双酚A和壬基酚长期暴露对斑马鱼繁殖的影响

内分泌干扰物的研究目前正逐渐成为国际研究新的热点，但长期暴露条件下对生殖情况的研究目前较少。本文研究了环境类雌激素双酚A（BPA）以及和壬基酚（NP）联合的作用，对暴露一个世代的斑马鱼（F1）（Danio rerio）的生殖情况、子代质量的影响。将健康的受精卵48h后暴露于200，500，1000μg·L^-1 BPA以及三个浓度与30μg·L^-1 NP的联合，同时设置空白对照和30μg·L^-1 NP对照。130dph将成鱼置于清水喂养20d，统计每天的产卵量、产卵次数、畸形率、孵化率、卵径、破膜时间以及耐饥时间。以此为终点评估暴露后斑马鱼的生殖能力和子代质量。与对照相比NP，BPA可以抑制斑马鱼的生殖，两种类雌激素联合作用可以极显著地抑制斑马鱼的生殖（P〈0．01）；BPA各浓度组导致F1代畸形率上升（P〈0．05）；高浓度BPA抑制产卵量（P〈0．05）；BPA200，500μg·L^-1会延长F1代的破膜时间和耐饥时间（P〈0．01），而BPA1000μg·L^-1则产生相反的效果（P〈0．05）。双酚A对斑马鱼的生殖有明显的抑制作用，双酚A和壬基酚有协同效应。长时间暴露于双酚A会影响斑马鱼的生殖能力和子代质量。     

Delivering oxytetracycline to first-feeding zebrafish, Danio rerio (Hamilton), and goby, Asterropteryx semipunctata Rüppell, larvae using lipid spray beads

Lipid spray beads (LSB) were evaluated for delivery of the low-molecular weight, water-soluble antibiotic, oxytetracycline·HCl (OTC) to fish larvae. Various OTC core-to-lipid ratios and OTC core concentrations were evaluated to maximize OTC delivery efficiency by LSB. Acceptability and digestion of LSB containing OTC and riboflavin by larval zebrafish, Danio rerio (Hamilton), and larval gobies, Asterropteryx semipunctata Rüppell, were also evaluated. Increasing LSB core-to-lipid ratios from 1:3 to 1:1 v/v resulted in an increase in encapsulation efficiency (EE) from 2.33 to 3.68% w/w. Increasing OTC concentrations of core solutions from 0.1 to 0.5 g OTC mL⁻¹ increased EE from 3.95 to 18.77% w/w, respectively. Although retention efficiency (RE) was unaffected by this increase, delivery efficiency was increased to 7.9 ± 0.7% w/w, after correcting for leakage losses because of the suspension of beads in water for 60 min. Consumption of LSB containing OTC by first-feeding zebrafish and goby larvae was confirmed by analysis of feeding incidence and gut fullness. Visual observations of larvae fed on LSB containing riboflavin indicated that larvae of both species digested LSB. Zebrafish larvae fed on OTC LSB contained 39.3 ± 2.5 ng OTC after purging LSB from their guts. Use of LSB provides an effective means of delivering therapeutics to fish larvae and could greatly reduce required doses compared with current methods of immersing larvae in solutions of therapeutic agents.     

Replacement of live food with a microbound diet in feeding Litopenaeus setiferus (Burkenroad) larvae

Microbound feeds have been well accepted by shrimps and farmers in many penaeid shrimp hatcheries. The present study focused on an adequate level of replacement of Artemia nauplii and microalgae by a microbound diet for rearing Litopenaeus setiferus (Burkenroad) larvae. A microbound diet (MBD) consisting of fishmeal, squid meal, shrimp meal, yeast meal and soybean meal was used. The first experiment was designed to obtain the optimum level of MBD to complete the live feeding schedule, from Protozoea (PZ_III) to Mysis (M_III). The experimental levels of the microbound diet tested were 2, 4, 6 and 8 mg MBD L^?1 day^?1. The next step was to determine the Artemia nauplii replacement level from PZ_I to M_III by MBD. These experiments were carried out either in the presence (Experiment 2) or in the absence of algae (Experiment 3). Four replacement levels were tested: 0% (4 mg MBD L^?1 day^?1: 1 Artemia nauplii mL^?1), 40% (5.5 mg MBD L^?1 day^?1: 0.6 Artemia nauplii), 60% (6.5 mg MBD L^?1 day^?1: 0.4 Artemia nauplii) and 100% (8 mg MBD L^?1 day^?1: 0 Artemia nauplii). In all experiments growth, survival, development, quality index (QI) and performance index (PI), were used to determine the optimum concentration of microbound diet. Results showed that 6 mg MBD L^?1 day^?1 can be recommended as a complement to live food for L. setiferus larvae from PZ_III to M_III. In the presence of algae, maximum growth and survival may be obtained in 40–60% (5.5–6.5 mg MBD L^?1 day^?1) of Artemia nauplii replacement levels. In the absence of algae, the Artemia nauplii replacement resulted in slower development, less salinity resistance, lower growth and lower survival than was obtained in larvae fed with algae.     

Early weaning of winter flounder (Pseudopleuronectes americanus Walbaum) larvae on a commercial microencapsulated diet

Like most small marine fish larvae, the stomachs of winter flounder Pseudopleuronectes americanus are undeveloped at first feeding and have relatively reduced digestive capacity. This work was undertaken to test whether larvae at the onset of stomach differentiation (larval size about 5.5 mm) could be early weaned onto a commercial microencapsulated diet. We assessed the effect of early weaning by first comparing growth performance (standard length, total protein content and age at metamorphosis) of larvae fed enriched live prey from first feeding to a size of 5.5 mm and then reared on three different feeding regimes until metamorphosis: (1) live prey (LP) as a control group; (2) mixed feeding of live prey and microencapsulated diet (LP‐ME); (3) exclusively microencapsulated diet (ME) after fast weaning over 4 days (to a larval size of 6.2 mm). No differences were observed between larval development in the two first groups, which began metamorphosis at 40 days old. The larvae of the third group showed significantly slower growth that resulted in a delay of 4 days in the onset of metamorphosis. Differences in live prey availability between the treatments and the short transition period to allow the larvae to adapt to the new diet were identified as possible contributing factors to the slower growth and to the delay in metamorphosis of early weaned larvae. In a second experiment, the transitional weaning period was increased until the larvae were 6.6 mm in length. Weaning at that size resulted in no slowing of growth or delay in metamorphosis, suggesting that the feeding schedule was adequate.     

Effects of Pro-Tex on zebrafish (Danio rerio) larvae,adult common carp (Cyprinus carpio) and adult yellowtail kingfish (Seriola lalandi)

Aquaculture practices bring several stressful events to fish. Stressors not only activate the hypothalamus–pituitary–interrenal-axis, but also evoke cellular stress responses. Up-regulation of heat shock proteins (HSPs) is among the best studied mechanisms of the cellular stress response. An extract of the prickly pear cactus (Opuntia ficus indica), Pro-Tex, a soluble variant of TEX-OE^®, may induce expression of HSPs and reduce negative effects of cellular stress. Pro-Tex therefore is used to ameliorate conditions during stressful aquaculture-related practices. We tested Pro-Tex in zebrafish (Danio rerio), common carp (Cyprinus carpio L.) and yellowtail kingfish (Seriola lalandi) exposed to aquaculture-relevant stressors (thermal stress, net confinement, transport) and assessed its effects on stress physiology. Heat shock produced a mild increase in hsp70 mRNA expression in 5-day-old zebrafish larvae. Pro-Tex increased basal hsp70 mRNA expression, but decreased heat-shock-induced expression of hsp70 mRNA. In carp, Pro-Tex increased plasma cortisol and glucose levels, while it did not affect the mild stress response (increased plasma cortisol and glucose) to net confinement. In gills, and proximal and distal intestine, stress increased hsp70 mRNA expression; in the distal intestine, an additive enhancement of hsp70 mRNA expression by Pro-Tex was seen under stress. In yellowtail kingfish, Pro-Tex reduced the negative physiological effects of transport more efficiently than when fish were sedated with AQUI-S^®. Overall, our data indicate that Pro-Tex has protective effects under high levels of stress only. As Pro-Tex has potential for use in aquaculture, its functioning and impact on health and welfare of fish should be further studied.     

重金属Cu~(2+)、Cd~(2+)、Hg~(2+)对斑马鱼胚胎发育的毒性效应

在半静态水的条件下,研究了铜离子(Cu~(2+))、镉离子(Cd~(2+))和汞离子(Hg~(2+))对斑马鱼(Danio rerio)胚胎发育的单一和联合毒性效应。采用48 h、72 h、96 h致死率和72 h、96 h孵化抑制率作为生理毒性终点,以其半数致死浓度(LC50)值作为毒性评价标准。结果表明:Cu~(2+)、Cd~(2+)和Hg~(2+)对斑马鱼胚胎的48 h-LC50分别为0.58 mg·L~(- 1)、0.72 mg·L~(- 1)和0.000 46 mg·L~(- 1),72 h-LC50分别为0.39 mg·L~(- 1)、1.15 mg·L~(- 1)和0.000 30 mg·L~(- 1),96 hLC50分别为0.08 mg·L~(- 1)、0.19 mg·L~(- 1)和0.000 18 mg·L~(- 1);Cu~(2+)、Cd~(2+)和Hg~(2+)的72 h孵化抑制率的LC50分别为0.05 mg·L~(- 1)、0.01 mg·L~(- 1)和0.000 04 mg·L~(- 1),96 h孵化抑制率的LC50分别为0.06 mg·L~(- 1)、0.05 mg·L~(- 1)和0.000 11 mg·L~(- 1);Cu~(2+)、Cd~(2+)和Hg~(2+)对斑马鱼胚胎毒性大小顺序为Hg~(2+)Cu~(2+)Cd~(2+)。Cu~(2+)、Cd~(2+)和Hg~(2+)对斑马鱼胚胎的致死率和孵化抑制率均具有明显的剂量-效应关系,72 h孵化抑制率可以作为斑马鱼胚胎最敏感的毒性终点指标。分别用单一毒性Cu~(2+)、Cd~(2+)和Hg~(2+)的96 h-LC50值,按毒性单位1∶1两两混合,对斑马鱼胚胎在第48、第72、第96小时的联合毒性均为协同作用;按毒性单位1∶1∶1混合共存时,对斑马鱼胚胎在第48、第72、第96小时的联合毒性均为拮抗作用。     

Fatty acid nutritional quality of sea urchin Paracentrotus lividus (Lamarck 1816) eggs and endotrophic larvae: relevance for feeding of marine larval fish

Sea urchin eggs and larvae have been suggested as potential live prey for marine fish larval feeding. This study evaluated the fatty acid composition of Paracentrotus lividus eggs, prisms and four-armed plutei, obtained from wild and captive broodstocks fed on raw diets: maize, seaweed and a combination of maize and seaweed. Amounts of essential fatty acids (EFA) for marine fish larvae [arachidonic acid (ARA), eicosapentaenoic acid (EPA) and docosahexanoic acid (DHA)] were determined in eggs and endotrophic larvae. ARA ranged from 3.93% in eggs from combination to 18.7% in plutei from maize diets. In any developmental stage, EPA amounts were always lower than 5% for the raw diets, and DHA showed null or trace amounts including the wild diet. Thus, broodstock-prepared diets had to be formulated based on different lipid sources (Algamac, linseed oil, cod liver oil and olive oil) in order to test eggs and larvae EFA enhancement. EFA improvement was possible for all tested prepared diets. Algamac diet lead to superior EFA enhancement mainly in DHA (7.24%, 4.92% and 6.09% for eggs, prisms and plutei, respectively) followed by cod liver oil diet. Only these two lipid sources should be considered for prepared broodstock diets in order to obtain suitable live prey for fish larval feeding.     

Natural food and artificial,dry starter diets: effects on growth and survival in intensively reared European grayling

Four artificial, dry feeds and two diets consisting of natural, live food were evaluated for use as starter diets in the intensive rearing of European grayling (Thymallus thymallus). Long-term effects of the different diets were also studied. The investigation was divided into two parts: a 14 day starter period and the subsequent 115 days, during which the long-term effects were studied. Diets differed between the different treatment groups during the initial 2 weeks, whereafter all fish were offered a single diet. At the end of the starter period, significant differences in weight (p<0.001) were obtained between natural (control) feed groups and three of the artificial feed groups. The highest growth rate (9.6% day^–1), and length increase (9 mm) as well as the lowest mortality rate (6.0%) were obtained with one of the artificial diets. Growth rate, length increase and mortality of grayling in the other diet groups varied from 3.6 to 7.7% day^–1, 5 to 7 mm and 7.0 to 9.1%, respectively. The switchover to a different dry diet resulted in an increase in mean daily mortality for all groups previously fed a dry diet. For the fish previously fed live food the mortality rate remained about the same. Groups of fish growing slowly during one period seemed to compensate for their initially low growth by increasing their growth rates during the next period. The highest mean daily mortality for all groups during the experiment was observed during the period with the highest water temperature. It was concluded that European grayling can be intensively reared during the start-feeding phase on artificial as well as natural diets, and that artificial dry diets can be used exclusively throughout the first summer.     

Feeding and digestion in the caridean shrimp larva of Palaemon elegans Rathke and Macrobrachium rosenbergii (De Man) (Crustacea: Palaemonidae) on live and artificial diets

Larvae of two caridean shrimp species, Macrobrachium rosenbergii (De Man) and Palaemon elegans Rathke, were fed live and artificial diets. P. elegans larvae fed exclusively live Artemia salina (15 nauplii mL^?1) developed into first postlarval stage (PL1) within 12 days at a temperature of 25°C and salinity 32.5 g L^?1. Their survival and mean total length at this stage were 88.5% and 6.7 mm respectively. M. rosenbergii larvae fed on 15 Artemia mL^?1 started to metamorphose into PLl within 24 days at 29–30°C and 12 g L^?1. Attempts to completely replace live Artemia for rearing P. elegans during early stages failed, and only a partial replacement was achieved for the larvae of both species. P. elegans larvae survived (49%) solely on a microgranulated diet (Frippak PL diet) from stage zoea (Z) 4–5 to PL1. Similarly, a microencapsulated diet (Frippak CD3) also sustained M. rosenbergii larvae from Z5–6 to PL1 with a 28% survival. Development of the larvae of both species was retarded by 2–3 days and their survivals were lower than those fed on the live diet. The inability of the early larvae of these caridean species to survive on artificial diets is attributed to their undeveloped guts and limited enzymatic capabilities. Trypsin activity in the larvae was determined for all larval stages. It was found that the highest trypsin activity, at stage Z4–5 in P. elegans and at stage Z5–6 in M. rosenbergii, coincides with a rapid increase in the volume of the hepatopancreas and the formation of the filter apparatus. These morphological changes in the gut structure appear to enable the larvae to utilize artificial diets after stage Z5–6. Low larval trypsin activities may be compensated by the easily digestible content of their live prey during early larval stages (Z1–Z4/5) and by longer gastroevacuation time (GET) and almost fully developed guts during later stages.