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.     

Determination of phosphorus saturation level in relation to clay content in formulated pond muds

An experiment was conducted to determine the amount of P needed to saturate simulated fish pond sediments, formulated to contain six levels of clay (0, 30, 41, 64, 73 and 81% by weight). A series of cylindrical cement tanks were filled to 20 cm depth with the six sediment types and triple superphosphate (TSP) solution was added to reach P saturation in sediment. Results showed that all sediment types reached constant inorganic-P concentration in the upper 5 cm after 12 weeks of TSP application, and P adsorption capacity of sediment increased with increasing clay content. Sediment P adsorption was slower and not significant (P > 0·05) below 5 cm depth except in the sediment type containing 0% clay. Regression analysis showed that the rate and adsorption capacity of P in sediment are primarily governed by clay content and its dominant minerals. While organic-P and loosely bound-P are commonly deposited in sediment, most inorganic-P is adsorbed by cations to form cation-P complexes. The linear relationship between cation-P saturation level and the percentage of clay in sediment is highly significant (r² = 0·84, P < 0·001) and, therefore, maximum adsorption capacity of cation-P in pond sediment can be approximated by Y = 0·019X (where Y represents the 100% saturation level in mg P g⁻¹ soil, and X is the percentage of clay in the sediment). In practice, the level of P saturation in sediment can be approximated by the initial cation-P and clay contents in the top 5 cm of pond mud using the equation: P saturation (%) = initial cation-P (mg g⁻¹ soil) × 100/P adsorption capacity (mg g⁻¹ soil).     

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.     

Quantitative and qualitative studies on bacterial flora of hybrid tilapia (Oreochromis niloticus × O. aureus) cultured in earthen ponds in Saudi Arabia

The bacterial flora of the rearing pond water and sediment as well as the gills and intestine of healthy hybrid tilapia cultured in Saudi Arabia was estimated quantitatively and qualitatively, the isolates being identified at genus or species level. Total viable counts of bacteria (measured as colony‐forming units, cfu) were in the range 5.6 ± 0.8 × 10³ to 2.4 ± 1.2 × 10⁴ cfu mL^?1 in pond water; 9.3 ± 1.1 × 10⁶ to 1.9 ± 1.5 × 10⁸ cfu g^?1 in sediment; 7.1 ± 0.7 × 10⁵ to 8.7 ± 1.1 × 10⁶ cfu g^?1 in the gills of tilapia; and 3.4 ± 1.8 × 10⁶ to 5.8 ± 0.4 × 10⁷ cfu g^?1 in the intestine of tilapia. In total, 15 bacterial genera and 18 species were identified. Pond water and sediment bacteria reflected the bacterial composition in the gills and intestine of tilapia. In contrast to gill bacteria, more diversification was observed in intestinal bacteria. Corynebacterium urealyticum, Shewanella putrefaciens and Aeromonas hydrophila predominated in all samples. In pond water, C. urealyticurn, S. putrefaciens, A. hydrophila, Flavobacterium sp. and Pseudomonas sp. were the most predominant bacterial species (prevalence > 10%), whereas A. hydrophila, C. urealyticum, S. putrefaciens and Escherichia coli were predominant in pond sediment, and C. urealyticum, S. putrefaciens and A. hydrophila were predominant in both the gills and intestine of tilapia.     

Streptococcus iniae infection and tissue distribution in hybrid striped bass (Morone chrysops×Morone saxatilis) following inoculation of the gills

This study was designed to test the possibility that Streptococcus iniae enters through the gills and causes infection in hybrid striped bass. To determine the dose response, four groups of fish were inoculated with S. iniae via the gills with a dose of 5.0×10⁵, 2.6×10⁶, 5.0×10⁶, or 1.0×10⁸ CFU/fish. One group of fish was inoculated with tryptic soy broth (TSB) via the gills to serve as controls. The cumulative percent mortality was 13%, 27%, 100% and 100% for 5.0×10⁵, 2.6×10⁶, 5.0×10⁶ and 1.0×10⁸ CFU/fish, respectively. We also examined the tissue dissemination of S. iniae at 0.5, 4, 8, 12, 24, 48 and 72 h after experimental gill inoculation. Fish were inoculated with 2.6×10⁶ or 5.0×10⁶ CFU/fish, which caused low and high mortality, respectively. Within 48 h, fish inoculated with the 2.6×10⁶ dose were culture positive on the gill surface, blood of the first and second gill arches, blood of the third and fourth gill arches and the nares. However, for the dose of 5.0×10⁶ CFU/fish, S. iniae was also isolated from the olfactory, optic and cerebellum regions of the brain, eye, head kidney, trunk kidney, spleen and liver at 48 h. For the 2.6×10⁶ dose, S. iniae was not isolated until 48 h post-inoculation, but was isolated at 12 h for the 5.0×10⁶ dose. The results of this study indicate that S. iniae can enter hybrid striped bass through the gills. However, mortality at similar S. iniae doses was lower than we previously observed by inoculation of the nares.     

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.     

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.     

Protection, immune response and side-effects in European whitefish (Coregonus lavaretus L.) vaccinated against vibriosis and furunculosis

As indicated by market demand and cultivation prospects, the whitefish (Coregonus lavaretus L.) has the potential to become an important alternative to rainbow trout in fish farming in Finland. The fish processing industry has called for a fish species having non-pigmented flesh, and there is a long tradition of whitefish farming for stocking purposes in Finland. However, cultivation in net cages in the brackish water of the coastal area exposes fish to vibriosis (Listonella (Vibrio) anguillarum) and furunculosis (Aeromonas salmonicida salmonicida). Hence, profitable whitefish farming requires the efficient control of both of these diseases. The efficacy of vaccination (Apoject 1800®) was studied in the laboratory using challenge tests and by monitoring specific antibody production (ELISA) and the blood leucocyte pattern. The fish (22 g) were vaccinated at 15°C and the immunity developed during 8 weeks prior to challenge with vibriosis (1.8×10⁵ cfu) and 12 weeks prior to challenge with furunculosis (4.9×10² cfu). The challenge was performed by i.p. injection. Side-effects around the injection site were studied 16 weeks post-vaccination. The immune system of the whitefish responded well and vaccination gave good protection against vibriosis and furunculosis. The RPS was 100% following the Vibrio challenge and 99% following the Aeromonas challenge. The antibody levels continued increasing in the vaccinated group throughout the study period. Lymphocyte and neutrophil counts were elevated in vaccinated compared to non-vaccinated fish at 8 weeks post-vaccination, but not after 16 weeks. Vaccination produced weak adhesions around the injection site in 42% and minor pigmentation in 9% of the 65 fish examined.     

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.     

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.     

Evidences of the intertidal red alga Grateloupia turuturu in turning Vibrio parahaemolyticus into non-culturable state in the presence of light

Grateloupia turuturu, previously known as Grateloupia doryphora, has been widely reported to be an invasive algal species. There are no studies to relate the impact of its existence on its surrounding environment. In this paper, we present our results to show that about 70% of individuals collected from the field could turn Vibrio parahaemolyticus into non-culturable state on both selective (TCBS) and non-selective (2216E) culture medium in 24 h in the presence of light in live algal culture. Total bacteria counts on TCBS and 2216E plates dropped from the initial 565 (± 174) and 1192 (± 60) cfu ml^− 1 respectively to zero in 24 h. This effect disappeared when the alga was grown in darkness. The same effect was not found in two other intertidal macroalgae Laminaria japonica and Palmaria palmata. Further tests showed that the settlement ability of bacteria in seawater was impaired significantly in the presence of this alga in comparison with three other algal species.     

Effects of various feed supplements containing fish protein hydrolysate or fish processing by-products on the innate immune functions of juvenile coho salmon (Oncorhynchus kisutch)

Immunomodulators administered to fish in the diet have been shown in some cases to enhance innate immune defense mechanisms. Recent studies have suggested that polypeptide fractions found in fish protein hydrolysates may stimulate factors in fish important for disease resistance. For the current study, groups of coho salmon were reared on practical feeds that contained either fish meal (Control diet), fish meal supplemented with cooked fish by-products, or fish meal supplemented with hydrolyzed fish protein alone, or with hydrolyzed fish protein and processed fish bones. For each diet group, three replicate tanks of fish were fed the experimental diets for 6 weeks. Morphometric measurements, and serologic and cellular assays were used to evaluate the general health and immunocompetence of fish in the various feed groups. Whereas the experimental diets had no effect on the morphometric and cellular measurements, fish fed cooked by-products had increased leucocrit levels and lower hematocrit levels than fish from the other feed groups. Innate cellular responses were increased in all feed groups after feeding the four experimental diets compared with pre-feed results. Subgroups of fish from each diet group were also challenged with Vibrio anguillarum (ca. 7.71×10⁵ bacteria ml⁻¹) at 15 °C by immersion. No differences were found in survival among the various feed groups.     

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.     

Effect of probiotics, Enteroccus faecium, on tilapia (Oreochromis niloticus) growth performance and immune response

The aim of this study was to analyze the effect of a probiotic bacterium, Enterococcus faecium ZJ4 on growth performances and immune responses of tilapia (Oreochromis niloticus). The tilapias were treated with E. faecium ZJ4 at a final concentration of 1 × 10⁷ cfu ml^− 1 in aquaria water every 4 days. Six aquaria with three replicates for treated and controls were used. After 40 days, the tilapias supplemented with the probiotic showed significantly better final weight and daily weight gain (DWG) than those fed the basal diet (Control) (P < 0.05). There was no remarkable difference (P > 0.05) in the total serum protein, albumin content, globulin concentration and A/G ratio between the treated and control tilapias. The result of lysozyme activity assay was similar to these biochemical indexes. However, the complement component 3 content, myeloperoxidase (MPO) activity and the respiratory burst activity of blood phagocytes were higher (P < 0.05) in E. faecium treated tilapias (trial 1) than the controls.     

Feed intake, growth and feed utilization of offspring from wild and selected Atlantic salmon (Salmo salar)

The aim was to assess whether selection for increased growth rate in Atlantic salmon (Salmo salar) is associated with increased feed intake and/or better feed utilization. Growth responses of offspring from selected and wild lines of Atlantic salmon (initial weight 814±14 g and 533±12 g, respectively) were tested in a 14-week experiment. Selected and wild salmon increased body weight by 79 and 39%, respectively, during the experiment. Relative feed intake (DFI), thermal growth coefficient (TGC) and feed efficiency ratio (FER) were significantly higher in the selected (DFI: 0.67% BW d⁻¹, TGC: 2.96×10⁻³ and FER: 1.16), than in the wild, line (DFI: 0.48% BW d⁻¹, TGC: 1.39×10⁻³ and FER: 0.93). FER was positively correlated with mean growth rate (r=0.90, n=6, P<0.05), and differences between the lines indicated a 4.6% increase in FER per generation of selection. Fish of the selected line had a significantly lower intake of protein and energy per kilogram gain, so the higher growth rate of the selected line was the result of both greater feed consumption and more efficient feed utilization for growth. This implies that selection for increased growth rate in Atlantic salmon may improve both of these traits.     

Aerobic Bacterial Flora of Common Carp (Cyprinus carpio L.) Cultured in Earthen Ponds in Saudi Arabia

ABSTRACT

Quantitative and qualitative estimation of bacterial flora present in pond water, sediments, gills, and intestine of healthy common carp Cyprinus carpio cultured in Saudi Arabia were performed and identified to species level where possible. Mean total viable bacterial counts in pond water ranged from 1.2?±?2.9?×?10⁴ to 2.5?±?3.5?×?10⁵ cfu/mL; in sediments, 9.3?±?2.1?×?10⁷ to 2.7?±?3.5?×?10⁹ cfu/g; in gills filaments, 4.3?±?2.9?×?10⁶ to 1.6?±?3.9?×?10⁷ cfu/g; and in intestine, 8.7?±?4.1?×?10⁹ to 5.4?±?3.2?×?10¹⁰ cfu/g. Gram-negative rod-shaped bacteria dominated (76%) the populations. In total, 12 bacterial genera and 15 species were identified. Pond water and sediment bacteria had the reflection on bacterial composition of gills and intestine of carp. Intestinal bacteria showed more diversification in contrast to gill bacteria. Aeromonas hydrophila, Shewanella putrefaciens, Vibrio cholerae, Corynebacterium urealyticum, Vibrio vulnificus, Vibrio sp., and Staphylococcus sp. were the common bacteria in all the populations. In pond water and carp intestine, A. hydrophila, S. putrefaciens, V. cholerae, and C. urealyticum were the most dominant bacteria (prevalence ≥ 10%) where pond sediments and the carp gills experienced with more one dominant bacterium V. vulnificus. Only the A. hydrophila covered one fourth (25%) of the total bacterial populations.     

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.     

Shipping studies with juvenile and adult Malaysian prawns Macrobrachium rosenbergii (de Man)

Four studies examined shipping factors of packing technique, density, duration, type of water and use of habitat material for shipping juvenile and adult prawns Macrobrachium rosenbergii. Prawns were shipped in double polyethylene lined 38 × 38 × 20 cm deep styrofoam boxes containing oxygenated water. At temperatures of 19–20°C, 17 g prawns could be shipped safely for 42 h at a density of 10–12 prawns per box (12–15 g liter⁻¹ shipping water). Juveniles, mean size about 6 g, could be shipped at a density of 40 per box (18 g liter⁻¹) for 24 h or 20–25 per box (9–11 g liter⁻¹) for 48 h. Use of mesh material to increase surface area in the box did not appear beneficial nor did shipping in brackish water (salinity8‰). Adults packed unrestricted resulted in survival rates substantially higher than those obtained from immobilized prawns wrapped in mesh.

During the shipment, pH and dissolved oxygen concentrations decreased whereas ammonia concentrations increased. The decreased pH levels may have reduced the ammonia toxicity by decreasing the amount of toxic unionized ammonia (NH₃) in solution. In general, dissolved oxygen concentrations appeared more closely related to survival rates than did other water quality parameters which were measured.     

A simple test to estimate the salinity resistance of fish with specific application to O. niloticus and S. melanotheron

In order to develop a simple and accurate index of the salinity resistance of tilapia, batches of 10 juveniles (5 to 20 g) of two different species Oreochromis niloticus and Sarotherodon melanotheron reared in freshwater were subjected to gradual increases in salinity until 100% mortality. Seven daily increments of salinity were tested with 4 replicates: 2, 4, 6, 8, 10, 12 and 14 g l⁻¹ day⁻¹, while control batches were kept in fresh water. The temperature was maintained at 27 °C. The concentration of oxygen, ammonia and the pH were not limiting factors. The mortality, monitored on a daily basis, appeared after 2–51 days and was spread out over 1–20 days, depending on the increment of salinity. The higher the daily rate in salinity increase, then the shorter the time lapse before total mortality occurred. The cumulative mortality as a function of salinity fit well with simple linear regressions. The criterion of the resistance to salinity was the index MLS (median lethal salinity) defined at each daily rate as the salinity at which 50% of fish died. For S. melanotheron, the mean MLS was 123.7±3.5 g l⁻¹ whatever the daily rate in salinity. For O. niloticus, the MLS was 46.3±3.4 g l⁻¹ for daily increases in salinity ranging from 2 to 8 g l⁻¹ day⁻¹ and decreased significantly (P<0.05) above this level. The MLS-8 g l⁻¹ day⁻¹ ,which takes into account the full capacity of the fish to adapt to the increasing salinity, appeared to be a simple, optimized and efficient criterion for assessing the resistance to salinity for O. niloticus and S. melanotheron. This criterion can be a useful tool for ranking the different parental strains and hybrids of different genus and species of tilapia used in programmes of genetic selection for growth and salinity tolerance.