Development of digestive enzyme activity in spotted rose snapper,Lutjanus guttatus (Steindachner, 1869) larvae

We describe digestive enzyme activity during the larval development of spotted rose snapper, Lutjanus guttatus. Trypsin, chymotrypsin, leucine aminopeptidase, pepsin, amylase, lipase, and acid and alkaline phosphatase activities were evaluated using spectrophotometric techniques from hatching through 30 days. The spotted rose snapper larvae present the same pattern of digestive enzyme activity previously reported for other species in which pancreatic (i.e., trypsin, chymotrypsin, amylase, and lipase) and intestinal (i.e., acid and alkaline phosphatases and leucine aminopeptidase) enzymatic activities are present from hatching allowing the larvae to digest and absorb nutrients in the yolk-sac and live prey by the time of first feeding. The digestive and absorption capacity of the spotted rose snapper increases during the larval development. A significant increase in individual activity of all enzymes occurs at 20 DAH, and around 25 DAH, the juvenile-type of digestion is observed with the appearance of pepsin secreted by the stomach, suggesting that maturation of the digestive function occurs around 20–25 DAH. Our results are in agreement with a previous suggestion that early weaning may be possible from 20 DAH. However, the patterns of enzymatic activities reported in our study should be considered during the formulation of an artificial diet for early weaning of the spotted rose snapper.     

Ascorbic acid requirement and histopathological changes due to its deficiency in juvenile spotted rose snapper Lutjanus guttatus (Steindachner, 1869)

A 13-week feeding trial was conducted to determine the optimal dietary ascorbic acid (AA) requirement of juvenile spotted rose snapper (Lutjanus guttatus). Six experimental diets containing 0, 7, 19, 29, 62 and 250 mg AA equivalent kg^?1 diet, supplied as l-ascorbyl-2-polyphosphate designated as VC0, VC7, VC19, VC29, VC62 and VC250, were fed ad libitum to triplicate groups of 20 juveniles [initial weight (IW) of 8 ± 1.85 g]. Nutrient deficiency resulting in fin erosion, dark skin, desquamation and erratic swimming was observed after 8 weeks in fish fed diets from 0 to 29 mg AA kg^?1. Fish that were fed all diets showed gill, kidney and brain alterations. The degree of tissue change values indicates that increasing vitamin C (VC) levels reduces these alterations. Fish fed diets VC29, VC62 and VC 250 showed a significantly higher growth rate in comparison with those fed the diets VC0, VC7 and VC17 (P > 0.05). The dietary requirement of VC in L. guttatus was estimated to be 29 mg kg^?1 of AA based on weight gain and specific growth rate, but more than 250 mg kg^?1 of AA is required to eliminate clinical signs and histopathological lesions.     

Effects of different feeding level on the growth, feed efficiency and body composition of juvenile mangrove red snapper, Lutjanus argentimaculatus (Forsskal 1775)

The effects of several feeding levels (1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4% and 4.5% of body weight per day, BW day⁻¹) on the growth, feed efficiency and body composition of juvenile Lutjanus argentimaculatus (body weight 27.1 g) were examined. Fish were fed a test diet (40% protein, 18.4% lipid and 13.4 kJ g⁻¹) for 75 days in three equal meals. Weight gain and specific growth rate increased with increasing feeding level up to 2.5% BW day⁻¹, after which no significant improvement in growth was observed. The feed efficiency, protein efficiency ratio, retention of protein and digestibility of nutrients did not differ for fish fed 1–2.5% BW day⁻¹, but decreased significantly when feeding levels were increased above 3% BW day⁻¹. The chemical compositions of whole fish or body organs were significantly affected by the feeding level. The condition factor, mesenteric fat, hepato- and viscerosomatic indices were higher in fish fed 2.5–4.5% BW day⁻¹. The cholesterol, triglycerides and haematocrit values were similar among treatments, except that high levels of plasma lipids were recorded in fish fed at 2.5% BW day⁻¹. Taking into consideration the growth, feed efficiency and body composition data attained in this study, a feeding level of 2.5% BW day⁻¹ is recommended for juvenile L. argentimaculatus weighing between 27 and 140 g.     

The effect of temperature and duration of incubation on the hatching of diapause eggs of Centropages hamatus (Lilljeborg) (Copepoda, Calanoida)

Diapause eggs of Centropages hamatus were used to investigate the effect of temperature and duration of incubation on egg hatching. Eggs were incubated for 10, 12, 14, 16, 20, 24, 28, 32, 36 and 40 h at 15°C and 14L–10D. After incubation for the designated period, eggs were transferred to 25°C and monitored periodically to determine egg hatching. Control eggs were incubated solely at 15°C and monitored for egg hatching. The greatest daily hatching success of eggs occurred within 1 or 2 days after transfer from 15°C to 25°C, while the controls required 3–4 days. The cumulative hatching success of eggs was significantly lower than the control, with the exception of eggs held for at least 36 h at 15°C before transfer to 25°C. These results indicate that overall time to hatching of diapause eggs of C. hamatus can be reduced by transferring the eggs to a higher temperature, for example, 25°C, following a minimum period of time (36 h) at reduced temperature, for example, 15°C. Exposure to 15°C for only 10 h does not appear to be sufficient to result in any subsequent hatching at higher temperature.     

Partial replacement of fishmeal by defatted soybean meal in formulated diets for the mangrove red snapper, Lutjanus argentimaculatus (Forsskal 1775)

This study was conducted to evaluate the effect on growth and feed efficiencies of the mangrove red snapper (Lutjanus argentimaculatus) when dietary fishmeal is partially replaced by defatted soybean meal (DSM). In the preliminary experiment, snapper (mean weight±SD, 58.22±5.28 g) were fed in triplicate with different dietary amounts of DSM (7.8–42.2%) that were formulated to be isonitrogenous and isocaloric. After 14 weeks, survival, growth and feed efficiencies, and hepatosomatic index (HSI) did not differ. Based on these results, a feeding trial was done using a positive control diet that contained 64% fishmeal, while the other four diets had DSM levels of 12%, 24%, 36%, and 48% that replaced fishmeal protein at 12.5%, 25%, 37.5%, and 50% respectively. All diets were formulated to have about the same protein level (50%), protein to energy ratio (P/E of 25‐mg protein kJ^?1), and dietary energy (19.8 MJ kg^?1). These were fed to triplicate groups of snapper (mean total weight tank^?1±SD, 73.19±1.2 g) at 15 fish (average weight, 4.88 g) per 1.5‐t tank for 19 weeks. Growth (final average weight and specific growth rate (SGR), feed conversion ratio (FCR), survival, and HSI were not significantly different (P>0.05) while protein efficiency ratios or PERs were similar in treatments with DSM. Among snapper fed DSM, haematocrit value was significantly lower in fish fed 48% DSM and not different with fish fed 36% DSM. Whole‐body crude fat of snapper fed 48% DSM was lowest while the crude protein and nitrogen‐free extract (NFE) levels were highest. Histopathological analysis showed that lipid vacuoles in livers of snapper were reduced in size as dietary DSM increased. There was slight lipid deposition in the liver of snapper at 36% DSM while at 48% DSM it was excessive and hepatocytes were necrotic. There were no differences in the histology of snapper intestine. Under the experimental condition of this study, DSM can be used in snapper diets at 24% (replacing 25% of fishmeal protein) based on growth, survival and feed efficiencies, and histology of liver and intestine. For a lesser diet cost, an inclusion level higher than 24% DSM is possible with a bioavailable phosphorus supplement.     

Effect of dietary protein and lipid levels and protein to energy ratios on growth, survival and body composition of the mangrove red snapper, Lutjanus argentimaculatus (Forsskal 1775)

The approximate levels of dietary protein and energy that would sustain good growth and survival of the mangrove red snapper Lutjanus argentimaculatus (Forsskal) were determined in two feeding experiments. In the preliminary experiment, six fish meal‐based diets were formulated to contain three protein levels (35%, 42.5% and 50%) and two lipid levels (6% and 12%) for each protein, with dietary energy ranging from 14.6 MJ kg^?1 to 20.5 MJ kg^?1. The protein to energy (P/E) ratios of diets ranged from 20.6 mg protein kJ^?1 to 27.5 mg protein kJ^?1. Diets were fed for 100 days to triplicate groups of snappers with an average initial weight of 24.8 ± 0.4 g. No significant interaction between different levels of protein and lipid was observed. Survival rates (93.8% to 100%), feed conversion ratios (FCR) (2.61–3.06) and condition factors (K) were not affected by different dietary treatments. Regardless of lipid level, fish fed 50% protein diets had a significantly higher specific growth rate (SGR) than fish fed the 35% protein diets, but not compared with the 42.5% diets (P < 0.05). Increasing lipid to 12% in all protein levels resulted in no improvement in growth over the 6% level. Fish body moisture did not vary while lipid levels based on dry matter were high (27.9% to 33.7%). Snapper appear to require more than 40% dietary protein and a high dietary energy level for good growth. In the second experiment, fish (21.1 ± 0.1 g) in four replicate groups were fed for 94 days with three diets (39%, 44% and 49% protein with P/E ratios of 21.1, 23.3 and 25.5 mg protein kJ^?1 respectively) containing similar dietary energy levels of about 19 MJ kg^?1. Average final weight, SGR and FCR were significantly higher in diets containing 44% and 49% protein diets (P > 0.05). There were no differences in survival rates, protein efficiency ratio (PER) and nutrient composition of snapper flesh. All fish had fatty livers. Results indicated that the diet containing 44% protein with a P/E ratio of 23.3 mg protein kJ^?1 was optimum for snapper growth under the experimental conditions used in the study.     

The effects of dietary protein and lipid on growth and body composition of juvenile and sub-adult red snapper, Lutjanus campechanus (Poey, 1860)

To allow for the initial identification of practical diet formulations for red snapper culture, the present study was conducted to evaluate the effects of feeding varying levels of dietary protein and lipid on growth and body composition of juvenile and sub‐adult red snapper. Twelve diets were formulated to contain varying levels of dietary protein and lipid. In trial 1, juvenile red snapper (initial mean weight 5.9 g) were offered diets with graded levels of dietary protein (32%, 36%, 40%, 44%) and practical energy to protein ratios. In trial 2, juvenile red snapper (initial mean weight 8.64 g) were offered isonitrogenous diets (44% protein) containing graded levels of dietary lipid (8%, 10%, 12%, 14%). Sub‐adult fish (initial mean weight 151.5 g) were used in trial 3 and maintained on diets similar to those of trial 1 (32–44% protein). Sub‐adult fish (initial mean weight 178.3 g) in trial 4 were offered isonitrogenous diets containing 32% dietary protein and graded levels of dietary lipid (6%, 8%, 10%, 12%). There were no significant differences in growth, feed efficiency ratio (FER) or survival in juvenile fish. Juvenile fish offered 32% dietary protein exhibited a significantly greater (P=0.0497) protein conversion efficiency (PCE) than fish offered a diet containing 44% dietary protein. Juvenile fish in trial 2 also had significantly higher (P=0.005) intraperitoneal fat ratios (IPFRs) at 14% dietary lipid than fish offered diets containing 8–10% dietary lipid, and displayed trends towards greater protein as a percent of whole‐body composition at 8–10% dietary lipid. Sub‐adult snapper in trials 3 and 4 showed no significant differences in growth, FER or survival. However, in trial 4 there was a general trend towards increased % weight gain (P=0.0615), FER (P=0.0601) and final mean weight (P=0.0596) with increasing levels of dietary lipid. Fish in trial 4 offered 6% dietary lipid also had significantly lower (P=0.0439) IPFR and PCE (P=0.0188) than fish offered 12% dietary lipid. Based on data obtained from these trials, inclusion of dietary protein at levels of 32–36% appears sufficient to support growth. For this level of protein, dietary lipid should be ～10% in order to meet the energetic demands of the fish and to spare dietary protein for growth.     

Effects of dietary astaxanthin concentration and feeding period on the skin pigmentation of Australian snapper Pagrus auratus (Bloch & Schneider, 1801)

A single‐factor experiment was conducted to investigate the effects of dietary astaxanthin concentration on the skin colour of snapper. Snapper (mean weight=129 g) were held in white cages and fed one of seven dietary levels of unesterified astaxanthin (0, 13, 26, 39, 52, 65 or 78 mg astaxanthin kg^?1) for 63 days. Treatments comprised four replicate cages, each containing five fish. The skin colour of all fish was quantified using the CIE L^*, a^*, b^* colour scale after 21, 42 and 63 days. In addition, total carotenoid concentrations of the skin of two fish cage^?1 were determined after 63 days. Supplementing diets with astaxanthin strongly affected redness (a^*) and yellowness (b^*) values of the skin at all sampling times. After 21 days, the a^* values increased linearly as the dietary astaxanthin concentration was increased before a plateau was attained between 39 and 78 mg kg^?1. The b^* values similarly increased above basal levels in all astaxanthin diets. By 42 days, a^* and b^* values increased in magnitude while a plateau remained between 39 and 78 mg kg^?1. After 63 days, there were no further increases in measured colour values, suggesting that maximum pigmentation was imparted in the skin of snapper fed diets >39 mg kg^?1 after 42 days. Similarly, there were no differences in total carotenoid concentrations of the skin of snapper fed diets >39 mg kg^?1 after 63 days. The plateaus that occurred in a^* and b^* values, while still increasing in magnitude between 21 and 42 days, indicate that the rate of astaxanthin deposition in snapper is limited and astaxanthin in diets containing >39 mg astaxanthin kg^?1 is not efficiently utilized. Astaxanthin retention after 63 days was greatest from the 13 mg kg^?1 diet; however, skin pigmentation was not adequate. An astaxanthin concentration of 39 mg kg^?1 provided the second greatest retention in the skin while obtaining maximum pigmentation. To efficiently maximize skin pigmentation, snapper growers should commence feeding diets containing a minimum of 39 mg unesterified astaxanthin kg^?1 at least 42 days before sale.     

Effect of carotenoids and background colour on the skin pigmentation of Australian snapper Pagrus auratus (Bloch & Schneider, 1801)

Three 2‐factor experiments were conducted to determine the effects of background colour and synthetic carotenoids on the skin colour of Australian snapper Pagrus auratus. Initially, we evaluated the effects on skin colour of supplementing diets for 50 days with 60 mg kg^?1 of either astaxanthin (LP; Lucantin^® Pink), canthaxanthin (LR; Lucantin^® Red), apocarotenoic acid ethyl ester (LY; Lucantin^® Yellow), selected combinations of the above or no carotenoids and holding snapper (mean weight=88 g) in either white or black cages. In a second experiment, all snapper (mean weight=142 g) from Experiment 1 were transferred from black to white, or white to white cages to measure the short‐term effects of cage colour on skin L^*, a^* and b^* colour values. Skin colour was measured after 7 and 14 days, and total carotenoid concentrations were determined after 14 days. Cage colour was the dominant factor affecting the skin lightness of snapper with fish from white cages much lighter than fish from black cages. Diets containing astaxanthin conferred greatest skin pigmentation and there were no differences in redness (a^*) and yellowness (b^*) values between snapper fed 30 or 60 mg astaxanthin kg^?1. Snapper fed astaxanthin in white cages displayed greater skin yellowness than those in black cages. Transferring snapper from black to white cages increased skin lightness but was not as effective as growing snapper in white cages for the entire duration. Snapper fed astaxanthin diets and transferred from black to white cages were less yellow than those transferred from white to white cages despite the improvement in skin lightness (L^*), and the total carotenoid concentration of the skin of fish fed astaxanthin diets was lower in white cages. Diets containing canthaxanthin led to a low level of deposition in the skin while apocarotenoic acid ethyl ester did not alter total skin carotenoid content or skin colour values in snapper. In a third experiment, we examined the effects of dietary astaxanthin (diets had 60 mg astaxanthin kg^?1 or no added carotenoids) and cage colour (black, white, red or blue) on skin colour of snapper (mean weight=88 g) after 50 days. Snapper fed the astaxanthin diet were more yellow when held in red or white cages compared with fish held in black or blue cages despite similar feed intake and growth. The skin lightness (L^* values) was correlated with cage L^* values, with the lightest fish obtained from white cages. The results of this study suggest that snapper should be fed 30 mg astaxanthin kg^?1 in white cages for 50 days to increase lightness and the red colouration prized in Australian markets.     

Habitat use, growth, and mortality of post-settlement lane snapper (Lutjanus synagris) on natural banks in the northwestern Gulf of Mexico

Three low-relief banks in the northwestern Gulf of Mexico were evaluated as nursery habitat of lane snapper (Lutjanus synagris). Trawl surveys were conducted in three habitat types (inshore mud, shell ridge, offshore mud) to quantify lane snapper distribution and abundance. Heald Bank and Sabine Bank were trawled in 2003 while Freeport Rocks was trawled in 2000 (Freeport A) and 2004 (Freeport B). Density of lane snapper varied among banks and years sampled: Sabine Bank (20.8 ± 2.8 ind ha⁻¹), Heald Bank (1.1 ± 0.4 ind ha⁻¹), Freeport A (12.7 ± 2.3 ind ha⁻¹), and Freeport B (3.0 ± 1.0 ind ha⁻¹). Habitat-specific differences in density were observed, although patterns were not consistent among banks. Otolith microstructure analysis indicated that post-settlement lane snapper ranged in age from 21 to 66 d, with hatch dates from 1 May to 31 August. Growth rates varied from 0.90 mm d⁻¹ at Heald Bank to 1.27 mm d⁻¹ at Sabine Bank, and habitat-specific differences in growth were negligible. Mortality of post-settlement lane snapper ranged from 15.2% d⁻¹ at Sabine Bank to 9.2% d⁻¹ at Freeport A. Our findings indicate that Heald Bank, Sabine Bank, and Freeport Rocks all serve as settlement habitat of lane snapper, which appear to be capable of successful settlement across a variety of habitats.     

The effect of light intensity on first feeding of the spotted sand bass Paralabrax maculatofasciatus (Steindachner) larvae

The effects of light intensity on feeding incidence and prey consumption at first feeding of spotted sand bass larvae (Paralabrax maculatofasciatus Steindachner), using four light intensity treatments (0, 100, 400, and 700 lx) were evaluated. Specimens were fed the rotifer Brachionus plicatilis at a density of 3 rotifers mL^?1. One hour after the addition of prey, 30±3 (mean±SEM) larvae were sampled from each treatment aquarium. Feeding incidence was evaluated as the percentage of larvae with prey in the digestive tract. Feeding intensity was measured as the number of prey in the digestive tract of the larvae. Histological analysis was carried out to describe the eye structure at the time of first feeding. Larvae fed in darkness (0 lx) had a significantly lower (P<0.05) feeding incidence (1.2±2.2%) and intensity (0.4±0.7 rotifers larvae^?1) than those larvae fed at 100 (28±11%, 1.8±0.2 rotifers larvae^?1), 400 (48±10%, 2.4±0.3 rotifers larvae^?1), and 700 lx (52±4%, 2.4±0.1 rotifers larvae^?1). Feeding incidence of the spotted sand bass larvae increased with light intensity while the feeding intensity showed no significant difference (P>0.05) between light treatments. Histological analysis of the eye structure showed that first feeding larvae had well‐formed lens along with a retina composed of pure single cones as photoreceptors.     

Effects of storage and incubation temperature on the viability of eggs, embryos and larvae in two strains of an African catfish, Heterobranchus longifilis (Siluriformes, Clariidae)

Two experiments, dealing with short‐term storage of ova and thermal conditions to optimize gamete and eggs management in hatcheries of the African catfish, Heterobranchus longifilis, were carried out. In the first experiment, ova collected by stripping from two strains of H. longifilis were stored for intervals up to 8 h at two temperature regimes: in a domestic refrigerator (3–5°C) and at ambient room temperature (20.5–22°C). In the second experiment, eggs were incubated from fertilization to hatching at different experimental temperatures (21, 25, 29, 32 and 35°C) to determine the effects of temperature on the kinetics of white egg appearance, hatching times and hatching quality. Gamete storage at warmer temperatures significantly prolonged viability irrespective of the strain used. In fact, the hatching rate for ova stored at 20.5–22 and 3–5°C for 5 h ranged between 75.2–79.3% and 6.5–9.4% respectively. Loss of viability was most noticeable after 6 h storage at ambient room temperature. Post‐storage viability significantly declined after 2 h exposure to the domestic refrigerator temperature. No hatching of normal larvae took place after 8 h post‐storage time. Results from the second experiment showed that time to maximum whitening of eggs was both strain‐ and temperature‐dependent. The time to maximum mortality of eggs was shorter in the Layo strain (LS) than in the Noun strain (NS), regardless of incubation temperature. The appearance of white eggs was shorter with increasing incubation temperatures. Hatching times decreased with increasing temperature, regardless of strain. Hatching took place from 21 to 27 h and 19 to 24 h after fertilization at temperature of 29°C, respectively, for NS and LS. The length of the hatching period was remarkably shorter for LS than NS at any tested incubation temperature, except 35°C. No hatching took place at 21°C. The highest proportion of normal larvae occurred at 25 and 29°C, respectively, for NS and LS. Hatching rate was highest at 25 and 29°C, respectively, for NS and LS. There was a significantly higher proportion of deformed larvae at 35°C regardless of the strain.     

Cloning and expression of the gene encoding an extracellular alkaline serine protease from Vibrio alginolyticus strain HY9901, the causative agent of vibriosis in Lutjanus erythopterus (Bloch)

A 750-bp internal fragment of the alkaline serine protease gene (asp) from the Vibrio alginolyticus strain HY9901 was amplified by polymerase chain reaction (PCR). The flanking sequences of the 5'- and 3'- ends of the asp gene were characterized by reverse and nested PCR. Sequence analysis showed that the asp gene contained an 1893-bp ORF encoding 630 amino acids. The deduced amino acid sequence of the ASP (alkaline serine protease) precursor showed significant homology with several bacterial alkaline serine proteases. Expression of the asp gene in Escherichia coli and activity tests of the ASP indicated that the N-signal peptide of the ASP precursor was essential to autocatalyse and fold correctly the enzyme to obtain activity. The purified ASP was lethal for Lutjanus erythopterus with an LD(50) of 0.25 microg protein g(-1) body weight.     

Investigation of the nutritional requirements of Australian snapper Pagrus auratus (Bloch & Schneider, 1801): effects of digestible energy content on utilization of digestible protein

This study used a curvilinear model to investigate the effects of different digestible energy (DE) levels on the digestible protein (DP) requirements of juvenile snapper Pagrus auratus. For each DE level (15, 18 or 21 MJ kg⁻¹), DP content was increased from about 210–560 g kg⁻¹ in seven evenly spaced increments by formulating a summit diet (highest DP content) and a diluent diet (lowest DP content) at the desired DE level and combining the summit and diluent diets in various ratios to achieve the desired DP content. This ensured the DE level remained relatively stable. Each of the 21 dietary treatments was fed to three replicate groups of snapper twice daily to apparent satiation for 57 days. At the completion of the trial, fish were weighed and killed for chemical analysis. Results indicated that the rapid growth of snapper weighing 30–90 g was highly dependent on the ratio of DP to DE and that optimum protein deposition did not occur until snapper were offered feeds with at least 350 g DPkg⁻¹, irrespective of DE level. According to the fitted models, diets formulated for snapper reared at temperatures from 20–25°C should contain approximately 23 g DP MJ DE⁻¹ to promote optimal weight gain and protein deposition. Based on the feeding regime used in this study, this could be achieved with practical diets containing a DP:DE ratio of 460:20, 420:18 or 350:15.     

Effect of combined photoperiod, water calcium concentration and pH on survival, growth, and moulting of juvenile crayfish (Procambarus clarkii ) cultured under laboratory conditions

The red swamp crayfish, Procambarus clarkii (Girard), is one of the most commonly farmed freshwater species in inland China due to its high market value and consumer demand. The aim of this study was to determine the optimum combinations of photoperiod, water calcium concentration and pH for juvenile survival, growth and moulting. In our orthogonal experiment, the three environmental factors were varied at three levels (photoperiod: 16L:8D, 12L:12D and 8L:16D; calcium concentration: 45.5, 65.5 and 85.5 mg L⁻¹; and pH: 6.8, 7.8 and 8.8). Range analysis showed that the maximum survival of juvenile crayfish occurred at photoperiods of 16L:8D or 8L:16D, water calcium concentration of 45.5 mg L⁻¹ and pH of 7.8; maximum weight gain at photoperiod 16L:8D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8; maximum length increase at photoperiod 16L:8D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8; and the highest moult frequency at photoperiod 12L:12D, water calcium concentration 65.5 mg L⁻¹ and pH 7.8. Analysis of variance indicated that photoperiod, water calcium concentration and pH significantly influenced only the weight gain of juvenile crayfish ( P <0.05). Taking growth into consideration, we suggest that a photoperiod of 16L:8D, calcium concentration of 65.5 mg L⁻¹ and pH 7.8 might be optimal conditions for rearing juvenile P. clarkii .     

Effect of cage colour and light environment on the skin colour of Australian snapper Pagrus auratus (Bloch & Schneider, 1801)

A two‐factor experiment was performed to evaluate the effects of cage colour (black or white 0.5 m³ experiment cages) and light environment (natural sunlight or reduced level of natural sunlight) on the skin colour of darkened Australian snapper. Each treatment was replicated four times and each replicate cage was stocked with five snapper (mean weight=351 g). Snapper exposed to natural sunlight were held in experimental cages located in outdoor tanks. An approximately 70% reduction in natural sunlight (measured as PAR) was established by holding snapper in experimental cages that were housed inside a ‘shade‐house’ enclosure. The skin colour of anaesthetized fish was measured at stocking and after a 2‐, 7‐ and 14‐day exposure using a digital chroma‐meter (Minolta CR‐10) that quantified skin colour according to the L^*a^*b^* colour space. At the conclusion of the experiment, fish were killed in salt water ice slurry and post‐mortem skin colour was quantified after 0.75, 6 and 22 h respectively. In addition to these trials, an ad hoc market appraisal of chilled snapper (mean weight=409 g) that had been held in either white or in black cages was conducted at two local fish markets. Irrespective of the sampling time, skin lightness (L^*) was significantly affected by cage colour (P<0.05), with fish in white cages having much higher L^* values (L^*≈64) than fish held in black cages (L^*≈49). However, the value of L^* was not significantly affected by the light environment or the interaction between cage colour and the light environment. In general, the L^* values of anaesthetized snapper were sustained post mortem, but there were linear reductions in the a^* (red) and b^* (yellow) skin colour values of chilled snapper over time. According to the commercial buyers interviewed, chilled snapper that had been reared for a short period of time in white cages could demand a premium of 10–50% above the prices paid for similar‐sized snapper reared in black cages. Our results demonstrate that short‐term use of white cages can reduce the dark skin colour of farmed snapper, potentially improving the profitability of snapper farming.     

Effects of photoperiod alterations on day-night variations in hypothalamic serotonin content and turnover,and monoamine oxidase activity in the female catfish,Heteropneustes fossilis (Bloch)

The effects of different photoperiod regimes, and total darkness on day-night variations in hypothalamic serotonin (5-HT) content and turnover index (TI), and monoamine oxidase (MAO) activity that occur exclusively during the gonadal preparatory phase (February–March) were investigated in female Heteropneustes fossilis. Exposure of the fish to long photoperiods (16L:8D; light between 06.00–22.00h, for 40 days elevated both the midphotophase and midscotophase values of 5-HT content and MAO, and abolished their day-night differences. The daily variations of 5-HT-TI was further intensified by the treatment. Under short photoperiods (4L:20D; light between 18.00–22.00h for 40 days), there was a significant decrease in both 5-HT content and TI, a reversal of the day-night variations of 5-HT content and MAO activity, and loss of the 5-HT-TI pattern. In fish maintained in total darkness the day-night variations were not found and there was a significant inhibition of 5-HT. Exposure of the fish to continuous light abolished the day-night variations of these correlates and elevated their values at both intervals. When the LD cycle was reversed, there was a corresponding shift in the day-night patterns. The gonadosomatic index (GSI) was significantly elevated in the long photoperiod and continuous illumination groups, significantly decreased in the short photoperiod and total darkness groups, and unchanged in the reversed LD cycle regime. The results indicate that the day-night variations of 5-HT and MAO are photoperiod-dependent and are controlled by the prevailing LD cycle during the gonadal preparatory phase of the annual reproductive cycle. The photoperiod effects on the gonadal activity may be mediated through the hypothalamic serotonergic system.     

Effects of dietary astaxanthin source and light manipulation on the skin colour of Australian snapper Pagrus auratus (Bloch & Schneider, 1801)

Two experiments were conducted with Australian snapper Pagrus auratus (Bloch and Schneider, 1801). The first was aimed at determining the dietary level of astaxanthin that improved skin redness (CIE a^*values) of farm‐reared snapper. Farmed snapper (ca. 600 g) fed a commercial diet without carotenoids were moved to indoor tanks and fed the same diet supplemented with 0, 36 or 72 mg astaxanthin kg^?1 (unesterified form as Carophyll Pink?) for nine weeks. Skin redness (CIE a^* values) continued to decrease over time in fish fed the diet without astaxanthin. Snapper fed the diet containing 72 mg astaxanthin kg^?1 were significantly more red than fish fed the diet with 36 mg astaxanthin kg^?1 three weeks after feeding, but skin redness was similar in both groups of fish after 6 and 9 weeks. The second experiment was designed to investigate the interactive effects of dietary astaxanthin source (unesterified form as Carophyll Pink? or esterified form as NatuRose?; 60 mg astaxanthin kg^?1) and degree of shading (0%, 50% and 95% shading from incident radiation) on skin colour (CIE L^*a^*b^*) and skin and fillet astaxanthin content of farmed snapper (ca. 800 g) held in 1 m³ floating cages. After 116 days, there were no significant interactions between dietary treatment and degree of shading for L^*, a^* or b^* skin colour values or the concentration of astaxanthin in the skin. Negligible amounts of astaxanthin were recovered from fillet samples. The addition of shade covers significantly increased skin lightness (L^*), possibly by reducing the effect of melanism in the skin, but there was no difference between the lightness of fish held under either 50% or 95% shade cover (P>0.05).     

Nutritional assessment of Australian canola meals. I. Evaluation of canola oil extraction method and meal processing conditions on the digestible value of canola meals fed to the red seabream (Pagrus auratus, Paulin)

This study assessed the nutrient and energy digestibility of a variety of canola protein products that were produced by processing canola meal under different conditions, using the red seabream, Pagrus auratus. The test canola protein products included solvent‐extracted canola meal, expeller‐extracted canola meal alone or subjected to one of two different heat treatments (120 or 150°C for 30 min), and expeller meal further processed to produce a canola protein concentrate (CPC). Solvent‐extracted soybean meal was also included in the study as a reference ingredient. Daily feed intake and blood thyroid hormone levels over the experimental period were also examined. The total digestible protein content of the expeller‐ and solvent‐extracted canola meals was 356 and 358 g per kg dry matter (g kg DM^?1) respectively. The total digestible energy content of the expeller‐ and solvent‐extracted canola meals was 14.23 and 8.60 MJ kg^?1 respectively. The organic matter digestibilities of the solvent‐extracted canola meal were poorer than noted for the expeller‐ and solvent‐extracted soybean meal. Notably, the two sources of canola meals used in this study (solvent and expeller meals) did not cause problems with declining feed intakes or changes to blood levels of thyroid hormones when included in the diets of the fish at a 30% level, and the diets were fed over a 3‐week period. The preparation of a CPC resulted in gains in total digestible energy, but a reduction in the amount of total digestible protein, relative to the expeller canola meal from which it was produced. However, the protein concentrating process marginally reduced the relative digestible value of the protein content. Heating expeller meal at 120 or 150°C for 30 min resulted in progressive reductions of all nutrient and energy digestibilities.