了生态风险管控策略。研究结果表明：不透水地表扩张、经济增长、人口增长均对生态风险有显著的驱动作用,其中不透水地表扩张的驱动作用占主导地位。经济增长和人口增长通过影响不透水地表率间接作用于生态风险,在2005年时,人口密度对生态风险的驱动作用大于GDP强度,在2010年及之后,GDP强度对生态风险的驱动作用则变得比人口密度的驱动作用更大。北京市的生态风险管控策略为：在不透水地表率介于40%—70%的区域降低不透水地表率和实施土地利用及景观格局优化;不透水地表率大于70%的区域应防止不透水地表进一步扩张;不透水地表率低于40%的区域需通过提升生态系统质量、强化风险防范、降低风险暴露等途径来管控风险。此外,应通过提高海淀、昌平、通州、顺义等辖区的人口和经济城市化集约水平来降低生态风险。本研究阐明了城市化对区域生态风险的驱动作用...     

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.     

Genetic use restriction technologies (GURTs): strategies to impede transgene movement

No clear consensus has emerged in the debate about the risks posed by transgenic crops and how to assess these risks accurately. In the meantime, interest is growing in strategies to impede transgene movement. This attention is being driven, in part, by expanding interest in using transgenic crops to produce pharmaceutical and industrial products. Potential strategies to impede transgene movement have been published in the scientific literature, and numerous patents have been submitted; however, the efficacy of such strategies has still to be evaluated in a field situation. In this review, we discuss some of the genetic strategies that could be used to restrict the spread of transgenes, although at present many of these technologies are still largely at a theoretical stage of development.     

Development of natural growth regimes for hatchery-reared steelhead to reduce residualism, fitness loss, and negative ecological interactions

Wild steelhead (Oncorhynchus mykiss) typically spend two or more years in freshwater before migrating to sea, but hatchery steelhead are almost ubiquitously released as yearlings. Their large size at release coupled with life history pathways that include both male and female maturation in freshwater present ecological risks different from those posed by hatchery populations of Pacific salmon. Yearling hatchery reared steelhead that fail to attain minimum thresholds for smoltification or exceed thresholds for male maturation tend to ‘residualize’ (i.e., remain in freshwater). Residuals pose ecological risks including size-biased interference competition and predation on juvenile salmon and trout. Three hatchery populations of steelhead in Hood Canal, WA were reared under growth regimes designed to produce a more natural age at smoltification (age-2) to aid in rebuilding their respective natural populations. Mean smolt sizes and size variability at age-2 were within the range of wild smolts for two of the three populations. The third population reared at a different facility under similar temperatures exhibited high growth rate variability and high male maturation rates (20% of all released fish). Experimentally comparing age-1 and age-2 smolt programs will help identify optimal rearing strategies to reduce the genetic risk of domestication selection and reduce residualism rates and associated negative ecological effects on natural populations. Investigations of Winthrop National Fish Hatchery summer-run steelhead will measure a) selection on correlated behavioral traits (‘behavioral syndromes’), b) degree of smoltification, c) changes in hormones that regulate gonad growth at key developmental stages, and d) conduct extensive post-release monitoring of fish reared under each growth regime.     

RNAi转基因作物安全评价研究进展

在秀丽隐杆线虫中首次发现双链RNA(dsRNA)能特异性地导致基因沉默(RNAi)现象后,人们开始大量地研究RNAi技术,并将其应用于功能基因的研究,来提高作物的抗性和改良遗传育种等。本文详细介绍了RNAi的技术原理,并且对RNAi技术与传统转基因技术的区别进行分析,阐述了该技术具有重要的生物学意义,以及在农作物害虫防治领域的占据独特优势。基于RNAi技术存在的潜在脱靶效应,从改良植物、靶标生物和生态环境的3个方面具体分析该技术可能存在的风险,为RNAi技术的风险评估提供参考。由于RNAi技术仍存在风险,为了维护生态多样性和保障人们的人身安全,应尽快建立起符合实际需求的安全性评价方法,本文针对RNAi转基因作物的环境安全和食用安全2个方面的评估方案进行概述。RNAi技术对减少害虫数量、提高水稻产量、降低种植成本以及减少化学农药污染、促进农业可持续发展来说具有重要意义,但该技术仍存在风险,需要进一步监管和研究,建立完善的生态评价系统,让RNAi技术在农业生产上发挥作用。     

Dispersal Potential is Affected by Growth-Hormone Transgenesis in Coho Salmon (Oncorhynchus kisutch)

Potential ecological consequences on the natural environment of fast‐growing transgenic fish with elevated intrinsic growth rates is an important question should such fish be allowed in commercial production systems. One trait that will strongly affect the spatial extent of consequences is the propensity of transgenic fish to disperse. We addressed this question in three experiments using different measures of spatial dispersal where we compared very young genotypically wild coho salmon with transgenic conspecifics in terms of: (i) group cohesion, (ii) exploratory behavior, and (iii) up‐ and downstream movements. Transgenic fish were more loosely aggregated, more likely to explore, and less likely to swim upstream, but equally likely to disperse downstream compared with genetically wild fish. These results show that dispersal behavior has been affected by transgenesis and that transgenic fish therefore may venture into habitats previously not used by wild fish. Given the importance of dispersal in ecological risk‐assessment of transgenic fish, continued work within this area is warranted and experimental habitats should mimic the potential receiving natural habitats to which transgenic fish are likely to escape or be released to.     

Assessing the ecological risks from the persistence and spread of feral populations of insect-resistant transgenic maize

One source of potential harm from the cultivation of transgenic crops is their dispersal, persistence and spread in non-agricultural land. Ecological damage may result from such spread if the abundance of valued species is reduced. The ability of a plant to spread in non-agricultural habitats is called its invasiveness potential. The risks posed by the invasiveness potential of transgenic crops are assessed by comparing in agronomic field trials the phenotypes of the crops with the phenotypes of genetically similar non-transgenic crops known to have low invasiveness potential. If the transgenic and non-transgenic crops are similar in traits believed to control invasiveness potential, it may be concluded that the transgenic crop has low invasiveness potential and poses negligible ecological risk via persistence and spread in non-agricultural habitats. If the phenotype of the transgenic crop is outside the range of the non-transgenic comparators for the traits controlling invasiveness potential, or if the comparative approach is regarded as inadequate for reasons of risk perception or risk communication, experiments that simulate the dispersal of the crop into non-agricultural habitats may be necessary. We describe such an experiment for several commercial insect-resistant transgenic maize events in conditions similar to those found in maize-growing regions of Mexico. As expected from comparative risk assessments, the transgenic maize was found to behave similarly to non-transgenic maize and to be non-invasive. The value of this experiment in assessing and communicating the negligible ecological risk posed by the low invasiveness potential of insect-resistant transgenic maize in Mexico is discussed.     

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。     

Factors that contribute to the ecological risks of salmon and steelhead hatchery programs and some mitigating strategies

State and federal agencies in the United States annually release millions of hatchery salmon and steelhead into public waters. Many of the hatchery programs are located in areas where the wild populations are now listed under the U.S. Endangered Species Act (ESA) (16 U.S.C. §§ 1531–1544). These hatchery programs pose genetic and ecological risks to wild fish populations. Genetic risks occur when hatchery and wild fish interbreed and usually occur within a taxonomic species. Ecological risks occur when the presence of hatchery fish affects how wild fish interact with their environment or with other species and may affect whole species assemblages. This paper reviews some of the factors that contribute to ecological risks. Important contributing factors include the relative abundance of hatchery and wild fish in natural production areas, hatchery programs that increase density-dependant mortality, residual hatchery fish, some physical advantages that hatchery fish can have over wild fish, and life history characteristics that may make some species especially vulnerable to the effects of ecological risks. Many of these risk factors can be mitigated by management activities that reduce the level of interactions between hatchery and wild fish. This paper concludes by recommending twelve mitigation strategies that may be useful when agencies need to bring hatchery programs into compliance with the take provisions of the ESA.     

转青鱼生长激素基因异源四倍体鲫鲤

生态安全性是转基因鱼走向市场的瓶颈,通过转基因四倍体鱼同转基因二倍体鱼杂交获得不育的转基因三倍体鱼是解决该问题的有效途径之一.本研究构建了青鱼β-actin基因启动子和青鱼生长激素（GH）基因精确连接的＂全鱼＂基因pbcAbcGHc;并采用显微注射法将pbcAbcGHc导入异源四倍体鲫鲤受精卵.对照养殖结果表明,150日龄的转基因异源四倍体鲫鲤原代（P0）的体重及体长明显大于对照组.选择60尾P0代转基因异源四倍体鲫鲤,采用PCR方法检测出外源青鱼GH基因在P0代转基因四倍体尾鳍基因组DNA中的整合率为90%;对20尾雄性P0代转基因四倍体精液样本的PCR检测发现,13个样本具有外源青鱼GH基因的整合.在一尾生长速度显著的P0代转基因四倍体鲫鲤的肌肉、肝脏、肾脏和卵巢组织中可检测到外源青鱼GH基因的转录.本研究成功获得了具有明显生长优势的P0代转青鱼GH基因异源四倍体鲫鲤,为建立转青鱼GH基因异源四倍体鲫鲤纯系和研制不育的转基因三倍体鱼奠定了基础.     

Disease resistance, stress response and effects of triploidy in growth hormone transgenic coho salmon

Diploid and triploid coho salmon Oncorhynchus kisutch transgenic for growth hormone (GH) and control coho salmon were compared for differences in disease resistance and stress response. Resistance to the bacterial pathogen Vibrio anguillarum was not affected in transgenic fish relative to their non‐transgenic counterparts when they were infected at the fry stage, but was lower in transgenic fish when infected near smolting. Vaccination against vibriosis provided equal protection to both transgenic and non‐transgenic fish. Triploid fish showed a lower resistance to vibriosis than their diploid counterparts. Diploid transgenic fish and non‐transgenic fish appeared to show similar physiological and cellular stress responses to a heat shock. These studies provide information useful for both performance and ecological risk assessments of growth‐accelerated coho salmon.     

Interface of biotechnology and ecology for environmental risk assessments of transgenic fish

Genetically engineered fish with enhanced phenotypic traits have yet to be implemented into commercial applications. This is partly because of the difficulties in reliably predicting the ecological risk of transgenic fish should they escape into the wild. The ecological consequences of the phenotypic differences between transgenic and wild-type fish, as determined in the laboratory, can be uncertain because of genotype-by-environment effects (GXE). Additionally, we are limited in our ability to extrapolate simple phenotypes to the complex ecological interactions that occur in nature. Genetic background can also shape the phenotypic effects of transgenes, which, over time and among different wild populations, can make risk assessments a continuously evolving target. These uncertainties suggest that assessments of transgenic fish in contained facilities need to be conducted under as wide a range of conditions as possible, and that efficacious physical and biological containment strategies remain as crucial approaches to ensure the safe application of transgenic fish technology.     

Ecological Risk Assessment of Organochlorine Pesticides in Surface Waters of Lake Taihu,China

The residues of organochlorine pesticides (OCPs) in surface waters from Meiliang Bay, Gonghu Bay, and Xukou Bay of Lake Taihu, China, were investigated, and their ecological risks were assessed using the risk quotient method and probabilistic risk assessment. Environmental concentrations of OCPs in surface water of these bays were relatively lower compared with other rivers or lakes in China. Calculation of risk quotient associated with taxonomic groups indicates moderate ecological risks from OCPs for crustaceans and insects in these bays, while the ecological risks were low for fish and negligible for phytoplankton. The ecological risk quotients associated with individual OCPs were lower than 0.01 in these bays, suggesting a negligible risk to aquatic organisms. Ecological risk from α-HCH was relatively lower compared with DDTs, endosulfans, and γ-HCH. The combined ecological risks were evaluated using probabilistic risk assessment for only eight OCPs owing to a lack of available toxicity data for β-HCH and δ-HCH. The percentage of species with the potential to be at risk from mixture of OCPs was lower than the criteria of 5% in each bay, indicating that the combined ecological risks were acceptable.     

我国转基因鱼研制的历史回顾与展望

中国自从诞生了首例转基因鱼以来,在后续30多年里取得了一系列重要研究进展。全球范围的转基因鱼研究包括多种养殖鱼类,目标性状涉及快速生长、抗病抗逆和品质改良。现在已经初步建成转基因鱼育种技术体系和安全评估体系,为转基因鱼产业化奠定了重要基础。本文以转基因黄河鲤育种研究为主线,简要回顾了转基因鱼研究的发展历程,并对转基因鱼育种面临的问题和发展前景进行了分析和展望。     

两种不同终止子在转基因鲤鱼中的促生长效应

转基因构建体中启动子的选择会直接影响转植基因的活性, 近年来有研究表明转基因构建体中终止子的选择会一定程度地影响转植基因的活性。为了更好地筛选转基因构建体和培育快速生长的转“全鱼”生长激素(Growth hormone, GH)基因鱼, 文章用鲤鱼β-actin基因终止子和生长激素基因终止子分别构建了转基因构建体, 显微注射得到转“全鱼”GH基因鱼P0代养殖群体, 比较两种不同终止子构建体的活性。统计分析发现, 生长激素基因终止子构建体的养殖群体的体重频率呈正态分布且平均体重显著高于β-actin基因终止子构建体的养殖群体, β-actin基因终止子构建体的养殖群体的体重频率呈右倾趋势的非正态分布。值得一提的是在混合养殖组中得到一条生长最为快速的鲤鱼证实为转基因阳性且为生长激素基因终止子构建体的转基因鲤鱼。该结果表明转“全鱼”生长激素基因鲤鱼可快速生长, 并能将转植基因向下代遗传。实验结果提示生长激素基因终止子构建体比β-actin基因终止子构建体表现的促生长活性要强。     

Risk Posed by Pesticides to Aquatic Organisms in Rivers of Northern Inland New South Wales,Australia

An ecological assessment was conducted to determine the risk posed by agricultural pesticides to inland rivers of north-west New South Wales (NSW), Australia. A preliminary screening of 30 pesticides provided a short-list of eight for further investigation (atrazine, chlorpyrifos, diuron, endosulfan, fluometuron, metolachlor, profenofos, prometryn). Selection was based on chemical characteristics, toxicity, detection frequencies and environmental concentrations. Hazard quotients were calculated for both spray and non-spray seasons. Where possible, hazard quotients were calculated for both acute and chronic exposures for crustaceans, insects, micro-organisms, molluscs, plants and vertebrates. Chlorpyrifos, endosulfan and profenofos posed a high hazard (HQ > 0.5). A probabilistic risk analysis indicated that chlorpyrifos, endosulfan and profenofos posed a risk from acute exposure during the spray season, while endosulfan also posed a risk from chronic exposure during the spray season. The risks posed by profenofos and chlorpyrifos were characterised by a low probability of detection, but these detections affected a high percentage of species. The risks posed by acute and chronic exposures of endosulfan were characterised by a high probability of detection, but only a limited number of these detections affected a high percentage of species. Risk during the non-spray season was not assessed, as the detection of pesticides was infrequent during this period.     

Vertical Position Reflects Increased Feeding Motivation in Growth Hormone Transgenic Coho Salmon (Oncorhynchus kisutch)

Growth hormone (GH) gene transgenesis has allowed the production of salmon with an inherently increased growth potential, on average two to threefold higher compared with daily specific growth rates observed in normal, non‐transgenic fish. This difference quickly results in animals of very different sizes at age, and is associated with specific morphological effects and enhanced appetites in transgenic animals. However, less is known of the feeding and antipredator behaviour of GH‐transgenic fish, information that can help with predictions of potential ecological consequences of release or escape of transgenic fish into the wild. In a series of experiments, transgenic (T) and normal (N) coho salmon of varying age and size (from 0.5 to 40 g, 3.5–21 mo) were studied singly, in pairs, and in groups during feeding and simulated predation threat. Vertical position generally did not differ between T and N fry, but at larger size (>4 g) T fish remained closer to the surface than N fish both during feeding and predatory attacks, probably as a consequence of inherent differences in feeding motivation and later reinforcement by associative learning. This difference in vertical position was not the result of competition as it remained even after either fish in the pair had been removed. In nature, where predators may attack from above (birds) or below (fish), this kind of behaviour may translate into higher risk of predation, which could increase mortality and lower the fitness of transgenic fish, unless their increased growth rate can compensate for the increased risk‐taking.     

The foraging and antipredator behaviour of growth-enhanced transgenic Atlantic salmon

Growth rate has been established as a key parameter influencing foraging decisions involving the risk of predation. Through genetic manipulation, transgenic salmon bred to contain and transmit a growth hormone transgene are able to achieve growth rates significantly greater than those of unmanipulated salmon. Using such growth-enhanced transgenic Atlantic salmon, we directly tested the hypothesis that relative growth rates should be correlated with willingness to risk exposure to a predator. We used size-matched transgenic and control salmon in two experiments where these fish could either feed in safety, or in the presence of the predator. The first experiment constrained the predator behind a Plexiglas partition (no risk of mortality), the second required the fish to feed in the same compartment as the predator (a finite risk of mortality). During these experiments, transgenic salmon had rates of consumption that were approximately five times that of the control fish and rates of movement approximately double that of controls. Transgenic salmon also spent significantly more time feeding in the presence of the predator, and consumed absolutely more food at that location. When there was a real risk of mortality, control fish almost completely avoided the dangerous location. Transgenic fish continued to feed at this location, but at a reduced level. These data demonstrate that the growth enhancement associated with the transgenic manipulation increases the level of risk these fish are willing to incur while foraging. If the genetic manipulation necessary to increase growth rates is achievable through evolutionary change, these experiments suggest that growth rates of Atlantic salmon may be optimized by the risk of predation. Copyright 1999 The Association for the Study of Animal Behaviour.