The effects of hydrogen peroxide treatment on different life-cycle stages of the salmon louse, Lepeophtheirus salmonis (Krøyer 1837)

The effects of hydrogen peroxide administered at a concentration of 1500 ppm for 20 min at 7.5 °C on different life-cycle stages of Lepeophtheirus salmonis were examined experimentally. The mobile adult and pre-adult stages of L. salmonis readily reattached to Atlantic salmon after hydrogen peroxide treatment. Adult female lice, but not adult males or pre-adults, reattached in significantly lower numbers than untreated controls. Survival of early chalimus stages (I and II) was not affected by hydrogen peroxide treatment, but their subsequent development to chalimus III and IV was delayed compared with untreated controls. Nauplii and copepodid larvae of L. salmonis were almost all dead by 1 hour post-treatment. A very few copepodids survived 24 h post-treatment. Egg strings of L. salmonis at an early stage of development failed to hatch after hydrogen peroxide treatment, but those with pigmented eggs did hatch, although in significantly reduced numbers. Treated egg strings did produce viable copepodids, although in significantly reduced numbers compared with controls.     

Early development of resistance to the salmon louse, Lepeophtheirus salmonis (Krøyer), in juvenile pink salmon, Oncorhynchus gorbuscha (Walbaum)

This study examined the effect of fish weight on the susceptibility of post-emergent pink salmon to Lepeophtheirus salmonis (Krøyer). Three trials were conducted, each with two stocks of pink salmon, Oncorhynchus gorbuscha (Walbaum), at starting weights of c . 0.3, 0.7 and 2.4 g, respectively. In each trial, duplicate tanks of fish were exposed to 0, 25 (only in Trial 1), 50 or 100 copepodids per fish. Mortality in Trial 1 was c . 37%, regardless of stock following exposures to 50 or 100 copepodids. Mortalities occurred up to 26 days after exposure, and more than 80% of the lice on the dead fish were chalimus stages. Infections with adult or preadult lice were observed on c . 35% of fish surviving to 37 days after exposure. Mortality was 5% in Trial 2 and there was no mortality in Trial 3. The abundance of L. salmonis was lower in Trial 3 compared with Trials 1 or 2. Histological changes in the skin coincident with fish growth included a thickening of the epidermis, infiltration of the dermis with fibroblasts by the end of Trial 1 and the first evidence of scales by the end of Trial 2; scales were evident throughout Trial 3. These results showed that the previously reported innate resistance to L. salmonis displayed by pink salmon develops in fish heavier than 0.3 g and appears to be functional by 0.7 g. This resistance coincided with changes to the epidermis and dermis, including the formation of scales. The present results indicate that elevated risk associated with L. salmonis infection among migrating post-emergent pink salmon may occur during a relatively brief period before the fish reaches 0.7 g.     

Experimental infections with Lepeophtheirus salmonis (Krøyer) on threespine sticklebacks, Gasterosteus aculeatus L., and juvenile Pacific salmon, Oncorhynchus spp

Experimental infections with Lepeophtheirus salmonis (Krøyer) were established on threespine sticklebacks, Gasterosteus aculeatus L., juvenile pink, Oncorhynchus gorbuscha (Walbaum), and chum, Oncorhynchus keta (Walbaum), salmon. The prevalence and abundance of infections were initially higher on sticklebacks than on either salmon species. The initial prevalence and intensity of infections on chum salmon were higher than those on pink salmon, and declined on both species during louse development. The rate of parasite development to adult stages was similar on all species although development beyond the preadult stage was not observed on sticklebacks. These results confirm previous field observations on the occurrence and development of L. salmonis on threespine sticklebacks.     

Spatial and temporal distribution of Lepeophtheirus salmonis (Kr?yer) larvae in a sea loch containing Atlantic salmon, Salmo salar L., farms on the north-west coast of Scotland.

A plankton survey investigated spatial and temporal distributions of larval sea lice in a sea loch on the north-west coast of Scotland. Plankton tows were collected approximately weekly at two depths (0 and 5 m) at five sample stations over a 2-year period. The survey began in March 2002 when Atlantic salmon, Salmo salar L., farms in the study area stocked with fish and finished in March 2004, after farms completed harvesting. Over 90% of lice recovered were Lepeophtheirus salmonis (Kr?yer); therefore, the results of this paper refer largely to that species. The data were analysed using generalized additive models with explanatory variables for time, depth and station. Greatest densities of nauplii were recovered at stations adjacent to farms indicating that the local salmon farms were a likely source of larvae at times during the production cycle. There were significant temporal trends in larval densities and the stocking and harvesting of farms can possibly account for these increases and decreases, particularly at the start and at the end of the production cycle. The distribution of copepodids was more widespread than that of nauplii, indicating that the larvae can be transported several kilometres from the point of release.     

Effect of geographic origin, temperature and timing of broodstock collection on conditioning, spawning success and larval viability of Ruditapes decussatus (Linné, 1758)

Culture of Ruditapes decussatus is clearly limited by the availability of seed, as this production proceeds almost exclusively from natural recruitment. Artificial spawning and larval rearing programs could provide an alternative source of spat. This study was designed to evaluate the effect of different conditioning temperatures on the broodstock maturation, spawning success and larval viability of two geographically (north and south of the Iberian Peninsula) distinct populations of European clam (R. decussatus) collected at different periods of the year in order to create “optimal” artificial spawning and larval rearing programs. Two batches of clams from each population were collected in October and February, and conditioned at 18 ± 1°C, 20 ± 1°C and 22 ± 1°C. Of the three variables analysed the timing of broodstock collection was the most determining factor for gametogenic development, spawning and larval rearing. Geographic origin and conditioning temperature also greatly affected the spawning. The results also showed that the February conditioning was more effective than October and that the best conditioning temperatures were 20 ± 1°C and 22 ± 1°C for the northern and southern populations, respectively. These results suggest that the efficient conditioning temperature for each population of the same species is related to the seasonal temperature regime from their geographic origin. Larval viability and growth performance seemed to be independent of the broodstock conditioning.     

Effect of temperature on α-tocopherol, fatty acid profile, and pigments of Diacronema vlkianum (Haptophyceae)

Diacronema vlkianum was grown in polyethylene bags at two different temperatures (18 and 26°C) in the laboratory. The biochemical composition level decreased when the temperature increased from 18 to 26°C. The maximum cell number at 18°C was 11.9 × 10⁶ cells ml⁻¹, while maximum cell number at 26°C was 1.6 × 10⁶ cells ml⁻¹. The maximum level of α-tocopherol was 257.7 ± 21.6 μg g⁻¹ dry weight (DW) at 18°C. The highest total carotenoids and chlorophylls were 6.5 mg g⁻¹ DW and 4.3 mg g⁻¹ DW, respectively, and the main pigments were determined as astaxanthin and lutein. Polyunsaturated fatty acids were found to be the predominant group, reaching 39.5% of the total fatty acids at 18°C. This comprised 20:5(n − 3) as the main polyunsaturated fatty acids (20.4%, at 18°C) followed by 22:6(n − 3) (4.8%, at 18°C). The results suggest that D. vlkianum can be successfully used as feed in shellfish hatcheries or aquaculture hatcheries, either as a substitute or in association with other microalgae, when this algae is cultured at 18°C.     

Effects of salinity, dietary level of protein and 17α-methyltestosterone on growth hormone (GH) and prolactin (tPRL177 and tPRL188) levels in the tilapia, Oreochromis mossambicus

In the present study, we examined the long-term effects of environmental salinity, diet (35% and 25% crude protein) and 17-methyltestosterone (MT) on corresponding levels of pituitary and serum growth hormone (GH) and prolactins (tPRL₁₇₇ and tPRL₁₈₈) in the tilapia (Oreochromis mossambicus). We observed no discernible patterns in serum GH that would suggest an effect of salinity, diet or MT. However, serum GH levels in all treatments declined at 1 and 3h after first feeding. Serum tPRL₁₇₇ and tPRL₁₈₈ were significantly higher in freshwater (FW) than in seawater (SW) and levels were significantly affected by dietary protein. tPRL₁₇₇ levels were higher in all groups fed a 35% protein diet, but tPRL₁₈₈ levels were higher only in the groups fed the MT-treated 35% protein diet; only serum tPRL₁₈₈ levels were affected by MT. Moreover, serum tPRL₁₇₇ and tPRL₁₈₈ increased throughout the sampling time-course. Subsequent work using fasted tilapia suggests that first feeding is likely to initiate the post-prandial suppression of serum GH levels. In contrast with the picture observed in blood, pituitary glands of SW animals showed higher levels of GH than FW fish. Pituitary GH was elevated by MT in both FW and SW. We also observed that pituitary tPRL₁₇₇ and tPRL₁₈₈ levels were higher in FW fish than in SW fish; tPRL₁₇₇ and tPRL₁₈₈ levels were elevated by MT only in FW animals. To assess the somatomedin activity of plasma from FW- and SW-reared tilapia, we measured [35S]-sulfate incorporation into ceratobranchial cartilage explants in vitro. Plasma from SW-adapted tilapia showed greater activity in this assay than plasma from FW-reared tilapia, suggesting that the GH-dependent IGF bioactivity of plasma is higher in SW-reared tilapia. Collectively, these studies suggest that the growth-promoting actions of SW rearing and of MT administration in tilapia may be linked to elevations in GH and/or prolactin (tPRL₁₇₇ and tPRL₁₈₈)levels.     

Effects of chronic exposure to γ-HCH (Lindane) on brain serotonergic and gabaergic systems,and serum cortisol and thyroxine levels of rainbow trout,Oncorhynchus mykiss

Rainbow trout (Oncorhynchus mykiss) were implanted intraperitoneally with 0.5 ml.100 g^–1 body weight of coconut oil alone (controls) or coconut oil contaning 0.05 mg of -HCH (Lindane). After 18 days, changes in brain serotonin and GABA metabolism, as well as in serum cortisol and thyroxine levels, were measured. A lower final body weight was observed in -HCH treated fish when compared with control fish. No significant differences were found for serum thyroxine levels between control and treated fish, but a significantly higher cortisol level was found in the -HCH-implanted trout. Although GABA levels did not differ significantly in any brain region in the two treatment groups, the activity of the serotonergic system was significantly altered by the pesticide in both the hypothalamus and the telencephalon.     

First results on migrating shad (Alosa fallax) and mullet (Mugil cephalus) echocounting in a lock on the Rhône River (France) using a split-beam sounder,and relationships with environmental data and fish caught

The application of fisheries acoustics in shallow waters opens new fields of observation and allows new methods for abundance estimates. In this paper we deal with echocounting of migrating fish crossing the Beaucaire-Vallabrègues lock, the first obstacle encountered on the Rhône, using a SIMRAD EY500 split-beam sounder, 70 kHz frequency. Two transducers sampled a section of water and evaluated approximately 35% of the lock section, based on a calibration sphere. The number of fish crossing the lock was calculated for each fish lock. Crossing data for the fish were recorded (dates, position, exit time). The present analysis did not allow the identification of the two major species that cross the dam: shad (Alosa fallax) and mullet (Mugil cephalus). The number of fish counted was positively correlated with the catches per unit effort (CPUE) by recreating fishermen below the dam and with increasing water temperature.     

Start-up of a “zero-discharge” recirculating aquaculture system using woodchip denitrification,constructed wetland,and sand infiltration

Recirculating aquaculture systems (RAS) discharge management limits the development of the aquaculture sector, because RAS do not automatically result in low nutrient emissions. Research has helped develop discharge management systems such as wetlands and woodchip bioreactors that have been adopted by Danish commercial model trout farms. To further develop the Danish concept, we have modelled and built a novel “zero-discharge” recirculating aquaculture system with an annual capacity of approximately 14 tonnes. The aim of this paper is to describe the entire concept and present the results from the start-up phase of the whole system. The concept includes the treatment of RAS effluent (overflow and sludge supernatant) using a hybrid solution of a woodchip bioreactor, constructed vertical wetland, and sand infiltration. Using this three-step process, the nitrate, phosphorus, and organic matter effluent are decreased to acceptable levels to reuse the water in the RAS process reducing the need for new raw water. In the first nine months of operation, a water treatment field was used as an end-of-pipe treatment to ensure the water was safe to recirculate for fish. During the winter, the water temperature dropped to 2.7 degrees in the sand filter, but the frost did not reach the water levels in any of the treatment processes. It therefore appears that a hybrid solution can operate sufficiently even in winter conditions. In the first year of operation, a woodchip bioreactor can remove 97 % of the nitrate, although the slow start-up of the RAS caused the bioreactor to be N-limited. On average, 79 % and 92 % of the inflow phosphate concentration was removed in the woodchip bioreactor and the entire hybrid treatment field respectively. The wetland and sand filter removed organic matter sufficiently (35 %), but because of the longer than designed actual water residence, it leached from the bioreactor more than was expected. Further experimentation is needed to identify the financial applicability and performance during higher feeding rates.     

Effects of 3,5,3′-triiodo-L-thyronine (T3) and 6-n-propyl-2-thiouracil (PTU) on growth of GH-transgenic coho salmon, Oncorhynchus kitsutch

GH-transgeniccoho salmon (Oncorhynchus kitsutch) juveniles were fed diets containing 3,5,3-triiodo-L-thyronine (T₃; 30 ng/g fish) or 6-n-propyl-2-thiouracil (PTU; 20 ug/g fish), to assess the effect of these drugs on the physiology, growthand survival in comparison with untreated transgenicand non-transgenic salmon. After 84 days, food intake, feed efficiency, survival, growth, hepato-somatic index (HSI), viscera-somatic index (VSI), plasma L-thyroxine (T₄), T₃and growth hormone (GH) levels,and cranial morphological abnormalities were determined. Growth of transgenic salmon was significantly faster than the nontransgenic salmon,and was increased by exogenous T₃and reduced by PTU. Food intake of transgenic salmon was higher than that of the nontransgenic group, but was reduced by exogenous PTU administration. Food conversion efficiency of transgenic salmon was lower than that of nontransgenic salmon,and also was increased by T₃ but reduced by PTU in the transgenic fish. The survival rate in all transgenic groups was significantly higher than that of nontransgenic,and transgenic T₃and PTU treatment groups showed higher survivals than the transgenic-control group. The HSIand VSI of the transgenic fish were higher than the nontransgenic fish;and both parameters in the transgenic salmon were increased by PTU, but reduced by T₃. The plasma T₄ level in transgenic salmon was approximately 1.5-fold higher relative to the nontransgenic fish, whereas no difference was observed among the transgenic groups. Plasma T₃ levels in transgenic salmon were also approximately 2-fold higher relative to the nontransgenic fish. However, the plasma T₃ level in transgenic animals was increased by exogenous T₃ administration, but was reduced by exogenous PTU to that observed in nontransgenic salmon. The plasma GH level of transgenic fish was higher than that of the nontransgenic salmon,and the level was increased by the exogenous T₃, whereas exogenous PTU did not reduce significantly GH levels in transgenic salmon. Transgenic fish also displayed cranium, jawand opercular abnormalities typical of the effects of this gene construct incoho salmon, indicating that some imbalance in growth processes has been induced. However, these abnormalities (especially cranial disruptions) were diminished by administration of exogenous PTU. In conclusion, exogenous T₃and PTU treatments can induce hyperthyroidismand hypothyroidism, respectively,and have inverse effects on growthand skeletal abnormalities of transgenic salmon constitutively expressing GH.     

Effect of diethylstilbestrol (DES) and 17 β-estradiol (E2) on growth, survival and histological structure of the internal organs in juvenile European catfish Silurus glanis (L.)

The effect of supplemented commercial diets with diethylstilbestrol (DES—15, 30 and 60 mg kg^?1) and 17 β-estradiol (E2—30 and 60 mg kg^?1), two chemicals commonly used in sex reversal procedure in fish, on survival and growth parameters of juvenile European catfish (Silurus glanis) was evaluated. During the two experiments, lasting 28 days each, fish were kept at temperature 25.2–26.5 °C, pH 7.4–9.3 and oxygen concentration 5.0–7.3 mg O₂ dm^?3. DES supplementation resulted in depressed growth rate of catfish. In experimental groups fed with E2, no negative effect on growth parameters was found. Both chemicals did not result in observed mortality. In all of the experimental DES groups, hepatosomatic index increased significantly, which suggests negative influence on physiological condition of catfish. DES supplementation significantly changed cytological factors of liver cells and caused hepatic alterations in parenchyma, such as vacuolization and blood congestion. Similarly, supplementation of E2 in food resulted in changes in cytological parameters of hepatocytes. However, E2 did not cause pathological changes within the liver tissue. Histological examination of the catfish gonads showed 19 and 38 % of sterile fish after treatment with 30 and 60 mg kg^?1 of DES, respectively. The results suggest that DES served in food could be ineffective in hormonal feminization process of European catfish. No disturbances of sex differentiation process after E2 treatment were observed. However, slight feminization effect in the highest level of E2 treatment group was recorded.     

Water quality,phytoplankton composition and growth of Litopenaeus vannamei (Boone) in an integrated biofloc system with Gracilaria birdiae (Greville) and Gracilaria domingensis (Kützing)

A 28-day indoor trial was conducted to evaluate the water quality, phytoplankton composition and growth of Litopenaeus vannamei in an integrated biofloc system with Gracilaria birdiae and Gracilaria domingensis. The experimental design was completely randomized with three treatments: control (shrimp monoculture); SB (shrimp and G. birdiae) and SD (shrimp and G. domingensis), all with three replicates. Random sampling was done (6 % of total population per experimental unit) to confirm white spot syndrome Virus (WSSV) infection using nested-PCR analysis due to suspicion of presence of the virus in the experiment (treatment and control groups). Shrimp L. vannamei (2.63 ± 0.10 g) were stocked in experimental tanks at a density of 425 shrimp m^?3, and the Gracilaria was stocked at a biomass of 2.0 kg m^?3. Shrimp mortality began in both the experimental and control groups at 10 days of culture. The integrated biofloc system (shrimp and seaweed) increased settleable solids (by 26–52 %); final weight (by 6–21 %); weekly growth (by 17–43 %); weight gain (by 17–43 %); specific growth rate (by 16–36 %); and yield (by 5–7 %) and decreased feed conversion ratio (by 21–28 %) and Cyanobacteria density about 16 % as compared to the control (shrimp monoculture). The use of red seaweed Gracilaria in an integrated biofloc system can enhance shrimp growth and reduce Cyanobacteria density in the presence of WSSV.     

An enzyme linked immunosorbant assay (ELISA) for testosterone, estradiol, and 17,20β-dihydroxy-4-pregenen-3-one using acetylcholinesterase as tracer: application to measurement of diel patterns in rainbow trout (Oncorhynchus mykiss)

A simple and rapid Enzyme Linked ImmunoSorbant Assay (ELISA) is described and validated for testosterone, estradiol, and 17,20-dihydroxy-4-pregnen-3-one (17,20P). A general procedure for preparation of the acetylcholinesterase labeled steroid is described which is applicable to any steroid. Use of acetylcholinesterase tracer increased the sensitivity of assay so that reliable measurements of each steroid could be achieved with only 10 l of plasma. The ELISA was applied to measurement of all three steroids every hour for over 24 hours in a female trout using cannulation of the dorsal aorta. This high sampling frequency revealed several short-term (<2 h) episodic pulses of testosterone and estradiol.     

Effect of long-term administration of dietary β-1,3-glucan on growth, physiological, and immune responses in Litopenaeus vannamei (Boone, 1931)

β-1,3-Glucan at different dietary doses was administered to enhance the growth, immunity, and survival against nitrite stress in Pacific white shrimp, Litopenaeus vannamei. Four different diets supplemented with 0, 250, 500, or 1,000 mg of β-1,3-glucan kg⁻¹ diets were fed to L. vannamei. Growth performance (weight gain and survival rate), physiological conditions (blood total protein, glucose, lactate, triacylglycerols, cholesterol levels) and immunological responses (superoxide dismutase, catalase, lysozyme, acid phosphatase, and alkaline phosphatase activities) of shrimp were recorded after 84-day feeding and 120 h after exposed to nitrite-N. After 84-day feeding, 250 mg kg⁻¹ β-1,3-glucan diet resulted in better weight gain (P < 0.05). Before the nitrite stress, blood lactate, triacylglycerols, and cholesterol level in shrimp fed with 250 mg kg⁻¹ β-1,3-glucan diet were significantly higher than those observed in shrimp fed with other diets (P < 0.05). Higher activities of catalase, lysozyme, and alkaline phosphatase were observed in shrimp fed with 500 or 1,000 mg kg⁻¹ β-1,3-glucan diet as compared to those obtained in shrimp fed with other diets (P < 0.05). After 120-h nitrite stress, blood protein, lactate, superoxide dismutase, catalase, and alkaline phosphatase activities in shrimp fed with 500 or 1,000 mg kg⁻¹ β-1,3-glucan were significantly higher than those observed in shrimp fed with other diets (P < 0.05). Glucose and triacylglycerol levels of shrimp fed with 500 or 1,000 mg kg⁻¹ β-1,3-glucan were significantly lower than those observed in other diets (P < 0.05). In shrimp fed with 500 and 1,000 mg kg⁻¹ β-1,3-glucan and 120-h after nitrite stress, the mortality was significantly lower than that observed in shrimp of control. Together, in this 84-day feeding trial, 250 mg kg⁻¹ β-1,3-glucan improved growth, whereas 500 mg kg⁻¹ β-1,3-glucan preferentially improved nitrite resistance, probably through accelerating energy metabolism and activating immune system.     

Effect of Neguvon® and Nuvan® treatment on crabs (Cancer pagurus,C. maenas), lobster (Homarus gammarus) and blue mussel (Mytilus edulis)

Survival tests were carried out with crabs, lobsters and blue mussels held in tanks of flowing seawater and subjected to Neguvon at 10, 1, 0.5 and 0.1 ppm and to Nuvan at 1, 0.01 ppm. Homarus gammarus was the most susceptible species; the animals died within 24 h at Neguvon levels of 0.5 ppm and higher and at Nuvan levels of 0.1 ppm and higher. The use of these chemicals in salmon farming appears to have deterious effects on marine invertebrate species.     

Suitability and optimization of FAO’s small-scale aquaponics systems for joint production of lettuce (Lactuca sativa) and fish (Carassius auratus)

Aquaponics is a developing technique that combines the simultaneous production of plants (hydroponics) and fish (aquaculture). With it, the use of resources (i.e., water, nutrients, land) is reduced whilst at the same time minimising residues’ discharge to the environment. Among its benefits, it allows the production of healthy vegetables and fish in reduced spaces by means of small-scale systems. In this work, three of them based on FAO models with different hydroponic subsystems (nutrient film technique -NFT-, floating raft, and vertical felt) are tested to produce lettuce (Lactuca sativa) and goldfish (Carassius auratus). Water parameters as well as the growth of plants and fishes were monitored in two different production cycles. The hydroponic subsystem that outperformed the best was the NFT, both in terms of crop production and water consumption. All systems showed similar results in fish production. Further research is needed to corroborate the outputs obtained when using other combinations of plants and fishes. Small-scale aquaponic systems are particularly interesting for self-production and even more so in urban environments with reduced available space.     

Isolation and characterization of Aeromonas schubertii from diseased snakehead, Channa maculata (Lacepède)

Pure bacterial cultures were isolated from diseased snakeheads, Channa maculata (Lacepède), suffering high mortality in a farm in Zhongshan, southern China. Three isolates, namely ZS20100725, ZS20100725-1 and ZS20100725-2, were identified as Aeromonas schubertii. All the isolates showed high 16S rRNA sequence similarities with A. schubertii. The isolates exhibited strong virulence to snakeheads in experimental challenges with LD(50) ranging between 1.4 × 10(4) and 6.4 × 10(6) CFU g(-1). Two of the isolates were positive for haemolysin, elastase, lipase and lecithinase by phenotypic determination, which was further confirmed by PCR amplification of the haemolysin and elastase genes. In sterile liquid medium, the best growth conditions of strain ZS20100725 were 30 °C, pH 7 and 0.5% salinity (w/v). Antibiotic susceptibility tests showed that strain ZS20100725 was susceptible to cefoxitin, cefoperazone and chloramphenicol. Furthermore, histopathology of diseased snakeheads infected with A. schubertii showed necrosis and congestion in liver, kidney and spleen and also damage to the cardiac muscle, intestine and gills.