大田软海绵酸对黑鲷胚胎发育及仔鱼的急性毒性

为了研究大田软海绵酸(okadaic acid,OA)对生物胚胎组织发育过程的毒性作用机制,采用静水和半静水实验法,用OA毒素与过滤海水按等对数间距分设5个浓度组,同时以过滤海水为对照组,对人工养殖的黑鲷鱼卵和仔鱼进行了毒性实验。实验结果表明:一定浓度的OA使鱼卵的死亡率和孵出仔鱼的畸形率升高,但对孵化率的影响较弱。OA对黑鲷仔鱼的24、48、72和96h的LC50分别为51.63、45.58、37.97和25.72nmol·L-1,仔鱼体内毒性蓄积程度也随时间逐渐减弱。OA毒素对仔鱼的谷胱甘肽过氧化物酶(GSH-PX)和酸性磷酸酶(ACP)的活性均有诱导作用,与对ACP的影响相比,OA毒素对GSH-PX活性的诱导作用更为突出。此外,OA毒素还会对黑鲷仔鱼的肝脏细胞的核酸代谢及DNA合成有一定的影响,使得RNA和DNA的含量比值发生显著变化。     

菹草生长对沉积物间隙水中各形态磷浓度的影响

通过人工构建湖泊生态系统,用湖泊沉积物的上层泥和下层泥分别代表清淤前后的2种基质,研究菹草(Potamogeton crispus)生长对沉积物不同层次间隙水中各形态磷浓度的影响.垂直方向上,将2种基质中的间隙水分为表层、中层和底层3个层次.结果表明,菹草生长能够不同程度地降低各层间隙水中磷浓度,上层泥和下层泥2种基质均表现为表层间隙水溶解性总磷(DTP)和溶解性活性磷(SRP)含量降低幅度最大,DTP含量分别降低68.97％和83.09％,SRP含量分别降低54.57％和96.02％.菹草的吸收作用使得各层间隙水中磷浓度均低于无沉水植物的对照组.试验结束时,2种沉积物中TP含量降低幅度差异不显著,说明疏浚前后种植菹草均可有效控制湖泊内源磷污染.     

富营养化水体中浮游动物对藻类的控制作用

采用室内受控生态系统进行实验.采用于桥水库原水接种后,人工设置营养盐质量浓度梯度,模拟富营养化程度不同的水体,以研究浮游动物的生长情况,及其控制藻类生长的作用.使用SPSS统计软件进行数据分析.结果显示:过高的营养盐浓度,可能会对轮虫的生长产生抑制作用,而甲壳类动物的总数量基本由营养盐质量浓度决定.在四个营养盐质量浓度梯度下,浮游动物与藻类之间均存在一定的显著或极显著相关关系,说明浮游动物均能够发挥一定的控藻作用.当营养盐ρ(TN)=3mg·L-1、ρ(TP)=0.02mg·L-1左右,浮游动物的控藻作用最明显.当营养盐质量浓度过高时,浮游动物的控藻作用受影响.     

吡唑醚菌酯对不同阶段斑马鱼的毒性效应评价

为评价吡唑醚菌酯对鱼类不同生命阶段的毒性效应,以斑马鱼(Brachydanio rerio)为测试生物,测定了其对斑马鱼胚胎、仔鱼(孵化3 d和孵化12 d)和成鱼的急性毒性及对胚胎和仔鱼(孵化3 d)的发育毒性。结果表明,吡唑醚菌酯对斑马鱼胚胎、仔鱼(孵化3 d)、仔鱼(孵化12 d)和成鱼的96 h-LC50分别为0.0578(0.0540~0.0631)、0.0299(0.0282~0.0320)、0.0330(0.0310~0.0350)和0.0756(0.0692~0.0862)mg·L-1,其毒性大小依次为仔鱼(孵化3 d)仔鱼(孵化12 d)胚胎成鱼。与对照组相比,吡唑醚菌酯0.101 mg·L-1处理,可对斑马鱼胚胎24 h自主运动产生显著抑制作用;0.0609、0.0777和0.101 mg·L-1处理,可对斑马鱼胚胎48h心跳速率产生显著抑制作用;0.0609和0.0777 mg·L-1处理,可对斑马鱼胚胎96 h孵化仔鱼体长产生显著抑制作用。在供试浓度下吡唑醚菌酯暴露96 h后,均可对斑马鱼仔鱼(孵化3 d)体重和体长产生明显的抑制效应,对斑马鱼仔鱼(孵化3 d)心跳未产生明显影响。在高浓度下,吡唑醚菌酯可诱导斑马鱼胚胎产生畸形,主要包括心包水肿、卵黄囊水肿等;而对仔鱼(孵化3 d)未观察到明显的致畸效应。     

铜、镉、敌敌畏和甲胺磷对南美白对虾的亚急性毒性作用

在恒温和充气条件下,研究了重金属(铜、镉)和有机磷农药(敌敌畏、甲胺磷)对南美白对虾的亚急性毒性作用及对虾体胆碱酯酶的影响.结果表明:南美白对虾幼虾在各浓度重金属(铜、镉)和有机磷农药(敌敌畏、甲胺磷)试液中经过60d的亚急性中毒试验,除铜在低浓度促进生长外,其他各组体长、体重及蜕皮次数均小于对照组,且随毒物浓度的增加而逐渐降低;有机磷农药对南美白对虾体内胆碱酯酶具有显著影响,虾体内胆碱酯酶含量随水体中有机磷农药浓度的升高而显著降低;重金属对胆碱酯酶也有一定影响,胆碱酯酶随重金属离子浓度升高也有逐渐降低的趋势,但重金属对胆碱酯酶的影响效果没有有机磷农药显著.     

海洋纤毛虫实验生态学研究I：不同浓度葡萄糖对种群增长的影响

为揭示不同碳氮比对纤毛虫种群增长的影响以及纤毛虫对碳源和能量来源的选择性 ,利用实验生态学方法 ,对海水养殖水体中 3种常见的纤毛虫原生动物 :扇形游仆虫 (Euplotesvannus)、海洋尾丝虫 (Uronemamarinum)和巨大拟阿脑虫 (Paranophrysmagna)在牛肉浸膏及梯度浓度葡萄糖培养基中的种群增长过程进行了初步的探讨 .数据分析表明 ,在牛肉浸膏培养基中加入不同浓度的葡萄糖对于纤毛虫种群增长影响显著 ,其中在 0 .0 5 gL-1葡萄糖浓度的培养基中 3种纤毛虫种群增长最佳 (P <0 .0 5 ) ,而在 0 .4 gL-1浓度时纤毛虫的生长繁殖均受到高度抑制 (P <0 .0 5 ) .这一结果显示 ,不同C N比对纤毛虫的种群增长有着直接的影响 .图 3表 3参 10     

投喂频率和饵料种类对黄颡鱼仔稚鱼生长和存活的影响

研究了以浮游动物为饵料日饱食投喂3次(F3)、2次(F2)、1次(F1)等不同投喂频率对3～32日龄黄颡鱼仔稚鱼生长和存活的影响,并比较了单独投喂浮游动物(Ⅰ组)、水蚯蚓(Ⅱ组)及两者混合投喂(Ⅲ组)对4～34日龄黄颡鱼生长和存活的影响.结果表明:(1)投喂频率对黄颡鱼仔稚鱼生长具有显著影响,F3组个体平均全长、体重显著高于F2、F1组;3～29日龄全长和体重特定生长率(SGR)与投喂频率间直线正相关(SGR1=1.0366D+3.2347,R2=0.9025;SGRW=3.00t3D+9.4829,R2=0.8833),投喂越多则生长越快.(2)17-25日龄时Ⅱ组生长显著低于Ⅰ、Ⅲ组;28日龄Ⅱ组生长仍显著低于Ⅲ组,但与Ⅰ组问没有显著差异;到34日龄,Ⅲ组生长最快,Ⅱ组生长超过Ⅰ组,3组间生长差异显著(P<0.05),表明28日龄后水蚯蚓为黄颡鱼更合适的饵料.(3)饵料种类对34日龄内的仔稚鱼成活率没有影响.试验表明,日饱食3次更有利于黄颡鱼仔稚鱼的生长;28日龄后投喂饵料应增加水蚯蚓;充足的适口饵料是影响黄颡鱼苗种生长的重要因素.图2表2参18     

饵料密度对南方鲶仔稚鱼生长、成活、残食的影响

以枝角类为饵料,分两阶段研究了饵料密度对2～30日龄(Days after-hatching,DAH)南方鲶仔稚鱼生长、成活、残食的影响.前期阶段:2～6DAH仔鱼,分为未投饵组和投饵组,投饵组的饵料密度设为250、500、1000、2000个/L;后期阶段:7～30DAH稚鱼,饵料密度设为250、500、1000、2000、4000个/L.结果表明:前期阶段,250个/L饵料密度组仔鱼生长最快,终体重为0.055g;未投饵组的仔鱼残食十分严重,残食率高达60%;500个/L饵料密度组的仔鱼成活率最高,达95.56%.后期阶段,2000个/L饵料密度组稚鱼生长最快,终体重为1.57g;250、500个/L饵料密度组稚鱼的残食较为严重,当饵料密度增加至2000个/L时,残食率下降至10.00%,成活率达到83.33%.随饵料密度的增加,两试验阶段仔稚鱼的特定生长率(Special growth rate,SGR)均呈先增加后降低的趋势,全长变异系数呈下降趋势.综合生长和成活两项指标,2～6DAH仔鱼的适宜饵料密度为500个/L,7～30DAH稚鱼的适宜饵料密度为2000个/L.饵料不足引发的严重残食,是南方鲶苗种成活率低的重要原因.适宜的饵料密度能够有效地控制残食.图2表2参27     

底泥浸出液对脊尾白虾抗氧化解毒酶及相关基因表达的影响

以脊尾白虾(Exopalaemon carinicauda)糠虾幼体为实验材料,研究底泥浸出液对抗氧化解毒酶和基因表达的影响。用长江口水域的底泥浸出液与过滤海水按等比例间距稀释成4个体积浓度水平(1∶4,1∶2,1∶1,1.5∶1),以过滤海水为对照组,每一个浓度组设3个平行样本,进行20 d的毒性实验。结果表明:实验进行20 d,底泥浸出液对糠虾幼体肌肉组织CAT酶活性有一定的诱导作用,浸出液浓度与CAT酶活性表现出正相关的剂量效应关系。实验开始10 d,不同浸出液胁迫下,GST酶活性均出现增加受到诱导,实验20 d后,GST酶活性下降受到抑制作用,但不同浸出液浓度胁迫下抑制和诱导效应关系不明显。虾类肌肉组织3种基因(CAT、GSTD和HSP70)表达量基本都表现为实验开始10 d时升高,而实验20 d后表达量下降,仅为10 d数值的百分之一左右,表明底泥浸出液对CAT、GSTD和HSP70的3种基因的mRNA转录水平表达在一定时间内有一定的诱导作用,但胁迫时间过长时,则产生较明显的抑制作用。     

水体生态系统对不同浓度水平苯酚污染的急性毒性响应

针对浙江某地苯酚泄漏事件,结合实地监测和实验室研究,探索苯酚的水体污染化学行为对水体生态系统的影响和急性毒性效应.本研究选取水生生态系统中明亮发光杆菌T3、优势藻四尾栅藻Scenedesmus quadricauda以及模式鱼斑马鱼Brachydanio rerio为研究对象,探索水体中生产者、消费者及分解者等整个生态系统对不同苯酚浓度胁迫下的生态响应.结果表明,水生生物对苯酚污染浓度的生态响应差异显著,虽然苯酚对发光细菌、四尾栅藻及鱼体都具有一定的毒性影响,但四尾栅藻抑制率与苯酚浓度之间并没有良好的线性关系.发光细菌的相对发光率与苯酚浓度的关系表明,0.2—25 mg.L-1苯酚浓度对水体会产生低毒作用,而30—100 mg.L-1苯酚浓度对水体会产生中毒作用.苯酚对四尾栅藻的影响主要表现为光合作用的暗反应破坏,高浓度苯酚处理组中四尾栅藻会将残余苯酚浓度作为自身碳源而短暂地促进其生长.斑马鱼在苯酚浓度高于15 mg.L-1时,会发生体重下降、死亡率上升、呼吸急促、游动迅速以及导致水质变坏等现象,并且在100 mg.L-1苯酚浓度处理下,鱼体内富集大量苯酚,通过食物链进而对人类健康产生影响.     

Growth inhibition in Japanese medaka (Oryzias latipes) fish exposed to tetrachloroethylene

A recent study in our laboratory has demonstrated that tetrachloroethylene (TCE) is acutely toxic to Japanese medaka (Oryzias latipes) larvae with a 96 hr-LC50 of 18 (17-19) mg/mL (Spencer et al., 2002). In the present study we hypothesize that TCE exposure induces a developmental effect in Japanese medaka. Growth and age specific sensitivity of Japanese medaka larvae were studied with four age groups (7, 14, 21 and 28 days old) to determine tetrachloroethylene effects on these parameters. The medaka larvae were exposed for 96 hours in a single concentration (10 mg/mL) of TCE. The toxic endpoints evaluated were larvae weight, length, water content and protein concentration. The study revealed that exposure of medaka larvae to this sub-acute concentration of TCE significantly reduced length and weight in the treated group. The difference in growth between control and treated groups was more obvious in age versus length, than in age versus weight. The dry weight-fresh weight ratio (dw/fw) was shown to be higher in the control group. Water content in TCE-treated medaka was higher than in the control group, and younger fry had more water content than older ones. A higher protein concentration was also observed in TCE-treated medaka compared to the control group. These results indicate that TCE has a profound effect on the growth and development of Japanese medaka larvae.     

Growth, carbon, nitrogen and caloric content of Solea senegalensis (Pisces: Soleidae) from egg fertilization to metamorphosis

Eggs and larvae of the Senegal sole, Solea senegalensis Kaup, were reared from fertilization until the end of metamorphosis, which occurs by Day 17 after hatching at 19.5 °C. Changes in energy content and biomass quality were studied in terms of dry weight and of carbon, nitrogen and energy content. S. senegalensis spawned eggs of about 1 mm diameter which hatched 38 h after fertilization. Average dry weight of individual eggs was 46 μg, the chorion accounting for about 18% of total dry weight. Gross energy of recently fertilized sole eggs was approximately 1 J egg⁻¹. From fertilization to hatching, eggs lost 8% of their total energy (chorion not included). After hatching, larvae lost 14% of their initial energy until the start of feeding which occurred about 48 h afterwards. The principal components catabolized during embryogenesis were carbon-rich compounds that decreased by 26%, while nitrogen-rich compounds decreased by only 10% and were practically unaltered from hatching to the start of feeding. Feeding larvae displayed constant growth during the period studied (specific growth rate on a dry weight basis was 0.26 d⁻¹). The relative proportion of carbon and nitrogen content revealed an accumulation of high energy compounds in the days before metamorphosis. By Day 14, the energy content reached values similar to those of recently hatched embryos, but decreased again during metamorphosis. Received: 10 June 1998 / Accepted: 28 January 1999     

Spatio-temporal modeling of striped-bass egg, larval movement, and fate in the San Francisco Bay-Delta

Most models developed for the movement and fate of eggs and larvae of aquatic species are based on a particle tracking approach. Although this method has many advantages due to its high flexibility, particle tracking may become computationally intensive for complex geometries and when large numbers of particles are needed to simulate the population properly. In continuous models based on advection and dispersion mechanisms, the computational burden is independent of the size of the population. We developed a continuous fate and transport model for striped bass eggs and larvae in the San Francisco Bay-Delta. The model predicts the concentration of eggs and larvae at any location over time. The method of moments was used to account for the effect of temperature and age on the transition of eggs to larvae and larvae to juveniles. Egg and larval mortality were represented as functions of temperature, and eggs also experienced settling mortality. The fate and transport model used the same one-dimensional spatial grid as the existing Delta Simulation Model II (DSM2) hydrodynamics model. DSM2 output of flow rates, water depths, and cross-sectional areas were inputted into the fate and transport model to determine transport. The model was applied to striped bass eggs and larvae data collected during years 1990-1994; agreement between the modeled and the measured data was acceptable in most cases. Exploratory simulations were performed to demonstrate how the model could be used to evaluate the effects on egg and larval survival and total juvenile production of water diversions for supply and agricultural use and changes in the long-term mean water temperature. The model can be further used to examine the impact of various operation strategies in the San Francisco Bay-Delta, where diversion losses of early life stages of fishes remain a major management issue.     

Laboratory culture of the lobate ctenophore Mnemiopsis mccradyi with notes on feeding and fecundity

Collection and culture of the large lobate ctenophore Mnemiopsis mccradyi Mayer is described, including the requirements for successful development of larvae. Particular attention must be given to the collection of these delicate animals, the handling and provision of live microzooplankton of suitable size for the larvae, and the provision of food densities for the adults which neither stimulate “wasteful” feeding nor limit their growth. Although these ctenophores will ingest detritus and algal cells in high concentration, they lost weight at the same rate as starved individuals unless provided with living zooplankton. Under optimum conditions, specimens would lay eggs within 13 days of their own birth. By the 17th day they laid eggs daily, and had produced an average of 8,000 eggs within 23 days after birth. The maximum number of eggs laid by a single wild individual within 24 h after being brought into the laboratory was 10,000. Their high fecundity, rapid generation time, and ability to self-fertilize help to explain their sudden appearance in bloom proportions at periods of high food concentration in the environment, often referred to in the literature.     

Effect of additions of dietary triacylglycerol microspheres on growth,survival, and settlement of mussel (<Emphasis Type="Italic">Mytilus</Emphasis> sp.) larvae

This study reports the effect of additions of dietary microspheres of triacylglycerol (TAG, extracted from the diatom Chaetoceros muelleri) on larval development and settlement of Mytilus sp. The first experiment showed that mussel larvae successfully ingested TAG microspheres as soon as they acquired the ability to feed from exogenous sources. In a second experiment, larvae were fed for 28 days on diets consisting of 0, 1, 20, or 50% TAG microspheres (based on the cell concentration of a full algal ration) added as partial replacements for a ration of Isochrysis sp. (T-ISO). Lipid content and growth of larvae fed on a diet composed of 20% TAG were higher than those of the control groups, whereas survival was negatively affected. No growth or survival effect was detected with larvae fed on a diet composed of 1% TAG, whereas high mortality after 14 days was observed for larvae fed on a diet composed of 50% TAG microspheres. In a third experiment, 22-day-old larvae were fed on rations of Isochrysis sp. supplemented with 0, 1, 10, 20, 50, and 100% TAG for 2 days and allowed to settle for a 7-day period. TAG content of the larvae increased with TAG added to the diet until a saturation threshold was reached between a 20 and 50% supplementation level. Similarly, free fatty acid (FFA) content increased with TAG level in the diet and was linearly correlated with TAG content of larvae. Increased levels of FFA in larvae were attributed to digestion of TAG supplements. Settlement success and survival of larvae were not affected by diet; however, regression analysis revealed that TAG level in pre-metamorphic larvae explained 28% of survival variability among cultures.Communicated by R.J. Thompson, St. Johns     

Food concentration and stocking density effects on survival and growth of laboratory-reared larvae of bay anchovy Anchoa mitchilli and lined sole Achirus lineatus

Bay anchovy (Anchoa mitchilli) eggs were stocked at densities from 0.5 to 32.0 l^-1 and larvae were fed on wild plankton (copepod nauplii) in concentrations that ranged from 50 to 5000 prey l^-1. Lined sole (Achirus lineatus) eggs were stocked at 0.5 to 16.0 l^-1 and larvae were fed wild plankton at concentrations from 50 to 1000 prey l^-1. Some larvae of each species survived at all stock and food levels to the transformation stage at 16 days after hatching. Survival rates for both species exceeded 40% when food concentration was 1000 l^-1 or higher. Growth and dry weight yields also increased significantly at the higher food concentrations. Effects of initial stocking density were not well defined, but both survival and growth decreased at the highest stocking rates. Standardized culture of bay anchovy and lined sole larvae can be based on a food concentration of 1000 copepod nauplii l^-1 to routinely produce healthy larvae.     

加拿大一枝黄花化感抑藻效应的初步研究

利用植物化感作用,可抑制有害藻类的生长,改善富营养化水体水质。文章选择我国重要的陆生入侵植物加拿大一枝黄花（Solidago canadensis L.）,系统研究了其根部、茎部和叶部水浸液对5种常见藻类：铜绿微囊藻（Microcystis aeruginosa）、斜生栅藻（Scenedesmus obliquu）、蛋白核小球藻（Chlorella pyrenoidosa）、水华鱼腥藻（Anabaena flos-aguae）和羊角月牙藻（Selenastrum capricornutum）生长的化感效应。研究结果表明：（1）加拿大一枝黄花水浸液对铜绿微囊藻的生长有强烈的化感抑制效应,7 d抑制率为90%左右,根、茎、叶部水浸液的72 h-EC50由高到低依次为34.84、12.54和9.57 g.L^-1,叶部对铜绿微囊藻的抑制作用最强;（2）加拿大一枝黄花叶部水浸液除了对水华鱼腥藻的生长无显著影响外,对其余4种受试藻均有明显的抑制作用,即加拿大一枝黄花叶部的化感抑藻效应较具广谱性;（3）相同质量浓度、同一部位的加拿大一枝黄花水浸液对不同藻种的化感作用类型和作用强度有所不同,即化感抑藻效应存在选择性,且这种选择性与藻类的分类学特征无关;（4）对同一种藻,加拿大一枝黄花根、茎、叶等不同部位水浸液的化感效应也有所差异。     

Effect of caffeine, norfloxacin and nimesulide on heartbeat and VEGF expression of zebrafish larvae

The use of pharmaceuticals during pregnancy may causes abnormalities to the embryo. Sometime the drug also effect to the new born if the drug transferred through lactation. We have used zebrafish model to see the effect of some pharmaceuticals on embryos and larvae. Three drugs, caffeine, norfloxacin and nimesulide, were used for this study to see the effect mainly the hatching rate of eggs, heart beat rate and the vascular endothelial growth factor (VEGF) expression of the larvae. VEGF is an important signaling protein that involved generating the new blood vessels during embryonic development. We have used 10, 20, 50, 100 microg ml(-1) concentrations of all the drugs to see the effect. No significant mortality or malformations were observed in zebrafish embryos. Hatching was stared from 60 hr. In control group, 91% hatching rate was observed. Lowest hatching rate was observed using highest concentration of norfloxacin (100 microg ml(-1)) and nimesulide (100 microg ml(-1)) i.e. 55 and 56% respectively. In control group, 110 to 115 heart beat rate was counted per minute. Significantly higher heart beat was observed in caffeine treated group which is 125 to 140 min(-1) Lower heart beat was noted in nimesulide treated group which is 100 min(-1). We have tried to observe the possible effect of VEGF of the larvae by these three drugs. Expression of VEGF was very low in caffeine treated group. Almost no VGF expression was observe in 100 microg ml(-1) caffeine treated group. These studies suggest that there is a possibility that high dosage of caffeine can harm the unborn baby or new born babies, if the mothers use caffeine.     

Laboratory spawning and rearing of a marine fish,the silverside Menidia menidia menidia

Adult silversides, Menidia menidia menidia (Linnaeus), were collected in early March, 1974 and maintained in 3 recirculating seawater tanks in the laboratory. Respective groups were fed Moore-Clark Fry Fine at 3, 7 and 10% of their body weight per day. The photoperiod (light intensity approximately 2000 lux) was increased in increments of 10 min/day from 12 h light to 14 h light. The water temperature was increased by 1C°/day from the ambient collection temperature, 14°C, to 22°C. Twenty-four days after beginning laboratory conditioning, fish in each tank were stripped. There was a significant increase (², =0.05) in the number of ripe males at all three feeding levels, compared to an initial field-collected group that was checked at the beginning of the conditioning period. Females also showed significant increases in ripeness at the 7 and 10% but not at the 3% feeding level. The gonadal indices (gonad weight expressed as percentage of body weight) of both sexes were significantly greater than those measured for the initial field-collected group, but did not differ from those of adults collected from the field at the time laboratory conditioning was terminated. Techniques for maintaining eggs from field-ripened adults in the laboratory have been developed, and the effect of salinity on the percentage emergence of larvae determined. The highest emergence rate of larvae was 61% when eggs were maintained at 30 S. Emergence was 56% at 20 S and 47% at 10 S. The effect of delayed feeding on survival and growth of larvae was determined at 20 and 30 S and 25°C. Survival and growth was best for larvae fed Artemia sp. nauplii immediately after emergence at 30 S.Contribution No. 252, Gulf Breeze Environmental Research Laboratory.Associate Laboratory of the National Environmental Research Center, Corvallis, Oregon, USA.     

An experimental evaluation of transgenerational isotope labelling in a coral reef grouper

Transgenerational isotope labelling (TRAIL) using enriched stable isotopes provides a novel means of mass-marking marine fish larvae and estimating larval dispersal. The technique, therefore, provides a new way of addressing questions about demographic population connectivity and larval export from no-take marine protected areas. However, successful field applications must be preceded by larval rearing studies that validate the geochemical marking technique, determine appropriate concentrations and demonstrate that larvae are not adversely affected. Here, we test whether injection of enriched stable barium isotopes (¹³⁵Ba and ¹³⁷Ba) at two dose rates produces unequivocal marks on the otoliths of the coral reef grouper Epinephelus fuscoguttatus. We also assess potential negative effects on reproductive performance, egg size, condition and larval growth due to injection of adult female fish. The injection of barium isotopes at both 0.5 and 2.0 mg Ba/kg body weight into the body cavities of gravid female fish was 100% successful in the geochemical tagging of the otoliths of larvae from the first spawning after injection. The low-dose rate produced no negative effects on eggs or larvae. However, the higher dose rate of 2 mg Ba/kg produced small reductions in yolk sac area, oil globule area, standard length and head depth of pre-feeding larvae. Given the success of the 0.5 mg Ba/kg dose rate, it is clearly possible to produce a reliable mark and keep the concentration below any level that could affect larval growth or survival. Hence, enriched Ba isotope injections will provide an effective means of mass-marking grouper larvae.