Bacterial diversity of tilapia (Oreochromis niloticus) cultured in brackish water in Saudi Arabia

The bacterial flora occurring in brackish pond water, sediment, gills and intestine of healthy tilapia cultured in Saudi Arabia were estimated both quantitatively and qualitatively, and the isolates were identified to genus or species level. Total viable count of bacteria ranged from 1.4±1.5×10³ to 8.6±2.7×10³ cfu ml⁻¹; 1.2±3.1×10⁶ to 7.3±1.1×10⁷ cfu g⁻¹; 8.7±1.9×10⁵ to 2.1±0.9×10⁶ cfu g⁻¹; and 2.8±2.4×10⁷ to 1.0±1.6×10⁸ cfu g⁻¹ in the pond water, sediment, gills and intestine of brackish water tilapia, respectively. In total, 19 bacterial species were identified. The bacteria were predominantly Gram-negative rods (87%). Pond water and sediment bacteria influenced the bacterial composition of gills and intestine of tilapia. In contrast to gill bacteria, more diversification was observed in intestinal bacteria. The predominant (prevalence >10%) bacterial species were Vibrio parahaemolyticus, Vibrio carchariae, Vibrio alginolyticus, Chryseomonas sp., Vibrio vulnificus, and Streptococcus sp. in all the populations with the exception of the sediment population where Streptococcus sp. was replaced by Shewanella putrefaciens. Vibrio spp. (58% of the total isolates) dominated the total bacterial population.     

Filtration of king scallops (Pecten maximus)

Filtration rates of hatchery-reared king scallop (Pecten maximus L.) juveniles, fed a single species alga diet (Pavlova lutheri (Droop) Green), were measured at a range of temperatures (6–21 °C). Weight specific filtration rate (ml min⁻¹ g⁻¹ (live weight)) of juveniles of a selected size range of 17–19 mm shell height (0.26–0.36 g live weight) increased with temperature above 16 °C and decreased below 11 °C, but was not significantly different between these two temperatures. Measurements at 16 °C using juveniles with a wider size range of 10–25 mm shell height (0.05–0.8 g live weight) gave the allometric equation: filtration rate (ml min⁻¹)=12.19×weight (g)^0.887. Filtration rate decreased significantly when the cell concentration was greater than 200 cells μl⁻¹ (4.25 mg (organic weight) l⁻¹). With six other algae food species, filtration rates similar to those with P. lutheri were only achieved with Chaetoceros calcitrans (Paulsen) Takano. All other algae species tested were cleared from suspension at significantly lower rates. Experiments with diet mixtures of P. lutheri and these other algae suggested that this was usually a reflection of lowered filtration activity, rather than pre-ingestive rejection of cells. In experimental outdoor nursery rearing systems, the filtration rate was inversely proportional to the concentration of cells in the inflow, in the range 5–210 cells μl⁻¹. It was not affected by flow rate (2–130 l h⁻¹, equivalent to 0.12–28.38 l h⁻¹ g⁻¹ (live weight)) with scallop juveniles stocked from 2 to 62 g l⁻¹. The results are discussed in relation to on-growing scallops at field sites.     

Marine shrimp aquaculture: a novel waste treatment system

Ecuadorian Penaeus vannamei were cultured in dirt ponds (each of approximately 163 m²) at four different stocking densities, i.e. 5 shrimp m⁻², 10 shrimp m⁻², 15 shrimp m⁻² and 20 shrimp m⁻². Experiments were carried out over three different periods during the year. Each experiment lasted for 11–14 weeks. No commercial feed was given to the shrimp. The only input to the ponds was about 30 kg of cattle manure per pond per week. Chemical composition of the cattle manure was analyzed. Water quality parameters such as temperature, pH, DO and turbidity were recorded twice daily for each experiment; nutrients (nitrite, nitrate, ammonium and phosphate), water ATP, sediment ATP, H₂S and chlorophyll were measured twice weekly for each experiment. Shrimp were sampled either weekly or bi-weekly for body weight measurements.

The results showed a negative correlation between stocking density and growth. Weekly growth ranged from 0·44 to 1·58 g week⁻¹. Survival was over 50% in all treatments and averaged at 70·8%. Under these stocking densities, shrimp production ranged from 4·4 to 18·8 kg ha⁻¹ day⁻¹. The stocking density of 15 shrimps m⁻² provides better production than the other stocking densities.

Water quality data did not relate to any shrimp growth. Water nutrient levels in pond discharge water were less than or equal to the nutrients in the incoming water in spite of the weekly addition of cattle manure and did not increase with the addition of cattle manure. No coliform bacteria were detected in any pond water samples through the study period. This indicates digestion of cattle manure in marine shrimp ponds would not pollute the environment with high concentrations of dissolved nutrients.

Thus, a marine shrimp pond can be considered a dissolved nutrient marine treatment plant converting unwanted cattle manure (1841 kg cattle manure ha⁻¹ week⁻¹ in this study) into a valuable commodity — shrimp.     

Observations of pond hydrodynamics

This paper provides a detailed observation of pond hydrodynamics. Bathymetry and aerator deployment largely control the hydrodynamic regime. Aerators drive pond circulation, which is resisted by bottom and bank friction. Hydrodynamic friction is characterized by sediment grain size. One particular pond is offered as a proxy representative of Australian mariculture ponds in general: Pond X. The pond had been stocked with P. monodon at densities in the range 25–35 m⁻² about ten times during the period from 1989 to 1995. Observations were made in the year 1996. The site was on the east coast of Australia at 18° south latitude. The location was sheltered by a mountainous offshore island, such that winds effecting the pond averaged 2.3 m s⁻¹. Water exchange occurred at a rate of about 8.3% of pond capacity per day. Pond X was found to be rectangular, 120 m long and 87 m wide. The banks were found to slope between 1:1.5 and 1:7.6, with an average slope near 1:3. The pond was shallowest in one corner, where depth was 834 mm, and drained diagonally with 1872-mm maximum water depth. Aeration was provided by a total of six 1.5-kW machines, comprised of two of the paddlewheel type and four of the propeller-aspirator type. Each machine was found to deliver 200 N of horizontal thrust into the pond water column. The combined effect of the aerators created a circulation current averaging about 11 cm s⁻¹ around the pond periphery. The central region of the pond was found to have a fluctuating velocity of less than 1 cm s⁻¹. The mass-average size of pond soil and sediment was found to vary from 25 μm in the central region to 250 μm at the banks and where the bottom was swept by paddlewheels.     

The effects of temperature and food ration on metabolite concentrations in newly settled king scallop (Pecten maximus) spat

Juvenile king scallop (Pecten maximus L.) between 0.2 and 1.5 g wet shell weight per individual, reared at 10, 14 and 17 °C, were fed either a full ration equivalent to 0.5 g (ash-free dry weight of algae) g^− 1 (live weight of spat) week^− 1 or a restricted algal ration of 0.07 g (ash-free dry weight of algae) g^− 1 (live weight of spat) week^− 1. The algal food was a mixture of two microalgal species: Chaetoceros ceratosporum (Droop) Green and Pavlova lutheri Ostenfeld. Measurements were made of tissue concentrations of adenylate tri-, di- and mono-phosphate, lactate, succinate, acetate, pyruvate and l-alanine. The metabolite measurements were used as indicators of viability in animals exposed to a matrix of temperature and food supply conditions. It was found that the variations in metabolic responses could be correlated with those observed in a previous study that measured body condition in relation to the same set of variables. It was also found that the data from the present study were most strongly influenced by temperature-related effects and that there was an optimal temperature range between 10 and 14 °C. The metabolic responses, measured in terms of metabolite concentrations, would seem to confirm the existence of a bi-stable physiology in P. maximus in which there is a switch in state between 10 and 14 °C.     

Metabolic scope, swimming performance and the effects of hypoxia in the mulloway, Argyrosomus japonicus (Pisces: Sciaenidae)

The culture of the mulloway (Argyrosomus japonicus), like many other Sciaenidae fishes, is rapidly growing. However there is no information on their metabolic physiology. In this study, the effects of various hypoxia levels on the swimming performance and metabolic scope of juvenile mulloway (0.34 ± 0.01 kg, mean ± SE, n = 30) was investigated (water temperature = 22 °C). In normoxic conditions (dissolved oxygen = 6.85 mg l^− 1), mulloway oxygen consumption rate (M·o₂) increased exponentially with swimming speed to a maximum velocity (U_crit) of 1.7 ± < 0.1 body lengths s^− 1 (BL s^− 1) (n = 6). Mulloway standard metabolic rate (SMR) was typical for non-tuna fishes (73 ± 8 mg kg^− 1 h^− 1) and they had a moderate scope for aerobic metabolism (5 times the SMR). Mulloway minimum gross cost of transport (GCOT_min, 0.14 ± 0.01 mg kg^− 1 m^− 1) and optimum swimming velocity (U_opt, 1.3 ± 0.2 BL s^− 1) were comparable to many other body and caudal fin swimming fish species. Energy expenditure was minimum when swimming between 0.3 and 0.5 BL s^− 1. The critical dissolved oxygen level was 1.80 mg l^− 1 for mulloway swimming at 0.9 BL s^− 1. This reveals that mulloway are well adapted to hypoxia, which is probably adaptive from their natural early life history within estuaries. In all levels of hypoxia (75% saturation = 5.23, 50% = 3.64, and 25% = 1 .86 mg l^− 1), M·o₂ increased linearly with swimming speed and active metabolic rate (AMR) was reduced (218 ± 17, 202 ± 14 and 175 ± 10 mg kg^− 1 h^− 1 for 75%, 50% and 25% saturation respectively). However, U_crit was only reduced at 50% and 25% saturation (1.4 ± < 0.1 and 1.4 ± < 0.1 BL s^− 1 respectively). This demonstrates that although the metabolic capacity of mulloway is reduced in mild hypoxia (75% saturation) they are able to compensate to maintain swimming performance. GCOT_min (0.09 ± 0.01 mg kg^− 1 m^− 1) and U_opt (0.8 ± 0.1 BL s^− 1) were significantly reduced at 25% dissolved oxygen saturation. As mulloway metabolic scope was significantly reduced at all hypoxia levels, it suggests that even mild hypoxia may reduce growth productivity.     

Comparison of nutrients release among some maricultured animals

Integrated mariculture is a feasible method to maintain sustainable and high productivity of aquaculture. The choice of cultured animals and biofilters in the integrated system has to be made on the basis of their nutrient release rates and the clearance rate of each component of the system. We are examining the nutrient release rates among fish (mangrove snapper, Lutjanus russeli, and sea perch, Abudefduf septemfasciatus), abalone (Haliotis diversicolor), scallops (Chlamys noblis), and green mussels (Perna viridis) in the laboratory. Fish feed is the major sources of inorganic nutrient input in fish farms. The orthophosphate and ammonia release rates of minced trash fish (1593 μg P g⁻¹ day⁻¹ and 150 μg N g⁻¹ day⁻¹) were respectively 6–12 times and 4–88 times higher than those of cultivated fish. Mangrove snapper had the overall highest nutrient release rate, followed by sea perch, abalone, scallops, and mussels for nitrite and nitrate; and followed by abalone, sea perch, mussels, and scallops for orthophosphate and ammonium. Among mollusks, abalone had the highest orthophosphate (162 μg P g⁻¹ day⁻¹), nitrate (1.4 μg N g⁻¹ day⁻¹), nitrite (1.6 μg N g⁻¹ day⁻¹) and ammonium (25.0 μg N g⁻¹ day⁻¹) release rates per gram wet weight per day. Abalone released large amounts of orthophosphate, nitrite and nitrate in the experiment. Scallops and green mussels had low nutrient release rates.     

Serotonin induces ovarian maturation in giant freshwater prawn broodstock, Macrobrachium rosenbergii de Man

This study investigated the effects of serotonin (5-hydroxytryptamine or 5HT) on ovarian development in Macrobrachium rosenbergii de Man. Adult female prawns at the ovarian stage I (spent) were injected with 5HT at 1, 5, 10, 20 and 50 μg g^− 1 body weight (BW) intramuscularly on days 0, 5 and 10, and sacrificed on day 15. The doses as related to the effect could be categorized into three levels: low (1 and 5 μg g^− 1 BW of 5HT), medium (10 and 20 μg g^− 1 BW of 5HT) and high (50 μg g^− 1 BW of 5HT). The low-dose, especially at 1 μg g^− 1 BW, caused prawns to exhibit a significant increase in ovarian index (ovarian weight/body weight × 100) (5.79 ± 0.09%) as compared to the control (1.49%). The ovaries of most of these prawns could develop to stage IV (mature) and contained synchronously mature oocytes while most of the control ovaries remained at stage I and II (proliferative), and contained only oogonia to previtellogenic (Oc1, Oc2) and early vitellogenic oocytes (Oc3). The medium- and high-dose treated prawns exhibited ovaries that could reach stages III and IV and contained various types of oocytes of different maturity. Pretreatment with 5HT receptor antagonist, cyproheptadine (CYP), at 10 μg g^− 1 BW before 5HT injection significantly suppressed the effect of 5HT. Intramuscular injection of the 5HT-primed thoracic ganglion culture medium into CYP-pretreated prawns resulted in the increase of ovarian index about 5–6 times more than in the control, and in the groups injected with 5HT-primed media from muscle strip, eyestalk and brain. The ovaries of most prawn could develop up to stage IV and contained synchronously developed vitellogenic (Oc4) and mature oocytes (Oc5). These findings suggest that 5HT indirectly induces ovarian development and oocytes maturation in M. rosenbergii, probably via a putative ovarian stimulating factor released from the thoracic ganglia.     

An improved enzyme preparation for rapid mass production of protoplasts as seed stock for aquaculture of macrophytic marine green algae

Preparation of protoplasts and their subsequent applications for both basic and applied research of marine macroalgae remains largely under developed due to lack of development of reliable methods with consistent yields of viable protoplasts. An improved enzyme preparation with a single commercial enzyme, e.g. 2% Cellulase Onozuka R-10 in 1% NaCl solution, was developed to produce protoplasts rapidly from different green algal genera of Ulva, Enteromorpha and Monostroma. The simple dissolution of enzyme powder in 1% NaCl resulted in exclusion of 2% Macerozyme R-10 from the mixture consisting of 2% Cellulase Onozuka R-10 with 3% NaCl earlier reported as superior for the same algae. Optimal conditions for the isolation of maximum yields of viable protoplasts were found to be with 2% Cellulase Onozuka R-10 incubated at 20 °C for 2 h in 1% NaCl solution with 0.8 M mannitol adjusted to pH 6.0. The protoplast yield with optimized enzyme mixture was as high as 102.8 × 10⁶ cells g^− 1 f. wt for M. oxyspermum while it was in the range of 74.4–88.6 × 10⁶ cells g^− 1 f. wt thallus for seven species of Ulva, and 82.5–95.4 × 10⁶ cells g^− 1 f. wt for three species of Enteromorpha. The regeneration rate of protoplasts isolated using this method ranged from 89 to 92% with normal morphogenesis. The seeding of nylon threads with isolated protoplasts of M. oxyspermum was successful and after 3–4 weeks the entire frame with nylon threads became thick green in color with tiny germlings in laboratory culture. Thus, the method described in the present study allow for rapid mass production of viable protoplasts that could be potentially used as a source for seed material for mariculture and for other applied phycological research.     

Effects of body weight, water temperature and ration size on ammonia excretion by the areolated grouper (Epinephelus areolatus) and mangrove snapper (Lutjanus argentimaculatus)

The effects of body weight, water temperature and ration size on ammonia excretion rates of the areolated grouper Epinephelus areolatus and the mangrove snapper Lutjanus argentimaculatus were investigated. Under given experimental conditions, L. argentimaculatus had a higher weight-specific ammonia excretion rate than E. areolatus. Weight-specific ammonia excretion rates of fasted individuals of both species showed an inverse relationship with body weight (W, g wet wt.), but a positive relationship with water temperature (t, °C). The relationships for total ammonia nitrogen (TAN) were: E. areolatus: TAN (mg N kg⁻¹ d⁻¹)=21.4·exp^0.11t·W^−0.43 (r²=0.919, n=60); L. argentimaculatus: TAN (mg N kg⁻¹ d⁻¹)=121.5·exp^0.12t·W^−0.55 (r²=0.931, n=60). Following feeding, the weight-specific ammonia excretion rate of E. areolatus increased, peaked at 2 to 12 h (depending on temperature), and returned to pre-feeding levels within 24 h. A similar pattern was observed for L. argentimaculatus, with a peak of TAN excretion being found 6 to 12 h after feeding. Stepwise multiple regression analysis indicated that weight-specific TAN excretion rates of both species increased with increasing temperature and ration (R, percent body wt. d⁻¹): E. areolatus: TAN (mg N kg⁻¹ d⁻¹)=22.8·t−28.8·R−378.2 (r²=0.832, n=24); L. argentimaculatus: TAN (mg N kg⁻¹ d⁻¹)=22.9·t−25.4·R−216.4 (r²=0.611, n=24). The effect of body weight on weight-specific postprandial TAN excretion was not significant in either species (p>0.05). This study provides empirical data for estimating ammonia excretion of these two species under varying conditions. This has application for culture management.     

Physiological changes in rainbow trout held under crowded conditions and fed diets with different levels of vitamins E and C and highly unsaturated fatty acids (HUFA)

Rainbow trout (Oncorhynchus mykiss) maintained in crowded (100 kg m^− 3) and uncrowded (20 kg m^− 3) conditions were fed 42 days with five experimental diets having different levels of vitamin E (25.6 and 275.6 mg kg diet^− 1), C (0 and 1000 mg kg diet^− 1) and HUFA (highly unsaturated fatty acids, 12.5 and 320.5 g kg diet^− 1): −E−HUFA, −E+HUFA, +E−HUFA, +E+HUFA, −C+E+HUFA. Cortisol, plasma metabolites, tissue glycogen, fish composition, and tissue fatty-acid profile were evaluated at the end of the experimental period. In general, no changes in cortisol levels were associated with crowding, although +E+HUFA and −C+E+HUFA fish showed higher levels (mean ± SE, 55.5 ± 11.1 and 78.0 ± 11.3 ng ml^− 1) as a consequence of a possible interaction between chronic crowding and diet composition. Protein and glucose con-centration in plasma displayed no effect of crowding, but liver glycogen showed a general tendency to decrease in −E−HUFA, −E+HUFA, +E−HUFA, +E+HUFA, −C+E+HUFA crowded groups (70.2 ± 2.1, 52.1 ± 2.5, 73.4 ± 7.4, 91.7 ± 3.3, 74.2 ± 8.4 mg g^− 1 tissue, respectively) compared to uncrowded groups (108.9 ± 14.2, 82.7 ± 8.8, 92.4 ± 10.7, 99.1 ± 10.0, 103.5 ± 15.6 mg g^− 1 tissue, respectively), thus proving significant in −E+HUFA fish. Variations in total lipids, triglycerides, total cholesterol and HDL as well as LDL cholesterol in plasma were manifested under crowding conditions, displaying a certain influence of vitamin E and HUFA dietary content. Final body composition, in general, showed no change attributable to fish density, but some differences associated with diet composition were found in lipid and moisture percentages of crowded fish. Liver and muscle fatty-acid profile revealed a clear effect of the dietary lipid source that was more evident in muscle than in liver at normal fish density, and in some cases this effect was modulated by dietary vitamin E and C content and fish-culture conditions.     

Effects of secondary crops on bacterial growth and nitrogen removal in shrimp farm effluent treatment systems

Secondary crops provide a means of assimilating some effluent nitrogen from eutrophic shrimp farm settlement ponds. However, a more important role may be their stimulation of beneficial bacterial nitrogen removal processes. In this study, bacterial biomass, growth and nitrogen removal capacity were quantified in shrimp farm effluent treatment systems containing vertical artificial substrates and either the banana shrimp Penaeus merguiensis (de Man) or the grey mullet, Mugil cephalus L. Banana shrimp were found to actively graze biofilm on the artificial substrates and significantly reduced bacterial biomass relative to a control (24.5±5.6 mg C m⁻² and 39.2±8.7 mg C m⁻², respectively). Bacterial volumetric growth rates, however, were significantly increased in the presence of the shrimp relative to the control (45.2±11.3 mg C m⁻² per day and 22.0±4.3 mg C m⁻² per day, respectively). Specific growth rate, or growth rate per cell, of bacteria was therefore appreciably stimulated by the banana shrimp. Nitrate assimilation was found to be significantly higher on grazed substrate biofilm relative to the control (223±54 mg N m⁻² per day and 126±36 mg N m⁻² per day, respectively), suggesting that increased bacterial growth rate does relate to enhanced nitrogen uptake. Regulated banana shrimp feeding activity therefore can increase the rate of new bacterial biomass production and also the capacity for bacterial effluent nitrogen assimilation. Mullet had a negligible influence on the biofilm associated with the artificial substrate but reduced sediment bacterial biomass (224±92 mg C m⁻²) relative to undisturbed sediment (650±254 mg C m⁻²). Net, or volumetric bacterial growth in the sediment was similar in treatments with and without mullet, suggesting that the growth rate per cell of bacteria in grazed sediments was enhanced. Similar rates of dissolved nitrogen mineralisation were found in sediments with and without mullet but nitrification was reduced. Presence of mullet increased water column suspended solids concentrations, water column bacterial growth and dissolved nutrient uptake. This study has shown that secondary crops, particularly banana shrimp, can play a stimulatory role in the bacterial processing of effluent nitrogen in eutrophic shrimp effluent treatment systems.     

Appetite, gut transit, oxygen uptake and nitrogen excretion in captive Atlantic halibut, Hippoglossus hippoglossus L., and lemon sole, Microstomus kitt (Walbaum)

Atlantic halibut, Hippoglossus hippoglossus L., eat larger satiation meals (mean 11.7% body weight) than lemon sole, Microstomus kitt (Walbaum), (2.6% body weight). Total gut clearance time was about 120 h for halibut and 72 h for lemon sole. There are marked differences in feeding behaviour between the two species; halibut feed in midwater and require several body lengths of approach swimming before taking large items of food, while lemon sole eat only off the bottom. In shared tanks, no aggressive interaction was observed. A duoculture system holding small numbers of lemon sole with the more valuable halibut is recommended as a means of minimizing food waste and tank fouling. Oxygen uptakes of 0.07-0.11 ml O₂ g fish wt⁻¹ h⁻¹ (depending on nutritional state) were recorded for the two species. Ammonia nitrogen outputs were also similar. Starved halibut excreted 2.32 μg N g⁻¹ h⁻¹, fed animals 5.08 μg N g⁻¹ h⁻¹. The corresponding values for lemon sole were 3.26 μg N g⁻¹ h⁻¹ and 6.37 μg N g⁻¹ h⁻¹, respectively.     

Effect of water temperature on the immune response of white shrimp Litopenaeus vannamei to Vibrio alginolyticus

White shrimp Litopenaeus vannamei held in 25‰ seawater at 27 °C or 28 °C were injected with TSB-grown Vibrio alginolyticus at 1 × 10⁴ colony-forming units (cfu) shrimp^− 1 or 1 × 10⁵ cfu shrimp^− 1, and then cultivated onward at water temperatures varying from 20 to 34 °C. Over 24–144 h, mortality of V. alginolyticus-injected shrimp held at 34 °C or 32 °C was significantly higher than that of shrimp held at lower temperatures. In a separate experiment, shrimp held in 25‰ seawater at 28 °C and then cultured onward at 20 to 32 °C were examined for immune parameters at 24–96 h. THC, phenoloxidase activity, respiratory burst, and SOD activity decreased significantly at 24 h after transfer to 32 °C. Shrimp held in 25‰ seawater at 27 °C and then cultured onward at 20 to 34 °C showed a significant reduction in phagocytic activity and clearance efficiency for V. alginolyticus at 24 h after transfer to 34 °C. It was concluded that transfer of shrimp from 27 or 28 °C to higher temperatures (32 and 34 °C) reduced their immune capability and decreased resistance to V. alginolyticus infection.     

Enrichment of Artemia with free methionine

Different enrichment procedures of the free amino acid (FAA) methionine were tested for Artemia nauplii. A direct enrichment protocol (methionine dissolved in the culture water) was compared with liposome enrichment protocols that varied in their membrane composition. During 16 h of direct enrichment in 5.3 mM methionine, the nauplii increased their content of free methionine between 20 and 30 times compared to the unenriched control (43.1±1.2 and 68.4±3.8 pmol·nauplius⁻¹ in two separate experiments vs. 2.4±1.0 pmol·nauplius⁻¹ in control). However, by encapsulating the identical amount of methionine into liposomes made from pure egg yolk phosphatidylcholine (PC) (>99% PC) and cholesterol, the nauplii content of free methionine reached 148.8±27.6 pmol·nauplius⁻¹, which is approximately 60 times more than in the unenriched control. Another liposome composition tested, made from crude egg yolk PC (>60% PC) and cholesterol, resulted in 90.5±4.1 pmol·nauplius⁻¹. The enriched nauplii still retained 80% of the free methionine after 8 h of incubation at conditions simulating feeding for Atlantic halibut larvae (13°C, 33.5 g·l⁻¹).

In conclusion: (1) Artemia nauplii can successfully be enriched with free methionine, (2) the high retention of free methionine in the Artemia nauplii following transfer to fish tanks shows that it is possible to offer fish larvae a feed with a high level of FAA, based on enrichment of Artemia nauplii.     

Modeling industry-wide sediment oxygen demand and estimation of the contribution of sediment to total respiration in commercial channel catfish ponds

Data collected from 45 commercial channel catfish, Ictalurus punctatus, ponds were used to develop empirical models predicting sediment oxygen demand (SOD). Seven acceptable models were combined with a Monte-Carlo sampling distribution to predict industry-wide sediment oxygen demand (SOD_i). The SOD_i values obtained from the best equation were used in simulations to assess the effect of diurnally varying water column dissolved oxygen (DO) concentrations on SOD and the effect of pond water depth on the contribution of SOD to overall pond respiration. Estimated SOD_i ranged from 62 to 962 mg m⁻² h⁻¹, with a mean of 478 mg m⁻² h⁻¹. There was a 95% probability of mean SOD_i being ≥700 mg m⁻² h⁻¹. The effects of diurnal variation in DO concentration in the water column on expression of SOD was modeled by combining maximum SOD_i, an empirical relationship between DO and SOD, and simulated pond DO concentrations. At DO concentrations >15 mg l⁻¹, diel SOD in catfish ponds exceeded 20 g O₂ m⁻² day⁻¹. But when average diel DO was <4 mg l⁻¹ and the range of DO concentration was 6–8 mg l⁻¹, SOD decreased to 13 g O₂ m⁻² day⁻¹ because DO availability limited the full expression of potential SOD. Respiration totals for sediment (average SOD_i), plankton, and fish respiration were calculated for pond water depths ranging from 0.25 to 4 m. Although whole-pond respiration increases as pond depth increases, the proportion of total respiration represented by sediment decreased from 48 to 10% by increasing water depth over this range. The results of these studies show that SOD is a major component of total pond respiration and that certain management practices can affect the impact of SOD on pond oxygen budgets. Mixing ponds during daylight hours, either mechanically or by orienting ponds for maximum wind fetch, will increase oxygen supply to sediments, thereby allowing maximum expression of SOD and maximum mineralization of sediment organic matter. Given a mixed condition caused by wind or other artificial means, the construction of deeper ponds increases the total mass of DO available for all respiration, causing nighttime DO concentrations to decline at a slower rate, reducing the need for supplemental aeration. Because a pond’s water volume decreases over time from sediment accumulation, annual aeration costs will increase with pond age. Constructing ponds with greater initial depth will therefore reduce long-term cost of aeration, allow more flexible management of pond water budget, and reduce the long-term expense associated with pond reconstruction.     

Furunculosis in Atlantic salmon (Salmo salar L.) is not readily controllable by bacteriophage therapy

The potential of bacteriophage therapy to control bacterial disease in farmed fish was tested using, as an example, furunculosis of Atlantic salmon, caused by Aeromonas salmonicida subsp. salmonicida.

In vivo testing with Atlantic salmon and rainbow trout (Oncorhynchus mykiss Walbaum) showed no adverse effects, with bacteriophage generally cleared within 96 h of administration by either intraperitoneal (i.p.) injection or oral in-feed.

Juvenile Atlantic salmon were administered a combination of bacteriophage O, R and B (1.9 × 10⁸ pfu fish^− 1) by i.p. injection, after they had been challenged with A. salmonicida subsp. salmonicida 78027, also by i.p. injection. The fish that were injected with bacteriophage immediately after challenge died at a significantly slower rate then those that were either not treated with bacteriophage, or treated 24 h post-challenge. However, the end result (100% mortality) was not affected.

In further experiments the effects of oral (1.88 × 10⁵ pfu g^− 1 fish^− 1 daily for 30 days), bath (1.04 × 10⁵ ml^− 1 daily for 30 days) and i.p. (6.25 × 10⁷ pfu fish^− 1) phage treatment to control furunculosis in experimentally infected Atlantic salmon were compared with antibiotherapy (treatment with 10 mg kg^− 1 bw^− 1 day^− 1 oxolinic acid for 10 days), using an indirect cohabitation challenge. No protection was offered by any of the bacteriophage treatments, compared to the positive challenge group, although significant protection was offered by the oxolinic acid treatment. Analysis of samples taken from the trials demonstrated that bacteriophage were correctly administered to the fish and, on occasion, were isolated from fish that had succumbed to furunculosis. It was also shown that bacteriophage resistant A. salmonicida subsp. salmonicida isolates could be recovered from mortalities in all the treatment groups.

The results suggest that, although there were no safety problems associated with the approach, furunculosis in Atlantic salmon is not readily controllable by application of bacteriophage.     

Effect of docosahexaenoic acid enrichment in Artemia on growth of Pacific bluefin tuna Thunnus orientalis larvae

Feeding enriched Artemia induces growth failure in Pacific bluefin tuna (PBT) Thunnus orientalis larvae; however, feeding of yolk-sac larvae of marine fish promotes larval growth. It is considered that this growth failure partly results from dietary docosahexaenoic acid (22:6n-3, DHA) deficiency. Therefore, we examined the effect of DHA contents in enriched Artemia on the growth of PBT larvae. Artemia nauplii were enriched with graded levels of DHA ethyl ester, and fed to PBT larvae for 9 days. Yolk-sac larvae of Japanese parrotfish Oplegnathus fasciatus were used as a reference diet. The DHA contents in Artemia increased with the enrichment from 0 mg g^− 1 dry weight basis (DW) to 25 mg g^− 1 DW, while the content in the reference diet was 21 mg g^− 1 DW. Feeding of enriched Artemia significantly improved the growth of PBT larvae. However, this improvement was negligible when compared with the excellent growth of the larvae that were fed the reference diet. PBT larvae that were fed the reference diet accumulated approximately twice or much higher levels of DHA in the neutral and polar lipids in the body when compared with the larvae that were fed enriched Artemia. These results show that PBT larval growth cannot be promoted by feeding enriched Artemia even if the DHA contents in Artemia are elevated to the same levels as those of yolk-sac larvae. The incorporation of dietary DHA into phospholipids in the fish body may be desirable for the normal growth of PBT larvae.     

A laboratory scale recycling water unit for tilapia breeding

The technical features of a laboratory scale water recycling unit for experimental small scale tilapia breeding are described. Two units (1 and 2) were operated during a 6 month period, carrying a similar fish load (7·5 kg) and feeding rate (2% fish body weight/day). Unit 1 received natural illumination, while unit 2 was artificially illuminated (14/10 - light/dark cycle). Both units were equipped with a biological filter bed (substrate surface area, 3500 cm²). In unit 1, total ammonium and nitrite concentrations ranged from 0·05 to 0·5 mg liter⁻¹, while nitrate varied between 10–40 mg liter⁻¹. In unit 2 corresponding values were 0·15-3 mg liter⁻¹, 0·05–0·8 mg liter⁻¹ and 10–40 mg liter⁻¹. Temperatures ranged between 20–29°C and pH values between 7·5–6·9 in both units. Dissolved oxygen concentrations decreased gradually from 5·6 to 3·4 mg liter⁻¹ in unit 1 and from 5·6 to 2·6 mg liter⁻¹ in unit 2. Twenty-six spawnings occurred in unit 1 in March and April, while only eight spawnings occurred in unit 2, possibly because of the absence of sunlight. The significance of these results are discussed.     

Harvesting marine microalgae species by chitosan flocculation

Marine microalgae are still an important larval feeding source. One of the most promising harvesting techniques of the algae produced appears to be chemical flocculation. We report results obtained with chitosan flocculation of five marine species of microalgae of importance to mariculture (Skeletonema costatum, Dunaliella tertiolecta, Thalassiosira nordenskoldii, Chlorella sp. and Thalassionema sp.). The algae were grown in the laboratory in 20-liter batch cultures under normal conditions in artificial seawater. Without pH control, a 100% flocculation efficiency was reached at fairly high chitosan concentrations (above 40 mg liter⁻¹). When the final pH was adjusted to around 7·8–8·0, a 100% flocculation efficiency was obtained with chitosan concentrations of 40 mg liter⁻¹ or more. However, when pH was adjusted to around 7 or less, prior to chitosan addition for S. costatum and Chlorella sp., the concentration of chitosan required to obtain a 95–100% flocculation efficiency was reduced to 20 mg liter⁻¹ for Chlorella and 2 mg liter⁻¹ for S. costatum. The results are discussed in the light of the currently accepted theories on flocculation.