Combined effects of rearing density and tank colour on the growth and welfare of juvenile white sea bream Diplodus sargus L. in a recirculating water system

Appropriate rearing conditions for successful farming of white sea bream Diplodus sargus L. have not yet been studied in depth, while one of the major problems is the species increased aggressiveness. Given the known effect of density and background colour on fish growth, welfare and social behaviour, the present study aimed to investigate whether the two factors combined could favour D. sargus performance. Juveniles (17.37±0.06 g) were reared in white, light blue and black tanks under low (7 fish tank⁻¹ or 1.96 kg m⁻³) and high (28 fish tank⁻¹ or 7.79 kg m⁻³) density for 87 days. Water quality was not affected by experimental treatments. The best overall performance (growth, food utilization, body protein content, liver fatty acids) was achieved under low density and white or light blue tanks. Increased incidence of social interactions was indicated in fish under high density or when reared in light blue and black tanks. The present results clearly suggested that the use of black tanks should be avoided. On the other hand, stocking D. sargus at a density of up to 7.79 kg m⁻³ may be feasible because productivity was greater even if growth was slower. In this case, however, the use of white tanks is highly recommended.     

Dietary supplementation of mannan oligosaccharide on white sea bream (Diplodus sargus L.) larvae: effects on development,gut morphology and salinity tolerance

The influence of dietary mannan oligosaccharide (MOS) on the development, gut integrity and quality (in respect of stamina and survivability) of white sea bream Diplodus sargus L. larvae was investigated. White sea bream larvae were held under appropriate rearing conditions and fed Artemia, enriched by A1 DHA Selco^? with the addition or absence of MOS (Bio‐Mos®). The results indicated that larval growth performance and survivability were not affected by the MOS supplementation. Light microscopy revealed that MOS supplementation significantly improved the intestinal morphology by increasing the villi surface area by over 12%. Transmission electron microscopy revealed that MOS supplementation increased the microvilli length by 26% compared with the control. Salinity challenge experiments showed that MOS significantly increased larval stamina and survival in both 0 and 60 mg L^?1 salinity water by 13% and 22.9% respectively. These improvements in the larval quality at the early stages of fish development are important for the efficiency of intensive hatchery production.     

Effects of rearing density on growth, brain neurotransmitters and liver fatty acid composition of juvenile white sea bream Diplodus sargus L.

White sea bream Diplodus sargus L. is a fish species that develops dominance hierarchies and aggressive behaviour against its conspecifics. On the other hand, stocking density is known to affect social interactions, while an appropriate density for D. sargus efficient farming remains to be elucidated. For this purpose, juvenile D. sargus (14.3 g) were reared in 88.4 L tanks, for 10 weeks, under six rearing densities (10, 15, 20, 25, 30 and 35 fish tank^?1). Water quality was not affected by rearing density. Best growth, in terms of final weight, specific growth rate, weight variation and food utilization, was achieved at 10 fish tank^?1. At this treatment the lowest body water content, hepatosomatic and splenosomatic indices, higher body and liver lipid content were also observed. Brain neurotransmitter results indicated increased dopaminergic activity in populations held at higher rearing densities, while serotonergic activity was not affected. Moreover, no significant differences were detected for liver fatty acids. Results of the present study, compared with previously reported data, suggest that, within the range tested, the lowest realistic rearing density for juvenile white sea bream would be 10 fish tank^?1. Higher rearing densities led to decreased growth, which seemed to be related to changes in intraspecific competition and social behaviour.     

Improved performance of gilthead sea bream, Sparus aurata, larvae after ozone disinfection of the eggs

Ozone (O₃) dissolved in seawater (ODS) was evaluated, as an egg disinfectant, on the spawn of captive gilthead sea bream, Sparus aurata, brood stock. Four contact times (CT) were tested (0.6, 1.2, 2.4 and 4.8 mg min L⁻¹) where CT was calculated by multiplying the dissolved O₃ concentration (0.3 mg L⁻¹) by different exposure periods (2, 4, 8, 16 min). There was also a disinfected seawater treatment that contained no O₃ or derived compounds (CT 0) and an untreated seawater control. All ODS treatments reduced egg surface bacterial counts to zero, which was significantly (P<0.05) lower than the CT 0 and the control groups (194 and 1320 plate⁻¹ respectively). Nevertheless, the hatching rate was high in the control and the CT treatments 0, 0.6 and 1.2 (88.7%, 87.3%, 89.5% and 83.7% respectively) while eggs exposed to a CT 2.4 and 4.8 hatched poorly (36.5% and 20.4% respectively), which was likely due, at least in part, to larvae unable to break the egg chorion successfully. Swim‐bladder inflation was significantly higher in the ODS groups (>97%) compared with the control and CT 0 treatments (ca. 70%). The results suggest that a 2‐min exposure of eggs to 0.3 mg O₃ L⁻¹ of ODS (CT 0.6) would improve current protocols in marine larviculture.     

Spawning season,spawning grounds,and egg production of red sea bream in Hiuchi-nada,Seto Inland Sea

The spawning season and grounds of red sea bream in Hiuchi-nada, the central part of the Seto Inland Sea, were described using a new method based on monoclonal antibodies for identifying Pagrus major eggs, and the daily egg production (standardized by the incubation time and survival rate) was estimated. At the peak of spawning (May), the ranges of sea temperature, salinity, and chlorophyll a where red sea bream eggs occurred were 14.8–17.4°C, 32.0–33.0, 0.5–4.4 μg/L, respectively. The main spawning grounds of the red sea bream were confirmed as being the areas near the Geiyo Islands, Misaki Peninsula, Saijyo, Niihama. The spatial spread of red sea bream eggs increased with the egg developmental stage. The ranges of daily egg production in 2005 and 2006 were 0.3–19.3 and 0.2–6.7 × 10⁹ eggs/day, respectively. In Hiuchi-nada, aquaculture farms are located close to the spawning grounds, and the potential spawning population from the aquaculture farms equaled or exceeded that of the estimated spawning population obtained by the egg production method. Red sea bream eggs in Hiuchi-nada might be produced by both wild and aquaculture-based spawning populations.     

Efficacy of formalin and povidone–iodine disinfection techniques on the eggs of three marine finfish species

Surface disinfection trials were performed on eggs from three marine finfish species: California yellowtail (CYT; Seriola lalandi), white seabass (WSB; Atractoscion nobilis) and California halibut (HA; Paralichthys californicus). All three species were spawned from captive populations maintained at the Hubbs‐SeaWorld Research Institute (HSWRI). Five disinfection treatments were used for each species; Treatment 1 included 100 mg L^?1 of formalin (F100) for 60 min (current HSWRI treatment), Treatment 2 included 1000 mg L^?1 of formalin for 15 min (F1000), Treatment 3 included povidone–iodine of 50 mg L^?1 for 15 min (PI50), Treatment 4 included povidone‐iodine of 100 mg L^?1 for 10 min (PI100) and Treatment 5 involved a control with no chemical treatment (CONT). For each treatment, the per cent egg hatching rate, per cent survival to first feeding and notochord length at the time of hatching to the nearest 0.1 mm were recorded. Bacteria were also cultured from eggs after treatment to determine the effectiveness of each treatment in reducing the bacterial counts (CFU mL^?1). Treatments F100, F1000 and CONT yielded the highest hatch rates for each species (70–80%), whereas treatments PI50 and PI100 yielded the lowest hatch rates (0–2%). There were no significant differences in survival to first feeding or notochord length, which suggests that the disinfection treatments did not have a negative effect on the yolk sac larvae. The PI50 and PI100 treatments had the lowest bacterial colony counts, showing almost zero bacterial growth. The highest bacterial growth occurred in the F100, F1000 and CONT treatments. Based on the results from this study, the F100 treatment provided the best balance of disinfection and larval health for CYT, WSB and HA.     

Improving cold storage of subitaneous eggs of the copepod Acartia tonsa Dana from the Gulf of Mexico (Florida – USA)

Developing methods to store copepod eggs is necessary to increase the availability of copepods as a live food for the aquaculture industry and aquarium trade, and also to allow the exchange of copepods between researchers. The present study, evaluated the effect of glucose and two antibiotics (kanamycin sulphate and oxytetracycline HCl) on extending the shelf life of cold-stored subitaneous Acartia tonsa eggs. Also, egg development effects on the survival of the eggs were tested. Glucose did not have any significant effects on the survival of the eggs. However, the addition of antibiotics to the storage vials resulted in an increase of the survival of the eggs. In the best case, the shelf life of the eggs was almost doubled. After 7 days, the kanamycin+glucose treatment led to a hatching success of 86±1% of the hatchable eggs, while the untreated eggs presented a hatching success of 47±6%. However, long exposure to high concentrations of antibiotics was lethal to the copepod eggs. After more than 30 days of exposure to 100 mg L⁻¹ of oxytetracycline, the survival of the eggs was lower than in the untreated samples. After 45 days, oxytetracycline-treated eggs (100 mg L⁻¹) presented a hatching success of 4–5% while the non-stored eggs still had a hatching success of 9%, and the eggs treated with a lower concentration of antibiotics (10 mg L⁻¹) showed a hatching success up to 21–23%. The size of the nauplii in all trials tended to decrease as the period of cold storage at 1°C increased. We consider that the use of antibiotics at the right dosage to be a means to increase the storage capacity of the Gulf of Mexico strain of A. tonsa eggs, which do not show any capacity to be stored for long periods of time, compared with some other strains. In addition eggs that were between 5 and 7 h old survived longer when stored in the cold than eggs, which were freshly spawned or closer to hatching.     

Effects of commonly used disinfectants on bacterial load,hatchability and survival of Bluefin Sea bream (Sparidentex hasta) eggs

The efficacy of four chemical reagents, iodophor, formalin, hydrogen peroxide and bronopol as fish egg surface disinfectants were evaluated in bluefin sea bream (Sparidentex hasta). Fertilized eggs were counted and subjected to a static bath dip treatment in different concentrations of the above chemicals for 4 min before being incubated at 20 ± 0.5°C for 40 h. Treatment efficacy of the different disinfectants was evaluated by assessing the bactericidal activity, egg hatch percentage and survival of larvae up to 3 days post hatch. Results showed that iodophor at medium concentrations (75 and 100 ppm) was the best of all tested disinfectants in bacterial killing ability (12% reduction in the bacterial counts), egg hatching per cent (99.8% and 99.6% respectively) and larval survival up to 3 days post hatch (50.8% and 54.8% respectively). Formalin was the second best disinfectant at levels of 100 and 150 ppm. Hydrogen peroxide gave good results compared with the control while, bronopol showed discouraging results. In conclusion, iodophor appeared to be suitable for bluefin sea bream eggs disinfection with a 4 min exposure to 75–100 ppm when applied 14–16 h after egg fertilization.     

Diluted rotifer, Brachionus plicatilis (Muller), extract as culture medium stimulates hatching of red sea bream, Pagrus major (Temminck & Schlegel)

Hatching rates and hatching times (50% hatch) of red sea bream, Pagrus major (Temminck & Schlegel), eggs in culture media that were prepared to dilute rotifer Brachionus plicatilis Muller extract at 4000-, 6000-. 8000- and 10 000-fold with artificial sea water (σ₁₅, = 24.0) were investigated. Although differences in hatching rate between the culture media and control artificial sea water were negligible, hatching time was significantly shorter in media at 4000- and 6000-fold dilutions than in media at 8000- and 10 000-fold dilutions and in the control artificial sea water, when the eggs immediately post spawning were incubated. The hatching rates of eggs at three developmental stages, immediately post-spawning, gastrula stage and heart-beat initiation stage, incubated respectively in the medium at 6000-fold dilution and the control, did not differ either, yet the hatching times at each egg stage in the medium were significantly shorter than that in the control. These results suggest that hatching enzyme secretion from the hatching glands is promoted to perceive chemical stimuli in the water-soluble fraction of rotifer extract via the embryo's chemoreceptors, from the heart-beat initiation stage to immediately before hatching, thus shortening the hatching time.     

Prothrombin time, activated partial thromboplastin time and fibrinogen values in Mediterranean marine teleosts

Coagulation profiles for marine fish were determined with the standard global screening assays, the prothrombin time (PT) and activated partial thromboplastin time (APTT) using homologous tissue thromboplastin and phospholipid extracts to investigate species-specificity of brain thromboplastins, and to establish a base-line coagulation profile for marine fish of economical importance for Mediterranean aquaculture (sea bream, Sparus aurata, sea bass, Dicentrarchus labrax, red porgy, Pagrus pagrus, common dentex, Dentex dentex). Sea bass and sea bream displayed the shortest PT (9.0–15.0 s) and APTT (22.1–35.8 s) in comparison to the other species (PT: 24.8–37.2 s, APTT: 39.6–66.4 s). Red porgy displayed the lowest fibrinogen value (1.2–2.3 g l^–1), while no differences were found among the other species (2.2–2.8 g l^–1). It is concluded that the interaction of the tissue factor and plasma clotting factor is relatively species specific and that the use of homologous brain extracts is preferable for haemostasis study in marine fish.     

Effects of adult stocking density on egg production and viability in cultures of the calanoid copepod Acartia tonsa (Dana)

The effect of stocking density of the calanoid copepod Acartia tonsa was evaluated in a 96 h rearing experiment. Possible density‐dependent egg production and egg viability were analysed at stocking densities of 100, 200, 300, 400 and 600 adults L⁻¹. Temperature, oxygen saturation and algal concentration were kept optimal. A non‐density‐dependent mortality rate of 15–19% day⁻¹ was documented. A non‐significant density‐dependent egg production was observed between 100 and 600 adults L⁻¹. The average egg production was 22.5±8.8 egg female⁻¹ day⁻¹ in all densities. The average egg hatching success was 84.7±4.8% and was never observed below 76.1%, with no significant differences across the stocking densities. Conclusively, as a practical recommendation for the aquaculture industry, copepod cultures with densities ranging from 100 to 600 adults L⁻¹ and presumably even more dense cultures are possible with the studied species obtaining a steady egg production and still high egg viability.     

真鲷与黑鲷杂交与多倍体育种系列研究-Ⅰ

真鲷属于鲷科,真鲷属,分布于我国四大海域。黑鲷属于鲷科,鲷属,我国沿海均有分布。真鲷与黑鲷杂交属于属间远缘杂交。本研究利用春季繁殖的真鲷种群与春季繁殖的黑鲷种群进行交配。用HCG与LHRH-A_2催产,网箱内自然受精。正交以真鲷为♀,黑鲷为♂;反交以黑鲷为♀,真鲷为♂。两种杂交组合的雌雄性比均为1:1.5。正交组合的受精率为73％,反交组合为67％。正交组合的孵化率为85％,反交组合为87％。该研究证明海水鱼属间远缘杂交可以成功,并为海水鱼杂交育种推广应用奠定了基础。     

Efficacy of formalin,iodine and sodium chloride in improvement of egg hatching rate and fry survival prior to the onset of exogenous feeding in yellow perch

Egg disinfection is considered the most important routine work in hatcheries to avoid fungal and/or bacterial infection of fish eggs. The aim of this study was to determine the effectiveness of three disinfectants: formalin, iodine and sodium chloride on the hatching success of yellow perch eggs. The disinfectants were tested in triplicate at different concentrations for 15 and 30 min bath treatments. Two experiments were conducted; formalin at five concentrations (25, 50, 100, 150 and 200 mg L^?1) and 25 mg L^?1 iodine were tested in the first experiment. The second experiment involved formalin at three concentrations (250, 500 and 1000 mg L^?1), iodine at three concentrations (50, 100 and 250 mg L^?1) and sodium chloride at three concentrations (500, 1000 and 3000 mg L^?1) were used. Iodine and sodium chloride‐treated eggs hatched earlier than formalin‐treated eggs. The highest mean percentage of eyed stage, hatching rate and survival to first feeding fry was observed at 200 mg L^?1 formalin for 30 min, 50 mg L^?1 iodine for 15 min and 500 mg L^?1 sodium chloride for 30 min. High concentrations of formalin (1000 mg L^?1), iodine (250 mg L^?1) and sodium chloride (1000 and 3000 mg L^?1) showed toxicity to yellow perch eggs, resulting in low hatching rate and survival to first feeding fry. We recommended formalin at a concentration of 150–200 mg L^?1 for 30 min as an effective, easily available and low‐cost disinfectant for routine use to improve yellow perch hatchability.     

Incubation of European Squid (Loligo vulgaris Lamarck, 1798) eggs at different salinities

Loligo vulgaris is a commercially important squid throughout the Mediterranean region and is a candidate species in biomedical and aquaculture research. Some loligo species (L. opalescens, L. forbesi, Sepiteuthis lessoniana) have now been cultured through some successive generations in closed, recirculating seawater systems. The effects of salinity on hatching European Squid (L. vulgaris Lamarck, 1798) eggs were investigated during November 2004. The egg capsules were incubated directly in salinity of 32, 34, 36, 38, 40, 42 and 37 g L^?1 (control group) at 19.8°C (SD 1.2°C), and a photoperiodicity of 12 h light:12 h dark for 16–23 days before hatching. In all treatments, the eggs were developed and hatched normally after 16–22 days at 32 g L^?1, 17–22 days at 34, 18–21 days at 42 g L^?1, 18–22 days at 36 and 40 g L^?1, 19–22 days at 37 g L^?1 and 19–23 h at 38 g L^?1. In the experiments, the highest hatching rate and hatching success (HS) of the eggs were obtained at 38 g L^?1 (hatching rate: 100% (SD 0%) and HS: 96.7% (SD 3.5%)) and the lowest hatching rate at 42 g L^?1 (hatching rate: 3% (SD 6%) and HS: 0%). Dorsal mantle lengths (DML) of new hatchlings ranged from 2.08 to 2.80 mm. The present study showed that salinity affects the hatching rate and HS of eggs and first hatching time and DML of paralarvae in L. vulgaris. The squid eggs at stage 11 (I) can tolerate 5 g L^?1 reduction and 3 g L^?1 increase in salinity.     

Comparison of the swimming performance of farmed and wild gilthead sea bream, Sparus aurata

Farmed gilthead sea bream, Sparus aurata, frequently escape from the sea cages and interact with wild populations. The impact of these interactions on the wild populations will depend, in part, on differences in performance of the bream. This study compared the swimming performance of the wild and farmed fish in a current channel. The absolute critical swimming speed (U_crit) increased with increasing size while the relative U_crit decreased. Even at the same length there were noticeable performance differences between the individuals. The wild sea bream have significantly higher (P<0.05) absolute U_crit performance (0.86±0.01 m s⁻¹) than the farmed fish (0.79±0.01 m s⁻¹) and significantly higher (P<0.05) relative U_crit performance (4.52±0.05 BL s⁻¹) than the farmed fish (4.21±0.05 BL s⁻¹). The present study suggests that cultured sea bream may not have the ability to compete with wild sea bream in native seawaters.     

Effects of four egg desticking procedures on hatching rate and further survival and growth of larvae in the tench (Tinca tinca L.)

Four desticking procedures for tench eggs (A: tannic acid solution (1 g L⁻¹) for 15 s; B: alcalase enzyme solution (8 mL L⁻¹) for 60 s; C: alcalase enzyme solution (15 mL L⁻¹) for 120 s; D: Woynarovich and Woynarovich (1980) solution for 58 min followed by tannic acid solution (1 g L⁻¹) for 15 s) were tested to obtain data about influence on embryo survival to hatching stage and further survival and growth of the larvae. In the tannic acid and Woynarovich and Woynarovich (1980) treatment (A and D) few eggs stuck together and some were adhered to the incubator walls, whereas in the alcalase treatments (B and C) eggs neither stuck together nor adhered to the incubator walls. Percentages of hatched larvae did not show significant differences (mean values ranged between 47.4% in treatment A to 37.0% in treatment C). Larvae deformities observed were <0.5% in all cases. There were no significant differences among survival and growth rates of the larvae from different egg desticking origin, reaching, after 30 days, mean survival values around 90% and total length and weight of 12.5 mm and 19 mg respectively.     

Preliminary Results on Rearing of Sparus Aurata × Pagrus Pagrus Hybrids. Performance Comparison With the Parental Species

Hybridization between gilthead sea bream, Sparus aurata, and red porgy, Pagrus pagrus, was undertaken and viable hybrids were produced by an artificial hormonal-induced cross. The hybrids, from the cross of female red porgy and male gilthead sea bream (Pp × Sa), survived through the yolk-sac larval stage. The hybrids produced from the cross of female gilthead sea bream and male red porgy (Sa × Pp) presented higher mortality in the larval stage (98.4%) compared to gilthead sea bream (80.0%) and red porgy (92.5%). Afterwards, the Sa × Pp hybrid was compared to the offspring of its parental species under the same rearing conditions (tanks of 500 l capacity, natural light, ambient temperature, self-feeding with commercial feed) in a 40–week experiment. In this comparative study, the Sa × Pp hybrid had intermediate mortality rate (6%), compared to gilthead sea bream (0%) and red porgy (21%). The growth profile of this hybrid resembled the slow growing phases of its parents (red porgy in warm period and gilthead sea bream in cold period). The feed conversion ratio between fish forms was not significantly different (1.54–1.84 in the Sa × Pp hybrid, 1.53–1.56 in the maternal species and 1.33–1.40 in the paternal one). The body weight variation in the Sa × Pp hybrid was quite high (31.7–37.9%), compared to that in gilthead sea bream (19.7–24.7%) and red porgy (18.0–31.3%). With respect to body coloration, the Sa × Pp hybrid was distinguishable and classified into groups of red porgy-like (55.1%), gilthead sea bream-like (40.6%), and intermediate forms (4.3%). The preliminary results of this study proved that the produced Sa × Pp hybrid had too low a performance for aquaculture use, but the interesting consequent hereditary potential could be a useful tool in fish hybridization practice.     

Surface disinfection of eggs from marine fish: evaluation of four chemicals

For surface disinfection of marine fish eggs Buffodine (1.06% free iodine), glutaraldehyde, chloramine-T and sodium hypochlorite (5% free chlorine) were tested using plaice (Pleuronectes platessa L.) as the main species for evaluation. Glutaraldehyde was the most promising candidate of the four chemicals tested. Good bactericidal effects without any documented negative effects on eggs and larvae were obtained at concentrations of 400–600 mg l^–1 and contact times of 5–10 min. Replicated experiments under identical disinfection conditions revealed a clear correlation between the degree of successful surface disinfection and the initial bacterial load of the egg batch.     

Chemical treatment of lobster eggs against epibiotic bacteria

Eggs of the European lobster, Homarus gammarus (L.), were exposed to malachite green (5, 10, 15 mg 1^–1: 10 min), glutaraldehyde (50, 100, 150 mg 1^–1: 3 min) and iodine as Buffodine^TM (50, 100, 150 mg 1^–1: 10 min). The efficiency of the treatments was tested by incubating eggs individually in wells of multiwell dishes with TSB agar for 14 days after exposure. In order to find any effect on viability, batches of 30 eggs from each of three females were incubated artificially in a recirculation system for 19 days and repeatedly exposed to the disinfectants. Iodine as Buffodine (150 mg 1^–1) was the only treatment that resulted in a significant decrease of the bacterial growth on lobster eggs, but the treatment also resulted in inhibited hatching compared with the control group. Thus, our results indicate that treatment with 150mg^–1 iodine as Buffodine could be a strategy for reducing bacterial growth on lobster eggs when massive egg mortality due to bacteria is otherwise unavoidable. The treatment could, however, lead to decreased viability of larvae due to inhibited hatching.     

Biological Performance of Red Porgy (Pagrus pagrus) Larvae under Intensive Rearing Conditions with the use of an Automated Feeding System

Pagrus pagrus is one of the promising species for the industry in the Mediterranean but its rearing is still far from satisfactory. In the present work, the conditions and results of larval rearing with the use of an automatic feeding system are presented. Eight populations were reared for 20 days with the pseudo green water methodology in two successive trials. Larvae were fed enriched rotifers during the entire rearing period, a mixed diet of rotifers and Artemia (Instar I when larvae reached 5 mm in length, replaced by Instar II after larvae reached 5.5 mm) and a formulated diet (after day 16 post hatching). A computerized system for feeding management was used. A total of 388,000 eggs were incubated and after 20 days, 237,973 larvae (TL = 6.96 ± 0.17 mm) were produced with a survival rate of 61.4 ± 6.3%. Individuals grew in terms of wet weight with an exponential rate of 0.167 ± 0.008 daily (R ²= 0.983) with no variations between replicates. Mean individual daily consumption at first feeding (day 4-post hatching), was 0.20 ± 0.06 mg of food (0.03 ± 0.01 dry weight), that is, 180–300 rotifers, while on day 20, consumption increased by 10 (2.081 ± 0.106 mg and 0.276 ± 0.014 in wet and dry weight, respectively). When compared with sea bream, consumption was higher by approximately 3–4-fold. The mean food conversion ratio of the experimental period was 4. In comparison with sea bream larviculture, food consumption of red porgy is higher, and a different feeding strategy is required to satisfy the requirements of the larvae.