Increasing survival and growth in larval leopard coral grouper (Plectropomus leopardus) using intensively cultured Parvocalanus crassirostris nauplii

Leopard coral grouper, Plectropomus leopardus are a heavily exploited, high-value fish commonly found in the Asian live reef food fish trade. In past decades, many attempts at the mass culture of various grouper species have been undertaken; however, their small mouth gape at first feed has resulted in very low survival when using traditional live feeds such as rotifers. The use of wild caught or extensively cultured copepods has yielded potentially promising increases in survival and growth, but overall survival to the juvenile stage remains low, making mass culture currently impractical. The current study sought to build on past developments in grouper culture and recent advancements in copepod culture technology by observing how growth and survival were influenced by the addition of intensively cultured copepods to the early diet of P. leopardus larvae. Six tanks of larvae, three replicates per treatment, were fed either eggs and nauplii of the calanoid copepod Parvocalanus crassirostris, at a starting density of 5 mL⁻¹, and the rotifer Brachionus rotundiformis, at a starting density of 10 mL⁻¹, or were fed only B. rotundiformis, at a density of 15 mL⁻¹, starting on the evening of 2 days post-hatch (dph) and continuing until 9 dph. After this initial period, all larvae were fed the same diet of rotifers, Artemia, and dry feed until the cessation of the trial at 21 dph. Larvae fed P. crassirostris in addition to rotifers had a significantly higher survival, 9.9 versus 0.5%, than those fed only rotifers. Growth was also significantly enhanced in larvae offered copepods. Larvae only fed rotifers were, on average, 1.5 mm shorter at 21 dph than those that had been fed copepods. More rapid development and the earlier onset of flexion were also noted in the larvae that were offered copepods. The use of intensively cultured copepods, in this study, increased survival tenfold over previous studies, with P. leopardus larvae fed wild-caught copepods. The application of intensively cultured copepods to the early diet of P. leopardus, along with future research to evaluate late-stage mortality issues, may facilitate commercial production of this species.     

Beneficial effects of thyme (Thymus vulgaris) extract on oxytetracycline‐induced stress response,immunosuppression, oxidative stress and enzymatic changes in rainbow trout (Oncorhynchus mykiss)

The aim of the present study was to investigate the effects of thyme extract (TE) and oxytetracycline (OTC) on rainbow trout (Oncorhynchus mykiss) stress response, immune parameters, oxidative stress and enzymatic changes. The fish were assigned into eight treatments receiving diets with OTC (0 and 2.5 g/kg) and TE (0, 5, 10 and 20 g/kg) for 2 weeks. Thereafter, serum characteristics and gut oxidative status were evaluated. OTC significantly increased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and cortisol and glucose levels; however, TE significantly suppressed these changes. OTC significantly decreased, but TE significantly increased gut catalase (CAT) and glutathione peroxidase (GPx) activities. OTC significantly decreased gut superoxide dismutase (SOD) and increased gut glutathione‐s‐transferase (GST) activities, malondialdehyde (MDA) content and blood monocyte percentage; TE significantly mitigated these changes. TE significantly increased serum total protein, total Ig, lysozyme and ACH50, and blood WBC; however, OTC significantly decreased serum lysozyme and ACH50 activities and blood WBC and neutrophil percentage. In conclusion, OTC administration causes immunosuppression, which might be related to deterioration of fish health. TE augments trout innate immune and antioxidant status and is capable to counteract OTC‐induced health and immune deterioration.     

Plasma growth hormone levels in fed and fasted rainbow trout (Oncorhynchus mykiss) are decreased following handling stress

Plasma growth hormone concentrations of rainbow trout,Oncorhynchus mykiss, fasted for six weeks, were significantly (p < 0.01) higher than in comparable fed animals; in the fasted fish, the levels fell progressively following acute stress (by displacing the fish within their home aquarium), with significant differences from pre-stressed fish evident between one and thirty-two hours after application of the stressor. Plasma growth hormone concentrations also fell significantly in the fed group, but differences were evident only between two and eight hours after stressor application.Plasma cortisol concentrations in pre- and post-stressed fed and fasted fish were similar. There was a bimodal response to stressor application in both groups, with significantly higher values relative to the pre-stressed sample evident one and eight hours after disturbance, but not after two, four or thirty-two hours. The changes in plasma cortisol levels between the initial (09:00h) sample and the sample taken eight hours later resembles the diet pattern seen in trout given access to self-demand feeders.Plasma glucose concentrations in pre-stressed fed animals were higher than in pre-stressed fasted fish. This relationship was also evident between one and four hours and thirty-two hours after stressor application. The post-stress rise in plasma glucose concentration was evident between one and four hours in the fed group, and between four and eight hours in fasted fish.The diel changes in plasma growth hormone and glucose concentrations could not be attributed to normal circadian patterns, and there was no apparent correlation between changes in plasma growth hormone and cortisol concentrations. There was a significant inverse correlation between plasma glucose and growth hormone concentrations when the total data set were analyzed, but these correlations were not apparent when the treatment groups were analyzed separately.     

Influence of dietary protein level on growth performance,digestibility and activity of immunity‐related enzymes of leopard coral grouper,Plectropomus leopardus (Lacépède, 1802)

An 8‐week feeding trial was conducted to assess the interaction between dietary protein levels and fish growth, digestibility and activity of immunity‐related enzymes of Plectropomus leopardus. Five diets with different protein levels (400 g/kg, 450 g/kg, 500 g/kg, 550 g/kg and 600 g/kg protein) were designed. P. leopardus fed with 500 g/kg, 550 g/kg and 600 g/kg dietary protein, showed higher weight gain rates than fish fed 400 g/kg and 450 g/kg dietary protein. Ingestion rate in fish fed with 500 g/kg dietary protein was significantly higher than those with other diets. P. leopardus fed with 500 g/kg, 550 g/kg and 600 g/kg dietary protein, showed that feed coefficients were significantly lower than those fed with 400 g/kg and 450 g/kg dietary protein. Net protein utilization was significantly lower in fish fed with 400 g/kg diet than those with other diets. Fish fed with 400 g/kg and 450 g/kg dietary protein had an apparent feed digestibility coefficient for dry matter that was significantly lower than that with other diets. Protease activity was highest in fish fed on 500 g/kg dietary protein. Fish fed with 500 g/kg dietary protein, had the highest superoxide dismutase activity. Fish fed with 600 g/kg dietary protein, had the highest alkaline phosphatase activity. Thus, a diet containing 500 g/kg protein is recommended for P. leopardus aquaculture.     

The role of temperature in the prey capture probability of drift‐feeding juvenile brown trout (Salmo trutta)

Abstract – Cold water temperatures are widely supposed to reduce the food intake of stream salmonids. Although cold temperatures have been documented to reduce swimming ability, digestion and gastric evacuation rates, little is known about how temperature influences the ability of fish to capture prey. We examined the effects of water temperature on the prey capture probability of drift‐feeding juvenile brown trout (Salmo trutta) in a laboratory stream. Temperatures ranged between 5.7 °C and 14 °C. We found significant effects of water temperature on prey capture probability and capture manoeuvre time. The mean capture probability dropped from 96% at 14 °C to 53% at 5.7 °C. At 8 °C and higher temperatures, foraging performances did not differ much among treatments. We suggest that reduced swimming ability could be one of the most important mechanisms for the observed pattern of reduced prey capture probability at cold water temperatures, but prey detection limitations and predator avoidance may play a role. Our results will be of use for bioenergetics‐based drift‐foraging models, which to date have not incorporated a temperature‐dependent prey capture function.     

Effects of iodized feed on stress modulation in steelhead trout, Oncorhynchus mykiss (Walbaum)

In the aquaculture industry, the physiological systems of fish can be chronically stressed by various biological, chemical and physical factors. Chronic stress leads to a decrease in the overall health and growth of the fish, making them more prone to diseases. Dietary iodine has been shown to reduce this stress response in chickens and increase disease resistance in dairy cattle, but the mechanisms by which iodine affects stress resistance and immunocompetency is not completely understood and has not been extensively studied in fish. This study investigated the effects of iodized feed as a nutritional supplement in relation to stress modulation in steelhead trout (Oncorhynchus mykiss). We investigated effects on primary, secondary and tertiary stress responses by measuring plasma levels of cortisol, glucose and thyroid hormones as well as haematocrit percentage and average growth rate in steelhead trout. Iodine‐supplemented fish, on average, had lower levels of plasma cortisol and glucose and lower packed cell volumes than fish fed with regular commercial feed (P<0.05). Iodine‐supplemented fish also showed higher levels of thyroid hormones and exhibited better growth over the period of the experiment (P<0.05). It was concluded that, used with other husbandry practices, use of iodine‐supplemented feed could lead to better host defence, growth and survival in fish raised in the aquaculture industry.     

有水与无水保活运输对大菱鲆生理应激及鱼肉品质的影响

为探究大菱鲆分别在有水及无水条件下运输过程中生理生化指标及肌肉品质的变化,为大菱鲆的保活运输提供参考.将大菱鲆进行暂养、降温、充氧包装后进行模拟运输,并在运输不同时间段取样,检测大菱鲆生理生化指标及肌肉指标.结果显示,模拟运输18 h后,有水运输组和无水运输组中的大菱鲆存活率仍为100％.大菱鲆肌肉中水分含量、粗脂肪、...     

The effect of water pH on swimming performance in rainbow trout (Salmo gairdneri,Richardson)

A Brett-type respirometer was used to measure the effect of water pH on swimming performance of rainbow trout (Salmo gairdneri). Variations in water pH between 6 and 9 had no measurable effect on maximum aerobic swimming speed. At water pH 4, 5, and 10, however, the critical velocity was only 55, 67, and 61% respectively of that recorded for fish in water of pH 7. Exposure to acid conditions increased coughing and breathing frequency. Acid exposure resulted in a decrease whereas alkaline exposure resulted in an increase in both whole blood and red blood cell pH. Blood gas and acid-base characteristics showed little change during swimming at 2.0 BL/second, but exhaustive swimming resulted in a marked and immediate drop in blood pH in fish in acid, alkaline and neutral water. The blood acid-base status was restored to resting levels after exercise in neutral and alkaline water, but the acidosis was maintained following exercise in acid water. Fatigue occurred earlier and blood lactate levels increased to a higher level in fish swum to exhaustion in acid or alkaline water, compared with fish in neutral water.     

Dietary nucleotide supplements influence the growth,haemato‐immunological parameters and stress responses in endangered Caspian brown trout (Salmo trutta caspius Kessler, 1877)

A feeding trial was conducted to evaluate the effects of Optimûn, a commercial nucleotide (NT) product, on the growth, haemato‐immunological and serum biochemical parameters and stress responses of Caspian brown trout fingerlings (average initial weight of 12.26 g). A basal diet was supplemented with levels of 0 (control), 1.5, 2.5, 3.5 and 5 g NT kg^?1 to formulate five experimental diets. After 8 weeks of feeding trial, fish fed diet with 2.5 g NT kg^?1 had the highest final weight compared with other treatments. The fish fed diets with 2.5 g NT kg^?1 had higher blood protein, albumin, albumin/globulin ratio, red blood cells, white blood cells and lymphocyte content and lower alkaline phosphatase. The lysozyme activity in serum was found to be significantly (P < 0.05) greater in fish fed diet with 2.5 g NT kg^?1. Concerning both stressors (confinement and salinity stress), fish fed diet with 2.5 g NT kg^?1 had lower plasma cortisol and glucose levels. Thus, administration of 2.5 g kg^?1 of the Optimûn dietary nucleotide formula is recommended to promote growth and immunity as well as to enhance stress responses of Caspian brown trout.     

Gill pathology caused by infestations of adult and preadult Dissonus manteri Kabata (Copepoda: Dissonidae) on coral trout, Plectropomus leopardus (Lacépède), (Serranidae)

Adult and preadult Dissonus manteri attached to the gills of Plectropomus leopardus cause significant pathology in the form of large hyperplastic nodules on the afferent (leading) edges of gill filaments. Nodules result from the dual actions of parasite attachment and feeding. The host response is characterized by severe epithelial hyperplasia, supplemented by fibroplasia and inflammation. Parasites attach close to the gill arch near the base of filaments. They have little effect on gill vasculature as the maxillipeds penetrate the filament superficial to the efferent filament artery and do not interfere with the blood vessels of the secondary lamellae. Tissue proliferation is limited to the wide portion of filament 'edge' epithelium in the proximal third and also does not extend to the secondary lamellae. Nodules are most numerous towards the ends of hemibranchs and are generally absent from the central regions. Leading hemibranchs bear significantly more nodules than their trailing counterparts. Of the total number of nodules, 20.5% are located on the pseudobranchs. Distribution patterns are considered to be primarily the result of D. manteri avoiding strong water currents, although this cannot explain the difference between numbers on leading and trailing hemibranchs.     

Commercial Bacillus probiotic supplementation of rainbow trout (Oncorhynchys mykiss) and brown trout (Salmo trutta): growth,immune responses and intestinal morphology

Probiotic administration is associated with the enhancement of host resistance to environmental and nutritional stressors, improving survival and growth rates. This study was carried out to evaluate the effects of dietary supplementation of Bacillus subtilis and Bacillus cereus toyoi on growth performance, innate immune responses and gut morphology of two trout species feeding a commercial diet high in soybean meal. A commercial probiotic (4.2 × 10⁹ CFU g^?1 of additive) was supplemented to the experimental diets at 0% (control), 0.03% (P₁; 6 × 10³ CFU g^?1 of diet) or 0.06% (P₂; 1.5 × 10⁶ CFU g^?1 of diet) and fed to brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) for 9 and 20 weeks respectively. Rainbow trout showed significantly better growth performance than brown trout, regardless of the dietary treatment. No effect of dietary probiotic supplementation was detected on growth performance, body composition or innate immune parameters (plasma lysozyme, alternative complement and peroxidase activities). In both species, after 9 weeks, intestinal lamina propria and submucosa were widened, with increased presence of inflammatory cells, significantly higher in groups fed probiotics. This inflammatory process, with villi and enterocytes noticeably damaged compared with the control group, was more pronounced in brown trout. Under the current trial conditions, the B. subtilis + B. cereus toyoi had no positive impact in either trout species, on the contrary a harmful effect was observed.     

The effect of different dietary inclusion levels of rutin (a flavonoid) on some liver enzyme activities and oxidative stress indices in rainbow trout,Oncorhynchus mykiss (Walbaum) exposed to Oxytetracycline

The study was conducted using a completely randomized design with five treatments each with three replicates. In each treatment, 30 fish (10 per replicate) with an average body weight of 30 ± 0.6 g were randomly allocated to 60‐L tanks. Treatments consisted of a control (a diet without rutin and oxytetracycline (OTC)), an OTC treatment (a diet free from rutin and containing 75 mg kg^?1 body weight OTC) and the (OTC + rutin) treatments including diets with different levels of rutin, 500 (R‐500 + OTC), 1000 (R‐1000 + OTC) and 2000 (R‐2000 + OTC) ppm and 75 mg kg^?1 body weight per day OTC which were fed to fish for 28 days. The aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in the blood serum after the OTC treatment were significantly (P < 0.05) higher than in the control. After the three (OTC+rutin) treatments, the AST and ALT activities were not significantly different from the control. The level of glucose in the blood serum was significantly lower (P < 0.05) in all treatments as compared to the OTC treatment. Superoxide dismutase (SOD) and catalase (CAT) activities in red erythrocyte lysates were significantly (P < 0.05) reduced after OTC treatment in comparison with the control. After the three (OTC+rutin) treatments, the values of these indices were significantly (P < 0.05) higher than the OTC treatment, in particular for the R‐1000 + OTC treatment. It seems that rutin, as a non‐enzymatic inhibitor, is capable of preventing damage to the liver tissue and reducing oxidative stress.     

The ability of clove oil and MS-222 to minimize handling stress in rainbow trout (Oncorhynchus mykiss Walbaum)

The effects of 60‐mg L^?1 clove oil and 60‐mg L^?1 tricaine methanesulphonate (MS‐222) on the blood chemistry of rainbow trout were compared after exposure to handling stress via caudal puncture blood sampling. Fish sampled by caudal puncture and subsequently exposed to anaesthetics showed a typical handling stress response over a 48‐h period. There were no significant differences between the responses of fish exposed to equal concentrations of clove oil and MS‐222, with the following exceptions: the blood glucose at full anaesthesia, and lactate at full recovery increased significantly in the clove oil‐exposed fish. In a subsequent experiment, the stress response observed in fish sampled by caudal puncture and exposed to clove oil and MS‐222 was compared with a non‐anaesthetized control group. The increases in plasma cortisol levels were significantly lower at recovery in fish treated with either anaesthetic compared with the control fish. Fish exposed to MS‐222 had significantly higher cortisol levels at 1 h. These findings show that few differences exist between the anaesthetic effects of clove oil and MS‐222 on the physiological response of fish to stress. However, clove oil is more effective at reducing the short‐term stress response induced by handling and blood sampling, and is recommended as an alternative fish anaesthetic.     

Physiological responses associated with capture and crowding stress in matrinxãBrycon cephalus (Gunther, 1869)

The dynamics of plasma cortisol, blood glucose, plasma chloride and liver glycogen were investigated in matrinxã (Brycon cephalus) submitted to capture and various periods of crowding. A total of 400 fish (700±22 g weight) were distributed in four ponds divided into four 50‐m² squares (25 fish/square, 350 g L^?1), where they were acclimated for 30 days. On the sampling day, after 24 h without food, all fish from three squares were transferred to the fourth square. Six fish were sampled before the procedure (control group, zero time) and 1, 3, 6 and 24 h after the capture and crowding. Each sampling was performed in a different pond to prevent additional stress. Fish were anaesthetized and blood and liver collected for biochemical analysis. Water temperature, pH, dissolved oxygen, alkalinity, ammonia and nitrite levels were within acceptable levels for matrinxã rearing. Slight but not significant increases were verified in plasma cortisol and blood glucose levels, as were decreases in plasma chloride and liver glycogen levels. The results suggest that matrinxã is highly tolerant to the procedures of capture and short‐term crowding.     

The physiological response to capture and handling stress in the Atlantic sharpnose shark, Rhizoprionodon terraenovae

Secondary effects of capture and handling stress in the Atlantic sharpnose shark, Rhizoprionodon terraenovae, were investigated in this study. Twenty-four sharks were serially bled and changes in several hematological parameters were monitored over a 1-h time period, following capture by hook-and-line. Blood samples were obtained from each shark at 0, 15, 30, 45 and 60 min. All 0-min (initial) blood samples were obtained within 3 min of capture. Blood glucose (9.2–13.1 mmol l^–1), lactate (1.5–28.9 mmol l^–1), and plasma osmolality (871–929 mOsm kg^–1) all increased after capture, whereas blood pH (6.86–6.78) declined. Hematocrit values (initial = 25.1%) remained unchanged throughout the 1-h stress period. Due to the short amount of time it took to obtain the initial sample and the lack of a significant relationship between the initial time and the initial parameter levels, all initial samples are considered the best approximation of the predisturbance resting levels. The use of repeated measures in this study enables us to describe the dynamics of the secondary stress response in the Atlantic sharpnose shark.     

Pathology of attachment and vascular damage associated with larval stages of Dissonus manteri Kabata, 1966 (Copepoda: Dissonidae) on the gills of coral trout, Plectropomus leopardus (Lacépède) (Serranidae)

Abstract. Tissue responses in Plectropomus leopardus gills infested with Dissonus manteri copepodid and ehalimus larvae were almost entirely proliferative, with little or no cellular infiltration. The pathological changes were related to attachment, feeding and parasite size, and comprised progressive mechanical erosion, epithelial hyperplasia and fibrosis. When present, infiltrating cells were associated with initial attachment by the copepodid and with the frontal filament basal plate. Approximately 2.4% of the respiratory area was lost as a result of direct vascular damage. Disruption of branchial blood flow resulted from the burrowing and feeding activities of the larvae and compression of adjacent vessels as the parasite increased in size. Chalimus stages were attached by a frontal filament which consisted of a stem with a terminal anchor, and a basal plate in contact with the cartilaginous filament ray. Following chalimus release, the branchial tissue appeared to repair rapidly. No frontal filament remains were found.     

Cellular responses to temperature stress in steelhead trout (Onchorynchus mykiss) parr with different rearing histories

This study compared the heat-shock response and metabolic energy status in hatchery-raised and two groups of wild-caught steelhead trout parr collected from the Navarro River watershed, California. Wild parr were from coastal and inland sites with different thermal regimes. Fish were exposed in the laboratory to 25 ± 0.2°C for 2-h and the heat-shock response was assessed in muscle tissue via induction of heat-shock proteins (hsps) 63, 72, 78, and 89. Metabolic measurements included muscle phosphocreatine (PCr), ATP, ADP, and AMP, and hepatic glycogen. Inland and coastal fish overlapped considerably with regard to their hsp responses and energetic endpoints, but hatchery fish were distinct in the biochemical patterns they exhibited. Hsp expression levels after temperature shock were significantly lower in hatchery than in wild fish. Hatchery fish also had significantly lower hepatic glycogen and higher muscle ADP, ATP, and PCr concentrations than wild fish. Coastal and inland steelhead did not differ significantly with regard to peak hsp72 and hsp89 levels or to concentrations of energy metabolites. However, fish from the warm-water, inland site expressed significantly less hsp63, maintained higher basal levels of hsp72, and induced hsp89 more slowly than fish from the cold-water, coastal site. Discriminant function analysis revealed that hatchery fish can be distinguished from wild Navarro River fish with 84.9% certainty using the following function: f(x) = − 43.6 + 0.14(Gly) + 4.1(PCr) + 186.4(AMP) + 80.8(ADP) − 0.14(hsp63) + 0.005(hsp72). This study demonstrates that within a single species, rearing conditions or genetic variation can influence an organism’s disposition and cellular response to thermal stress. Extrapolation of results from laboratory studies on hatchery fish to wild fish may therefore not be possible, and caution must be used when interpreting hsp data obtained for wild fish with different thermal histories.     

Effects of size and fragmentation of marine reserves and fisher infringement on the catch and biomass of coral trout, Plectropomus leopardus, on the Great Barrier Reef, Australia

Abstract  A spatially structured simulation model of the population dynamics and line fishing exploitation of common coral trout, Plectropomus leopardus Lacepède, was used to evaluate the effects of infringement and different amounts and arrangements of marine reserves on the Reef Line Fishery of the Great Barrier Reef, Australia. With no marine reserves and under a constant future effort level equal to that for 1996, the size of the population was reduced and the biomass stabilised at about 40% of pre-exploitation levels. Marine reserves were ineffective at conserving biomass when limited infringement was allowed throughout an entire reserve. When infringement was absent altogether or limited to the edges of reserves, larger marine reserves lead to lower total catches and higher overall biomass. When infringement was limited to reserve edges, a single large closure was more effective at conserving biomass than more fragmented arrangements. Simulations suggested that marine reserves might lead to better conservation of a fishery-targeted species if infringement is negligible or limited to reserve margins. Even where infringement occurred only at the edges of reserves, a network of small reserves may be less effective at conserving a targeted species than a smaller number of larger reserves.     

Cortisol,glucose and hematocrit changes during acute stress,cohort sampling,and the diel cycle in diploid and triploid brook trout (Salvelinus fontinalis Mitchill)

In this study, groups of diploid (control) and triploid brook trout were subjected to a 5 min acute handling/confinement stress. Blood samples were collected from equal numbers of diploid and triploid fish at regular predetermined intervals, beginning prior to confinement and continuing for up to 3h after confinement. These blood samples were used for the measurement of hematocrit and plasma cortisol and glucose concentrations. The effects of cohort sampling and the diel cycle on these variables were also analysed. Except for minor differences, both groups responded similarly to the acute stress, with short-term elevations in plasma cortisol and hematocrit levels, and a more sustained increase in plasma glucose levels. No significant differences were found between diploid and triploid fish in their reactions to cohort sampling. However, the act of removing an individual from a tank caused significant changes in plasma cortisol levels in cohorts remaining in that tank (p < 0.001). Significant diel effects were observed for both ploidy groups in plasma cortisol (p < 0.05) but not in plasma glucose or hematocrit. To whom correspondence should be addressed