Genetic diversity and relatedness estimates for captive barramundi (Lates calcarifer,Bloch) broodstock informs efforts to form a base population for selective breeding

Aquaculture of barramundi or Asian seabass (Lates calcarifer) is growing in both Australia and Southeast Asia and there is substantial interest to improve production efficiency through selective breeding. The establishment of a large and genetically diverse base population is a prerequisite for a sustainable and long‐term productive breeding program. Before selective breeding programs can begin for Australian barramundi it is important to assess the overall genetic diversity of current captive broodstock populations. To address this question, 407 captive barramundi broodstock from eight separate Australian broodstock populations were genotyped using 16 polymorphic microsatellite DNA markers. A Bayesian STRUCTURE analysis indicated that captive Australian broodstock are broadly divided into two genetic stocks. Multivariate analysis between broodstock individuals and pairwise F_ST between broodstock populations also supported the existence of two stocks. Comparisons with data obtained from natural stocks suggested that hatchery individuals were either sourced from the two stocks or represented an admixture between them. Genetic diversity was low within each broodstock population (allelic richness ranged from 2.67 to 3.42 and heterozygosity ranged from 0.453 to 0.537) and relatedness estimates within hatcheries were generally low (average r was equal to 0.141). We recommend sourcing captive individuals according to high levels of neutral genetic diversity and low levels of relatedness for the establishment of a base population. We also make recommendations about including genetically diverse wild individuals.     

Dietary administration of natural immunostimulants on growth performance,haematological, biochemical parameters and disease resistance of Asian Sea bass Lates calcarifer (Bloch, 1790)

This study was aimed to address the promising evaluation of Cissus quadrangularis plant (stem) and lipopolysaccharide (LPS from bacteria) supplemented diets on innate immune response in Lates calcarifer fingerlings against Aeromonas hydrophila infection. Fingerlings were fed supplemented diets containing four different concentrations of C. quadrangularis (0.5, 1.0, 1.5 and 2.0 g kg^?1 feed), LPS (25, 50, 75 and 100 mg kg^?1 feed) and control (normal formulated diet) for 60 days. The fish fingerlings fed supplemented diet displayed significant differences (P < 0.05) in specific growth rate (SGR) and relative percentage survival compared to the control group fed without C. quadrangularis and LPS‐supplemented diet. Fingerlings were injected intraperitoneally with 100 μL lethal dose of A. hydrophila containing 1 × 10⁶ CFU g^?1. Supplementation of C. quadrangularis and LPS diet significantly increased biochemical profile such as protein, lipid and carbohydrate content, haematological parameters of L. calcarifer fingerlings in different experimental periods when compared with the control group. Dietary doses of C. quadrangularis and LPS‐supplemented diet significantly influenced growth performance and increased survival rate in L. calcarifer fingerlings against A. hydrophila infection.     

Effect of dietary chitosan on haematology,innate immunity and disease resistance of Asian seabass Lates calcarifer (Bloch)

This study was performed to evaluate the effect of dietary chitosan on haematology, innate immunity and protection against Vibrio anguillarum in Asian seabass, Lates calcarifer. A basal diet supplemented with 0, 5, 10 and 20 g chitosan kg^?1 diet was fed to the four different groups for 60 days. The haematological (total erythrocyte count, total leucocyte count, total serum protein, albumin, globulin and albumin‐globulin ratio) and innate immune parameters (phagocytic ratio, respiratory burst, serum lysozyme and serum bactericidal activities) were monitored at fortnight interval to assess the effect of chitosan feeding in Asian seabass. All the studied haematological and innate immune parameters were increased significantly (P ≤ 0.05) in chitosan‐fed groups in comparison with control. However, the group fed diet containing 10 g chitosan Kg^?1 feed showed highest haematological and innate immune parameters on 45^th day in comparison with other groups. Moreover, the fish fed the diet containing 10 g chitosan Kg^?1 feed had significantly higher post‐challenge survival (75.56 ± 4.44%) on the 30th day following V. anguillarum challenge. Therefore, this study suggests that chitosan at 10 g kg^?1 diet could be used as prophylactic in Asian seabass culture to enhance the protection against any possible infection by V. anguillarum.     

Dietary influence of Fucoidan supplementation on growth of Lates calcarifer (Bloch)

Sulphated polysaccharides exhibit a range of activities which promote health and growth benefits to aquaculture species. One class of sulphated polysaccharides derived from marine algae is fucoidan, which is suggested to exhibit an affinity to the negative muscle growth inhibitory protein myostatin. Addition of fucoidan to the diet of fish therefore may lead to enhancements in growth through inactivation of the myostatin protein. To examine the growth enhancing potential of this sulphated polysaccharide, fucoidan extracts from Undaria pinnatifida were added to the diet of juvenile barramundi, Lates calcarifer. The fucoidan extract was of 86% purity and included in two treatment diets at 5 g kg^?1 (0.5%) and 10 g kg^?1 (1%) respectively. Fish were fed the diet for 52 days. At the end of the feeding trial, barramundi fed with the 1% fucoidan inclusion diet exhibited enhanced growth with significantly increased lengths, weights and muscle fibre area (hypertrophy) when compared to the 0.5% fucoidan inclusion diet and the non‐fucoidan control diet. This research suggests that addition of fucoidan to the diet of fish may have growth promoting effects.     

Dietary administration of astaxanthin improves feed utilization,growth performance and survival of Asian seabass,Lates calcarifer (Bloch, 1790)

This study was undertaken to assess the impacts of dietary astaxanthin supplementation on growth performance, feed utilization, survival, and serum growth hormone (GH) availability of Asian seabass, Lates calcarifer, with special reference to dose–response relationships and variations during different feeding phases (short‐term, medium‐term and long‐term). Fish were fed the following diets in triplicate for 90 days: the control (CD), AX50 (50 mg astaxanthin/kg diet), AX100 (100 mg astaxanthin/kg diet) and AX150 (150 mg astaxanthin/kg diet). The findings revealed that fish exhibited significant linear increments (p < .05) in specific growth rate (SGR), weight gain, feed utilization efficiency and survival when fed various diets with escalating levels of astaxanthin. Supplementation with dietary astaxanthin significantly augmented (p < .05) GH levels in fish. Significant positive associations (p < .05) were observed between circulating serum GH levels and SGR of fish from all groups following three consecutive feeding phases, denoting a robust cause‐and‐effect relationship. Circulating GH concentrations were considered as a sensitive biomarker of growth performance in Asian seabass. This study illustrated that supplemental astaxanthin could be administered in culture protocols to improve the growth rate and commercial hatchery production of Asian seabass, and possibly other teleost species.     

Combined or individual effects of dietary magnesium and selenium nanoparticles on growth performance,immunity, blood biochemistry and antioxidant status of Asian seabass (Lates calcarifer) reared in freshwater

This study was conducted to investigate effects of and interactions between nanoselenium (NanoSe) and nanomagnesium (NanoMg) on growth, humoral immunity, serum biochemistry and antioxidant capacity of juvenile Asian seabass Lates calcarifer reared in freshwater. Four groups of fish with an average weight of 32.78 ± 2.23 g were fed one of the experimental diets for 6 weeks: (a) control (basal diet); (b) NanoSe (basal diet + 4 mg NanoSe/kg diet); (c) NanoMg (basal diet + 500 mg NanoMg/kg diet); and (d) combination (basal diet + 4 mg NanoSe/kg diet + 500 mg NanoMg/kg diet). Fish fed with NanoSe‐supplemented diets (NanoSe and combination) showed higher weight gain, specific growth rate and feed intake. The combination of NanoSe and NanoMg enhanced the immune response. Also, fish fed on combination diet showed higher serum alanine aminotransferase and aspartate transaminase levels but lower glucose concentration. The activities of liver superoxide dismutase, glutathione peroxidase and catalase were not different among all groups, but liver malondialdehyde level was lower in fish fed diets supplemented with NanoSe and/or NanoMg. It could be concluded that NanoSe improved growth performance; the combination of both NanoSe and NanoMg enhanced humoral immunity; and NanoSe and/or NanoMg improved antioxidant capacity in Asian seabass.     

Development of two cell culture systems from Asian seabass Lates calcarifer (Bloch)

Two new cell culture systems namely epitheloid cells of Lates (LCE) and fibroblastic cells of Lates (LCF) have been developed from fry and fingerling of the economically important brackishwater fish Lates calcarifer. Primary cultures were initiated by explant technique using caudal fin of fingerling and whole body tissue of the fry. The nutritional requirements and the growth pattern in response to different culture environment were similar for the two cell cultures. The culture medium used was Leibovitz‐15 supplemented with 20% fetal bovine serum (FBS) and 1% fish serum. The LCE comprised of epithelioid cells and LCF cells were fibroblastic. With a split ratio of 1:2, the confluency of cells was attained in 8–10 days at an incubation temperature of 28°C. The cells were found to grow well in a wide range of temperature (24–32°C) and stable at 20 and 36°C. The growth rate of LCF and LCE cells increased proportionately with the concentration of FBS from 5% to 20%. A decrease of serum level to 10% after eight subcultures produced no apparent change in cell morphology and growth rate. The viability of cells was found to be 70% when revived after a month of storage in liquid nitrogen (?196°C).     

Vibrio alginolyticus infection in Asian seabass (Lates calcarifer,Bloch) reared in open sea floating cages in India

Asian seabass (Lates calcarifer) has been designated as a candidate species for open sea cage culture in India. A case of vibriosis in Asian seabass reared in open sea floating cages is reported. Haemorrhage and ulcer were observed grossly in the diseased fish. The pathogen, identified as Vibrio alginolyticus based on biochemical and molecular characterization, was isolated from liver, gill, kidney, brain and blood. Histological examination of the diseased fish showed congestion, haemorrhage and necrosis in vital organs. The LD₅₀ studies showed that the organism was virulent to Asian seabass, and the LD₅₀ value was 10^3.2 CFU g^?1 fish.     

Visual thresholds for feeding and optimum light intensity for larval rearing of Asian seabass,Lates calcarifer (Bloch)

This study examined the retinomotor responses and prey ingestion rates of 10‐, 15‐, 20‐ and 30‐day‐old Asian seabass Lates calcarifer under different light intensities from 0 to 1000 lx to determine the visual thresholds. Subsequently, two age groups of seabass larvae were reared under light intensities of 10, 100 and 1000 lx to determine the optimum illumination in hatchery tanks. Retinomotor response was absent in 10‐day‐old larvae, but quite marked in 15‐ and 20‐day‐old seabass at 1 lx and higher. Ingestion of Artemia nauplii by 10‐day‐old larvae was almost zero at <1 lx, increased significantly at 1 lx, and was maximal at 10–100 lx. Artemia ingestion under dim light <1 lx improved with age, and older larvae took more prey in complete darkness due to the presence of rod cells (and also free neuromasts). Larvae from 13 to 26 days group had similar survival and growth at 10–1000 lx, however, from 5 to 10 days group showed similar survival rate with highest weight gain at 100 lx. Therefore, we recommend that hatchery rearing tanks be illuminated such that the larvae in the water are exposed to approximately100 lx.     

Effect of temperature and salinity on sex inversion in Asian Seabass (Lates calcarifer): relationship with plasma sex steroids concentration and aromatase activity of gonad and brain

Asian Seabass which shows precocious sex inversion in cultured environment is a major impediment to maintain a proper broodstocks. Seabass grown commercially in freshwater were transported to the research facility and held in freshwater at 29°C for 4 weeks and acclimatized to the experimental conditions. Fish were daily fed with a commercial pellet (50% protein, 18 MJ kg^?1) to satiety. Blood, brain and gonad collected before and at the end of the experiment were analysed for sex steroids level and aromatase activity. There was an increase in Plasma E₂ levels with temperature in fish at 34°C whereas no significant difference was observed at 24 and 29°C although the highest plasma T level was detected in fish at 34°C which had significantly lower level than at the beginning, except those held at 24°C in freshwater. Plasma 11‐KT was significantly greater in fish at 24°C compared with 29 or 34°C which had clear opposite to the E₂. Aromatase activity in brain was higher at 29°C than at either 24 or 34°C, whereas gonadal aromatase recorded highest at 34°C. These findings concluded that high temperature in culture facility induces sex inversion of Asian Seabass.     

Production of protein hydrolysates from skipjack tuna (Katsuwonus pelamis) viscera as feeding attractants for Asian seabass (Lates calcarifer)

Skipjack tuna viscera hydrolysate (TVH) was produced by enzymatic hydrolysis using alcalase. A study of the effect of time and enzyme concentration was performed and formed the basis for preparing TVH with the desired degree of hydrolysis (DH). An enzyme to substrate ratio of 5 g kg⁻¹ (TVH A1) and 16.4 g kg⁻¹ (TVH A2) at pH 8.0 with a temperature of 50 °C, and hydrolysis time of 60 min were found to be optimal conditions to obtain hydrolysis of 42.9% and 62.9%, respectively. Both TVH A1 and TVH A2 were analysed for chemical and amino acid composition and peptide size. The effectiveness of TVH at different DH as feeding attractants was estimated in a feeding trial at different inclusion levels. Ten experimental diets , including a fish meal‐based reference diet, were fed to Asian seabass for 28 days. Fish fed the diet supplemented with 30 g kg⁻¹ of TVH A2, and the reference diet showed the highest feed intake (P < 0.05). The growth performance of fish fed diets supplemented with TVH was slightly higher than those of fish fed the control diet, but the reference group showed a significantly higher growth rate (P < 0.05). These findings suggest that the supplementation of feed with TVH that has been processed under suitable conditions can improve feed palatability and growth performance when utilizing alternative protein sources in Asian seabass diets.     

Growth,enzymatic glutathione peroxidase activity and biochemical status of juvenile barramundi (Lates calcarifer) fed dietary fermented lupin meal supplemented with organic selenium

To investigate the effects of high level of lupin meal (LM) supplemented with organic selenium (OS) on the growth and blood biochemistry of barramundi (Lates calcarifer), four isocaloric and isonitrogenous diets were prepared, containing either non‐fermented or fermented LM, and either supplemented with 2 mg OS/kg (LM, LM_OS, FLM and FLM_OS), or not. A fishmeal (FM)‐based diet formulated for juvenile barramundi was used as a control diet. Fish (initial mean weight of 5.88 g) were triplicated and fed the test diets for 75 days. The findings demonstrated that growth performance of fish fed with the FLM and FLM_OS diets were similar to fish fed with the FM diet (p > .05). The antioxidant glutathione peroxidase (GPx) activity, and haemoglobin (Hb) of fish fed with the FLM_OS diet were significantly higher than that of FM‐fed fish (p < .05). Plasma alanine aminotransferase (ALT) activity was significantly increased in fish fed with non‐fermented diets (LM and LM_OS) than in those fed with fermented LM diets (FLM and FLM_OS) (p < .05). However, there were no significant differences in ALT activity among LM_OS, FLM, FLM_OS and FM diets. There was an interaction between the LM and OS on plasma CK activity; the CK of fish fed with diets supplemented with OS was higher in non‐fermented LM diets but lower in fermented LM diets (p < .05). This study suggests that fermented LM have an obvious potential to substantially replace 75% FM protein in the diets of barramundi.     

Susceptibility of freshwater rearing Asian seabass (Lates calcarifer) to pathogenic Streptococcus iniae

Asian seabass (Lates calcarifer) has been recognized as an economically important aquaculture species which can be adapted to and cultivated in wide range of salinities. The number of freshwater intensive seabass farms in Thailand is increasing annually. Here, we first describe the susceptibility of Asian seabass, which were cultured in freshwater, to Streptococcus inae (SI) and their pathological changes. Three isolates of putative SI were identified using a combination of standard biochemical assays and species‐specific PCR prior subjected to in vivo challenge. Accumulated mortalities of the fish which received 10⁷ CFU fish^?1 of either SI1J, SI SGSA or SI2J were 90%, 90% and 100% at 7 days‐post infection (dpi), respectively, and mortalities increased sharply between 3 and 5 dpi. Clinical signs such as erratic swimming and opaque eyes were identified from a few infected fish, while most died rapidly without any abnormal signs. Histopathological manifestations were observed in the multiple organs (kidney, liver and brain). Haemorrhage, hyperhemia, cellular degeneration and inflammatory cells infiltration were commonly found within the internal organs. Notably, the formation of numerous encyst‐like lesion aggregated by eosinophilic cells, resembling macrophages, were typically found in the brain of the infected fish. Summarily, this study first revealed that freshwater reared Asian seabass is highly susceptible to SI infection and haemorrhagic septicaemia was a major pathological change that could be found in the infected fish.     

Effect of dietary mannan oligosaccharide on growth,body composition,haematology and biochemical parameters of Asian seabass (Lates calcarifer)

A feeding trial was conducted to study the effect of mannan oligosaccharide (MOS) supplementation on growth, body composition, haematology, biochemical parameters and histology of Asian seabass (Lates calcarifer) fingerlings (8.13 ± 0.06 g average weight). Mannan oligosaccharide was supplemented at five different concentrations viz., 0%, 0.5%, 1%, 1.5% and 2% levels in the diet (40% protein and 9% lipid) of L. calcarifer. The results of the 60 days feeding trial showed that there was a significant (P < 0.05) increase in the final body weight, weight gain (WG), WG%, average daily gain, survival, specific growth rate, daily growth coefficient, hepatosomatic index and viscerosomatic index in the fish fed diet containing 1% MOS. Whole body composition of post‐fed animals showed non‐significant differences (P > 0.05) among the various treatment groups. The analysis of haematological parameters showed that there was no significant (P > 0.05) differences among different treatments but for the haemoglobin content which was significantly (P < 0.05) higher in the fish fed 1% MOS. Biochemical parameters revealed that glucose, urea, cholesterol and triglyceride content showed significant (P < 0.05) difference between control and MOS‐supplemented group. Histological observations of post‐fed animals revealed that MOS supplementation resulted in increased absorptive surface area of the intestine and increased glycogen deposition in liver. The result of this experiment infers that MOS supplementation has got a beneficial effect in the diet of seabass and supplementation at 1% level is optimal for improving the growth.     

Prediction of fillet yield in hatchery populations of Asian sea bass,Lates calcarifer (Bloch, 1790) using body weight and measurements

International Aquatic Research - The purpose of this study was to develop and validate prediction equations for fillet traits in Asian sea bass. Ninety-day old fish (average...     

Dietary butyric acid improved growth,digestive enzyme activities and humoral immune parameters in Barramundi (Lates calcarifer)

A six‐week feeding trial was conducted to determine the effect of butyric acid (BA) inclusion in diet on performance of Barramundi (Lates calcarifer). Fish (12.0 ± 0.2 g) were fed with four experimental diets contained 0.0 (control), 2.5, 5.0 and 10.0 g BA/kg in triplicate. Fish fed on 5 and 10 g BA/kg diets had higher growth and total alkaline protease and lipase activities than the other treatments. Fillet protein content in BA‐supplemented groups was higher than the control. Liver catalase activity was highest in 2.5 g BA/kg group compared with the other groups. The values of liver superoxide dismutase activity and serum total protein concentration in 5 and 10 g BA/kg treatments were higher than those in the control (p < .05). Moreover, the highest and the lowest levels of serum lysozyme activity were noticed in the 10 g BA/kg and the control groups, respectively (p < .05). Also, the highest respiratory burst activity was observed in fish fed 5 g BA/kg feed. Supplementing diet with 2.5 or 5 g BA/kg resulted in higher serum haemolytic activity in fish compared with the other groups. The number of red and white blood cells increased in fish fed BA‐supplemented diets compared with the control (p < .05). In conclusion, the results of this study showed inclusion of BA at 5 g/kg diet in L. calcarifer juveniles improved growth and upgraded the general health condition by enhancing fish antioxidant enzyme activities and haemato‐immunological responses.