Effect of stocking density on survival and growth performance of pikeperch, <Emphasis Type="Italic">Sander lucioperca</Emphasis> (L.), larvae under controlled conditions

The effect of stocking density on the survival and growth of pikeperch, Sander lucioperca (L.), larvae was examined in two consecutive experiments. In experiment I, 4-day-old larvae [body wet weight (BW): 0.5 mg; total body length (TL): 5.6 mm] were reared in 200-l cylindro-conical tanks in a closed, recirculating system (20 ± 0.5°C) at three stocking densities (25, 50 and 100 larvae l⁻¹) and fed a mixed feed (Artemia nauplii and Lansy A2 artificial feed) for 14 consecutive days. At densities of 25 and 100 larvae l⁻¹, growth rate and survival ranged from 2.7 to 1.9 mg day⁻¹ and from 79.2 to 72.3%, and fish biomass gain ranged from 0.6 to 2.0 g l⁻¹, respectively. There were two periods of increased larval mortality: the first was at beginning of exogenous feeding and the second during swim bladder inflation. In experiment II, 18-day-old larvae (BW: 35 mg; TL: 15.6 mm) obtained from experiment I were reared under culture conditions similar to those of experiment I, but at lower stocking densities (6, 10 and 15 larvae l⁻¹). The fish were fed exclusively with artificial feed (trout starter) for 21 consecutive days. At densities of 6 and 15 larvae l⁻¹, the growth rate and fish biomass gain ranged from 28.8 to 23.1 mg day⁻¹ and from 2.0 to 3.3 g l⁻¹, respectively. The highest survival (56.5%) was achieved at a density of 6 larvae l⁻¹. Mortality at all densities was mainly caused by cannibalism II type behaviour (27–35% of total). In both experiments, growth and survival were negatively correlated and fish biomass gain positively correlated with stocking densities. The present study suggests that the initial stocking density of pikeperch larvae reared in a recirculating system can be 100 individuals l⁻¹ for the 4- to 18-day period post-hatch and 15 individuals l⁻¹ for the post-19-day period.     

The effect of different diets on the growth and mortality of juvenile pikeperch (<Emphasis Type="Italic">Sander lucioperca</Emphasis>) in the transition from live food to formulated feed

The efficiencies of four feeding methods were compared in the transition period from live food to dry feed in pikeperch (Sander lucioperca). Pond-nursed fry were trained to artificial feed without a transition (Group P) and with a transition, using a combination of dry feed and different kinds of live food—Chironomus larvae (Group C), Tubifex (Group T) and Daphnia (Group D)—during a 12-day-long trial. The ratio of dry feed for the transition groups was increased from 0 to 100% in 3-day intervals. The highest specific growth rates and condition factors were detected for Group C (4.2%, 1.46 ± 0.5) and Group T (4.2%, 1.48 ± 0.6), respectively. The survival percentages of the groups were as follows: 86.7 ± 9% (Group C), 78 ± 4% (Group T), 52 ± 1.4% (Group D) and 41 ± 9% (Group P). The rate of cannibalism ranged between 6.3 and 13.7% during the transition period. Larger-sized fish seemed to learn more rapidly to accept dry feed than their smaller counterparts.     

Masculinization of Convict Cichlid (<Emphasis Type="Italic">Cichlasoma nigrofasciatum</Emphasis>) by immersion in <Emphasis Type="Italic">Tribulus terrestris</Emphasis> extract

We have examined the effects of Tribulus terrestris (TT), a non-toxic herb, on sex reversal in the Convict Cichlid Cichlasoma nigrofasciatum with the aim of introducing a new environmentally friendly method for masculinization in C. nigrofasciatum. TT is a natural plant product that elevates the testosterone levels in humans and animals. Different concentrations (0.0, 0.10, 0.20 and 0.30 g/l) of TT extract were tested for their effect on sex reversal in C. nigrofasciatum by immersing newly hatched offspring once weekly for 2 months in TT extract. Of the dosages used in the present study, 0.30 g/l TT was the most effective in terms of masculinization, resulting in a maximum male ratio of 87.23% (P < 0.001). Sex ratios of 79 and 85% at 0.10 and 0.20 g/l TT, respectively, were also significantly different from the expected 1:1 ratio (P < 0.001). Histological examinations revealed that the testes of fish treated with TT extract contained all stages of spermatogenesis, clearly demonstrating that the administration of TT extract to C. nigrofasciatum stimulated spermatogenesis. Total survival rates in all treatments and the control were uniformly high, ranging from 88.57 to 90% (P > 0.05). We concluded that TT had no negative effect on the survival rate of C. nigrofasciatum. In addition, all groups of TT-treated fish exhibited successful growth acceleration compared to the control group, but only the 0.30 g/l TT treatment significantly improved the growth rate of C. nigrofasciatum. (P < 0.01). Sex reversal in C. nigrofasciatum demonstrated that TT-treated 0-day-old larvae showed successful sex reversal, spermatogenesis and a better growth rate than untreated progenies.     

Partial characterization of the digestive enzymes of Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> under culture conditions

The digestive enzyme activities of Pacific bluefin tuna Thunnus orientalis were evaluated for specific activity and characterized for pH and temperature optima in crude extracts of stomach, caecal mass, and proximal, middle and distal intestine. A higher level of alkaline proteolytic activity was detected in the caecal mass than in the proximal intestine. Total alkaline proteases, trypsin, chymotrypsin and leucine aminopeptidase (LAP) were tested. The temperature and pH analyses showed that proteolytic activity as well as lipase were maximal in the alkaline range, with a maximum at pH 9.0 and at temperatures between 35 and 60°C, except for the pepsin, which showed maximum activity at the same temperatures but in the acid range (pH 3.0). The α-amylase activity showed a broader range in activity, both for pH and temperature, with higher activity over the alkaline pH values and higher temperature. The lipase activity seems to be nondependent on bile salts under our assay conditions, resulting in a significant activity reduction in the presence of bile salts. This knowledge will allow the development of a gastrointestinal model (everted intestine) where food or feed will be hydrolysed with the fish’s own enzymes, a project that is being undertaken in our laboratory as a contribution to the development of novel diets for tuna fish.     

Conditions for the induction of some selective enzymes from <Emphasis Type="Italic">Bacillus subtilis</Emphasis> and their hydrolysis ability against mackerel and gracilar (asparagus, <Emphasis Type="Italic">Gracilaria verrucosa</Emphasis>)

ABSTRACT:   This study was conducted to optimize the cultivation conditions for Bacillus subtilis to produce proteases, amylases and cellulase, and to further investigate the hydrolysis ability against mackerel and asparagus. The extracellular enzymes from B. subtilis after 2 and 4 days incubation in a modified medium, containing 1% skim milk, 1% soya meal, 0.25% starch, 0.25% K₂HPO₄, 0.5% NaCl and 0.05% MgCl₂ were collected for the hydrolysis of asparagus and minced mackerel, respectively. Except for the α,α-diphenyl-β-picrylhydrazyl (DPPH) scavenging ability of the hydrolyzed asparagus, the trolox equivalent antioxidation capacity and DPPH scavenging ability of both samples increased significantly ( P  < 0.05) after 1 h hydrolysis and further increased during elongated hydrolysis at 50°C. Sodium dodecylsulfate–polyacrylamide gel electrophoresis indicated severe degradation of muscle proteins during hydrolysis. Changes in reducing sugar, soluble proteins and peptides before/after hydrolysis suggested the extracellular enzymes from B. subtilis could effectively hydrolyze the mackerel or asparagus, and subsequently improve their antioxidation ability.     

Effects of heavy metals in river waters in Japan on immobility and mortality of <Emphasis Type="Italic">Daphnia magna</Emphasis> and <Emphasis Type="Italic">Oryzias latipes</Emphasis> larvae

ABSTRACT:   Samples of river waters containing high concentrations of zinc and other heavy metals but low concentrations of other anthropogenic contaminants were collected to investigate the relationship between toxicity of heavy metals and naturally present organic matters or hardness, as well as the effects of heavy metals on aquatic organisms. Acute toxicity tests were conducted for the water samples using Daphnia magna and medaka Oryzias latipes . Almost all the D. magna died in river waters containing high concentrations of zinc, but O. latipes in the same waters were hardly affected. Since the test organisms were not only exposed to zinc but also other heavy metals in the river waters, we examined the toxicity using toxic units composed of zinc, copper, lead, and cadmium. The results of a bioassay with the river waters showed that the mortality of D. magna did not depend solely on the total number of toxic units of heavy metals. The organic matters and the hardness of the river waters could decrease the acute toxicity of zinc and other heavy metals to D. magna .     

Fortification of <Emphasis Type="Italic">niboshi</Emphasis> with histidine prepared from <Emphasis Type="Italic">katsuobushi</Emphasis> protein digest

ABSTRACT:   Free and total histidine contents in the muscle of various fishes and invertebrates caught in the Seto Inland Sea were estimated. These contents were higher in the muscles from skipjack, yellowfin tuna, yellowtail and mackerel than in those from lizard fish, sea bream and hairtail. Invertebrate muscles, except squid, contained low free and total histidine. Boiled and dried anchovy ( niboshi ) contained lower free histidine than that of dried anchovy (suboshi). To prepare free histidine-rich extracts, the waste residue of soup stock from smoke-dried and shaved skipjack ( katsuobushi ) was digested with proteases. The extract of the enzyme digest contained high free histidine and was used to fortify niboshi with free histidine. If taken, the fortified niboshi may suppress food intake, and prevent obesity.     

Determination of the quality parameters of pike perch <Emphasis Type="Italic">Sander lucioperca</Emphasis> caught by gillnet,longline and harpoon in Turkey

ABSTRACT:   The effects of the different catching methods (gillnet, longline, harpoon) on sensory, chemical (pH, total volatile base nitrogen, K -value) and microbiological (total viable count [TVC]) changes in pike perch Sander lucioperca stored in ice were investigated. The same soaking time was used for both gillnet and longline fishing. The catching method had considerable influence on the freshness quality of pike perch. The acceptable shelf life was 15 days for pike perch caught by gillnet, and 22 days for longline and harpoon. The initial concentrations of inosine monophosphate (2.4 μmol/g) in pike perch caught by gillnet were significantly lower ( P  < 0.05) than longline (4.1 μmol/g), and especially by harpoon (16.7 μmol/g). However, the initial K -values for fish caught by harpoon were significantly ( P  < 0.05) lower (24.36%) than fish caught by longline and gillnet (57.69%, 64.41%, respectively). The average K , Ki, G and H -values at rejection day in terms of sensory assessment were approximately 90, 98, 156 and 40%, respectively, for all catching methods during ice storage. However, TVC reached 7.0 log cfu/g after approximately 11 days of storage for fish caught by gillnet, 19 days for fish caught by longline and 8 days for fish caught by harpoon. The result of this study suggests that the best catching method for preserving the freshness of pike perch is longline, based on the data obtained from the sensory and microbiological analysis.     

Chemical composition of dried eyeballs from <Emphasis Type="Italic">Euphausia superba</Emphasis> and <Emphasis Type="Italic">Euphausia pacifica</Emphasis>

ABSTRACT:   Dried krill eyeballs were obtained from Euphausia superba and Euphausia pacifica by using a patented industry method and their chemical compositions were examined. Crude protein content was 77.7% and 80.8% of the dry matter of E. superba and E. pacifica , respectively. The dominating amino acids in both krill were glutamic acid, aspartic acid, glycine, arginine, leucine and tyrosine. Crude fat content was 10.9% and 5.4% of dry matter of E. superba and E. pacifica , respectively. The main lipid class of the extracted lipids was phospholipids at 88.5% in E. superba and 96.4% in E. pacifica . The dominating fatty acids in both krill were 22:6 ( n- 3), 20:5 ( n- 3), 16:0 and 18:1 ( n- 9). Astaxanthin (3, 3'-dihydroxy-β, β-carotene-4, 4'-dione) content of E. superba and E. pacifica was 566 mg/100 g and 252 mg/100 g of dry matter, respectively. Retinol of E. superba and E. pacifica was 153.0 mg/100 g (510 000 IU/100 g) and 57.6 mg/100 g (192 000 IU/100 g) of dry matter, respectively. The lipophilic extract of E. superba by using n -hexane contained 1923 mg/100 g of astaxanthin at approximately four times higher than the dried eyeballs.     

Sterols from bivalves <Emphasis Type="Italic">Calyptogena soyoae</Emphasis> and <Emphasis Type="Italic">Bathymodiolus septemdierum</Emphasis> living in deep sea

From the two species of bivalves, Calyptogena soyoae around a cold seep and Bathymodiolus septemdierum near hydrothermal vents in the sea, sterols were isolated using high-pressure liquid chromatography analysis of the lipid fraction guided by characteristic ¹H NMR signals of the sterol skeleton. The minor sterol composition of C. soyoae included 24-methylenecycloartanal, cycloeucalenol, and obutusifoliol, which are known phytosterols. From B. Septemdierum, lathosterol and cholesterol as main sterols together with more diverse sterols were obtained. The difference between these species and their sterol contents is most likely because of feeding modes and metabolism of nutrients from their habitat.     

Effects of fish CYP inducers on difloxacin <Emphasis Type="Italic">N</Emphasis>-demethylation in kidney cell of Chinese idle (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>)

A drug–drug interaction occurs when the effect of one drug is altered by the presence of another drug which is generally associated with the induction of cytochrome P450s (CYPs) activity. Thus, unexpected treatment failures often happen resulting from inappropriate coadministration in fisheries. However, little information is available about CYP induction in fish. The reaction of difloxacin (DIF) biotransformation to sarafloxacin (SAR) belongs to N-demethylation catalyzed mainly by CYP(s). In order to supply useful information on CYP induction, the present study assessed the effects of fish-specific CYP inducers on DIF N-demethylation and enzyme kinetics in kidney cell of Chinese idle (CIK; grass carp (Ctenopharyngodon idellus)) by RP-HPLC. Results demonstrated that the amounts of SAR formation and enzymatic parameters Clint and Vmax were significantly increased due to β-naphthoflavone (BNF) pretreatment. Therefore, we suggest that CYP1A may be involved in DIF N-demethylation in CIK. This study provides instructive information to ensure treatment success via avoiding CYP induction in fisheries.     

Effect of dietary <Emphasis Type="SmallCaps">l</Emphasis>-tryptophan on cannibalism,survival and growth in pikeperch <Emphasis Type="Italic">Sander lucioperca</Emphasis> (L.) post-larvae

The effect of supplemented commercial diets with crystalline l-tryptophan (TRP—5, 10 and 20 g TRP kg^?1) on cannibalism, survival and growth parameters of pikeperch post-larvae (Sander lucioperca) was evaluated. Fifteen-day-old pikeperch larvae (mean weight—6.8 mg) were reared during the next 28 days (20.5 °C, 16L:8D) in glass aquaria in a recirculating aquaculture system. The enzyme-linked immunosorbent assay showed that TRP-supplemented diets were effective in increasing the levels of serotonin (5-HT) in the body tissue of pikeperch. TRP supplementation resulted in a slight decrease in both types of cannibalism, although the reduction in cannibalism impact did not amount to more than a few percent. TRP treatment had no significant influence on the final survival of pikeperch post-larvae (ranged from 20.1 ± 12.4 to 29.0 ± 12.9 %). However, contrary to the earlier studies conducted on other fish species, no significant difference in the growth rates and feeding behavior of pikeperch between TRP-fed and control group were found. The final body weight and growth rate ranged from 0.211 ± 0.014 to 0.243 ± 0.016 and from 12.19 ± 0.38 to 12.76 ± 0.35 % day^?1, respectively. To our knowledge, this is the first study reporting the effects of TRP supplementation on the cannibalism–survival–growth relations in fish in the post-larval stage.     

Optimum temperature and salinity conditions for growth of green algae <Emphasis Type="Italic">Chlorella ellipsoidea</Emphasis> and <Emphasis Type="Italic">Nannochloris oculata</Emphasis>

ABSTRACT:   The effects of temperature and salinity on growth of green algae Chlorella ellipsoidea and Nannochloris oculata were determined to compare the optimum culture conditions. A four-temperature (15, 20, 25, and 30°C) × three-salinity (10, 20, and 30) factorial design with triplicates was applied. Specific growth rate (SGR), maximum density, and duration to reach maximum density of C. ellipsoidea were significantly affected by both temperature and salinity. The highest SGR was observed in C. ellipsoidea at 25°C and salinity 10, but the maximum density was very low. The highest maximum density was achieved in C. ellipsoidea at 15°C and 10. The slope constant of the linear relationship between semilogarithmic growth of C. ellipsoidea and day of culture was highest at 15°C and 10. The SGR and duration to reach maximum density of N. oculata were significantly affected by both temperature and salinity. However, maximum density of N. oculata was significantly affected by temperature, but not salinity. The highest maximum density was achieved in N. oculata at 25°C and 30, but SGR was significantly lower than that of N. oculata at 25°C and 10. The slope constant of the linear relationship between semilogarithmic growth of N. oculata and day of culture was highest at 25°C and 30. Based on these results, the condition of 15°C and salinity 10 seemed to be optimal for maximum density of C. ellipsoidea , and the condition of 25°C and 10 and 30 for SGR and maximum density for N. oculata , respectively.     

Effects of illumination on early life development and digestive enzyme activities in common pandora Pagellus erythrinus L. larvae

The activities of main digestive enzyme (proteases, amylase and lipase) and animal husbandry (mainly growth and survival) were studied in common pandora Pagellus erythrinus larvae until 30 days after hatching (DAH). Three different illumination levels (10, 30 and 100 lx) were compared in triplicate and green water technique was carried out. At the end of the experiment, illumination did not affect neither survival nor growth except 10 lx treatment. Similarly, specific growth rate (SGR) was different in 10 lx treatment (p < 0.05), although no differences were found in group 30 and 100 lx treatments (p > 0.05). In all groups, trypsin and chymotrypsin specific activities were firstly detected on day 3 related with mouth opening and slightly increased until 20 and 25 DAH respectively, and after this date specific activities of those decreased. Although, there was a significant difference between 10 lx treatment and other experimental groups (p < 0.05), no differences were found in other treatments (p > 0.05). Pepsin was firstly detected on day 25 related with stomach formation and sharp increase was observed until 30 DAH and then slight decrease was measured from this date and no differences were found between all groups. Amylase was firstly determined on day 2 and increased to day 5. After this date, slight decreases were measured in all groups and continued until end of experiments. The highest specific activity of amylase was determined in 30 lx treatment and no significant differences were found between groups (p > 0.05). Lipase was firstly detected on day 4 and increased to day 10. Then, activity of lipase decreased until day 15 and increased again until 25 DAH. Slight decreases were found in all groups until day 30 and continued to end of experiments. No significant differences were detected among groups (p > 0.05).

Finally, the significant improvement in survival, larval development and specific enzyme activities of larvae were determined in 30 lx treated group. It is thought that this phenomenon is related to optimal keeping conditions provided by the medium illumination level for Pagellus erythrinus larvae.     

Dynamics of Total Body Fatty Acids During Early Ontogeny of Pikeperch (<Emphasis Type="Italic">Sander lucioperca</Emphasis>) Larvae

The fatty acid composition of pikeperch (Sander lucioperca) was determined according to their physiological status, during starvation (10 days) and feeding (28 days). In starved larvae, polyunsaturated, monounsaturated and saturated fatty acids were utilized as metabolic substrates until day 9. At day 10, all fatty acid levels remained stable or, at least, increased in larval body. Among fatty acids, docosahexaenoic acid 22:6 n-3 was used preferentially (20.3% from total fatty acids utilized) followed by palmitoleic acid 16:1 n-7 (13.9%) and then by oleic 18:1 n-9 (12.3%), linoleic 18:2 n-6 (10.1%), linolenic 18:3 n-3 (9.7%) and eicosapentaenoic 20:5 n-3 (9.1%) acids. On the other hand, arachidonic acid 20:4 n-6 was utilized very lowly (0.3%). In fed larvae, no utilization of body fatty acids was observed during the experiment. It seems that energy requirements (and others) of fed larvae were satisfied by feed.     

Suspension feeding and growth of ark shell <Emphasis Type="Italic">Anadara granosa</Emphasis>: comparison with ubiquitous species <Emphasis Type="Italic">Scapharca subcrenata</Emphasis>

The ark shell Anadara granosa is a species peculiar to the Ariake Sound. To determine why distribution of A. granosa in Japan is largely confined to this area, we examined feeding and growth of A. granosa as functions of environmental and biological variables. The results were compared with those of another ark shell Scapharca subcrenata, which is ubiquitous in Japan. Feeding experiments indicated that A. granosa is eurythermal and euryhaline, as is S. subcrenata, but is adapted to temperature slightly higher than S. subcrenata. Weight-specific clearance rate (CR) of A. granosa as a function of soft-body dry-weight (w) followed the power function of w (CR=2.7×w ^−0.37), with coefficient and exponent very close to those for S. subcrenata. Growth rate of A. granosa increased linearly with daily ration, similar to S. subcrenata. Thus, feeding and growth characteristics of A. granosa were comparable to those of S. subcrenata and no ‘peculiarities’ of the former were detected. Therefore, the factors that make A. granosa a species restricted to the sound are probably not directly related to feeding or growth characteristics.     

Tench (<Emphasis Type="Italic">Tinca tinca</Emphasis> L.) larvae rearing under controlled conditions: density and basic supply of <Emphasis Type="Italic">Artemia</Emphasis> nauplii as the sole food

After artificial reproduction of tench, larvae must be maintained indoors, and studies on rearing conditions are needed, focussing on the reduction of labour and costs. Three experiments on larvae (5th day post-hatch) were conducted for 25 days using Artemia nauplii as the sole food in order to determine basic feeding and density conditions during the first rearing period. Tench were maintained in 25 l fibreglass tanks, supplied with an artesian water flow throughout of 0.2 l min⁻¹. Water temperature was 22.5 ± 1°C, and the photoperiod was natural. Larvae fed on a restricted amount of nauplii reached high survival rates, even with the minimum of 50 nauplii larva⁻¹ day⁻¹. This amount of food may be sufficient at least for the first 25 days of exogenous feeding if fast growth is not the priority, and high densities can be maintained with good survival rates (over 90% up to 160 larvae l⁻¹ and 77% with 320 larvae l⁻¹). When food was supplied in excess once a day, high survival rates were achieved (91–97%), without differences among the densities tested. Animals at a density of 100 l⁻¹ reached the highest length (15.57 mm) and individual weight (46.8 mg). This growth is greater than those reported in studies feeding several times a day. It could be deduced that, while live food remains available for tench, it is not necessary to feed so frequently. Considering the relationship among the initial number of animals, final survival and growth and ration supplied, the new data reported here are useful to establish suitable stocking densities under both culture and experimental conditions.     

Insights into kisspeptin- and leptin-signalling on GnRH mRNA expression in hypothalamic organ cultures of immature pikeperch <Emphasis Type="Italic">Sander lucioperca</Emphasis>

Two types of gonadotropin-releasing hormones (GnRH) were identified as gnrh1 and gnrh2 in pikeperch Sander lucioperca. The administration of rodent leptin on hypothalamic organ cultures of immature pikeperch resulted in significantly elevated levels of gnrh2, but not in gnrh1 mRNA, whereas kisspeptin-10 administration did not affect gnrh1 or gnrh2 expression. These results represent preliminary insights into leptin-GnRH-signaling on a hypothalamic level in fish, potentially coupling fat metabolism and the activation of the reproductive axis during puberty. Mammalian leptin and kisspeptin-10, however, failed to initiate a consistent response in pikeperch and their use cannot be recommended.     

Blood vessel melanosis: a physiological detoxification mechanism in Atlantic cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>)

Melanin deposits in white muscle represent a potentially serious quality problem for farmed cod. Black lines of deposited melanin are associated with blood vessels and consist of layers of melanin-filled cells. Despite numerous anecdotal reports of melanosis in farmed cod no detailed investigations into the underlying mechanisms have been undertaken. In this study we have confirmed that melanosis is present in both farmed and wild cod. However, the incidence of melanosis in wild fish is much lower than that in farmed fish. In addition, we have demonstrated that tyrosinase, a copper-dependent metalloenzyme that is the primary enzyme of melanin synthesis, is present in cod tissues. Copper levels are significantly different in the white muscle of farmed (0.5 ± 0.03 mg/kg wet weight) and wild (0.34 ± 0.01 mg/kg wet weight) cod (P < 0.05) and in the blood vessels of farmed (2.23 ± 0.37 mg/kg wet weight) and wild (0.32 ± 0.02 mg/kg wet weight) cod (P < 0.05). Similarly, differences in the levels of tyrosinase activity were observed in both muscle and blood vessels. Melanised tissues contain higher copper concentrations than tissue containing little or no melanin. Furthermore, we have shown that the addition of copper to protein extracts from fish muscle causes an increase in tyrosinase activity in vitro. Our results suggest that overabundance of copper in commercial feeds leads to increased melanin synthesis via increased tyrosinase activity. We propose that melanin is deposited at sites of high copper exposure and subsequently sequesters copper as a means of reducing the overload stress on other homeostatic mechanisms.