Feeding gelatinized carbohydrate in the diets of magur,Clarias batrachus (Linnaeus, 1758): Effects on growth performance,enzyme activities and expression of muscle regulatory factors

This study was conducted to illustrate the effect of dietary gelatinized starch (GS) on the growth performance, enzyme activities and expression of MyoD and Myf5 in magur, Clarias batrachus fingerlings. Four iso‐nitrogenous (37%) and iso‐lipidic (6%) diets containing 15% (D‐1), 25% (D‐2), 35% (D‐3) or 45% (D‐4) GS were fed to 240 fingerlings (2.5 ± 0.5 g) in triplicates groups for 8 weeks. The maximum weight gain %, specific growth rate, protein efficiency ratio, lower FCR and higher mRNA expression of MyoD or Myf5 were found in the D‐3 group fed with 35% GS. Higher hepatosomatic index, viscerosomatic index, body lipid and lower moisture content were found in the D‐4 group. Aspartate transaminase and alanine transaminase activities were found to be higher in the D‐1 group. Amylase, glucose‐6‐phosphate dehydrogenase and blood glucose were higher in the D‐4 group. Superoxide dismutase, catalase and hexokinase activities remain unaffected by the dietary GS levels. Hence, the overall results indicate that 35% GS can improve growth performance and upregulate myogenic regulatory factors, but 45% GS level will favor lipogenesis and compromise growth. Furthermore, Myf5 gene showed more immediate response than the MyoD to the dietary carbohydrate in magur.     

Induction of fertilizable eggs by conspecific vitellogenin implantation in captive female walking catfish,Clarias batrachus (Linn.)

The synthesis of vitellogenin (Vg) is induced by conspecific Vg (Vg1 and Vg2) and estradiol‐17β (E₂) as demonstrated by the pattern of ³H‐serine incorporation in the liver and plasma proteins. The incorporation studies indicated that the label was first incorporated into the liver after which it appeared in the blood in both E₂‐ and Vg‐treated male catfish. Since Vg was capable of inducing its own synthesis, experiments were conducted in females during preparatory–prespawning period (March–May) to make them gravid by implanting Vg pellets. Two implantations of 4 mg Vg1 pellets into female catfish with an interval of 15 days, followed by laboratory maintenance for 45 days of initial implantation showed a significant increment in ovarian weight with concomitant formation of yolky oocytes through synthesis and incorporation of Vg, whereas Vg2 implantation was not effective in this regard. Histological observation of yolky oocytes in Vg1‐treated group showed the peripheral migration of germinal vesicle (eccentric germinal vesicle), which indicates the onset of maturation. On 45th day, third implantation with 2 mg Vg pellets was performed and after 15 days, fish were hormonally induced with a single injection of hCG (2,000 IU/kg fish). Six groups were considered such as initial control, BSA‐implanted control, Vg1‐implanted, Vg2‐implanted, catfish collected from the field on the last day of the experiment and catfish collected during spawning period in this experiment with 3–7 fish in each group. Each of the experimental fish was sexually mature and the body weight was between 100 and 125 g. The percentage of ovulation and fertilization in the eggs of Vg1‐implanted group was 91% and 78%, respectively, which was almost similar to that of gravid female catfish collected during breeding period (July). The breeding performance in BSA‐ and Vg2‐treated females was very poor. The fertilized eggs were hatched in the laboratory conditions. Thus, in the female catfish, Vg1 not only induces vitellogenesis but also makes the oocytes viable for fertilization.     

Food consumption and digestive enzyme activity of Clarias batrachus exposed to various temperatures

The effect of temperature on the food consumption rate and the digestive enzyme activities of Clarias batrachus (80.60 ± 5.34 g) were evaluated. Fish were exposed to six different temperatures of 10, 15, 20, 25, 30 and 35 °C following an acclimation temperature of 25 °C. The rate of temperature change was 2 °C day^?1. Highest food consumption was recorded at 25 °C. It gradually reduced with decreasing water temperature. Food consumption rate was significantly (P < 0.05) lower at 10 °C compared with other treatments. Hence, 46.67, 8.20–23.58 and 1.02–6.15% reduced food consumptions were recorded in groups exposed at 10, 15 and 20 °C temperatures, respectively, compared with the 25 °C. The consumption rate was not affected in fish exposed at 30 and 35 °C. Total protease, trypsin and chymotrypsin activities were significantly (P < 0.05) higher in fish exposed at 25 °C compared with others. Lipase activity was significantly (P < 0.05) higher in fish exposed at 30 °C compared with others. Lowest enzyme activities were recorded at 10 °C. Water temperature below 25 °C affected the food consumption and digestive enzyme activities in fish that served as indicators of stress in fish.     

Dietary vitamin C mitigates monocrotophos toxicity in the freshwater fish Clarias batrachus

The amelioration effect of balanced diet with supplementation of vitamin C against stress was evaluated in freshwater paddy field fish, Clarias batrachus, exposed to organophosphate insecticide, monocrotophos (MCP). Three groups of fishes, that is, (E₀) Control, (E₁) MCP (2.14 mg L^?1) exposure and E₂) Vitamin C, MCP (0.2 g kg^?1 body weight + 2.144 mg L^?1) were selected for the sublethal study. The effect of MCP on protein metabolism in physiological sensitive tissues, namely gills, kidney, liver and muscle of the fish has been determined. Stress on Clarias batrachus was evident that the total protein, amino acid and ammonia contents were decreased, and urea and glutamine levels were elevated in all tissues for 30 days. Similarly, the activities of protease, aminotransferases and phosphatases were elevated significantly in all the tissues during the same period. Fish fed with a diet supplemented by vitamin C (0.2 g kg^?1 body weight) could significantly counter these effects.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Nigella sativa seed protects against 4‐nonylphenol‐induced haematotoxicity in Clarias gariepinus (Burchell, 1822): Oxidant/antioxidant rebalance

4‐Nonylphenol (NP) is an emerging concern contaminant which is widely spread in the aquatic ecosystem. Nigella sativa seed (NSS) has multifaceted therapeutic values. This study aimed to give insight into the potential protective effect of NSS on NP‐induced haematotoxicity in Clarias gariepinus through evaluation of haematological parameters, oxidant/antioxidant balance of blood lysate and histopathological investigation of blood smear. One hundred and fifty fish were divided into five groups (30/group). First group served as control which did not received NP exposure and fed basal diet without NSS supplementation. The other four groups were exposed to NP at a dose of 0.1 mg L^?1 and fed diets supplemented with NSS at levels of 0, 10, 25 and 50 g/kg diet, respectively. Macrocytic hypochromic anaemia, thrombocytopenia, leucopenia, neutrophilia, lymphopenia, monocytosis and eosinophilia were observed following NP exposure together with increase in morphological erythrocyte alterations and micronuclei formation. Elevation in total peroxide and malondialdehyde and depletion in total antioxidant capacity of blood lysate were reported. We concluded that supplementation of NSS markedly ameliorated the previously listed manifestations, and the most effective doses were 25 and 50 g/kg feed.     

Mortality and pathology of hybrid catfish,Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), associated with Edwardsiella ictaluri infection in southern Thailand

Enteric septicaemia of catfish (ESC) caused by Edwardsiella ictaluri is becoming an increasing problem in aquaculture and has been reported worldwide in a variety of fish species. This study reports ESC in hybrid catfish, Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), cultured in southern Thailand. The bacteria were identified as E. ictaluri by conventional and rapid identification systems, as well as by genetic and phylogenetic characterization. Analysis of 16S rRNA indicated 100% homology to the 16S rRNA sequence of several E. ictaluri strains in GenBank. Plasmid profiles demonstrated 4.0‐ and 5.6‐kb plasmids, compared with the 4.8‐ and 5.6‐kb plasmids in the US isolates, and representative genes of three of the four known pathogenicity islands of US isolates were present. Serologically, lipopolysaccharide (LPS) purified from the Thai isolates was not recognized by a monoclonal antibody against the LPS of US isolates. Fish experimentally infected with E. ictaluri showed 23–100% mortality within 14 days with a 168‐h LD₅₀ of 6.92 × 10⁷ CFU mL^?1 by immersion and a 96‐h LD₅₀ of 1.58 × 10⁶ CFU fish^?1 by intraperitoneal injection. Examination of tissue sections obtained from both naturally and experimentally infected fish indicated that infection of hybrid catfish with E. ictaluri produced lesions in several organs including liver, kidney, spleen, heart and brain. Histopathology findings included cellular necrosis, focal haemorrhage, infiltration of lymphocytes and multifocal granulomatous inflammation in the infected organs.     

Pre‐challenge and post‐challenge haemato‐immunological changes in Oreochromis niloticus (Linnaeus, 1758) fed argan oil against Lactococcus garvieae

The present study investigated the effects of argan oil, obtained from Argania spinosa, on pre‐ and post‐challenge immuno‐haematological and biochemical responses of Nile tilapia, Oreochromis niloticus. For this purpose, the fish were fed diets containing 0, 0.5%, 1% or 2% argan oil for 45 days. Following 45 days of feeding, fish were challenged with Lactococcus garvieae and mortality was recorded for 15 days. During the pre‐challenge period, significantly higher respiratory burst activity, total white blood cell (WBC), serum lysozyme activity and myeloperoxidase activity were determined in the argan oil‐fed groups. The serum glucose and cholesterol levels decreased whilst total protein and albumin did not change in the groups fed with argan oil‐supplemented diets. After challenge with Lactococcus garvieae, the percentage survival (%) was found to be the highest in the 1% and 2% argan oil‐supplemented feeding groups. Also, there was a significant increase in weight gain, specific growth rate and feed conversion ratio in those fish fed argan oil. The results of this study indicated that after the supplementation of fish diets with argan oil, especially at 1% and 2% concentrations, the immunological, haematological and biochemical values remained similar in both the pre‐ and post‐challenge periods and the immune response against L. garvieae in Nile tilapia was modulated.     

The Three‐spined Stickleback,Gasterosteus aculeatus Linnaeus 1758, plays a minor role as a host of Lepeophtheirus salmonis (Krøyer 1837) in the Gulf of Maine

The sea louse, Lepeophtheirus salmonis (Krøyer 1837), is a significant parasite of farmed salmon throughout the Northern Hemisphere. Management of on‐farm louse populations can be improved by understanding the role that wild fish play in sustaining and providing refuge for the local population of sea lice. In this study, 1,064 sticklebacks were captured. Of these animals, 176 individuals were carrying a total of 238 sea lice, yielding a prevalence and intensity of 16.5% and 1.4 lice per fish, respectively. Detailed examination of the sea lice on the three‐spined sticklebacks captured in Cobscook Bay found two L. salmonis individuals using three‐spined sticklebacks as hosts. A 2012 survey of wild fish in Cobscook Bay, Maine, found multiple wild hosts for Caligus elongatus (von Nordmann 1832), including three‐spined sticklebacks (Gasterosteus aculeatus L.), but no L. salmonis were found in this earlier study.     

A production season of turbot larvae Scophthalmus maximus (Linnaeus, 1758) reared on copepods in a Danish (56°N) semi‐intensive outdoor system

Turbot were reared from yolk sack larvae to juvenile in an outdoor semi‐intensive system. Three production cycles were monitored from May to September. A pelagic food chain was established with phytoplankton, copepods and turbot larvae. Abiotic and biotic parameters of lower trophic levels together with turbot larval survival, development, prey electivity and growth were monitored. A decreasing larval survival from 18.4% in May to 13.6% in July and just 7.0% in September was observed. The overall phytoplankton and copepod abundance decreased during the productive season. The turbot larval growth showed significant differences between larvae below (isometric) and above (allometric) 7 mm. Larval fish gut content showed no differences with available prey between production cycles. Therefore, it appears that the available prey concentration is governing their growth in this outdoor system. First‐feeding turbot larvae exhibited active selection for nauplii whereas developed larvae switched to copepodites and adult copepods. Although developing turbot larva exhibited active selection towards copepod size classes, there was no evidence of selective feeding on either of the two dominant copepod species. The turbot larvae's prey ingestion was modelled together with the standing stock of copepod biomass. The model results indicated that the estimated need for daily ingestion exceeded the standing stock of copepods. Hence, the initially established food web was unable to sustain the added turbot larvae with starvation as a consequence. We therefore suggest several solutions to circumvent starvation in the semi‐intensive system.     

A cost‐effectiveness analysis of live feeds in juvenile turbot Scophthalmus maximus (Linnaeus, 1758) farming: copepods versus Artemia

The biological benefits of copepods as live feed for marine finfish larvae have already been well established in the literature. Copepods have better biochemical compositions that improve growth, reduce malpigmentations and allow successful farming of ‘new’ marine finfish species. However, their current usage is quite limited. One of the reasons has been lack of economic knowledge concerning the cost‐effectiveness of copepod application compared to other commonly used feed items such as the brine shrimp Artemia. In this study, a cost‐effectiveness analysis is made on two alternative live feed items (copepods and Artemia) in juvenile turbot farming. Unit cost of production and profit are compared between the two feeding regimes using a unique data set from an existing turbot fry production facility in Denmark. The result reveals that copepods are not only biochemically superior but they are also economically a cost‐effective alternative. Thus, a commercial use of copepods will significantly reduce the production costs for turbot. Furthermore, the unexploited economic potential can be utilized for the successful farming of other high‐valued marine finfish species such as tuna, flounders, cod, sole and halibut. Generally, the biochemical superiority coupled with economic benefits can lead to the commercial utilization of copepods as complementary live feed in the short run and in some situations as a substitute in the long run.     

Effect of different culture conditions on the structural diversity of prokaryote communities in the sediment of earth ponds stocked with gilthead seabream Sparus aurata (Linnaeus, 1758)

In this study, we evaluate the use of polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE) for monitoring the effect of different aquaculture practices on sediment prokaryote (Archaea and Bacteria) communities. The effect of initial fish (gilthead seabream Sparus aurata) stocking density on the structural diversity of prokaryote communities of earth ponds bottom sediments was evaluated using PCR‐DGGE after a 5 month grow‐out period. An identical approach was used to monitor the effect of supplying different fish feeds [commercial feed (CD) versus an ecofeed (ECO)]. One additional variable was the use of copper sulphate (CuSO₄) as an algicide in some of the experimental rearing tanks. The statistical analyses of prokaryote community profiles showed that the presence of fish in earth ponds significantly influenced the structure of sediment prokaryote communities, when compared with earth ponds without fish, independently of the stocking density. Our results also indicated that the structure of the prokaryote communities of earth ponds supplied with the ECO feed shared a strong similarity with that fed CD. Curiously, the use of CuSO₄ in ponds receiving the ECO feed promoted significant differences on the structural composition of the bacterial community, but not on the archaeal community. DGGE molecular fingerprints are suitable for fast evaluation of new management practices in food‐fish production on earth ponds by monitoring shifts on microbial communities in bottom sediments.     

Dietary levamisole modulates the immune response and disease resistance of Asian catfish Clarias batrachus (Linnaeus)

In order to determine the immunomodulatory effect of dietary levamisole in Asian catfish (Clarias batrachus), fish were fed four different diets for 10 days: a formulated diet as control and the same diet supplemented with 50, 150 or 450 mg levamisole kg^?1 feed. The serum bacterial agglutination titre against Aeromonas hydrophila as a measure of specific immunity, serum haemagglutination titre, natural haemolytic complement activity (ACH₅₀), myeloperoxidase and lysozyme activities, total protein level and oxidative radical production by neutrophils as a measure of non‐specific immunity as well as disease resistance against A. hydrophila challenge to separate vaccinated and non‐vaccinated groups were evaluated at 0, 1, 2 and 3 weeks after last administration of levamisole. Levamisole supplement at the lowest level (50 mg kg^?1) significantly enhanced oxidative radical production and serum myeloperoxidase (MPO) content immediately after 10 days of feeding, which reached peak values after 3 and 2 weeks of feeding respectively. Haemolytic complement and haemagglutination titre were significantly enhanced after 3 and 1 weeks respectively. Haemolytic complement activity and MPO activities were significantly raised to 150 mg kg^?1 after 3 and 2 weeks, respectively. At the highest level of levamisole feeding (450 mg kg^?1) significant decreases in superoxide production and complement activity were measured immediately after levamisole feeding, which returned to the normal level after 1 week post‐ feeding. Fish were challenged with a virulent strain of A. hydrophila at 0, 1, 2 and 3 weeks after levamisole feeding, and the cumulative per cent survival was recorded over 10 days. Feeding levamisole at 50, 150 or 450 mg kg^?1 increased per cent survival in vaccinated fish immediately after levamisole feeding, and survival was significantly higher at 450 mg kg^?1. There was no difference in mortality patterns in non‐vaccinated fish. The results support the use of levamisole at 50 mg kg^?1 feed for 10 days as an immunostimulant in Asian catfish farming.     

Sesbania aculeata leaf meal as replacer of de‐oiled rice bran in aquaculture feed: Growth,IGF‐1 expression,metabolic and biochemical responses in Cyprinus carpio (Linnaeus 1758)

A feeding trial was conducted to evaluate the effect of inclusion of Sesbania leaf meal in the diet of Cyprinus carpio as a replacer of de‐oiled rice bran (DORB). Bacillus subtilis was used to ferment the Sesbania leaf meal (SLM) to produce FSLM (fermented sesbania leaf meal). Fishes were fed with five isonitrogenous (30% CP) and isocaloric diets for 60 days and treatments were viz., C (30% DORB, 0% SLM), R15 (15% DORB, 15% SLM), R30 (0% DORB, 30% SLM), F15 (15% DORB, 15%FSLM) and F30 (0% DORB, 30% FSLM) by replacing DORB with SLM. A similar growth performance with control was observed in 15% FSLM group while there was a growth reduction in all the raw leaf meal fed groups. Similarly, the expression of insulin‐like growth factor‐1 (IGF‐1) was significantly (p < .05) higher in control and 15% FSLM fed groups. Growth and metabolic enzymes such as protease, amylase, ALT and AST activities were significantly higher in the fermented leaf meal fed groups compared to their non‐fermented counterparts. Antioxidant enzymes, serum transaminase enzymes (SGOT, SGPT) and total bilirubin (TBR) were found to be increased by enhanced level of RSLM, however, reduced by inclusion of FSLM. However, 30% inclusion level of FSLM even after fermentation showed detrimental effect on growth and metabolic response at cellular and molecular level. Thus, the results concluded that fermented Sesbania leaf meal (FSLM) using B. subtilis could replace 50% DORB (15% FSLM group) in the diet of C. carpio without affecting the growth and other physiological responses.     

Conditioning of the European flat oyster (Ostrea edulis,Linnaeus 1758): effect of food ration

To establish a reliable protocol for the broodstock conditioning of the oyster Ostrea edulis (Linnaeus 1758) in hatcheries is important to obtain the best number of viable larvae all year even outside the natural season for maturation and spawning. Hatchery production of bivalves depends on their initial gonadal maturation state, which is linked to season and several other factors, such as temperature, daylight, salinity and food supply. Ration and diet composition are important in broodstock conditioning because food availability affects fecundity and gametogenic development. In this work, the effect of food ration on O. edulis conditioning was studied in autumn and winter. The results of this study showed that the broodstock conditioned with a food ration of 6% and 9% produces the highest number of larvae in autumn and winter. These food rations produced rapid gonadal development and larval release in a short period with a similar larval production. The oysters conditioned with a food ration of 3% delayed larval release and produced a significantly lower number of larvae in autumn. In the 9% ration treatment, the percentages of mortality were significantly higher in both autumn and winter. Thus, a 6% food ration is likely to be the most suitable for flat oyster conditioning combined with a gradient of temperature (14–18°C) and a gradient of daylight (8–16 h).