Effects of dissolved oxygen on hemolymph parameters of freshwater giant prawn, Macrobrachium rosenbergii (de Man)

Profiles of changes in physiological parameters of freshwater giant prawns, Macrobrachium rosenbergii, exposed to various dissolved oxygen (DO) levels of 7.75, 4.75, 2.75, and 1.75 mg l⁻¹ are reported. The parameters involved in osmoregulation and oxygen transport were monitored for a 6-day period. Notable depressions in hemolymph osmolality, Na⁺, K⁺, and Cl⁻ contents were observed within 24 h after exposure to hypoxia at 2.75 and 1.75 mg O₂ l⁻¹, and thereafter remained at rather steady levels, which were significantly lower than those under normoxic conditions (4.75–7.75 mg O₂ l⁻¹). The extent of depression of osmotic-related constituents, hemolymph osmolality and Cl⁻ in particular, increased with decreased DO.

Oxyhemocyanin constituted 65.46–65.84% of total hemolymph proteins under the various DO levels examined; both hemolymph oxyhemocyanin and proteins showed notable elevations 24 h after exposure to hypoxic conditions, and reached the highest and constant level by 48 h after exposure. The compensatory responses of prawns to reduced O₂ were manifested by increased O₂ uptake through augmentation of hemocyanin, which results in enhancement of oxygen binding capacity of the hemolymph. In the same period, a significant surge of the respiratory products, PCO₂ and HCO₃⁻, was also demonstrated 6 h after exposure to hypoxic conditions which resulted in hemolymph alkalosis. These processes likely resulted in an increase in water influx and consequent declines in hemolymph osmolality and ion composition. Furthermore, hyperventilation and respiratory alkalosis, indicated by increased oxyhemocyanin and pH, respectively, were found to be predominant responses of M. rosenbergii to hypoxic stress.     

Energy balance of Litopenaeus vannamei postlarvae fed on animal or vegetable protein based compounded feeds

L. vannamei postlarvae are normally raised with a protein dense diet (50% protein) rich in fishmeal. Part of the protein is utilized for energy purpose instead of protein synthesis. Based on a previous energy partitioning study, the effects of two isoenergetic compounded feed treatments – animal protein (AP) and vegetable protein and carbohydrates (VPC) – upon growth efficiency and energy budget of shrimp postlarvae and early juveniles were determined. Recovered energy (RE) or production (P) after 50 days trial was similar (2 J day^− 1) in both treatments, from PL₁₄ to PL₁₉. However, early juveniles discriminated between animal protein (116 J day^− 1) and vegetable protein and carbohydrates (88 J day^− 1). The difference in respiration indicated a higher heat increment with AP compared to VPC. At maintenance level, energy used was lower with AP than VPC treatment. Postlarvae and early juveniles employed protein as a main energy substrate (O:N < 20). Differences in the efficiencies observed in the calculated energy budget were attributed to the presence of carbohydrates in diet and not to the protein source. The advantage of incorporating vegetable protein source in the diet of harvesting shrimp may eventually contribute towards a reduction of fishmeal costs and waste products as well as to achieve sustainable shrimp farming.     

Effect of dietary vitamin C supplementation on the oxygen consumption, ammonia-N excretion and Na/K ATPase of Macrobrachium nipponense exposed to ambient ammonia

The 96-h LC₅₀ of ammonia-N and the effects of dietary vitamin C on oxygen consumption, ammonia-N excretion and Na⁺/K⁺ ATPase activity of Macrobrachium nipponense exposed to ambient ammonia were investigated. The results showed that the 96-h LC₅₀ of ammonia-N was 36.6 mg l⁻¹ for the freshwater prawn, M. nipponense, at pH 8.0. When prawns were exposed to high ambient ammonia-N concentrations, the oxygen consumption rate increased and ammonia excretion decreased. Dietary vitamin C supplementation led to higher oxygen consumption and lower ammonia excretion. Na⁺/K⁺ ATPase activity increased with increased ambient ammonia-N exposure in the range of 0–18.3 mg l⁻¹, and then was reduced at ambient ammonia-N 36.6 mg l⁻¹. Na⁺/K⁺ ATPase activities of prawns fed a vitamin C-supplemented diet were significantly lower than those of prawns fed a diet which was not supplemented with vitamin C.     

Air bubbling results in carbon loss during microalgal cultivation in bicarbonate-enriched media: experimental data and process modeling

Turbulence is a major variable controlling growth of microalgae. When turbulence is provided by means of air bubbling, mass transfer phenomena take place between the culture medium and the air bubbles. A mathematical model has been developed to evaluate the effects of air bubbling in bicarbonate-enriched media for microalgae cultivation, and in cultures of the halotolerant alga Dunaliella viridis. Simulations have been made using a four order Runger–Kutta method combined with Marquardt's algorithm to establish the parameters of the system. The model permits: (i) simulation of the experimental behavior of organic (cellular) and inorganic (CO₂ + H₂CO₃ + HCO₃⁻ + CO₃²⁻) carbon in the cultures; (ii) performance of a sensitive analysis of the effects of varied operative conditions and (iii) development of strategies for optimizing the use of culture media and subsidiary energy. Simulation of the model demonstrated that in air-bubbled HCO₃⁻ enriched media there is a loss of CO₂ from the medium to the air bubbles, which escapes from the culture vessel, and no transference of CO₂ from the air bubbles to the water occurred. Validation of the model has been carried out by comparison of experimental and predicted values under different conditions.     

Effect of hyperoxygenation and low water flow on the primary stress response and susceptibility of Atlantic salmon Salmo salar L. to experimental challenge with IPN virus

Intensive salmon smolt production normally includes reduced water flow and hyperoxygenation (added oxygen) of remaining water. There is little information on how different water quality parameters influence the fish health and the susceptibility to infectious diseases. The current experiment was carried out to evaluate if the combination of hyperoxygenation and reduced water flow (hyperoxic) can act as a chronic stressor to salmon in freshwater (FW) in such a way that it increases the susceptibility to IPN virus (IPNV) following seawater transfer. In FW, after 22 days of hyperoxic exposure plasma ion, TBARS and cortisol were measured. The cortisol levels were significantly (p = 0.011) higher in the hyperoxic group compared to controls maintained under normal oxygen saturation and water flow (normoxic), indicating chronic stress. Hyperoxygenation in FW caused decreased plasma [Cl⁻] compared to the normoxic group (p = 0.037), while [K⁺] tended to be higher in the hyperoxic group (p = 0.088). No significant differences were observed in plasma [Na⁺], total osmolality, TBARS or hematocrit, but there was a tendency towards a lower hct in the hyperoxic compared to the normoxic group. In SW the mortality was higher in the hyperoxic group challenged with IPNV (34%) compared to the normoxic group challenged with IPNV (20%) (p = 0.02), and no mortality was observed in the PBS injected fish. The challenged fish showed an overall increase in plasma cortisol day 8, 10, 12 and 14 post-challenge (p = 0.015, p = 0.000, p = 0.046 and p = 0.022 respectively). After SW transfer and challenge, plasma [K⁺] was elevated in both challenged groups, but no consistent trends were found for plasma [Cl⁻], [Na⁺] or total osmolality during the SW phase. There were no significant differences in the gene expression level of IFN 1, Mx and IL 1β prior to challenge, suggesting that the basic expression level of these genes were not affected by hyperoxygenation. IPNV was detected in kidney and pylorus, by immunohistochemistry, cell culture, and RT-PCR in head kidney. This experiment indicates that chronic stress induced by a combination of low water flow and hyperoxygenation increases the susceptibility to IPNV challenge.     

Hemolymph Oxyhemocyanin and Protein Levels and Acid-Base Balance in the Tiger Shrimp Penaeus monodon Exposed to Copper Sulfate

Abstract.— Subadult tiger shrimp Penaeus monodon (30.15 ± 2.19 g) were exposed individually in sea water of 25%o to 0 (control), 1.11, 4.76, 10.06 and 19.09 mg/L copper for 24 h. Hemolymph pH, PO₂ (partial oxygen pressure), PCO₂ (partial carbon dioxide pressure), HCO₃, oxyhemocyanin and protein levels were determined. Hemolymph PO₂ increased, whereas hemolymph pH, PCO₂, HC0₃-, oxyhemocyanin. protein and the ratio of oxyhemocyanidprotein levels decreased with increasing concentrations of ambient copper in the range of 1.11 to 19.09 ma/L. It is concluded that P. monodon following 24-h exposure to ambient copper as low as 4.76 mg/L shows a reduction of oxyhemocyanin, protein, and the ratio of oxyhemocyanin to protein, and develops metabolic acidosis in the hemolymph.     

Hemagglutinating activity and electrophoretic pattern of hemolymph serum proteins of Penaeus monodon and Macrobrachium rosenbergii to white spot syndrome virus injections

The giant fresh water prawn Macrobrachium rosenbergii is known to be highly tolerant to white spot syndrome virus (WSSV) infections when compared to the widely cultured marine tiger shrimp Penaeus monodon. At present, the exact mechanism of tolerance by M. rosenbergii to WSSV is not known. In this study, we attempt to study the effect of WSSV injections on the hemagglutination activity of the hemolymph serum of both P. monodon and M. rosenbergii and look for changes if any, on their hemolymph serum protein electrophoretic patterns. Our results show that M. rosenbergii had significantly (p < 0.05) higher hemagglutinating activity against mouse erythrocytes when compared to P. monodon. As the infection progressed to 48 h there was a further increase (p < 0.05) in the hemagglutination activity in M. rosenbergii, while it decreased in P. monodon. 12% SDS-PAGE analysis of the hemolymph serum of M. rosenbergii infected with WSSV did not show any new protein bands, whereas few bands with decreased intensity was observed in moribund P. monodon where the hemagglutinating activity was also observed to be decreased. The results indicate that hemolymph hemagglutinin levels are modulated in crustaceans as a response to viral infections.     

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.     

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.     

Reactive nitrogen and phosphorus removal from aquaculture wastewater effluents using polymer hydrogels

We have developed poly(allyl amine hydrochloride) (PAA · HCl) polymer hydrogels, that efficiently remove nitrate (NO₃⁻), nitrite (NO₂⁻), and orthophosphate (PO₄³⁻) nutrient anions from the aquaculture wastewater. The hydrogels were prepared by chemically crosslinking linear PAA · HCl chains with epichlorohydrin (EPI). The anion binding capacity of the pH sensitive polymer gels was measured in standard solutions and studied as a function of gel synthesis parameters. Equilibrium NO₃–N, NO₂–N, and PO₄–P loading of 15, 1.6, and 17 mg/g of dry gel, respectively, were calculated from the measurement of decrease in anion concentration in aqueous solutions using UV–vis spectrophotometry. Batch experiments showed that nutrient concentrations in aquaculture wastewater effluents decreased with regard to PO₄–P by 98+%, NO₃–N by 50+% and NO₂–N by 85+% within 3 h of reaction. The regeneration of the hydrogels was demonstrated by the release of bound nutrient anions upon washing the gels with a 1 N NaOH solution. These results have demonstrated that the hydrogels are appropriate materials for treating aquaculture wastewater effluents, and reducing the nutrient anion concentrations to levels, less than 10 mg/l NO₃–N, 0.08 mg/l NO₂–N, and 0.3 mg/l PO₄–P, suitable for discharge into natural surface waters.     

Effects of elevated atmospheric CO2 on growth, photosynthesis and nitrogen metabolism in the economic brown seaweed, Hizikia fusiforme (Sargassaceae, Phaeophyta)

Hizikia fusiforme (Harv.) Okamura (brown seaweed) was cultured using aeration with two CO₂ conditions: outdoor air (actual atmospheric CO₂ concentration, averaging 360 μl l^− 1) and CO₂-enriched air (averaging 700 μl l^− 1), to investigate the possible adjustments of elevated atmospheric CO₂ to the growth, photosynthesis and nitrogen metabolism in this mariculture species. Aeration with CO₂-enriched air reduced the pH in the culture medium in comparison with aeration with air. The mean relative growth rate was enhanced when H. fusiforme was grown at high CO₂ with respect to normal CO₂. There was little change in the rate of light-saturated photosynthesis, dark respiratory rate and apparent photosynthetic efficiency, measured in natural seawater, between thalli grown in high and normal CO₂ contents. However, both the mean nitrate uptake rate and the activity of nitrate reductase at light period were increased following culture at high CO₂, indicating an enhanced nitrogen assimilation of H. fusiforme thalli with the CO₂ enrichment in culture. It was proposed that the intensive cultivation of H. fusiforme would remove nutrients more efficiently with the future elevation of CO₂ levels in seawater, which could be a possible solution to the problem of ongoing coastal eutrophication.     

Pharmacokinetics and the active metabolite of enrofloxacin in Chinese mitten-handed crab (Eriocheir sinensis)

The pharmacokinetics and active metabolite of enrofloxacin were estimated after single intramuscular administration (10.0 or 20.0 mg/kg body weight) to the Chinese mitten-handed crab (Eriocheir sinensis) in fresh water at 25.0 ± 1.0 °C. Levels of enrofloxacin and its metabolite ciprofloxacin in the main tissues (hemolymph, hepatopancreas, muscle, ovary and spermary) were simultaneously detected by HPLC. Enrofloxacin concentration–time profiles for the hemolymph in both tests were described by a two-compartment open model with first-order absorption. Distribution half-time (T_1/2), elimination half-time (T_1/2β), body clearance (CL/F), mean residence time (MRT_0–∞), area under the concentration–time curve from 0 to ∞ h (AUC_0–∞) and apparent volume of distribution (Vd/F), which derived from the pharmacokinetic model, were 0.427 h, 21.3 h, 0.133 l/h/kg, 60.0 h, 96.9 μg/ml/h and 4.08 l/kg, respectively, at a dose of 10.0 mg/kg body weight, and 0.216 h, 12.3 h, 0.189 l/h/kg, 85.8 h, 187 μg/ml/h and 3.35 l/kg, respectively, at a dose of 20.0 mg/kg body weight. Similarities were found between the hemolymph concentration–time curves of the two tests; for example, instant absorption process followed by the distribution phrase, and a second absorption peak at 6 h post-treatment. After intramuscular administration of 10.0 mg/kg body weight, absorption of enrofloxacin was observed in the main edible tissues (hepatopancreas, muscle, ovary and spermary), and the drug residue was the highest in the hepatopancreas, where the ‘drug sink’ phenomenon occurred. Comparative pharmacokinetics showed fast absorption, broad distribution and fast elimination of enrofloxacin in E. sinensis after intramuscular dosing. Regarding ciprofloxacin, the main active metabolite of enrofloxacin, though relatively low levels were detected in all the main tissues of the crab, its kinetics in the hemolymph in the two tests were not described by a one- or two-compartment open model.     

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.     

Design and operation of nitrifying trickling filters in recirculating aquaculture: A review

This review deals with the main mechanisms and parameters affecting design and performance of trickling filters in aquaculture. Relationships between nitrification rates and easily accessible process parameters, like bulk phase concentration of TAN, O₂, organic matter (COD), nitrite, temperature, HCO₃⁻, pH and the hydraulic loading of the trickling filter, are discussed in relation to the design and operation of such filters. Trickling filter design procedures are presented and one of them, a model describing the nitrification performance of trickling filters by plug-flow characteristics, is discussed in greater detail. Finally, practical aspects in relation to filter design and operation are presented.     

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.     

Age, growth, and reproduction of silky sharks, Carcharhinus falciformis, in northeastern Taiwan waters

To examine the age, growth and reproduction of silky sharks, Carcharhinus falciformis, in the waters off northeastern Taiwan, 469 specimens (213 females and 256 males) were collected from August 2000 to January 2002 at the Nanfanao fish market, northeastern Taiwan. The relationship between body weight (W) and total length (TL) for both sexes combined was expressed as: W = 2.92 × 10⁻⁶ TL^3.15 (n = 469, p < 0.01). The relationship between TL and vertebral centrum radius (R) for both sexes combined data was estimated as: TL = 25.979 + 18.197R (n = 250, p < 0.01). Growth bands (including translucent and opaque zones) in precaudal vertebrae formed once a year between December and January and were counted up to 11 and 14 for females and males, respectively. The von Bertalanffy growth function (VBGF) was used to model the observed length at age data. The sexes combined VBGF predicted an asymptotic length (L_∞) = 332.0 cm TL, growth coefficient (k) = 0.0838 year⁻¹, age at zero length (t₀) = −2.761 year (n = 250, p < 0.01). Size at 50% maturity for males was estimated to be 212.5 cm based on the logistic curve, which corresponded to 9.3 years. Females matured at 210–220 cm, which correspond to 9.2–10.2 years. The length at birth was estimated to be 63.5–75.5 cm TL. The number of embryos per litter was 8–10 and sex ratio of embryos was 1:1.     

Effects of starvation and recovery on the survival, growth and RNA/DNA ratio in loliginid squid paralarvae

The ability of Loligo opalescens paralarvae to resist and recover from starvation was examined by measuring their survival, growth rate and RNA/DNA ratios during starvation and refeeding. Paralarvae were fed Artemia sp. nauplii, zooplankton and mysid shrimp. Fourteen days after hatching they were separated into five feeding treatments: a control treatment (food was always available) and treatments starved for 2, 3, 4 and 5 days, and then refed. Each day, 5–7 paralarvae from each treatment were anesthetized to measure mantle length and wet weight (WW), and then RNA and DNA were extracted using an ethidium bromide fluorometric technique. Paralarvae did not survive 4 and 5 days of starvation, showing that at 15 days of age and at 16 °C the limit to recovery was 3 days of starvation. Paralarvae starved for 2 and 3 days showed compensatory growth that mitigated the effects of starvation, in that at the end of the experiment (10 days), they attained mean final body weights similar to the control treatment. Differences in the RNA/DNA ratios between control and starved paralarvae were detected within 2 days of food deprivation. For paralarvae starved 2 and 3 days, it took 1 day after refeeding to attain RNA/DNA ratios not significantly different from the control treatment. Additionally, RNA/DNA ratios were highest during the day (0800, 1200, 1600 h) and lowest at night (0000, 0400 h), suggesting daytime feeding activity. Growth rates ranged from − 14% to 21% WW day^− 1 and the resulting equation between RNA/DNA ratio and growth rate (GR) of paralarvae was GR = 1.74 RNA/DNA − 11.79 (R² = 0.70). After starvation, there was a reduction in growth variability in all starved treatments, while growth variability remained high in the control treatment. Findings from the present study indicate that nucleic acids are a valid indicator of nutritional condition and growth in squid paralarvae.     

Spawning of ocean pout (Macrozoarces americanus L.): evidence in favour of internal fertilization of eggs

Sperm physiology, in vivo artificial insemination and spawning of the ocean pout (Macrozoarces americanus L.), a marine bottom fish, were studied. Milt was collected from the reproductive tract of mature males by suction using a catheter. The uncontaminated milt, having a very low sperm concentration, contains highly motile spermatozoa and sperm motility was retained in vitro at 4 °C for at least 24 h in both seminal plasma and ovarian slime collected from the oviduct of pre-spawning females. Instead of activating sperm, dilution in sea water instantly immobilized the spermatozoa of ocean pout. Osmolarity and pH of ocean pout seminal plasma were in the ranges 365–406 mOsM and 7.2–7.5, respectively. A study of the ionic composition of ocean pout seminal plasma demonstrated the presence of various ions including Na⁺, K⁺, Ca²⁺, Mg²⁺, and Cl⁻, with a remarkably lower K⁺ concentration compared to that from other fish species. Since injections of milt containing motile sperm into the ovaries of pre-spawning females, which spawned in the absence of males, yielded fertilized ocean pout eggs, it is concluded that the ocean pout exhibits internal fertilization. The larvae hatched after 3 months of egg incubation in ambient sea water (9–10 °C). With proper timing of in vivo artificial insemination of mature females, fertilized ocean pout eggs can be obtained from fish reared in captivity.     

Dopamine induces transient modulation of the physiological responses of whiteleg shrimp,Litopenaeus vannamei

Levels of glucose, lactate, Na⁺, K⁺, Cl⁻, protein, and oxyhemocyanin in the hemolymph and its osmolality were measured when the whiteleg shrimp (Litopenaeus vanammei; 20 ± 1.5 g), were individually injected with saline or dopamine at 10^− 8, 10^− 7, or 10^− 6 mol shrimp^− 1. Results showed that elevations of hemolymph glucose and lactate occurred at between 2 and 4 h, increases in hemolymph osmolality, Cl⁻, Na⁺, and total protein occurred at 2 h, and a reduction in hemolymph oxyhemocyanin occurred at 4 h after the dopamine injection. All physiological parameters except K⁺ had returned to the control values 8–16 h after receiving the dopamine. The injection of dopamine also significantly decreased the oxyhemocyanin/protein ratio of L. vannamei at 2 h as a result of the elevation of hemolymph protein. These results suggest that stress-inducing dopamine causes a transient period of modulation of energy metabolism, osmoregulation, and a respiratory response in L. vannamei in adapting to an environmental stress.     

Substitution of Chaetoceros muelleri by Spirulina platensis meal in diets for Litopenaeus schmitti larvae

The nutritional response of Litopenaeus schmitti larvae to substitution of Chaetoceros muelleri by Spirulina platensis meal (SPM) was evaluated. The substitution levels (S) were 0%, 25%, 50%, 75% and 100%, dry weight basis. Final larval length (FL) ranged from 1.98 to 3.16 mm for the different substitution levels. There was a significant relationship between S and FL, described by the following quadratic equation: FL = 2.853 + 0.01598S − 0.000233S². The substitution level (S) yielding maximum FL was 34.2%. Development index (DI) values ranged from 2.84 to 3.93 and were dependent on substitution level. The corresponding equation was DI = 3.799 + 0.00945S − 0.000189S² (P < 0.01). Maximum DI was obtained at 25.0% substitution. Survival was high (82–87%) and no significant differences were found between treatments. Protein digestibility of either microalgae was high, with 92% for SPM and 94% for C. muelleri, with no significant differences between them. The results in this study indicate that an adequate balance of nutrients in relation to the requirements of the species is critical. To simultaneously improve FL and DI, a 30% substitution of C. muelleri by SPM is suggested. This is equivalent to feeding 0.15 mg larvae^− 1 day^− 1 dry weight basis of a 70% C. muelleri/30% SPM diet, representing 0.078 mg protein larvae^− 1 day^− 1, 0.026 mg lipids larvae^− 1 day^− 1 and 2.732 J larvae^− 1 day^− 1.