Trypsin activity and physiological aspects in larval rearing of European sea bass (Dicentrarchus labrax) using live prey and compound diets

The aim of this study was to evaluate the potential of a compound diet as a live prey substitute for feeding European sea bass larvae ( Dicentrarchus labrax L.). The effect of a commercial diet (Nippai ML feed) and live prey ( Artemia nauplii) on tryptic enzyme activity, protein content, growth (standard length) and survival rates of sea bass larvae were tested during a 27-day rearing experiment. Sea bass larvae were divided into two groups. The live food group (control group) was fed exclusively on newly hatched Artemia nauplii (Inve AF grade), the test group was fed exclusively with the compound diet from day 15 onwards. As trypsin has been demonstrated to be a useful indicator for evaluating digestibility of food and the nutritional condition of fish larvae, individual tryptic enzyme activity was determined in both feeding groups. Larvae older than 14 days after hatching and fed on live food showed a significantly higher tryptic enzyme activity than larvae fed the compound diet. A similar relationship between tryptic activity and standard length in both test groups was detected only in small larvae (standard length < 7 mm). The usefulness of proteolytic enzyme activity measurements in larval fish research, as well as its use in aquaculture nutrition research, was confirmed. Protein content, increase in length and survival rates of the sea bass larvae were additionally determined in order to evaluate an influence on the diet. The protein content of larvae fed the Artemia nauplii was higher and the growth of larvae fed the compound diet was reduced. Larval mortality was not affected by the diet given.     

Effects of experience on feeding success by larval white seabass, Atractoscion nobilis

Laboratory experiments on larval white seabass, Atractoscion nobilis , showed that first feeding success on mixed zooplankton was zero, and less than 5% on Artemia nauplii , for larvae of different sizes. Experience increased success from 20% after 2 days to 69% after 14 days when larvae fed on Artemia nauplii , and to 42% after 4 days of feeding on zooplankton. This increase was attributed to learning in larvae less than 5·0 mm s.l. , and in part to size-related factors in larvae greater than 5·0 mm s.l.     

Growth of Perca fluviatilis larvae fed with Artemia spp. nauplii and the effects of initial starvation

Experiments were performed to test the growth and survival of perch larvae fed with Artemia spp. nauplii and to determine the most advantageous time of first feeding. During a first experiment, perch larvae were divided into groups according to their hatching dates. Daily ingestion of Artemia spp. nauplii began significantly 2 days after mass larvae hatching. By day 7, the daily prey acceptance was over 75% among the separate larvae groups. During a second 14-day feeding experiment, larvae fed after a 2- or a 3-day delay showed the best growths: 12.5 mm and 13.7 mg. There was no significant difference (P > 0.05) among survival rates (85.3%) of perch larvae fed after a 1-, 2- or 3-day delay.     

Use of the rotifer, Brachionus calyciflorus Pallas, in freshwater ornamental fish larviculture

The Brachionus calyciflorus used in this study were produced by batchculture using Chlorella spp. as feed. Larviculture experiments in indoor10-l and 200-l tanks revealed that, compared with egg yolk, the rotifersused as starter food significantly improved the growth and survival of DwarfGourami larvae (Day 2–12). These beneficial effects also extended tothe subsequent Artemia feeding phase (Day 13–32), suggesting that thequality of starter food is crucial to later development. At metamorphosis,the overall survival rate of larvae fed on rotifers in indoor tanks(65.1–74.5%) was about four times of that obtained in extensiveculture in open ponds (17.5%). In Discus, larvae are dependent on thebody slime of their parent as a nutrient during the first two weeks ofexogenous feeding. Our observation demonstrated that Brown Discus larvaecould be raised in the absence of the parent fish by using rotifers asstarter food followed by Artemia nauplii. Their growth and survival ratewere comparable to those on parental feeding. The artificial feeding wouldeliminate the risk of larvae being eaten by the parent fish and shorten thebrooding interval of the spawners, thereby leading to higher yield of fry.This feeding protocol is less tedious and more practical for use incommercial farming of Discus than the existing strategies of smuggling thebatch of larvae to foster parents or feeding the larvae with egg food. Theuse of rotifers would enable freshwater larviculture to improve larvalperformance, increase yield, and facilitate breeding of new fish specieswith small larvae.     

Growth and physiological changes during metamorphosis of Senegal sole reared in the laboratory

The metamorphosis of Solea senegalensis was studied in larvae reared at 20° C and fed four different feeding regimes. A, Artemia (4 nauplii ml⁻¹); B, Artemia (2 nauplii ml⁻¹); C, mixed diet (2 nauplii ml⁻¹ and 3 mg ml⁻¹ microencapsulated diet); and D, microencapsulated diet (3·7 mg ml⁻¹). Rotifers were also supplied in all cases during the first days of feeding. These feeding regimes supported different growth rates during the pre-metamorphosis period (regime A, G=0·376 day⁻¹; regime B, G=0·253 day⁻¹; regime C, G=0·254 day⁻¹; regime D, G=0·162 day⁻¹). Larvae started metamorphosis 9 days after hatching (DAH) when fed the regime A, 13 DAH with regime B, 11 DAH with regime C and 15 DAH with regime D. A minimum 5·6–5·9 mm L_T was required under all feeding regimes to initiate the metamorphosis. Eye translocation was completed when the larvae reached 8·6–8·7 mm L_T (regimes A, B and C), but only 7·3 mm L_T with regime D. 4·4–6·2 days were required to complete eye migration under the regimes A, B and C, and 18·3 days under the regime D. This transformation is concomitant with changes in body reserves, and with the pattern of some digestive enzymes.     

Growth and survival of tench larvae fed under different feeding strategies

Two feeding experiments were conducted to evaluate the growth and survival rates of tench Tinca tinca L. larvae when initially fed with a combination of three different brands of Artemia nauplii under two conditions: (A) in the laboratory and (B) on a commercial farm. At the same time, a protozoan culture of the freshwater ciliate Colpoda cucullus was additionally tested in one of the experiments to possibly enhance the initial hunting behaviour of the larvae. The larvae were fed every 4 h from the onset of exogenous feeding up to 14 days of age. Three types of commercial Artemia products, mainly differing in size and high unsaturated fatty acid (HUFA) content, were used. One group was fed with C. cucullus as a starter. The different combinations of Artemia nauplii were used to evaluate possible effects on larval growth. The final growth at 26.3 °C, expressed in length and weight, did not show significant differences, suggesting the use of the most economically feasible Artemia strain studied. The experiments confirmed that using smaller prey during the first 2 days of feeding increases their survival rate, although the mean final survival rate was high (89%). In the experiment carried out at the commercial fish farm facilities, experimental groups were also fed with Artemia nauplii, using the EG type either enriched or not enriched with HUFA. Again, one of the groups was offered the ciliate C. cucullus as initial feed. Final growth showed significant differences when using Colpoda culture as a starter feed, although this test resulted in the lowest survival rate (69%), indicating that further studies on the management of its culture should be undertaken to improve the applicability of the technique. The mean final survival rate was 83%.     

4种生态因子对虎斑乌贼幼体生长与存活的影响

旨在研究温度、盐度、饵料种类、投饵量对虎斑乌贼（Sepia pharaonis）幼体生长与存活率的影响,以确定其生长发育的最佳生态条件,为人工育苗提供理论依据。在室内控制条件下,采用单因子试验研究了不同温度（18、21、24、27、30和33℃）、不同盐度（18、21、24、27、30和33）、不同饵料种类（虾糜、活糠虾、卤虫无节幼体、桡足类、死糠虾、虾糜＋强化卤虫后无节幼体、强化卤虫后无节幼体）、活糠虾不同投饵量（0、2、4、6、8和10 g/d）对虎斑乌贼幼体生长发育的影响。结果显示：不同温度对虎斑乌贼幼体生长影响显著（P〈0.05）,最适温度为24-27℃,在最适温度下,存活率为84.4%-91.1%,特定生长率为4.82%-6.13%,存活率（y）与温度（x）的函数关系为y=-0.15x3＋30.637x-447.002（r2=0.923）。幼体适宜盐度为24-33,最适盐度为27,在最适盐度条件下,其存活率为（90.0±5.29）%,特定生长率为（3.71±0.34）%。投喂7种开口饵料,以活糠虾效果最佳,存活率为（96.7±2.94）%,特定生长率为（3.77±0.23）%;强化卤虫后无节幼体效果次之,存活率为（95.6±2.31）%,特定生长率为（2.54±0.15）%,其余各组培养效果均不理想。投喂活糠虾,随着投饵量的增加,其存活率随之提高,摄食量（y）与个体重（x）的回归方程为y=0.227x-0.063（r2=0.921）。     

Food selection and growth of young snakehead Channa striatus

Food selection and growth of young snakehead Channa striatus were studied in the laboratory and in a field trial. In the laboratory, first-feeding snakehead larvae of 6–7 mm total length (TL) with a mouth opening of 0.55 mm selected for Artemia nauplii, and against formulated feed. Fish began feeding on formulated feed at 12mm TL when their mouth width reached 1.0mm. In both laboratory and field trials, snakehead diets changed as fish size increased. For fish 15–20 mm TL, cladocerans and copepods were 96.5% of their diet. With fish 30–40 mm TL, zooplankton consumption was greatly reduced while benthic organism consumption increased. Fish 45–50 mm TL fed exclusively on benthic invertebrates. Diet shift from zoo-plankton to benthic invertebrates was not due to reduced zoo-plankton availability, but was instead related to changes in gill raker structure. Low density of benthic invertebrates in the field trial caused reduced fish growth rates when fish switched diets from zooplankton to benthos. Our results indicate that snake-head can take Artemia nauplii as a larval starter food, then accept formulated feed at ≥12 mm TL. Zooplankton can serve as food for snakehead < 40 mm, but formulated feed should be provided for larger fish which are unable to catch zoo-plankton.     

不同开口饵料对西伯利亚鲟仔鱼生长、存活和体成分的影响

分别采用水丝蚓（Limnodrilus sp.）、卤虫无节幼体（Artemia nauplii）、枝角类（Moina sp.）和人工饲料饲养西伯利亚鲟仔鱼30 d,研究不同开口饵料对西伯利亚鲟仔鱼生长、存活率和体成分的影响.结果表明：卤虫无节幼体为西伯利亚鲟最适开口饵料,仔鱼的存活率最高（96.67%）; 投喂水丝蚓组生长速度最快;而人工饲料组生长速度明显低于其他组,且成活率最低.不同开口饵料组间仔鱼体成分差异显著,人工饲料组水分含量最高,且粗蛋白和粗灰分含量最低.采用卤虫无节幼体为西伯利亚鲟仔鱼开口饵料,然后采用水丝蚓进行强化培育,能获得较好的生长速度和存活率.     

Sibling cannibalism in dorada under experimental conditions.

Cannibalism among embryos and larvae of Brycon moorei (Characidae) occurs during daytime and night-time, and persists under permanent darkness. Embryos and larvae of dorada provisioned with formulated feed over the first week of exogenous feeding did not survive, except for those exerting cannibalism. When oered alternative fish prey [embryos of Prochilodus magdalenae (0·5–0·8 mg) and Oreochromis niloticus (9·10 mg)], 1-day-old embryos of dorada preferred preying on these, thereby reducing early cannibalism. However, this promoted depensatory growth and more intense cannibalism later in the larval stage. Dorada provisioned with Artemia nauplii in excess showed more homogeneous growth and higher survival, most cannibalistic acts being restricted to the first 24 h of exogenous feeding, just after oral teeth were fully developed (21 h after hatching). Provisioning dorada with Artemia nauplii a few hours before their oral teeth were fully developed reduced early cannibalism from 41 to 15%. High proportions of deformed fish caused higher mortality, both directly and indirectly, as they promoted early cannibalism, depensatory growth and more intense cannibalism among larvae. The initial sorting of embryos, based on their occupation of the water column improved survival significantly during the first week of exogenous feeding, up to 52% in progenies containing <10% of deformed fish. Size-grading of larvae and young juveniles over the next 2 weeks reduced cannibalism to 2·6 and 1·9% day ⁻¹, in the first and second weeks, respectively. These results indicate that cannibalism in dorada can be mitigated eciently through appropriate rearing procedures, and open promising perspectives for the intensive culture of this fast-growing tropical species.     

Development of larval diets for milkfish (Chanos chanos)

This study aimed to develop nutritionally balanced and cost-effective processed diets for milkfish larvae ( Chanos chanos Forsskal). Two larval diets (feed A and feed B) were formulated and prepared to contain 45&percnt| protein and 10&percnt| lipid. Several larval diet preparations were tried such as microbound/ unpelleted (freeze-dried), microbound/pelleted (oven-dried) and microbound/flaked (drum-dried) and assessed in terms of feed particle size and buoyancy, water stability and feed acceptability. The preparation that gave the best particle size and buoyancy as well as good water stability was prepared as the microbound diet (using K -carrageenan as a binder) and flaked using a drum drier. A series of feeding experiments were conducted to determine the growth and survival of milkfish larvae reared on various feeding schemes using these processed larval diets which were fed either solely or in combination with live feed. Larvae in control treatments were reared on live foods such as Brachionus plicatilis and Artemia nauplii. Larvae were observed to ingest the diets, indicating that the feeds had suitable physical characteristics and were attractive to the larvae. The overall results of the feeding trials showed that the artificial diets could be fed to milkfish larvae in combination with Brachionus rotifers starting on day 2 or day 8, and could be fed alone starting from day 15. These promising results would reduce the dependence of milkfish larvae on live feed and would have significant economic benefits in the form of simplified milkfish hatchery procedures.     

The use of live and frozen Artemia salina nauplii enriched with fluorochromes for mass‐marking vendace Coregonus albula (L.) larvae

The aim of this study was to investigate the use of Artemia salina nauplii enriched with chosen fluorochromes for mass marking vendace Coregonus albula (L.) larvae. In the experiment, vendace larvae (6 DPH) were fed with live or frozen A. salina nauplii immersed in tetracycline hydrochloride (TC), calcein (Cal) or alizarin red S (ARS) for four subsequent days. More successful effects (marking otoliths) were obtained by feeding the fish with live nauplii enriched with fluorochromes, regardless of the dye type. The highest percentage of marked otoliths (100%) was recorded in the group fed with live nauplii immersed in TC. In the groups fed live or frozen nauplii enriched with Cal and ARS, a lower percentage of marked individuals (63.3%–73.7% and 56.7%–63.3%, respectively) were recorded. The survival rate of vendace larvae in particular groups oscillated between 93.7% and 95.7%. There were no significant differences in the total body length and body weight of the reared vendace larvae among different groups. In conclusion, for mass marking of vendace larvae using a feeding method, fish fed A. salinalive nauplii enriched with TC at a dose of 600 ppm is recommended for fishery practice.     

Mass marking of Leuciscus idus larvae using Artemia salina as a vector of fluorescent dyes

A method for the mass marking of ide Leuciscus idus larvae by feeding them Artemia salina nauplii that were immersed in different solutions of alizarin red S, tetracycline hydrochloride and calcein was tested. The best quality marks were obtained after feeding fish for 4 days with nauplii that had been immersed in 200 mg l^?1 alizarin red S.     

Ammonia toxicity as a criterion for the evaluation of larval quality in the prawn Macrobrachium rosenbergii

The feasibility of a short-term ammonia toxicity test as an evaluation criterion for larval quality was assessed in three trials. In each one, Macrobrachium rosenbergii larvae originating from the same spawn were nutritionally differentiated in two groups by feeding them either a nutrient-rich (Artemia nauplii enriched for 24 h with n-3 highly unsaturated fatty acids (HUFA) and ascorbic acid (AA)) or a nutrient-poor diet (Artemia nauplii starved for 24 h). Throughout their development, larvae from both treatments were exposed during 24 h to six concentrations of total ammonia (NH₄⁺+NH₃) and a control (no ammonia added). Based on mortality rates, the median lethal concentration for 50% of the population (LC₅₀) was estimated. As expected from earlier work, larvae fed the optimal diet presented higher n-3 HUFA and AA contents as well as higher growth and metamorphosis rates. From the moment the effect of diet quality was analytically detectable in the tissues of the larvae, the ammonia test was able to distinguish both groups of larvae. Differences in ammonia tolerance were observed as early as larval stage 4 and remained evident throughout larval development. The short-term ammonia toxicity test proved to be a valuable, sensitive and reproducible criterion for the establishment of larval quality.     

Preliminary investigation of feeding performance of larvae of early red-spotted grouper, Epinephelus coioides, reared with mixed zooplankton

Larvae of red-spotted grouper, Epinephelus coioides, were reared inoutdoor tanks with nauplii of copepods (mainly Pseudodiaptomus annandaleiand Acartia tsuensis) and/or rotifers, Brachionus rotundiformis. Grouperlarvae successfully started feeding on early stage nauplii even though theirabundance was as low as approximately 100 individuals l^–1 andshowed better survival and growth thereafter compared to those fed withrotifers only. Incidence of feeding reached 100% on day 4 whennauplii were available and only on day 9 when rotifers were given alone.Larvae seemed to be poor feeders at the onset of feeding, attempting tocapture any food organisms in the tank water. Selective feeding ability oflarvae started from day 4 and the larvae then preferred to feed on medium-and large-size nauplii rather than on rotifers as they grew. Larvae appearedto have a better chance at surviving in the presence of early stage nauplii,which were probably caught more easily than rotifers.     

The significance of diet in the growth and development of larvae of the blue crab,Callinectes sapidus rathbun,under laboratory conditions

The following protistan diets were tested on blue crab larvae: the algae Isochrisis galbana Parke, Monochrisis lutheri Droop, Dunaliella sp., and an unknown mixture; and the ciliated protozoans Euplotes vannus Muller and Parauronema virginianum(2/1) Thompson. None of these diets resulted in development past the first zoea stage, although some apparently were ingested and delayed mortality as compared to unfed controls.The rotifer Brachionus plicatilis Müller sustained good survival through early zoea development; however, rotifer-fed larvae did not metamorphose to the megalopa. Larvae of the polychaete Hydroides dianthus (Verrill) sustained crab larvae throughout zoea development, resulting in 17% survival to metamorphosis. The percentage mortality per stage was significantly lower in polychaete-fed larvae when compared with rotifer-fed larvae during zoea stages III, VI, and VII. Mean intermolt duration varied between diet treatments during the first three stages, but showed no differences during later zoea development. In tests on groups of late stage sibling larvae, Artemia salina L. nauplii gave development to metamorphosis, whereas rotifers did not.All the diets so far tested on blue crab larvae are classified according to their ability to sustain development. It is demonstrated that the two diets which allow completed development, Hydroides dianthus larvae and Artemia salina nauplii, contain 2–3 times as much lipid per dry weight as do rotifers. A metabolic requirement for lipid late in development may be indicated. Invertebrate larvae derived from yolky telolecithal and centrolecithal eggs may be an important dietary component for brachyuran larvae.     

Larval rearing of burbot (Lota lota L.) using Brachionus calyciflorus rotifer as starter food

Burbot Lota lota L. is one of the endangered freshwater fish species in western Europe for which the development of controlled larval rearing procedures could produce enough material for stock enhancement. The suitability of the freshwater rotifer Brachionus calyciflorus as a start food for larviculture of burbot was investigated. After yolk‐absorption, the larvae were stocked in 40‐L tanks under different feeding conditions: clear water rearing conditions with rotifers (Brachionus calyciflorus) for 10 days (R), green water conditions (Chlorella sp.) with rotifers offered for 10 days (MALR), green water conditions (Chlorella sp.) for 3 days followed by clear water in combination with rotifer feeding for 7 days (AL3R), and clear water conditions with Artemia nauplii offered for 10 days (Art). After the 10‐day feeding, all groups received Artemia nauplii up to 35 days post‐hatching. Larval survival was counted at day 10 and at the end of the 35‐day rearing experiment. At day 35, a significant survival difference was noted between the groups where rotifers were supplemented with algae vs only Artemia. At the end of the experiment, the highest survival rate (69.20%) was obtained with larvae receiving only algae in the first 3 days of feeding. Lowest survival rate (24.90%) was obtained with larvae receiving only Artemia for 35 days. This indicates that smaller prey are essential for burbot at first feeding. Larval length and wet weight were measured at the time of mouth opening, at days 7, 10, and 21, and at the end of the experiment (day 35). On day 35, mean length of the larvae varied significantly between the treatments. However, the final wet weight of the larvae did not vary significantly between the treatments.     

不同饵料对稀有鲫生长和繁殖的影响

饵料对鱼类生长发育和繁殖具有重要影响。为了筛选稀有鲫(Gobiocypris rarus)幼鱼和成鱼阶段最适投喂方式, 实验将出膜5周末(日龄为35 day after hatching)的幼鱼随机分为5个组: A组投喂丰年虫; B组每周前6d (days)投喂丰年虫, 后1d投喂商业化微颗粒S3饲料; C组每周前3.5d投喂丰年虫, 后3.5d投喂饲料; D组每周前1d投喂丰年虫, 后6d投喂饲料; E组一直投喂饲料; 各组均采用饱食投喂策略。每2周统计生长、存活指标, 直至第21周(147 dah), 在17周(119 dah)取材用于观察性腺发育程度。在产卵后统计各组产卵量、孵化率和子代畸形率。结果显示: (1)E组存活率和特定生长率显著低于其他组(P<0.05); (2)从产卵量、孵化率和子代畸形率上看, B组产卵量显著高于其他组(P<0.05); (3)从性腺组织学上看, 不同投喂方法对精巢的成熟度无显著影响, 但投喂过丰年虫的稀有鲫卵巢发育成熟度显著优于E组。研究结果提示:适量加入丰年虫比单一投喂活饵或饲料更有利于稀有鲫的生长和繁殖。建议在标准化养殖过程中, 幼鱼和成鱼期的稀有鲫采取丰年虫与饲料投喂频次比值为6﹕1的方式最佳。     

Consumption and gut evacuation rate of laboratory‐reared spotted seatrout (Sciaenidae) larvae and juveniles

The temperature and mass dependence of maximum consumption rate was measured for larval and early juvenile spotted seatrout Cynoscion nebulosus . Maximum consumption ( C _MAX) estimates were obtained from feeding and gut evacuation experiments on larvae (3·8–19 mm standard length, L _S) at three temperatures (24, 28 and 32° C), and maximum consumption experiments on juveniles at three temperatures (20, 26 and 32° C). Feeding levels were determined for larvae fed live prey ( Brachionus plicatilis and Artemia salina ) ad libitum . The midgut and total evacuation times were estimated for fish feeding continuously and discontinuously using alternate meals of tagged and untagged live prey. Temperature and fish size had significant effects on gut evacuation and consumption. The gut evacuation time increased with increasing fish size, and decreased with increasing temperatures. Mass‐specific midgut contents increased for small larvae <0·156 mg dry mass ( M _D)( c . 4 mm L _S), and decreased for larger larvae and juveniles. Maximum consumption was modelled by fitting a polynomial function to a reduced dataset of individuals feeding at high levels. The C _MAX model predicted an initial increase in specific feeding rate from 70 to 155% M _D day⁻¹ for small larvae, before declining for larger larvae and juveniles.