Growth performance and biochemical composition of the oysters Crassostrea sikamea,Crassostrea angulata and their hybrids in southern China

The Kumamoto oyster (Crassostrea sikamea) and the Portuguese oyster (C. angulata) are important aquaculture species which naturally coexist along the southern coast of China. To understand the potential feasibility of hybridization between the two species, we conducted two‐by‐two factorial cross‐experiments in Beihai (Guangxi province), and also compared the survival and growth of the hybrids to that of the two parental progenies during the grow‐out period from July 2014 to July 2015. Genetic analysis confirmed that the hybrid spats were true hybrids. Additionally, the biochemical composition of the 1‐year‐old oyster progenies was determined. In July 2015, the mean shell height of the hybrids was 42.98 ± 6.29 mm, which was higher than that of the Kumamoto oyster progeny. The cumulative survival rate of the hybrids was 26.37 ± 1.32%, which was higher than that of the progeny of the Portuguese oyster. Mean lipid content of the hybrids was 13.65 ± 1.63% of dry weight, which showed obvious heterosis compared to those of the two parental progenies. Observation of gonads revealed that all hybrids were completely fertile. Furthermore, relative expression of the lipid homeostasis genes, SREBP (sterol regulatory element‐binding proteins), PPARα (peroxisome proliferator‐activated receptor) and INSIG (insulin‐induced gene) were found to vary between parental progenies and the hybrids, thus providing a possible reason for difference in the lipid contents of these experimental groups. Overall, the hybrids were viable, rich in lipid and completely fertile and thus could serve as a promising aquaculture stock for oyster breeding in southern China.     

Phenotypic trait of Crassostrea hongkongensis♀× C. angulata♂ hybrids in southern China

The Hong Kong oyster (Crassostrea hongkongensis) and the Portuguese oyster (Crassostrea angulata) are important aquaculture species in southern China. To understand the potential feasibility of hybridization between the two species, two‐by‐two factorial cross‐experiments were conducted in Shenzhen (Guangdong province). An asymmetry in fertilization was revealed: C. hongkongensis eggs could be fertilized by C. angulata sperm, but the reciprocal cross was not possible. The fertilization and hatching success in the C. hongkongensis females × C. angulata males (HA) cross was lower than the two intraspecific crosses. Moreover, hybrid larvae had a slower growth rate than those from parental progenies due to genome incompatibility. However, the size of the spat and adult growth were greater than those of the two parental progeny, with obvious heterosis during the grow‐out stage. The survival advantage of hybrids was examined at all times. The surprising finding was that all hybrids were completely fertile, meaning they could produce normal and functional gametes. Genetic analysis by molecular markers was applied to confirm that the HA spat were true hybrids. Our results revealed that the interspecific hybridization between C. hongkongensis and C. angulata is completely successful in one direction. Furthermore, hybrids are viable, fast growing and completely fertile, which provides a promising method for the genetic improvement of oysters as a new aquaculture stock in southern China.     

Tetraploid induction in tropical oysters,Crassostrea belcheri (Sowerby) and Crassostrea iredalei (Faustino)

Tetraploid induction has been conducted on temperate oysters but not on tropical oysters. In this study, different heat shocks (32, 35 and 38°C) and cold shocks (1, 4 and 7°C) were used to induce tetraploidy in two tropical oyster species, Crassostrea belcheri and Crassostrea iredalei, through meiosis I inhibition. Temperature shocks were applied on the newly fertilized eggs at 8–10 min post fertilization and terminated when second polar bodies began to form in the control eggs. The ploidy of the larvae and spat was determined via direct chromosome count. The percentage of larval survival until Day 20 was low (between 0.4% and 42.9%) for both temperature shocks and oyster species. No surviving larva was recorded for induction at 1, 4 and 38°C. Tetraploid spat was only recorded in C. iredalei but the percentage is low through heat shock induction of 32 and 35°C. This study shows that the tetraploid induction success rate was slightly higher in C. iredalei compared to C. belcheri. No surviving tetraploid spat were recorded for both oyster species through the cold shock method. This study shows that heat shock can be used to inhibit meiosis for the production of tetraploids but more experiments need to be conducted to determine the optimum temperature when dealing with tropical oysters.     

Hybridization between Crassostrea hongkongensis and Crassostrea ariakensis at Different Salinities

To evaluate the survival and growth performance of hybrid larvae and juveniles, we conducted hybridization experiments between Crassostrea hongkongensis and Crassostrea ariakensis at three salinities (15, 23, and 30 ppt). Hybrid fertilization occurred in one direction (C. hongkongensis ♀ and C. ariakensis ♂), and the fertilization rates did not differ significantly among the three trial salinities. Molecular genetics analysis revealed that hybrid progenies were true hybrids. Hybrid juveniles survived better than C. hongkongensis juveniles at a salinity of 30 ppt. Hybrid larvae and juveniles showed intermediate growth between C. hongkongensis and C. ariakensis progenies. The growth of hybrid juveniles was significantly greater than that of C. hongkongensis juveniles at a salinity of 30 ppt. Overall, C. hongkongensis has a high economic value but favorable hatchery and farm of this species is restricted to lower salinity sites. Because the hybrids were shown to have greater growth and survival characteristics at high salinity, hybrids may be a more desirable candidate to be cultured in those areas of China with higher salinities, such as the Bohai Sea of China where the salinity is usually 30 ppt.     

The effect of temperature on physiological energetics of a fast‐growing selective strain and a hatchery population of the Pacific oyster (Crassostrea gigas)

Growth is an important aquaculture trait, which has been one of the most important traits for genetic improvement in aquatic species. Numerous fast‐growing domesticated aquaculture organisms are being cultured, but the physiological processes underlying growth remain largely unexplored. The Pacific oyster (Crassostrea gigas) is one of the most widely farmed bivalves. In order to improve the productivity traits, a fast‐growing selective strain of C. gigas was established. In this study, we compared physiological energetics of the fast‐growing selective strain and an unselected hatchery population of C. gigas under different conditions of temperature. Components of energy gain (clearance rates, ingestion rates, absorption efficiency and absorption rates), energy loss (metabolic rates) and resulting energy budget (scope for growth) were analysed at four temperatures (15, 20, 25 and 30°C). The results indicated that selective breeding oysters had a higher energy gain compared to unselected oysters, but the selected strain and hatchery population of oysters did not significantly differ in metabolic rates at all tested temperatures (p > 0.05). The enhanced energy gain resulted in almost two times higher scope for growth in selective breeding oysters compared with unselected oysters. Physiological characteristics of C. gigas were greatly affected by temperature and showed a fluctuation with the increase in temperatures. The selective breeding oysters outperformed those of hatchery population in growth performance by an increased energy gain along with a similar energy loss. This study provided insight into physiological energetics accounting for growth improvements in fast‐growing selective strain of C. gigas.     

Cytogenetic analysis of hybrids and hybrid triploids between the river puffer,Takifugu obscurus,and the tiger puffer,Takifugu rubripes

Cytogenetic analyses of the river puffer, Takifugu obscurus, the tiger puffer, Takifugu rubripes and hybrids produced between female T. obscurus and male T. rubripes and their hybrid triploids (produced by cold shock treatment at 4°C) were performed. T. obscurus had 2n = 44 chromosomes and 1.84 ± 0.019 pg DNA/cell, T. rubripes had 2n = 44 and 2.64 ± 0.015, the hybrids had 2n = 44 and 2.15 ± 0.010 and the hybrid triploids had 3n = 66 and 3.22 ± 0.010. The erythrocyte values of the hybrids were more similar to those for T. obscurus, whereas the hepatocyte, midgut and proximal tubule kidney cell values of the hybrids fell down between those for the parental species (p < .05). The erythrocyte, proximal tubule, hepatocyte and midgut epithelial cell sizes for the hybrid triploids were 1.5‐fold larger than those for the hybrids (p < .05). The thickness of retina and each layer in the hybrid triploids were 1.1‐fold larger than those of the hybrids (p < .05) and did not differ significantly among T. obscurus, T. rubripes and the hybrids (p > .05); however, the hybrid triploids had fewer cell nucleus outer layers than the hybrids (p < .05). Gonad development in the hybrids and hybrid triploids was less matured than in T. obscurus and T. rubripes. The metaphase nucleolus organizer regions (NORs) and the gill cells of T. obscurus, T. rubripes and the hybrids contained two satellite telocentrics, whereas the hybrid triploids contained three satellite telocentrics.     

Different responses between orange variant and cultured population of the Pacific oyster Crassostrea gigas at early life stage to temperature‐salinity combinations

Although breeding of rare shell colour variants has drawn widespread attention from shellfish breeders, the potential disadvantages of their adaptive capacity have been ignored in practice. To explore the difference in adaptive capacity between orange shell variant (OSO) and commercially cultured population (CPO) of the Pacific oyster Crassostrea gigas at early life stage, the development to D‐larvae and larval survival and growth (just 23 and 30°C for larval experiment) of them were compared under different temperature (16, 23 and 30°C) and salinity (17, 25 and 33 psu) combinations. In this study, at 23°C and 25 psu, for both OSO and CPO there was no difference in fertilization rates and survival (p > .05) (mean percentages of D‐larvae after fertilized 40 hr ≥ 95.00%; mean larval survival rates on day 10 > 80.00%). However, the percentage of D‐larvae of CPO at 40 hr was significantly (p < .05) higher than OSO at temperatures of 16 and 30°C and 25–33 psu and 17 psu at 23°C. Similarly, CPO has a better larval survival on day 10 and growth than OSO at salinities of 17 and 33 psu at 23°C. Overall, our results indicate that OSO can have an equally good performance like CPO at early life stage under optimal condition (23°C; 25 psu), but the potential disadvantages in adaptive capacity will be shown at suboptimal conditions. These findings can guide future hatchery breeding of OSO, and suggest the potential disadvantages in adaptive capacity in rare colour variants need more attention in further breeding.     

Effects of temperature,salinity and body size on the physiological responses of the Iwagaki oyster Crassostrea nippona

The physiological responses of the juvenile Crassostrea nippona in terms of filtration, oxygen consumption and ammonia excretion to changes in temperature (16–32°C), salinity (15–35 psu) and body size (small, medium and large) were investigated. In this study, the values of filtration rate (FR), oxygen consumption rate (OCR) and ammonia excretion rate (AER) increased with temperature rising from 16°C to 24°C, reaching the highest values at 24°C and 28°C; with any further increase in temperature above this limit, these values decrease drastically (p < .05). The highest Q₁₀ coefficients were 2.75 for large, 3.54 for medium at 16–20 and 3.47 for small size at 20–24°C respectively. Moreover, the responses of FR and OCR were found to be influenced significantly by salinity, tending to increase concomitantly with salinity up to 25–30 psu, though the values of these parameters were diminished dramatically (p < .05) above this level, showing a reverse pattern from that observed in AER, which firstly decreased to the lowest level at 25 and 30 psu, and then severely (p < .05) increased to the highest level at 35 psu. In addition, the low O:N ratios of all sizes of C. nippona at 16°C and 30–35 psu were indicative of a protein‐dominated catabolism, whereas the O:N ratios of large size at 20–32°C and all sizes at 20–30 psu, indicating that the metabolic energy from protein diminished and lipid and carbohydrate were used as the energy substrates. Physiological rates of C. nippona were well correlated with its size. The average values of mass exponents (b‐values) estimated in the present study were 0.657 for OCR and 0.776 for AER at different temperatures, and 0.647 for OCR and 0.767 for AER at varying salinities, signifying that physiological process of C. nippona becomes relatively slower with increasing body size regardless of temperature or salinity. Finally, our results confirm that the optimal temperature and salinity for juvenile C. nippona lie within 24–28°C and 25–30 psu respectively. The results of physiological traits in response to environmental factors of this species are informative in site selection for the cultivation.     

Enhancing lipid nutritional quality of oysters by hybridization between Crassostrea gigas and C. angulata

In the present study, hybrid and inbred oysters were produced by factorial hybridization between two closely related species, the Pacific oyster Crassostrea gigas and the Portuguese oyster C. angulata. Tissues samples were used for the analysis of total lipid content (TLC) and fatty acid composition (FAC). In general, FAC but not TLC in hybrid oysters was significantly different from inbred oysters. Hybrid oysters have significantly lower ∑SFA composition but higher ∑PUFA composition than inbred oysters, with heterosis for ∑SFA, ∑MUFA and ∑PUFA of ?6.03%, ?6.53% and 14.31% respectively. Moreover, hybrid oysters have significantly higher EPA + DHA content and n3/n6 PUFA ratio than inbred oysters, indicating higher lipid nutritional quality than inbred oysters. Among them, the hybrid ♀ C. gigas × ♂ C. angulata appears to be the most nutritious. These results indicated that the lipid nutritional quality of oysters can be improved by hybridization.     

Characterization of triploid hybrid groupers from interspecies hybridization (Epinephelus coioides ♀ × Epinephelus lanceolatus ♂)

Interspecies hybridization is widely used in aquaculture as a beneficial strategy. Diploid and triploid hybrids have been detected from the interspecies hybridization of Epinephelus coioides ♀ × Epinephelus lanceolatus ♂. This is the first report of triploidization through hybridization in grouper. Confirmation has been obtained through flow cytometry, karyotyping and erythrocyte nuclei measurement. The chromosome numbers of E. coioides, E. lanceolatus, diploid hybrid grouper are 48 and triploid hybrid grouper are 72. Measurements of erythrocyte nuclei indicate that triploid fish have a larger nuclear surface than the diploid groupers, and the average ratio of triploid to diploid surface area is 1.59. During the first 1.5 years, triploid hybrid groupers grow faster than diploid hybrid groupers or either parent species. The average growth rate of triploid hybrids is 1.61 times greater than that of diploid hybrids at 6 months of age and 1.43 times greater at 18 months of age. The triploid hybrid groupers are inferior in gonadal development, with no primary‐growth‐stage oocytes appearing in the gonads at 18 months of age. Morphological studies indicate that triploid hybrid groupers have distinctive differences in snout length, eye diameter, body trunk shape, and tail shape development compared with diploid hybrid groupers. Triploid hybrid groupers have an advantage in growth ability, and artificial breeding of triploid groupers might be of great potential use in the grouper aquaculture industry.     

Triploidization in rohu × mrigal hybrid and comparison of growth performance of triploid hybrid

The present study was aimed at the identification of treatment optima to induce triploidy in ‘Labeo rohita (rohu) × Cirrhinus cirrhosus (mrigal)’ hybrid using heat shock treatment. The eggs were exposed at four different temperature regimes viz., 38, 39, 40 and 41°C for 1–3 min, applied 3–5 min after fertilization. After 4 min of fertilization, heat shock treatments for 1 and 1.5 min durations were found the best inducing triploidy up to 100% and 96% respectively. Survival rates upto yolk sac absorption were found to be 73% and 71% in rohu and mrigal, 68% and 67% in the reciprocal diploid hybrids and 61% and 60% in the reciprocal triploid hybrids (RTH). Triploidy was confirmed by chromosome counting that revealed the diploid chromosome number of rohu and mrigal at 2n = 50 and in their triploid hybrid chromosome number was found to be 3n = 75. Growth rate of the RTH showed a significant difference (P < 0.05) from the single species and the diploid hybrids. Triploids also showed higher survival rate over the diploids.     

The induction of Hsp70 synthesis by non‐lethal heat shock confers thermotolerance and resistance to lethal ammonia stress in the common carp,Cyprinus carpio (Linn)

Exposure to heat‐shock protein (Hsp) stimulating factors induces Hsp accumulation and confers tolerance to lethal ammonia stress on the common carp Cyprinus carpio. This study investigated whether a non‐lethal heat shock bestowed similar protective effects against ammonia and induced thermotolerance, both thought to be rendered by increased amounts of Hsps. The 30‐min lethal temperature (30 min LHT) and 1‐h lethal ammonia concentration (1 h LCT) for this species occurred at 41°C and 14.2 mg/L NH₃ respectively. Heating juvenile carp (5 cm) from 28°C to 32, 34 and 38°C, with a subsequent 8‐h recovery period augmented tolerance to lethal heat and ammonia perturbation by two to threefold as compared with animals held at 28°C. Protection occurred in conjunction with Hsp70 accumulation in gills, substantiating the role of this Hsp in enhancing the stress tolerance of common carp.     

Embryonic and larval development of a hybrid between kelp grouper Epinephelus moara ♀ × giant grouper E. lanceolatus ♂ using cryopreserved sperm

Hybridization was used to take advantage of desirable traits in offspring. In the present study, we applied the cryopreserved Epinephelus lanceolatus sperm into interspecific hybridization with E. moara. Successful hybridization between these two species was achieved and cultured in 23–24°C seawater (34‰). There was no difference in survival rate between hybrid (E. moara ♀ × E. lanceolatus ♂) and non‐hybrid (E. moara ♀ × E. moara ♂) at 24 hrs post hatch (HPH), but less hybrid (14.35 ± 8.02%) hatched than non‐hybrid (93.60 ± 1.65%), which might be due to irregularity cell cleavage and skeletal deformities from the formation of embryonic body to the later embryonic development. Similar phenomenon was found in hybrid embryos from fertilization with fresh sperm, indicating that species variations between parents, rather than cryopreserved sperm, resulted in deformities in embryos. Mean hatch time of the hybrid was 1 hr faster than that of E. moara. The hybrid was 1.95 ± 0.06 mm in total length when newly hatched and reached 38.00 mm at 58 days post hatch (DPH), which showed faster growth than E. moara (recorded in the previous study). Considering its faster growth, E. moara × E. lanceolatus hybrid was a potential breeding production in aquaculture. Over 212,000,000 larvae have been produced and launched in the market since 2015. The results of this study also shed some lights on further comparative studies in grouper hybrid performance.     

Changes in volatile compounds of living Pacific oyster Crassostrea gigas during air-exposed storage

Fisheries Science - The changes in volatile compounds of living oysters Crassostrea gigas with shells under air-exposed storage for 7&nbsp;days at 5 and 20&nbsp;°C were investigated...     

Hot water immersion lowers survival,shell growth rate and lysosomal membrane stability of oysters Crassostrea virginica (Gmelin)

This study compares the effect of two anti‐fouling treatments, hot water immersion (15 s at 60°C) and air drying (72 h) on the physiological status of the Eastern oyster Crassostrea virginica. The negative impact of hot water immersion was greater than that of air drying, but varied depending on the initial size of the oysters (40 vs. 60 mm shell height) and the time of the year (June vs. August). Groups treated with hot water exhibited a higher proportion of haemocytes with destabilized lysosomal membranes (HDLM; 47.5 ± 3.1%) than those exposed to air drying (37.5 ± 2.9%). This suggests that the oyster immunocompetency may be lowered by hot water immersion. Overall, the large oysters had lower HDLM values (32.9 ± 3.5%) than the small individuals in June (45.7 ± 2.8%) but similar values in August (46.6 ± 3.5%). Small oysters subjected to hot water immersion in June exhibited a 50% reduction in shell growth and a 50% mortality rate after one month. Our results indicate that air drying is more suitable than hot water immersion as an anti‐fouling treatment for <45 mm oysters.     

Growth performance and disease resistance against Vibrio vulnificus infection of novel hybrid grouper (Epinephelus lanceolatus × Epinephelus fuscoguttatus)

Groupers are economically important for aquaculture in Thailand. A novel hybrid grouper (Epinephelus lanceolatus × Epinephelus fuscoguttatus) has been successful cross‐bred; therefore, the present work aimed to assess the hybrid's traits. The growth performance, strength and tolerance to a pathogenic bacterial infection of this hybrid were compared with its parent species, tiger grouper and giant grouper. The results of all measured growth parameters indicated that the hybrid strain grew fastest followed by giant and tiger grouper respectively. The expressions of the growth‐related genes, insulin‐like growth factor (IGF) I and II, were also analysed in fish muscle and liver which are the main target organs in fish growth regulation. Among tested species, similar expression patterns of IGF‐I and IGF‐II were detected in both organs. The levels of these genes in liver and muscle of hybrid and giant grouper were higher than those of tiger grouper comparable with the growth manner. After challenge with Vibrio vulnificus, the immunological parameters, clearance time of Vibrio in haemolymph and survival was measured to verify the fish immunity. Leucocyte number, lysozyme activity and the ability to eliminate the pathogen were very high in hybrid and giant grouper while these parameters were lower in tiger grouper. Correspondingly, the mortality rate of tiger grouper was higher than others and % survival at the end of observation time (15 days post challenge) was lowest in infected tiger grouper. Altogether, the results suggested that the hybrid grouper has desirable traits that will improve cultured grouper.     

Insights on the association between somatic aneuploidy and ostreid herpesvirus 1 detection in the oysters Crassostrea gigas,C. angulata and their F1 hybrids

Cytogenetic abnormalities associated with viral infections, including from viruses of the Herpesvirales order, have been reported in vertebrate species. Ostreid herpesvirus 1 (OsHV‐1) has been detected worldwide during mortality outbreaks of the Pacific oyster Crassostrea gigas. On the other hand, a high proportion of aneuploid cells in somatic tissues have been observed in C. gigas. In this study, we analysed the putative association between aneuploidy levels and the detection of OsHV‐1 in gills of C. gigas, the Portuguese oyster C. angulata and their F1 hybrids cultured in Ria Formosa (Portugal). OsHV‐1 was detected by PCR in 5.4% of the total of oysters analysed (n = 111) namely in 11.1%, 8.0% and 1.7% of C. gigas, C. angulata and F1 hybrid respectively. Sequencing analysis of a viral fragment amplified with the C2/C6 primer pair revealed a high similarity with the OsHV‐1 reference type. Moreover, in situ hybridization confirmed the presence of OsHV‐1 in gill tissue. Oysters where OsHV‐1 was detected had a significantly higher mean percentage of aneuploid cells (25%) than the ones where the virus was not detected (18%). However, the overall low percentage of positive samples contrasted with the high mean percentage of aneuploidy observed, with 50% of the oysters analysed showing a percentage of aneuploid cells between 20% and 30%. We hypothesize that somatic aneuploidy may adversely affect oysters making them more prone to OsHV‐1 infection, but the virus is unlikely to be the cause of somatic aneuploidy.     

Production of inbred larvae through self‐fertilization using oocytes and cryopreserved sperm from the same individuals after sex reversal in eastern oyster Crassostrea virginica

The eastern oyster Crassostrea virginica can change sex which makes self‐fertilization possible if sperm can be cryopreserved. In this study, small (~1 year old) and large (~2–3 years old) oysters were biopsied for sperm collection. Survival of the biopsied oysters after 1 year was 50% for small oysters and 17% for large oysters. Oocytes were collected from sex‐reversed females, and self‐fertilized with cryopreserved sperm. Of the 24 cryopreserved samples, 14 individuals had ≤1% fertility when crossed with oocytes from unrelated females, indicating that the cryopreserved sperm had reduced fertility. The other 10 individuals had a fertility of 39 ± 25% when crossed with oocytes from unrelated females (non‐selfing), but showed a significantly lower success of self‐fertilization (12 ± 16%) (P = 0.008), while aliquots of the same oocytes had a fertilization of 83 ± 11% when crossing with fresh sperm. Larvae were produced at day 3 in the self‐fertilized families (12–94% of the fertilized oocytes), and survived to eyed‐larvae stage at days 11–14. Genotyping with 9 microsatellite markers confirmed that the larvae resulted from self‐fertilization in four families. This study demonstrated the feasibility of creating self‐fertilized inbred lines of oysters by use of non‐lethal sperm collection and cryopreservation.     

Effects of rhythmic temperature change on the growth,body composition and energy budget of hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀)

Effects of rhythmic temperature change on the growth, body composition and energy budget of hybrid grouper (Epinephelus lanceolatus ♂ × E. fuscoguttatus ♀) were investigated. Nine groups of fish received repeating cycles of low‐temperature manipulation (22°C) for 1, 2 and 4 days followed by recovery temperature (28°C) for (3, 7 and 11 days), (6, 14 and 22 days) and (12, 28 and 44 days) respectively, designated as L1R3, L1R7, L1R11, L2R6, L2R14, L2R22, L4R12, L4R28 and L4R44 respectively. In the control group (C), fish were reared at 28°C throughout the whole experiment. After 96‐day feeding trail, the final weight, relative weight gain rate, specific growth rate, food conversion efficiency and apparent digestibility coefficient of fish in L4R28 were significantly higher than those of control (p < .05). The crude protein contents of fish in L2R14, L2R22, L4R12, L4R28 and L4R44 were significantly higher compared to the control (p < .05). According to energy budget, fish in L4R28 exhibited significantly higher proportion of food energy assimilated into growth and lower proportion consumed for excretion than the control fish (p < .05). These results indicated the occurrence of complete or over growth compensation in hybrid grouper, and the use of repeated cycles of low temperature (22°C) for 4 days followed by recovery temperature (28°C) for 28 days could obtain enhanced growth, that is overcompensation, with improved apparent digestibility coefficient, food conversion efficiency and energetic efficiency into growth. This technique would be more practically applicable in indoor culture for this species.