Effects of dietary factors, stocking biomass and domestication on the nutritional and technological quality of the Eurasian perch Perca fluviatilis

The ante mortem determinism of the quality components in fish is multivariate. Among the various influencing factors, the effects of (i) rearing biomass, (ii) dietary features and (iii) domestication process on the technological and nutritional variables in perch Perca fluviatilis here were studied using two-levels fractional factorial design 2^4-1 (resolution IV). This work allowed identifying two main factors, i.e. domestication and diet composition, which influence both technological and nutritional variables. Domestication was the parameter showing the greatest effect on both filleting yield (FY) and total saturated fatty acids (SFA) content, whereas diet composition chiefly influenced viscerosomatic index (VSI) and total poly- and mono-unsaturated fatty acids (PUFA and MUFA) contents. Domesticated fish exhibited higher FY (42.8%) and total SFA (25.1% of total fatty acids or FA) content than wild fish (FY = 40.1%, total SFA = 22.9%). Concerning the effect of food composition, diet including only fish oil as lipid source (FO) induced higher VSI than diet including vegetable oil (FV) VSI = 8.8% vs 7.4%, respectively). Moreover, FV diet induced higher n-6 PUFA but lower n-3 PUFA contents in fillets than did the FO diet (total n-6 PUFA = 6.3% vs 5.4% and total n-3 PUFA = 52.6% vs 55.2% of total FA respectively). FV fish fillets had significantly higher total MUFA (17.7% of total FA) contents in muscle than FO fish (14.8%). Concerning this last variable, domestication played only a role in interaction with diet composition. With FO diet, no difference in total MUFA content between wild and domesticated perch was noticed. However, total MUFA content increased with FV diet and was significantly higher in fillets of wild fish (18.7% vs 16.6% of total FA for domesticated). No effect of either feeding rate or initial biomass on the body composition was noticed. In addition, perivisceral fat content was not influenced by any treatment.     

Sexual dimorphism of fast muscle fibre recruitment in farmed Atlantic halibut (Hippoglossus hippoglossus L.)

Commercially farmed Atlantic halibut (Hippoglossus hippoglossus) were reared at the Aga Marin's facility located on Dønna (Norway) under ambient environmental conditions in duplicate 15 × 15 × 8 m netpens (May 2004 to May 2005). Twenty fish were sampled five times over a twelve month production period during which time the average body weight increased from 1.26 to 2.08 kg (n = 100 fish). Body mass, fork length (L_F), and the number and size distributions of fast muscle fibres were determined in male and female fish. All males matured during the autumn whereas no maturation was observed in females. From the point of maturation females had superior growth performance to males and body mass and the total cross-sectional area of muscle were 1.4-fold (P < 0.05) and 1.3-fold higher (P < 0.01) respectively by May 2005. The total number of fast muscle fibres per trunk cross-section at 0.55 L_F was 24.5% higher in females (7.58 × 10⁵) than males (5.80 × 10⁵) prior to sexual maturation. In females, muscle fibre recruitment slowed with short days and low water temperatures (< 6.5 °C), but had increased to 1.01 × 10⁶ by May 2005 (P < 0.001). In contrast, there was no growth and no increase in muscle fibre number in males following the onset of maturation. The distribution of muscle fibre diameters prior to maturation in males was also significantly different between male and female fish matched for L_F. The results illustrate a sexual dimorphism of muscle fibre recruitment patterns in Atlantic halibut and highlight the adverse affects of sexual maturation in males on muscle growth.     

The reproductive biology of the baby clam, Marcia opima, from two geographically separated areas of India

A study of the reproductive cycle of the baby clam, Marcia opima (Gmelin) was conducted at two sites along the southeast (Tuticorin Bay) and southwest (Ashtamudi estuary) coast of India from December 1998 to January 2000. Histological evidence showed that baby clam from both the sites showed two spawning seasons. First spawning season was recorded from May to July in the southeast coast. In the southwest coast, the first spawning season was observed through March to May. A second spawning season from September to December was recorded in both the coasts. In the southeast coast, minimum and maximum condition indices were obtained in January 2000 and April 1999 respectively and decreased from May to June and November to January, signifying the two spawning periods. Most of the animals were either in spent or indeterminate gonadal stage from May to August and November to January indicating the active spawning during that months in the southeast coast.     

Influence of dietary soybean meal levels on growth, feed utilization and gut histology of Egyptian sole (Solea aegyptiaca) juveniles

The present study was undertaken in order to determine the effect of a dietary incorporation of soybean meal (SBM) on growth performance, nutrient utilization and gut histology of Egyptian sole Solea aegyptiaca. This species, which is genetically and morphologically close to Solea solea, is currently present in the Mediterranean Sea and reared in some Italian farms, using the same techniques for S. solea production and it could represent a promising new species for Mediterranean aquaculture.Three isoproteic and isolipidic extruded diets (crude protein, 47%, crude fat, 20%) were formulated containing 0 (diet 1), 18 (diet 2) and 30% SBM (diet 3). 540 juveniles (initial body weight of 6.9 ± 0.2 g) were equally distributed into nine 150 l squared tanks (bottom surface: 520 cm²) connected with a recirculation system (temperature 20 ± 1 °C and dissolved oxygen above 6 ppm). Animals have been hand-fed twice a day (at 9.00 a.m. and 5.00 p.m.) at a daily ratio of 1.5% body weight^− 1. Diets were tested in triplicate for 87 days. The fish were weighed at days 28, 57 and 87, and they were sampled at days 0, 57 and 87 for nutrient retention determination. One-way ANOVA with Newman-Keuls' post-test were used to analyze data (P ≤ 0.05).No differences in terms of palatability were observed and any diet has been consumed completely all throughout the experiment. At day 87, fish fed diet 1, 2 and 3 reached weights of 25.8 ± 0.8 g, 27.5 ± 0.9 g and 26.1 ± 1.5 g, respectively and no differences in performances or nutrient retention were found.Histopathological gut examinations have revealed no noticeable differences in the appearances of the intestines between any of diet groups. Intestinal mucosal cells presented well vacuolated upper, with well defined microvillar brush borders. On the basis of the results of this trial, SBM seems to be a good protein source for Egyptian sole and can be added in the diet up to 30% without any reduction in growth rate and no adverse effect on gut histology.     

Seasonal changes in muscle structure and flesh quality of 0+ and 1+ Atlantic salmon (Salmo salar L.): impact of feeding regime and possible roles of ghrelin

Growth performance, muscle cellularity, flesh quality, and plasma ghrelin were investigated in 0+ and 1+ farmed Atlantic salmon (Salmo salar L.) from 40 g to 4.3 kg. Reduced meal frequency was introduced in both smolt groups from ~1.5 kg; one meal per second day (<5 °C) to one daily meal (>5 °C), while control groups were fed one to three daily meals. Results show that 0+ salmon had higher final fibre number and density, pigment content, red colour intensity, firmer flesh, and lower fillet fat content than 1+ salmon at 4.3 kg, affected by season and smolt type. Muscle fibre recruitment was an important determinant of fillet firmness and colour, possibly influenced by the prenatal temperature regime. Fish fed reduced meal frequency showed temporal reduced feeding ration, but growth performance was not compromised in any smolt groups at harvest. However, fillet fat, gaping, and colour decreased by less frequent feeding, with permanent effects in 1+ salmon for gaping and fat. Reduced meal frequency is therefore considered to be a promising tool for managing important flesh quality attributes in salmon without compromising growth performance. It is also suggested that ghrelin stimulates short‐term appetite, and perhaps also in the longer term.     

Seasonal adjusted diets to Atlantic salmon (Salmo salar): Evaluations of a novel feed based on heat-coagulated fish mince, fed throughout 1 year in sea: Feed utilisation, retention of nutrients and health parameters

The present paper reports the results of a 1 year experiment using Atlantic salmon with a starting weight of 1.7 kg, and final weight of 7.4 to7.8 kg dependent of diet group. The experiment aimed to evaluate a direct production method of feed where landings from fishing vessels are used directly in feed processing. The dietary compositions followed the natural rhythm in lipid level of the catches of herring in the North Sea (Catch group) adjusted to be stable throughout one season only, and compared to a control diet (constant composition throughout the year) and a designed diet (opposite to the natural biorhythm of salmon lipid stores) (diet group Design). Four seasons were defined: spring (March, April, May), summer (June July August), autumn (September, October, November), winter (December, January, February). All diets varied in protein and lipid due to the adjustment according to season. The experiment was run in triplicate sea cages for each diet, using natural light and temperature regimes (21°N). All diet groups showed equal growth throughout each season, except for the Catch group during autumn showing lower SGR-values. SGR values varied significantly also between seasons for all diet groups, ranging from 0.29 during spring to 0.81 during autumn. Feed conversion ratio varied dependent on season and the dietary protein to energy ratio, at all seasons except winter. Quite a large variation between diet groups and seasons were registered in condition factor. Whole body lipid varied between seasons, as a consequence of dietary lipid levels, and increased as fish weight increased. After autumn growth only, whole body protein levels were significantly lower in the Catch group compared to the two other diet groups. Muscle lipid levels did not vary as a consequence of fish size alone, but varied substantially between seasons in both the Catch and Control groups, but not in the Design group. Muscle alpha-tocopherol and fatty acid profile varied due to diet in spring and autumn seasons, but with minor variations due to diet in summer and winter periods. Only minor influences from diet were registered in liver and heart compositions. Haematological parameters and plasma nutrient values ranged within normal values for salmon, and together with close to no mortality throughout the experiment, fish health was concluded to be good in all diet groups at all seasons. However, plasma asparagine aminotransferase (ASAT) values were quite high during summer and autumn, especially in the control group. These results point to the need of optimizing antioxidant composition of diets during the summer and early autumn season. Overall results show the possibility to obtain acceptable production results when feeding Atlantic salmon a diet adjusted to each season, as long as the fish protein to energy (P:E) requirements are met, further that total body lipid stores varied more than muscle lipid concentrations, and that the lipid levels were highly dependent on season, diet composition and fish size.     

The effects of fish hydrolysate (CPSP) level on Octopus maya (Voss and Solis) diet: Digestive enzyme activity, blood metabolites, and energy balance

As has been demonstrated in previous studies, Octopus maya can be fed on artificial diets. In the present study six different diets were assayed. Five diets were designed to test the effect of percentage of inclusion of fish protein concentrate (CPSP: 0, 5, 10, 15, and 20%) and were offered to octopuses as a specifically designed artificial diet. The sixth diet consisted of frozen crab (Callinectes spp) and was used as control diet. Blood metabolites and energy budget of octopuses were evaluated to determine how CPSP levels modulate the digestive capacity and allow retaining energy for growth. Wild animals (316.4 ± 9.8 g) were used in the study. Results showed that CPSP produced a positive specific growth rate (SGR, % day^− 1) with high value in octopuses fed 15% CPSP level. A maximum growth rate of 0.86% day^− 1 was recorded in these animals, a value that is extremely low when compared with the SGR obtained when animals were fed fresh crab (3.7% day^− 1). In general, blood metabolites were affected by diet composition, indicating that some metabolites could reflect the nutritional and/or physiological status of octopus. Preliminary reference values for O. maya fed crab were found for glucose (0.09 ±0.02 mg/ml), lactate (0.004 ± 0.002 mg/ml), cholesterol (0.16 ± 0.02 mg/ml), acylglycerol (0.14 ± 0.01 mg/ml), protein (0.37 ± 0.04 mg/ml), hemocyanin (1.85 ± 0.04 mmol/l), and digestive gland glycogen (1.86 ± 0.3 mg/g). Total energy content can be used as an indicator of tissue metabolic reserves. In the present study, higher energy content in the digestive gland and muscle was observed in octopuses fed crab, followed by animals fed 15% CPSP. Results from the digestive gland indicated that the retained energy derived from glycogen, suggesting that lipids and protein were the main sources of variation linked with energy content. In general, digestive gland proteases activity and trypsin were induced in octopuses fed 15% CPSP. The capacity of O. maya juveniles to adjust their digestive enzymes to different types of food was evidenced. Essential amino acid content (EAA) of the diet was not a limiting factor. When dietary EAA profiles were compared with O. maya EAA profiles, all dietary EAA resulted in a higher concentration than whole body octopus composition. In the present study, all experimental groups ingested between 3300 and 4106 kJ wk^− 1 kg^− 1 without statistical differences among treatments, indicating that experimental diets were as attractive as crab. Differences were recorded in the proportion of absorbed energy (Ab, %) between CPSP-based and crab meat diets, suggesting digestion limitations associated with artificial diets. The present results indicate that the 15% CPSP diet had characteristics that stimulate digestive enzymes and reduce energetic costs associated with its digestion (HiE or SDA), channeling more biomass production than the other experimental diets.     

Successful culture of larvae of Litopenaeus vannamei fed a microbound formulated diet exclusively from either stage PZ2 or M1 to PL1

In three separate experiments, harpaticoid copepods Tisbe monozota (alive and dead) and a microparticulate microbound diet were evaluated as alternatives to live Artemia nauplii as food, beginning at either stage PZ2 or M1, in the larval culture of Litopenaeus vannamei. Larvae were cultured in 2 L round bottom flasks at a density of 150 L^− 1 (Experiment 1) and 100 L^− 1 ( 3.2 and 3.3) at 28 °C, 35‰ salinity and 12:12 LD photoperiod, and fed 4×/day^- 1. Larvae were initially fed a mixture of phytoplankton to stages PZ2 or M1 and then fed either live Artemia, live or dead copepods, or a microparticulate microbound diet. The experiments were terminated and all larvae were harvested when more than 80% of larvae had molted to postlarvae 1 (PL1) within any flask representing any of the treatments. The comparative value of the different diets and feeding regimes was determined by mean survival, mean dry weight and total length of individual larva, and percentage of surviving larvae that were PL1. Trypsin activity of samples of larvae from each treatment was also determined. The microparticulate microbound diet effectively served as a complete substitute for Artemia nauplii when fed beginning at stage M1. When fed at the beginning of the PZ2 stage, survival was comparable to that of larvae fed Artemia, but mean dry weight, mean total length, and percent of surviving larvae that were PL1 generally were significantly less. Responses to the feeding of copepods, whether fed dead or live, as a substitute were generally significantly less than those of larvae fed either the Artemia nauplii or the microparticulate diet. Values of trypsin activity (10^− 5 IU/μg^- 1 dry weight) corresponded to the relative proportions of the different larval stages within a treatment, with higher activity being characteristic of early stages. Previously demonstrated successful results with another species of crustacean suggest that the microparticulate microbound diet has characteristics that should be effective in the culture of the carnivorous stages of other crustacean and fish larvae that are currently fed live Artemia nauplii.     

Evaluation of three seaweeds Gracilaria bursa-pastoris, Ulva rigida and Gracilaria cornea as dietary ingredients in European sea bass (Dicentrarchus labrax) juveniles

The aim of this study was to evaluate the inclusion of three seaweeds Gracilaria bursa-pastoris (GP), Ulva rigida (UR) and Gracilaria cornea (GC) as dietary ingredients on the performance, nutrient utilisation and body composition of European sea bass juveniles. Six experimental diets were formulated to replace 5% (GP-5, UR-5, and GC-5 Diets) and 10% (GP-10, UR-10 and GC-10 Diets) fish protein hydrolysate (CPSP) by each of the three seaweeds. A control diet was used, without inclusion of any seaweed. Diets were fed to duplicate groups of 25 European sea bass (Dicentrarchus labrax) juveniles (IBW = 4.7 g) for 10 weeks. Growth performance was only significantly reduced (P < 0.05) in fish fed the GC-10 diet, whereas the feed conversion ratio increased significantly in those fish. The apparent digestibility coefficients of dry matter and lipid were significantly lower in fish fed diet GC-10 relative to those fed the control diet. Carcass composition was similar among treatments, although fish fed GC-10 exhibited significantly higher ash content.The results obtained in this study suggest that the inclusion of G. bursa-pastoris (GP) and U. rigida (UR), up to 10%, can be considered as very interesting ingredients in diets for sea bass juveniles, as no negative consequences on growth performance, nutrient utilization or body composition were observed. On the other hand, the inclusion of G. cornea (GC) should be limited to 5% of the diet.     

Growth and gonadal development in diploid and triploid turbot (Scophthalmus maximus)

This study determined the effect of triploidy on the survival, growth and gonadal development of turbot from 6 to 48 months of age. From 6 to 24 months of age (first sexual maturity), survival was similar in both ploidies (P > 0.05). From 24 to 48 months of age, after the first sexual maturity, survival was 91.9% in diploids and 100% in triploids, which did not exhibit the post-spawning-associated mortality. Growth was similar for both ploidies during the first year of life. After that, triploids grew significantly (P < 0.05) more that diploids, with more marked differences after each spawning season. From 24 to 48 months, the average weight difference between both ploidies was 11.4 ± 1.9%, ranging from 4.3 to 23.0%. At 47 months of age, the biomass of triploids was 10.3% greater in total weight and 14.3% greater in eviscerated weight. Gonads of triploid males were similar to that of diploids, whereas in triploid females, they were significantly smaller and rudimentary. A histological analysis carried out at 47 months of age showed complete sterility of triploids in both sexes. Sex ratio was 1 male (M):0.6 female (F), for diploids, significantly (P < 0.05) different from 1:1, and 1 M:3.3 F for triploids, significantly (P < 0.05) different from 1:1 and from the diploids. Since females grow more than males, culture of triploids benefited from the high female ratio, which helped to reduce size dispersion. In addition, their sterility allowed better performance by avoiding the reduction in growth that takes place during the spawning periods. Together, these observations indicate that triploidy induction can be an interesting option for turbot aquaculture, especially for the production of large-size fish of more than 2 years of age.     

Automated image analysis as a tool to quantify the colour and composition of rainbow trout (Oncorhynchus mykiss W.) cutlets

The goal of this paper is to propose and evaluate automated image analysis methods for describing muscle cutlets in rainbow trout. The proposed automated image analysis methods were tested on a total of 983 scanned images of trout cutlets, and included quality traits such as fat percentage, flesh colour and the size of morphologically distinguishable subparts of the cutlet. A sub-sample of 50 images was randomly selected for manual segmentation of the cutlet, the dorsal fat depot and the red muscle and regions. The identification of these regions by manual and automatic image analysis correlated strongly (r = 0.97, r = 0.95 and r = 0.91, respectively). The estimated fat percentage obtained from image analysis, based on the area of visible fat and the colour of the cutlet flesh, correlated well with chemical fat percentage measured by mid-infrared transmission spectroscopy (MIT) (r = 0.78). The automated image analysis methods are therefore a reliable means of predicting the fat percentage of trout cutlets. Principal component analysis (PCA) loading plots were used to identify subsets of variables from the image analysis of special significance for further studies; cutlet area, dorsal fat depot area, red muscle area, back height, cutlet width, and width of left and right abdomen wall were among the variables selected. PCA loading plots of different colour variables indicated that simple statistical coefficients such as percentiles and mean values can be used to quantify different aspects of flesh colour. In conclusion, the methods presented here provide a powerful toolbox for describing important morphological structures and quality traits of trout cutlets.     

An insulin-like system involved in the control of Pacific oyster Crassostrea gigas reproduction: hrIGF-1 effect on germinal cell proliferation and maturation associated with expression of an homologous insulin receptor-related receptor

The putative involvement of insulin-like peptides in the control of the reproduction of the Pacific oyster Crassostrea gigas was investigated using different approaches. In conjunction with a monthly histological analysis of the oyster reproductive cycle, in vitro biological effects of the human recombinant IGF-1 (hrIGF-1) on dissociated germinal cells were mesured over 1 year using [³H]-thymidine and [¹⁴C]-amino acid mixture as tracers for DNA and protein synthesis. DNA synthesis was stimulated by hrIGF-1 in November (114 ± 11% for 10^− 7M), December (46 ± 6% for 10^− 7 M) and January, which was identified as the highest gonial mitosis period. A clear dose-effect was observed in January with a maximum activation of 68 ± 7% for 10^− 12 M. Germinal cell protein synthesis was also stimulated in March (20 ± 1% for 10^− 10 M), April (41 ± 5% for 10^− 13 M), May (25 ± 4% for 10^− 13 M), and by almost all of hrIGF-1 doses in June (21.5 ± 2% for 10^− 13 M) and July (34 ± 1% for 10^− 13 M). This suggests the involvement of insulin-like substances in gonadal tubule rebuilding (December), as well as in the development of germinal cells (March, April), and in the summer maturation of gametes (May, June, July). These insulin-like effects conform with the expression pattern of the recently identified C. gigas insulin receptor-related receptor (CIR): It appeared highly expressed in the gonadal area during gonial mitosis phase, but also in maturating oocytes, suggesting the involvement of an insulin-like system in gonial proliferation and maturation. Moreover, CIR showed differential expression during embryogenesis and larval developmental stages. The expression of maternal CIR during the embryonic and early larval development, followed by the increasing zygotic CIR expression from D larvae to 11-day-old veliger larvae, then a decrease until metamorphosis, also suggest that insulin-like peptide is involved in organogenesis.     

Effects of dietary canola oil level on growth performance, fatty acid composition and ionoregulatory development of spring chinook salmon parr, Oncorhynchus tshawytscha

This study assessed the potential of refined canola oil (CO) as a source of supplemental dietary lipid for pre-smolt spring chinook salmon (Oncorhynchus tshawytscha) over a period of 30 weeks. Triplicate groups of 320 fry (∼ 0.80 g), reared in flow-through well water (8-11.5 °C), were fed one of four steam-pelleted dry diets with equivalent gross energy (24.3 MJ/kg), protein (∼ 51.3%) and lipid (∼ 21.6%) content on a dry-weight basis. CO furnished either 0%, 33%, 67%, or 100% of the supplemental dietary lipid, with the remainder from a commercial blend of 1:1 anchovy oil and poultry fat (APF). Thus, CO comprised either 0% (dAPF), 25% (CO25), 49% (CO49), or 72% (CO72) of total dietary lipid content. Overall fish growth rate, feed intake, feed efficiency, protein utilization, percent survival, and terminal whole body proximate constituents were unaffected by diet treatment. Dietary lipid compositions reflected the ratios of CO and APF in the supplemental lipid and their respective fatty acid compositions. Whole body fatty acid compositions mirrored those of diet treatments. However, some essential fatty acids, namely, arachidonic acid (20:4n-6; AA), and docosahexaenoic acid (22:6n-3; DHA) were conserved in fish regardless of dietary CO level. Direct relationships were found between dietary and whole body concentrations of: 18:2n-6 (R² = 0.94; slope = 0.72), 18:3n-3 (R² = 0.99; slope = 0.58), 20:4n-6 (R² = 0.84; slope = 0.42), 20:5n-3 (R² = 0.99; slope = 0.43) and 22:6n-3 (R² = 0.82; slope = 1.25). High dietary levels of 18:3n-3, 18:2n-6, and 20:5n-3 may have been utilized for energy or converted to more unsaturated derivatives. Overall development of ionoregulatory ability, as assessed by 24-h seawater challenge tests, was unaffected by diet. However, whole body chloride content was generally inversely related to dietary CO level during early development. Our findings suggest that there is excellent potential for long-term replacement of fish oil with canola oil in the diet of pre-smolt spring chinook salmon, provided that some marine oil is present to ensure that the essential fatty acid needs of the fish are met.     

Development of phytoplankton communities and common off-flavors in a biofloc technology system used for the culture of channel catfish (Ictalurus punctatus)

The use of biofloc technology production systems continues to increase in the aquaculture industry worldwide. Recent research demonstrated that outdoor biofloc systems can be used to produce high yields of channel catfish (Ictalurus punctatus). However, studies have not yet been performed to determine the development and composition of phytoplankton communities and related off-flavor problems in these biofloc production systems. In this study, water samples were collected biweekly from May to November and channel catfish samples were collected during harvest in November from nine 18.6 m² biofloc culture tanks. Water and fillet samples were analyzed for levels of the common off-flavor compounds geosmin and 2-methylisoborneol (MIB). The development and composition of phytoplankton communities in each culture tank was also monitored. In addition, water and biofloc samples were evaluated to assess the microbial sources of geosmin and MIB within the culture tanks. Phytoplankton (including algae and cyanobacteria attached to bioflocs) biomass, as determined by concentrations of chlorophyll a in the water, gradually increased in all tanks over time. Phytoplankton communities that developed in the culture tanks were dominated by fast-growing, unicellular and small colonial types of green algae (chlorophytes) and diatoms (bacillariophytes) and slower growing, small colonial types of cyanobacteria (cyanophytes). A positive correlation (p < 0.05) between cumulative feed addition and chlorophyll a concentration was found. Although geosmin and MIB were present in the culture water of each tank during most of the study, levels were typically low and only one tank yielded catfish with geosmin and MIB in their flesh at levels high enough to be designated as off-flavor. A positive correlation (p < 0.05) between cumulative feed addition and MIB concentrations in the water of culture tanks indicates a greater potential for MIB-related off-flavor problems at high feed application rates. The microbial sources responsible for production of geosmin and MIB in the culture tanks remain unknown.     

Effects of dietary vitamin C on survival, growth, and immunity of large yellow croaker, Pseudosciaena crocea

An 8 weeks growth experiment was conducted to determine the effects of dietary vitamin C on the survival, growth, tissue ascorbic acid contents and immunity of large yellow croaker (Pseudosciaena crocea) with initial weight of 17.82 ± 0.68 g. Seven practical diets were formulated to contain 0.1, 12.2, 23.8, 47.6, 89.7, 188.5 and 489.0 mg ascorbic acid equivalent kg^− 1 diet, supplied as l-ascorbyl-2-polyphosphate (LAPP). Each diet was fed to triplicate groups of fish in seawater floating cages (1.5 × 1.5 × 2.0 m), and each cage was stocked with 120 fish. Fish were fed twice daily (05:00 and 17:00) to apparent satiation for 8 weeks. The water temperature fluctuated from 19.5 to 25.5 °C, the salinity from 25 to 28‰ and dissolved oxygen content was approximately 7 mg l^− 1 during the experimental period. Results showed that the specific growth rate (SGR) (from 1.80 to 1.96% d^− 1) had an increasing trend with the increase of dietary vitamin C, but no significant difference was observed among dietary treatments. No gross deficiency signs were observed in any of the experimental fish. Survival rate, however, significantly increased with increasing dietary vitamin C (P < 0.05). The vitamin C contents in liver and muscle correlated positively with the vitamin C in diets. The vitamin C requirement was estimated to be 28.2 mg kg^− 1 based on survival, and 87.0 mg kg^− 1 on liver content of vitamin C. The activities of serum lysozyme and alternative complement pathway (ACP), phagocytosis percentage (PP) and respiratory burst activity of head kidney significantly increased with increasing dietary vitamin C. The challenge experiment with Vibrio harveyi showed that fish fed the diets with supplementation of vitamin C had significantly lower cumulative mortality compared to the control group (66.7%), and the cumulative mortality (16.7%) in fish with 489.0 mg kg^− 1 ascorbic acid was significantly lower than that (41.7%) in fish with 23.8 mg kg^− 1 ascorbic acid. These results suggested that vitamin C significantly influenced the immune response and disease resistance of large yellow croaker.     

Histopathology and a real-time PCR assay for detection of Bonamia ostreae in Ostrea edulis cultured in western Canada

Bonamia ostreae is an intracellular haplosporidian parasite in European flat oysters Ostrea edulis that occurs on both coasts of the United States and causes significant mortality in Europe. Canada was considered free of B. ostreae until 2004, when it was first detected in O. edulis obtained for laboratory study from a western Canadian oyster farm. Bonamia ostreae was confirmed in O. edulis at the index farm in November 2004 using histopathology, conventional polymerase chain reaction (PCR) assays, restriction fragment length polymorphism (RFLP) analysis, and sequencing of the PCR product. Archived samples of European flat oysters obtained from the index farm between 1999 and 2004 (n = 343) were re-examined and all samples collected before 2003 (n = 306) were confirmed negative for B. ostreae by histopathology (n = 306) and PCR (n = 62). In archived samples from 2003, B. ostreae was detected in 3 of 37 O. edulis by histopathology (n = 2) and/or PCR (n = 2). Also, records indicate that B. ostreae was not detected in O. edulis (n = 348) from five other locations in western Canada between 1986 and 2000. To better understand the distribution and prevalence of B. ostreae in western Canada, 607 oysters from the index farm and 2 additional farms were sampled in the summer of 2005. All 3 farms had been stocked with O. edulis spat from the State of Washington, USA, where B. ostreae is endemic. Samples were analyzed by histopathology and a new real-time PCR that amplifies a 68-bp target DNA fragment. B. ostreae was detected in all three locations, with prevalence ranging from 0.5 to 11.1%. Diagnostic sensitivity of the real-time PCR method was consistently greater than histopathology. Also, preliminary evidence supports the conclusion that real-time PCR on paraffin sections is more sensitive than histopathology; B. ostreae DNA was confirmed in 4 oysters by real-time PCR on paraffin-embedded tissues (and independently confirmed on unfixed tissues) that was not detected by histopathology. As a result of these findings, O. edulis spat are no longer allowed to be imported from endemic areas into Canada.     

The effects of algal diets on population growth and egg hatching success of the tropical calanoid copepod, Acartia sinjiensis

As natural diets of fish larvae, a number of calanoid copepod species are being investigated for use as live prey in aquaculture hatcheries. One of these, the tropical calanoid copepod, Acartia sinjiensis, has good potential as a live feed for tropical reef fish larvae. However, the rearing techniques for A. sinjiensis require further development to improve productivity. This study was carried out to investigate the population growth and egg hatching success of A. sinjiensis when fed a range of mono-and binary algal diets, including algae in the form of frozen paste.For the population growth experiment, the final A. sinjiensis population, including eggs, nauplii, copepodites and adults, was determined after feeding eight algal diets (two frozen algae, four live monoalgal and two live binary algal diets) for 8 days at temperature 28 ± 1 °C; salinity 34 ± 1 psu and photoperiod 12 L:12 D. Five replicates, with an initial 12 adult A. sinjiensis per replicate, were set up for each treatment. In a separate experiment, effects of diets on egg hatching success were examined after 48 h incubation of eggs produced by A. sinjiensis fed the same eight diets.The results showed that diet significantly affected both population growth and hatching success of A. sinjiensis. Of the diets tested, the binary algal diets were more successful than monoalgal diets, while the frozen algae had little dietary value. The highest population growth was recorded on A. sinjiensis fed a binary diet of Tetraselmis chuii and the Tahitian strain of Isochrysis sp. (T-ISO) (final population: 1091 ± 80), which was significantly higher (P < 0.001) than all other diets tested except for the other binary diet of Nannochloropsis sp. and T-ISO (final population: 897 ± 123). Diet also had a significant effect (P < 0.001) on egg hatching rate, though the highest hatch rate was recorded with eggs produced by A. sinjiensis fed binary diet Nannochloropsis sp. and T-ISO (88.1 ± 2.1%), this was not significantly different from that of eggs produced by A. sinjiensis fed either T-ISO alone (88.0 ± 1.7%) or the binary algal diet of T. chuii and T-ISO (76.4 ± 7.1%). The results of this study suggest that among the diets tested, a combination of live T. chuii and T-ISO was the best for the culture of A. sinjiensis.     

Effects of the partial substitution of dietary fish meal by differently processed soybean meals on growth performance, nutrient digestibility and activity of intestinal brush border enzymes in the European sea bass (Dicentrarchus labrax)

This study evaluated the effects of replacing fish meal (FM) in the diet of European sea bass (Dicentrarchus labrax) with differently processed soybean meals on growth performance; apparent digestibility coefficients (ADCs); the activity of alkaline phosphatase and the brush border digestive enzymes: leucine amino peptidase, maltase and γ-glutamyl transpeptidase; and the fish morphometric and flesh quality. European sea bass with an initial average body weight of 187.8 ± 1.4 g, were fed either a FM based (control) diet or a soy replacement isoproteic, isolipidic and isocaloric diet. The treatments consisted replacing FM protein by: 25% toasted, dehulled and solvent-extracted soybean meal (SE25); 50% dehulled and toasted soy seeds subjected to dry extrusion and mechanical oil extraction (ME50); 50% enzyme-treated soybean meal (ET50) and an inclusion of 60% composed of 30% toasted, dehulled and solvent-extracted soybean meal and 30% enzyme-treated soybean meal (SE + ET/60). ADCs values of dry matter, crude protein, crude lipid and gross energy did not significantly differ between ME50 and SE + ET/60 but both of them were inferior to the other treatments (P < 0.05). Compared with the control FM-based diet, only the group that consumed the ME50 diet had lower specific growth rate, feed conversion ratio and gross protein retention efficiency values (P < 0.05). The protein efficiency ratio did not differ between the FM-based control group and all other soy derivate groups (P > 0.05). In order to gain more insight on the effects of the diet on fish growth, activity of the brush border enzymes was measured. No significant differences were found in leucine amino peptidase and maltase activities, between the control group and the other soy derivate groups. The activity of γ-glutamyl transpeptidase was significantly higher in the upper section in fish that consumed the control diet compared with fish that consumed the soy derivate diets. Yet, the main activity of this enzyme was found in the lower intestinal section. In all the soy derivate diets except for the SE25 diet, the activity of alkaline phosphatase was significantly lower than its activity in the control diet. Feeding diets that include various types and levels of soybean derivatives did not affect the whole body composition and slaughter yield but significantly reduced liver weight. The results show that in methionine supplemented diets, FM can be replaced in terms of protein by 25% SE, 50% ET or 60% soy composed of 30% SE and 30% ET without hampering fish performance. The inclusion of 50% protein from ME resulted in significantly lower growth performance and feed utilization.     

Growth, immune function, and disease and stress resistance of juvenile Nile tilapia (Oreochromis niloticus) fed graded levels of bovine lactoferrin

Juvenile Nile tilapia (Oreochromis niloticus) were fed nutritionally complete, practical basal diets supplemented with bovine lactoferrin (Lf) at 0, 200, 400, 800, or 1600 mg/kg diet to apparent satiation twice daily for 8 weeks. After the feeding trial, the effect of dietary Lf on growth performance, immune function, and resistance to Streptococcus iniae challenge and low-water stress was determined. Dietary Lf did not affect growth performance (weight gain, feed intake, feed efficiency ratio, or survival) or haematological parameters (haemoglobin, white and red blood cell counts, or haematocrit) (P > 0.05). Crowding stress produced significant increases in plasma cortisol, glucose, lactate, and osmolality from baseline values (P ≤ 0.001), but dietary Lf did not affect plasma glucose, osmolality, or cortisol concentrations (P > 0.05). The level of Lf in diet had a significant impact on survival following S. iniae challenge with fish fed the 800 mg/kg Lf diet having significantly higher survival than control fish (P ≤ 0.05). There was not a corresponding increase in activity of non-specific or specific immune parameters (plasma lysozyme and spontaneous haemolytic complement activities or agglutination antibody titer against S. iniae) with addition of Lf to diets (P > 0.05), but plasma iron decreased and total iron binding capacity (TIBC) increased significantly with increasing concentration of Lf in diet (P ≤ 0.05). The ability of Lf to sequester iron, an essential nutrient required for the growth of bacteria, is regarded as one of its key antibacterial properties. The increased survival with increasing dietary concentration of Lf seemed to correspond with a decrease in plasma iron concentration and not enhancement of non-specific or specific immune functions.     

Oxytetracycline (OTC) accumulation and elimination in hemolymph, muscle and hepatopancreas of white shrimp Litopenaeus vannamei following an OTC-feed therapeutic treatment

The white shrimp, Litopenaeus vannamei, has become a very important species for the development of shrimp aquaculture in Northwest Mexico. However, viral and bacterial diseases are considered a major threat to the development of this industry. In the present study a trial was conducted to evaluate the tissue distribution, maximum concentration, and elimination of the widely used antibiotic oxytetracycline (OTC) in L. vannamei using indoor tanks under laboratory-controlled conditions. OTC was given to shrimp simulating a therapeutic treatment through medicated feed for 14 days followed by a period of feeding without antibiotic for another 14 days to evaluate the elimination pattern. Samples of hemolymph, muscle, and hepatopancreas were taken from medicated animals every two days for 28 days. All tissues were removed and frozen immediately in liquid nitrogen. OTC levels were analyzed by High Performance Liquid Chromatography (HPLC). Results showed an important OTC increase during consumption of medicated feed in all examined tissues. OTC maximum concentrations were 33.54 ± 11.19, 194.37 ± 16.11, and 18.79 ± 5.87 µg g^− 1 for muscle, hepatopancreas and hemolymph, respectively. Although the highest OTC level was found in the hepatopancreas, it required only two days after the start of dosing to reach this value, whereas the maximum OTC for muscle and hemolymph was detected after eight days of dosing. Ten days after the cessation of medicated feeding, the drug content in the shrimp tail muscle was under the detectable limit for the method (0.01 µg g^− 1 of OTC).