Genetic comparison of experimental farmed strains and wild Icelandic populations of Atlantic cod (Gadus morhua L.)

Microsatellite DNA loci and the Pantophysin locus (Pan I) were used to investigate levels of genetic diversity within farmed strains of Atlantic cod Gadus morhua and to compare them with the wild source population. A total of 282 farmed samples originating from a spawning ground off the south-west coast of Iceland were sampled in the years 2002 and 2003, and 258 wild cod were collected at the same spawning ground in the same years. The farmed strains exhibited a lower mean number of alleles and allelic diversity than the wild samples at the microsatellite loci. Significant differences were observed between wild and farmed samples both in allele and genotype frequencies at the Pan I locus. We argue that the genetic divergence of wild and farmed samples of Atlantic cod may be due to a small number of effective founding breeders contributing to the genetic variation of the farmed strains, inducing a reduction in allelic diversity. We discuss the potential effect of breeding practices on the genetic diversity of Atlantic cod.     

Growth variation and fin damage in Atlantic cod (Gadus morhua L.) fed at graded levels of feed restriction

Atlantic cod (initial weight 55, 250 and 450 g) were deprived of feed for 1 month or fed one of two diets differing in crude protein and lipid levels at rations corresponding to about 25, 50, 75 or 130% of group satiation for two months. The fish were individually weighed at the start, mid and end of the experiment and growth variation and fin damage were registered. The fin damage patterns differed between size groups; 55 g cod had most wounds on the dorsal fins whereas the pectoral fins were the most damaged in the 250 g fish. The incidence of damage in these groups was high and increased significantly when feeding was restricted. In the 450 g cod there was little fin damage and the incidence did not seem to depend on diet treatment.In the 55 and 250 g cod groups, fast growing fish had lower incidence of fin damage than fish that grew slowly, suggesting that the fish that received most aggression were prevented from feeding. However, a similar trend was registered in non-fed fish, showing that the recipients of aggression also suffered other disadvantages.The variation in individual growth rates increased when feeding was restricted and the distribution was differently skewed depending on feeding level. The data provide evidence that competition is an important factor limiting growth of individual cod held in groups in culture.     

Characterisation of the microbiota associated with intestine of Atlantic cod (Gadus morhua L.): The effect of fish meal, standard soybean meal and a bioprocessed soybean meal

Populations of aerobic heterotrophic bacteria present in the gastrointestinal (GI) tract of healthy Atlantic cod (Gadus morhua L.) fed three different diets, fish meal, standard or a bioprocessed soybean meal (BPSBM), were estimated using the dilution plate technique. A total of 944 isolates were characterised by biochemical and physiological properties and 425 isolates were identified further by sequencing the 16S rRNA genes. Our results showed that gut microbiota were affected by dietary manipulation. The GI tract of fish fed fish meal was dominated by Gram-positive bacteria of the genera Brochothrix and Carnobacterium. The Gram-negative bacteria Chryseobacterium spp. and Psychrobacter glacincola, and Gram-positive bacteria belonging to Carnobacterium, dominated in the digestive tract of fish fed soybean meal. In contrast to these results, genus Psychrobacter dominated in the GI tract when fish were fed BPSBM.Until recently, it was generally suggested that the gut microbiota of fish were less diverse than in homoeothermic animals. However, the present study identified several “new” bacterial species isolated from the alimentary tract of Atlantic cod. These “new” bacterial species are not normally isolated from the GI tract of fish. Based on our finding we suggest that the GI tract microbiota of fish might not be as simple as believed.Antagonistic activity of carnobacteria regarding inhibition of growth of two fish pathogens (Aeromonas salmonicida ssp. salmonicida and Vibrio anguillarum) was observed. However, some difference in the antibacterial activity of Carnobacterium spp. was observed. Whether this antagonistic activity has any effect in challenge studies will be discussed, especially in relation to the finding that the digestive tract is one of the major infection routes in fish.     

Short-term crowding stress in Atlantic cod, Gadus morhua L. modulates the humoral immune response

Considering overcrowding as a stressor, the effects of a short-term exposure on the immune responses of Atlantic cod, Gadus morhua L. were determined at 2, 24 and 72 h post-crowding. While plasma glucose and total antioxidant capacity were quantified as stress responses, immune responses were based on humoral indices. Bactericidal capacity of plasma was also assessed. There was a significant increase in plasma glucose and total antioxidant capacity at 2 h post-crowding, however the former returned to pre-stress levels 24 h later and the latter remained elevated until 72 h. The plasma protein did not reveal any significant change, but there was a transient upregulation of globulin at 24 h post-crowding. The myeloperoxidase activity increased at 2 h after the stress, but returned to its pre-stress levels at 24 h. The lysozyme activity was upregulated even from the first time point after stress, although the differences were significant only at 24 and 72 h compared to the activity prior to stress. At 72 h post-crowding there was a significant increase in alkaline phosphatase activity while haemolytic activity showed a significant decrease. An increase in the proliferation of the bacterial pathogens, Vibrio anguillarum and Aeromonas salmonicida was seen from 24 h in the plasma of the stressed fish. Thus, our finding suggests that short-term crowding stress modulates the immune responses in Atlantic cod and therefore husbandry procedures should consider these effects while addressing better management strategies.     

Digestibility of diets containing different soybean meals in Atlantic cod (Gadus morhua); comparison of collection methods and mapping of digestibility in different sections of the gastrointestinal tract

The purpose of the experiment was to evaluate effects of partial replacement of fish meal with soybean meal on digestibility in Atlantic cod and effects of biotechnological processing of soybean meal. Other objectives were to find out if apparent digestibility in cod was affected by age, temperature and/or faecal sampling methods, and to map digestion and absorption patterns along the intestinal tract for different age classes and diets. A control diet with fish meal as the main protein source (FM), a diet with 24% of crude protein from conventional extracted soybean meal (SBM), and a diet with 24% of crude protein from a bioprocessed soybean meal (BP-SBM) were used. Yttrium oxide was added as an inert marker for digestibility determination. The diets were fed to duplicate groups of 534 g (1-year-old) and 1750 g (2-year-old) cod in seawater. Faeces were stripped from the distal-most section (distal chamber) of the intestine after 48 and 84 days of feeding. In addition, intestinal contents were removed by dissection after 84 days. The dissected intestine was divided into 5 sections and contents were analysed separately. Dietary soy did not alter the viscosity of the digesta. The replacement of fish meal protein with soybean protein reduced the apparent digestibility of crude protein, fat, energy and almost all amino acids, independent of type of soybean meal processing. Larger fish had higher digestibility of starch, ash and several amino acids than smaller fish. A temperature decrease from 9.1 °C at feeding day 48 to 6.8 °C at day 84 resulted in lower digestibility of starch, ash and several amino acids. Absorption of amino acids was quantitatively terminated in the distal part of the mid intestine when feeding the soy-containing diets, but continued into the distal chamber when feeding the FM diet. Absorption of fat was completed in the proximal part of the mid intestine when feeding the soy-containing diets, but continued into the middle part of the mid intestine when feeding the FM diet. Soy did not affect absorption pattern of starch along the intestinal tract. Faecal sampling by dissection resulted in higher apparent digestibility of macronutrients than stripping in the 2-year-old cod, and higher apparent digestibility of protein in the 1-year olds.     

Growth, feed efficiency, digestibility and nutrient distribution in Atlantic cod (Gadus morhua) fed two different fish meal qualities at three dietary levels of vegetable protein sources

Atlantic cod were fed six diets where the ratio of protein from fish meal to vegetable protein were varied from 91%, 67% to 46%, respectively. The experimental groups were performed in triplicate in a 20 week growth trial, increasing fish weight from about 167 g to 690 g. The vegetable protein sources constituted full-fat soybean meal and corn gluten meal at a fixed ratio of 1:2, and replaced either a high quality fish meal (DC_Mink 92.3%) or a lower quality fish meal (DC_Mink 85.6%) in the diet. All diets were extruded and balanced to be equal in gross energy, crude protein, lipid, carbohydrate, lysine and phosphorus. No difference in growth (SGR 1%) was observed comparing the two fish meal qualities. However, feed intake was significantly higher (9%) and feed efficiency lower (10%) for lower quality fish meal compared to high quality fish meal. Protein and amino acid digestibility was significantly reduced in the lower quality fish meal, while unaffected by vegetable protein inclusion for both fish meal qualities. Protein utilization as measured by protein efficiency ratio and net protein value was not affected by fish meal inclusion, except in the diet using lower quality fish meal and high vegetable protein inclusion. Lipid and energy digestibility was significantly reduced by vegetable protein inclusion for both fish meals. The lower fish meal quality increased lipid deposition in the liver and affected slaughter quality of cod by increasing HSI and dressing out percentage at low and intermediate substitution levels. Muscle composition showed small dietary changes except for arginine, while liver fatty acid composition clearly reflected fatty acid profile of full-fat soya in the diets. Corn gluten meal and full-fat soybean meal (2:1) can replace approximately 50% of dietary protein without affecting feed intake, growth, protein digestibility or slaughter quality of cod when exchanging high quality fish meal. For the lower quality fish meal diets some lower inclusion of vegetable protein sources seem to be acceptable.     

Digestive function and intestinal integrity in Atlantic salmon (Salmo salar) fed kernel meals and protein concentrates made from yellow or narrow-leafed lupins

This study assessed the effects of yellow lupin (Lupinus luteus) and narrow-leafed lupin (L. angustifolius) kernel meals and protein concentrates on the gastrointestinal integrity, capacity for digestive hydrolysis, and digestibility of nutrients in Atlantic salmon. A basal diet (FM) was made from fish meal, wheat, and fish oil. Six additional diets were formulated by replacing 30% of the FM diet with lupin kernel meal made from L. l. cv. Wodjil (LKM), L. a. cv. Belara (BKM), and L. a. cv. Myallie (MKM), lupin protein concentrates made from the same L. l. (LPC) and L. a. cv. M (MPC), or extracted soybean meal (SBM). All diets were extruded. Each diet was fed to three groups of 176 g salmon kept in 1 m² tanks with 5.6 °C saltwater for 3 weeks prior to sampling of blood, intestinal organs, digesta, and faeces. Inclusion of lupin meals in the diets resulted in harder and more condensed feed particles. Ulcer-like lesions were observed in the stomach of fish from all feeding groups, and this was worsened by lupin in the diet, but did not appear to be pellet hardness related. No consistent altered morphology was observed in the distal intestine (DI) of fish fed the FM and lupin diets, while the DI of fish fed SBM showed consistent and typical soybean meal-induced pathomorphological changes. Plasma cholesterol was higher when feeding MKM and LKM than when feeding FM, MPC, and LPC, with intermediate levels when feeding BKM and SBM. Feeding LKM and LPC resulted in a higher weight of the GIT when related to body weight. Trypsin activity and bile acid concentration were generally higher in digesta from the pyloric (PI) and mid (MI) intestine when feeding FM and lupin diets than when feeding SBM, while the opposite was seen for trypsin activity in digesta from DI. There were no effects of diet on leucine aminopeptidase (LAP) and maltase activity in PI and MI, but in DI the activity of these brush border enzymes were significantly lowered when feeding SBM. SBM in the diet resulted in watery faeces and lowered apparent digestibility of lipid, but this was not observed when feeding the lupin diets. To conclude, the tested lupin kernel meals and protein concentrates did not alter the intestinal function in Atlantic salmon when included at 30% of the diet. Dietary lupin was, however, involved in the worsening of ulcer-like gastric lesions.     

Resistance of aquaculture net cage materials to biting by Atlantic Cod (Gadus morhua)

Cod bite on aquaculture net cages has resulted in damages like frayed netting and holes, which in part can explain why cultured cod have escaped more frequently than salmon over the last years. We describe damages found on various netting materials subjected to cod bite through field experiments at commercial cod farms. Further, a method to test local cod bite resistance of traditional netting structures is suggested and initial results from a test jig prototype are given. Results from field experiments indicated that cod may have been attracted by types of netting that made it possible to draw filaments out of the twine, while stiff, coated netting structures and thick filaments showed no sign of bite damage during the test period. We concluded that netting materials for cod aquaculture must be resistant to cod bite or be repellent or uninteresting for cod. Based on the present findings, the better choice among the traditional netting materials seemed to be hard-laid netting materials, preferably with a primer that glues the filaments together.     

Effect of replacing fish meal with soybean meal on growth, feed utilization and carcass composition of cuneate drum (Nibea miichthioides)

The effect of replacing fish meal with soybean meal (SBM) in practical feeds for cuneate drum was evaluated in an 8-week net pen trial. The cuneate drum fingerlings (initial body weight 29.8 ± 1.3 g fish^− 1) were fed six isonitrogenous and isocaloric feeds containing 39% digestible protein and 16 MJ kg^− 1 digestible energy. The control feed was formulated to contain 40% herring meal, whereas in the other five feeds SBM was included at 11.3, 22.5, 33.8, 45.0 and 56.3% to replace 20, 40, 60, 80 or 100% of the fish meal. There were no significant differences in feed intake between fish fed the control feed and feeds in which SBM replaced 20 to 80% of the fish meal, but fish fed the fish meal free feed had higher feed intake than the other treatments. Weight gain linearly declined with the decrease of fish meal level. Final body weight (FBW) of fish fed the feeds in which SBM replaced 20% of the fish meal did not significantly differ from fish fed the control feed. Replacing 40 to 100% of the fish meal resulted in lower FBW and nitrogen retention efficiency (NRE), and higher feed conversion ratio (FCR) than those of fish fed the control feed. Fish fed the feeds in which SBM replaced 60 to 100% of the fish meal had lower condition factor and hepatosomatic index than those of fish fed the control feed. No significant differences in carcass protein content was found among the treatments, but fish fed the feeds in which SBM replaced 60 to 100% of the fish meal had higher moisture and lower lipid content in carcass than those of fish fed the control feed. Results of the present study appear to indicate that cuneate drum has a limited ability to utilize SBM as a protein source in practical feeds.     

Variation in fish condition between Atlantic cod (Gadus morhua) stocks, the effect on their productivity and management implications

A comparative analysis of the fish condition (Fulton’s K) of 10 cod stocks in the north Atlantic in relation to the temperature of their habitat, growth rates and their reproductive potential is presented. It is shown that the cod stocks in the north Atlantic display different levels of mean condition, which is partly due to the different temperature regimes of their habitats. Cod living in colder waters, e.g. Greenland, Labrador and Grand Bank stocks, were found to be in poorer condition than cod living in warmer waters, e.g. North Sea and Irish Sea stocks.

Poor condition causes reduced productivity in terms of slow growth and low recruitment potential. Stocks in better condition display significantly higher weights at age 4 than stocks in poor condition. The a coefficients (function’s slopes) obtained from standardised Ricker’s recruitment–spawning stock biomass (SSB) relationships were defined as indicators for the recruitment potential of stocks. These a coefficients were found to be positively correlated with the mean condition factor of the 10 stocks analysed. This indicates that stocks consisting of individuals in poor condition appear to be very susceptible to reduced recruitment at low SSB, while the stocks that consist of fish in good condition seem to behave more robustly with a higher probability of good recruitment at low SSB. The positive effect of the cod condition on their reproductive potential generally implies that the stocks in good condition in the temperate regions of the northeast and west Atlantic can sustain higher exploitation rates than stocks in poor condition in the colder regimes of the northwest Atlantic (Greenland, Labrador and Grand Bank). This is confirmed by the positive relationship established between the estimated biological management reference points F_med and the mean cod condition factors, as well as by the recent status of these stocks.     

Apparent protein and energy digestibility of common and alternative feed ingredients by Atlantic cod, Gadus morhua (Linnaeus, 1758)

Studies were conducted with Atlantic cod, Gadus morhua (L.), to determine the apparent digestibility coefficients (ADCs) of protein and energy and the digestible energy (DE) content in feed ingredients widely available in Canada. We also tested the assumption of “independency” used in digestibility studies. The feed ingredients included two fish meals (herring, anchovy), three crustacean by-product meals (whole krill, crab, shrimp), two animal by-product meals (poultry by-product, hydrolyzed feather), six oilseed meals (soybean, soy protein concentrate, soy protein isolate, canola, canola protein concentrate, flaxseed), two pulse meals (white lupin, pea protein concentrate) and two cereal grain meals (corn gluten, wheat gluten). Protein ADCs were high for wheat gluten meal (99.9%), soy protein concentrate (98.6%), soy protein isolate (97.4%), whole krill meal (96.3%), herring meal (93.3%), soybean meal (92.3%), anchovy meal (92.2%), pea protein concentrate (89.8%), white lupin meal (89.7%), crab meal (89.4%), canola protein concentrate (88.8%) and corn gluten meal (86.3%); mid-range for poultry by-product meal (80.2%) and canola meal (76.0%); and low for shrimp meal (66.7%), hydrolyzed feather meal (62.4%) and flaxseed meal (50.2-55.0%). Energy ADC was high for whole krill meal (96.3%), wheat gluten meal (95.4%), soy protein concentrate (94.9%), herring meal (92.8%), soy protein isolate (92.1%), soybean meal (88.1%) and anchovy meal (86.4%); mid-range for canola protein concentrate (83.3%), corn gluten meal (82.7%), crab meal (82.4%), pea protein concentrate (76.7%) and white lupin meal (75.3%); and low for poultry by-product meal (71.0%), canola meal (60.6%), hydrolyzed feather meal (58.9%), shrimp meal (41.4%) and flaxseed meal (21.2-37.4%). From the protein ADC data, results clearly showed that the basal diet and test feed ingredients were digested independently of one another in nearly all cases, the only exceptions being for those diets containing test ingredients of very high (> 99%, wheat gluten) or very low (< 67%, hydrolyzed feather and flaxseed) protein ADCs. In the case of DE, the basal diet and test feed ingredients were digested independently in all test diets without exception.     

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 variations in muscle growth dynamics and selected quality attributes in Atlantic halibut (Hippoglossus hippoglossus L.) fed dietary lipids containing soybean and/or herring oil under different rearing regimes

Atlantic halibut (Hippoglossus hippoglossus), 1630 ± 510 g at the start of the experiment were reared in tanks for 1 year and subjected to three different environmental regimes; Continuous light and Constant temperature (CC), Natural light and Constant temperature (NC) or Natural light and Ambient temperature (NA). Two diets were fed, where 50% of the dietary lipids were either herring oil or soybean oil supplemented. The fish were sampled in May, August and November the first year, and February and May the following year. The CC groups grew better than the NC and NA groups from May to November, and as a result had a significantly (p < 0.05) higher weight by the end of the experiment. This difference did however not affect muscle fibre size distribution, or the chosen muscle quality attributes texture, fat content, water or fat loss of the fillet. These quality attributes were influenced by season and body weight, whereas diet had no significant effect. The NA group had lower fat content of the fillet than the other environmental regimes in August, November and May_end, and fat content increased with increasing fish weight. Water and fat losses were lowest in November and February. The diet influenced the fatty acid profile of the musculature; the triacylglycerol fraction reflecting the dietary treatment while the polar fraction was less influenced by diet. The hardness of the fillet was not influenced by the environmental regimes, but by season, and hardness was at its lowest in August, and increased towards the following spring. The muscle fibre distribution varied only with season, and neither treatment nor diet had any effect on the muscle growth dynamics. Recruitment of new fibres seemed mainly to occur during winter, signified by maximum occurrence of fibres < 500 μm² by the end of the experiment in May.     

Growth and lipid composition of Atlantic cod (Gadus morhua) larvae in response to differently enriched Artemia franciscana

Considerable progress has been achieved in the intensive culture of Atlantic cod (Gadus morhua). However, there is little information concerning optimum live-feed enrichments for cod larvae, since many of the techniques used during the larviculture have been borrowed from other fish species and adapted for the production of Atlantic cod. The present study compared four different protocols for the enrichment of Artemia to be used as live feed for cod larvae. The protocols tested were: (1) AlgaMac 2000, (2) AquaGrow Advantage, (3) Pavlova sp. + AlgaMac 2000, and (4) DC DHA Selco + AlgaMac 2000. Larvae were fed differently enriched Artemia between 37 and 59 days post hatch. At the end of the experiment, larvae from treatment 1 [specific growth rate (SGR) = 10.4 ± 0.4% day⁻¹] grew faster than larvae from treatments 3 (SGR = 6.9 ± 0.2% day⁻¹) and 4 (SGR = 4.9 ± 0.4% day⁻¹, P < 0.0001). However, treatments 3 and 4 resulted in better larval survival at the end of the experimental period, estimated to be 3 on a scale from 1 to 5, whereas the survival estimates for the two other groups were 2. The treatments affected the fatty-acid composition of Artemia and of cod larvae. Larvae from treatment 1 had a higher percentage of AA (20:4ω6, P < 0.0001) and ω6DPA (22:5ω6, P < 0.0001) than the other larvae. Levels of DHA (22:6ω3) were similar in larvae from treatments 1 and 4, and higher than in the other larvae (P < 0.0001). Our results suggest that Artemia containing a DHA/EPA/AA ratio of 7/2/1 result in good larval performance. Joseph A. Brown—Deceased September 2005.     

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.     

Effects of soybean meal based diet on growth performance, gut histopathology and intestinal microbiota of juvenile rainbow trout (Oncorhynchus mykiss)

Effects of soybean meal (SBM) based diet on growth performance, histology of the intestinal epithelium and on the gut microbiota of juvenile rainbow trout (Oncorhynchus mykiss) were investigated on two trials lasting for 8 weeks (Trial I) and 18 weeks (Trial II). The microbiological characterization was done both with conventional plating techniques, biochemical profiling and length heterogeneity analysis of PCR amplified 16S rDNA (LH-PCR).Typical histological changes were found after 18 weeks of SBM feeding (Trial II). Mean height of simple foldings were significantly higher within the group fed with fish meal (FM) based diet. These changes were not, however, reflected in the appearance, behavior and weight gain of the fish, although the feed conversion rate (FCR) was higher, compared to fish fed with the traditional FM feed.In both SBM and FM groups the counts of cultivable intestinal bacteria dropped within the first 4 weeks of feeding (Trial I). Subsequently, the bacterial numbers increased in the FM group, but not in the SBM group by the 8th week of the trial. The LH-PCR data suggested also that diet-related qualitative changes in the microbiota had occurred (Trial II). The main identified bacteria were mainly representatives of the genera Aeromonas, Sphingomonas and Chryseomonas, and among the lactic acid bacteria, Lactococcus and Lactobacillus.Future research will be directed to the identification and eventual elimination the SBM-associated factors involved in the inflammatory responses and unfavorable FCR, in particular by modifying the intestinal microbiota.     

Comparison of conventional and molecular techniques to investigate the intestinal microflora of rainbow trout (Oncorhynchus mykiss)

The concept of the use of probiotic organisms or prebiotic compounds to modify the fish gut microflora is becoming a popular topic for investigation. A major flaw in many such studies is a failure to consider fully the nature of the established microflora, which is to be modified pre-, or probiotically. Since it is widely accepted that a large proportion of bacteria are non-culturable, the use of conventional bacteriological (culture) techniques alone to investigate fish intestinal microflora may be expected to bias results. We report a study designed to investigate the normal intestinal microflora of rainbow trout (Oncorhynchus mykiss) using both conventional bacteriological and molecular methods.Over an 18 month period, the intestinal microflora of a single population of laboratory-raised rainbow trout was investigated. Bacteria isolated using bacteriological techniques were identified using the BiOLOG system and 16S rRNA gene sequencing. Dominant bacteria consistently were Aeromonas sp. and Carnobacterium piscicola, demonstrating that the microflora is stable in fish kept in defined conditions. Restriction Fragment Length Polymorphism (RFLP) and 16S rRNA gene sequence analysis was used to investigate anaerobic and non-culturable bacteria. An obligate anaerobe, Clostridium gasigenes, was shown to be among the dominating intestinal microflora.     

Effects of diets containing a bacterial protein meal on growth and feed utilisation in rainbow trout (Oncorhynchus mykiss)

Triplicate groups of rainbow trout with initial weight 361 g were fed either a fish meal based control diet or diets containing 9, 18 or 27% bacterial protein meal (BPM) or 9% of an autolysate (AU) of the BPM. No significant treatment effects were found on specific growth rates (SGR), feed intake, feed efficiency ratio (FER), or retention of nitrogen, amino acids or energy. The apparent digestibility coefficients (ADC) of nitrogen, energy and most indispensable amino acids decreased when BPM was included in the diet. The ADC of lipid, sum of amino acids, arginine, lysine, threonine and most of the dispensable amino acids were reduced at 27% BPM inclusion compared to the control, 9% and 18% BPM diets. None of the ADC estimated was different in the 9% AU diet compared to the 9% BPM diet. The loss of nitrogen and energy in faeces per kg gain increased as the dietary BPM or AU levels increased, and the energy used for activity and maintenance was higher in fish fed the 27% BPM diet than in fish fed the other diets (P < 0.05). There were no significant differences in the urea concentrations in plasma, liver and muscle, whereas the uric acid level in plasma was elevated in trout fed the 27% BPM diet. Histological evaluation of tissue from the stomach, pyloric caeca, mid-intestine and distal intestine did not reveal any diet-related morphological changes.In conclusion, no significant differences in growth and feed efficiency were found in the rainbow trout fed diets containing up to 27% BPM, and the AU did not increase fish performance compared to the BPM. Based on the data from this study, at levels up to 27% dietary inclusion, BPM is a good replacement for fish meal in diets for rainbow trout.     

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.     

Effect of soybean meal,raffinose and stachyose on the growth,body composition,intestinal morphology and intestinal microflora of juvenile allogynogenetic silver crucian carp (Carassius auratus gibelio♀×Cyprinus carpio♂)

The study was conducted to investigate the effects of soybean meal (SBM), raffinose and stachyose on juvenile crucian carp (Carassius auratus gibelio♀ × Cyprinus carpio♂). The experimental diets consisted of one control diet based on fish meal (FM), one diet containing 300 g kg^?1 SBM and four FM‐based diets with the addition of either 6.7 g kg^?1 raffinose (Raf), 33.9 g kg^?1 stachyose (Sta), a combination of raffinose and stachyose (Raf?Sta) and finally a Raf?Sta diet supplemented with 2.5 g kg^?1 saponins (Raf?Sta?Sap). After 3 weeks of feeding, the relative gut lengths of SBM‐fed fish and the fish fed stachyose‐containing diets were shorter than those of the FM‐fed fish; further, more SBM‐fed fish showed fissures on the tips of the intestinal folds. After 8 weeks of feeding, the growth of SBM‐fed fish was significantly lower than that of FM‐fed fish (P<0.05). The fish fed Raf?Sta?Sap had a low relative gut length (P<0.05). In comparison with the other fish, the SBM‐fed fish had a higher number of large‐sized homogeneous vacuoles in the cytoplasm of epithelial cells and shorter microvilli. No significant difference was observed in body composition or intestinal microflora. The results indicated that raffinose and stachyose played no or only minor roles in the development of soybean‐induced growth reduction.