Several micronutrients in the rotifer Brachionus sp. may not fulfil the nutritional requirements of marine fish larvae

The current best practice intensive culture of larval Atlantic cod includes feeding rotifers from onset of exogenous feeding until 25–30 days after hatching. These larvae grow considerably slower and develop higher frequencies of deformities than larvae reared in semi‐extensive systems, using copepods as feed. The present study compares the micronutrient concentrations in rotifers with those of copepods, with the aim of identifying nutrients that may be limiting for normal growth and development of cod larvae. An additional criterion used is the nutrient requirements given for fish in general, by NRC (1993) , as nutrient requirements of cod remains to be determined. Rotifers were fed on four different diets, consisting of baker's yeast with cod liver oil (3.3 : 1 dry weight (DW)/v), baker's yeast with Algamac 2000^TM (3.5 : 1 DW), baker's yeast with live algae Chlorella (4.1 : 1 DW), and Culture Selco 3000^TM (CS). CS was a complete commercial diet for rotifers while the other diets are considered as based on raw ingredients. Compared with copepod nutrient levels, rotifers grown on yeast‐based diets supplemented with either cod liver oil, Algamac 2000 or Chlorella were apparently sufficient for covering the requirements in cod larvae for all the B‐vitamins, except thiamine. Rotifers cultured on the CS diet also had sufficient amounts of thiamine. Of the minerals, only calcium and magnesium were sufficient, using this criterion while iron was on the borderline. However, with reference to the requirements given for larger fish ( NRC 1993 ), only thiamine, vitamin A, manganese, selenium and perhaps copper, appear too low in the rotifers cultured without extra micronutrient supplementation. The other nutrients were present at levels intermediate between copepod and fish requirement levels. This study suggests that it is necessary to develop enrichment techniques to produce rotifers with sufficient amounts of all micronutrients. Such techniques will also be important tools for determining which nutrients are present at levels below the actual requirements in cod larvae.     

A comparison of fatty acid composition and quality aspects of eggs and larvae from cultured and wild broodstock of common sole (Solea solea L.)

Eggs from a F₁ cultured broodstock of sole were compared with eggs from wild‐caught breeders throughout one spawning season, to evaluate if egg quality may be affected by culture‐related conditions. Fourteen batches of eggs from cultured broodstock and 17 batches from wild‐caught sole were compared with respect to fatty acid (FA) composition, egg size, fertilization rate and hatching rate. Based on a multivariate analysis of the FA profiles, it was possible to discriminate between culture and wild inheritance. Eggs from cultured broodstock had high levels of C20:1(n‐9), C18:2(n‐6) and C18:3(n‐3), whereas eggs from wild fish had high levels of C16:1(n‐7), C20:4(n‐6) and C20:5(n‐3). Differences in FA profiles were most likely related to dietary differences. Fertilization and hatching rates were generally low and lowest in eggs from cultured broodstock, but not related to FA composition. Larval growth of one batch from each group was compared. Larval growth was not correlated to broodstock origin, FA composition or egg or larval size. However, larval survival was significantly lower for larvae from cultured broodstock.     

Egg quality determination based on the shape of the lipid vesicle in common dentex, Dentex dentex

The shape of the lipid vesicle (the ratio between the maximal and the minimal diameter and the shape coefficient) of the eggs of Diplodus puntazzo and Sparus aurata is correlated with larval survival rate (%) and can be used as an egg quality marker as reported previously by Lahnsteiner and Patarnello (2005) . In the present study, this method for egg quality determination has also been validated for common dentex, Dentex dentex, after introducing several modifications in the calculation and evaluation procedure in order to obtain higher sensitivity. In the present study, it could be demonstrated that the shape of the lipid vesicle is not only correlated with the hatching rate but also with the survival rate of 1, 2 and 3 days post hatch larvae and therefore it can also be used as an indicator of larval viability at early stages of development.     

匙吻鲟仔鱼期饵料的研究

鱼类早期生长的细微差别对其存活以及整个生活史阶段的生长，有着不可估量的影响，而摄食是影响仔鱼期生长的最重要的外界影响因子之一。在水温19．5～20．5℃下，对匙吻鲟（Polyodon spathula）仔鱼的开口时间、适宜开口饵料和投饵率进行了研究。结果表明，匙吻鲟仔鱼在6日龄初次摄食，摄食率在7—8日龄迅速升高，9—10日龄达到100％，11-13日龄略有降低，14-15日龄迅速降低，不可逆点（PNR期）出现在15日龄；以轮虫作为开口饵料，仔鱼的存活率和生长优于丰年虫和颗粒饲料组；养殖水体中轮虫密度在120—150个／mL可满足仔鱼的生长需求。     

Characterization of two microparticle types for delivery of food to altricial fish larvae

The characteristics of two microparticulate types were determined for delivery of nutrients to fish larvae. A yeast-based commercial diet for larval fish (Microfeast Plus L-10, Salt Creek, Inc., Salt Lake City, UT, USA) was encapsulated within cross-linked protein-walled capsules (CLPWC) or gelatin-alginate beads (GAB). Microparticle stability was determined by encapsulating a nontoxic red dye (Poly R 478) within CLPWC or GAB. The stabilities of both particle types suggested that high molecular weight, water-soluble nutrients could be delivered to freshwater fish larvae without major losses from leaching. Acceptability experiments included determination of gut fullness and feeding incidence of zebrafish Brachydanio rerio larvae fed on the two particle types. Acceptability of GAB by first-feeding zebrafish larvae was significantly greater than that for CLPWC. Acceptability of CLPWC increased in larger larvae and was similar to that for GAB. Both particle types were broken down by first-feeding zebrafish larvae. Growth experiments with CLPWC showed that up to 40% substitution of Artemia nauplii could be accomplished without reduced growth and survival of zebrafish larvae after a feeding period of 8 days. Twenty per cent substitution of Artemia nauplii could be achieved with GAB without reduced growth and survival.     

北部沿海人工繁殖长江蟹试验

首次在北部沿海进行了长江蟹人工繁殖试验，报告了亲蟹促产，越冬培育，饵料投喂，水质管理，幼体孵化，幼体培育等全套工艺流程，交配抱卵率达８１％，越冬成活率８４．６％，大眼幼体育成率２２．５％，认为长江蟹，黄河蟹幼体培育方法一致，培养结果，育成率无显著差异，但越冬水深应保持１．５ｍ以上，亲蟹须先进行１５￣２０天强化培育。     

A method to determine protein digestibility of microdiets for larval and early juvenile fish

A method to evaluate protein quality using in vivo methods was developed for larval fish. FluoSpheres^® fluorescent microspheres (10 μm) were incorporated into two test diets, our standard zein microdiet (ZMD) and a microdiet with identical ingredients except for the replacement of high quality fish meal with the same product cooked for 24 h at 80 °C (ZMD-CF). Several trials were performed to design a reliable method to test digestibility using FluoSpheres^® as a marker. The developed in vivo technique was tested on 35 days posthatch (dph) larval Atlantic cod ( Gadus morhua L.) and two tropical fish species in the early juvenile stage. The method took into account loss of total protein to the faecal pellet and water column. Apparent digestibility of protein in larval cod fed ZMD was significantly higher than that of larvae fed ZMD-CF ( P  < 0.05). A growth study to validate differences between the two diets showed significant differences in growth and survival of larvae fed ZMD versus ZMD-CF ( P  < 0.05). Further validation of our results was indicated through the use of a pH-stat method using enzymes extracted from 35 dph larval cod guts. This novel technique will be advantageous for researchers to evaluate feed ingredients for larval marine fish and is adaptable to many different areas of larval fish nutrition.     

The risk of parasite transfer to juvenile fishes by live copepod food with the example Triaenophorus crassus and Triaenophorus nodulosus

Natural zooplankton is a potential food resource for juvenile fish in fish farms as it is a good source of fats, carbohydrates, and protein. However, it is also a potential source of parasites and pathogens. The present study was conducted (1) to estimate the risk of parasite transfer by live copepod food under intensive farming conditions using the parasites Triaenophorus crassus and Triaenophorus nodulosus as example and (2) to look for strategies to avoid parasite infestation of juvenile fish.An easy and routinely applicable aniline blue staining method was developed to check the infestation levels of copepods with procercoids during the on-growing season of juvenile fish. From the copepod species occurring in zooplankton Cyclops spp. was infested frequently (9.4 ± 12.0%, maximum 38.7%), Diaptomus spp. infrequently (1.5 ± 1.6%, maximum 3.2%), Daphnia spp. never. Juvenile grayling, Thymallus thymallus, and corgonids, Coregonus sp., which had been fed with natural zooplankton revealed infestations rates with T. crassus of circa 5%, with T. nodulosus of circa 10%. As the occurrence of procercoids in the zooplankton was temporary limited, Triaenophorus infestation can be avoided by using artificial food instead of live copepods during the risky season.To prevent parasite infestation of juvenile fish methods were investigated to eradicate procercoids from copepods by chemical treatment (sodium chloride, hydrogen peroxide, citric acid treatment) and by freeze-thawing methods. Chemical methods failed to remove procercoids. In fish, which had been fed with frozen thawed zooplankton, no Triaenophorus spp. infestation was observed. Their survival rates were similar as in the control fed with live zooplankton, however the weight of the fish was significantly lower.     

Protein quality of larval feed with increased concentration of hydrolysed protein: effects of heat treatment and leaching

Four heat coagulated early weaning diets with increasing concentrations of pepsin hydrolysed protein, were investigated with regard to the change in protein quality during feed production and exposure to leaching. Water-soluble N, trichloroacetic acid-soluble N and amino acid (AA) profiles were determined in finished diets and in diets leached for 6 min. In vitro diet digestibility was measured and related to increasing inclusion of hydrolysed protein and N leakage. Seventeen to 47% of soluble N in the feed ingredients was made insoluble by heat denaturation during feed production, but the concentration of peptides and free amino acids (FAA) were not influenced. All peptides/FAA and 70–80% of water-soluble protein were lost after exposure to leaching. Increased inclusion of hydrolysed protein increased the loss of crude protein (15–30%). All taurine and 30% of histidine was lost during leaching, no other major changes in AA profile were found. There was no difference in digestibility between diets exposed to leaching. However, leached diets showed reduced digestibility as compared to diets that had not been exposed to leaching. In conclusion micro-bound type diets as used in this study have a low efficiency in delivering soluble N to fish larvae and should be carefully considered for this purpose.