Stress mitigating and growth enhancing effect of dietary tryptophan in rohu (Labeo rohita,Hamilton, 1822) fingerlings

An experiment was conducted to study the stress mitigation and growth enhancing role of dietary l-tryptophan (TRP) under thermal stress in rohu, Labeo rohita fingerlings for 45 days. Seven hundred and twenty fishes were distributed in three major groups that are ambient temperature (26 °C), 34 and 38 °C in triplicate following a complete randomized design. Acclimation of fishes to 34 and 38 °C over average ambient temperatures were carried out at 1 °C/day. Each group was fed with a diet supplemented with 0, 0.36, 0.72 or 1.42 % l-TRP. Results showed that blood glucose and serum cortisol level were found to be significantly higher (p < 0.05) in the higher temperature groups than the ambient temperature group. Similarly, aminotransferase, lactate dehydrogenase, malate dehydrogenase, CAT, superoxide dismutase activities were found to be significantly higher (p < 0.05) in the control groups (0 % l-TRP) and decreasing activities of these enzymes were observed with the increasing level of dietary l-TRP. In different temperature groups, l-TRP-supplemented groups were found to have higher (p < 0.05) growth, RGR and PER. The results obtained in the present study indicate that dietary l-TRP mitigates thermal stress and enhances growth. From the present study, we can conclude that dietary supplementation of l-TRP at the 0.72 % level in the diet is found to be optimum to reduce thermal stress even up to 38 °C in rohu, L. rohita. The baseline data obtained here could be useful for the farmers to formulate feeds to culture the fish in different agro-climatic zones.     

Detection of the first lectin with antimicrobial activity present in serum of the Amazonian fish tambaqui Colossoma macropomum

Tambaqui Colossoma macropomum is the most important Amazonian native species in South American aquaculture. Innate immunity at least partially depends on the recognition of pathogen-associated molecular patterns by receptor-recognizing pathogens (PRRs). Some PRRs have been characterized in fishes, and several studies have focused on the role of lectins in the immune system of various fishes. Lectins are proteins that specifically recognize carbohydrates and which have important biological functions. Tambaqui serum lectin (ComaSeL), which was identified on the basis of its hemagglutinating activity, was pre-purified, biochemically characterized, and used in assays of antibacterial activity against pathogenic bacteria in freshwater fishes. A study of the seasonality of this lectin was performed. Comasel activity was stable at a pH between 4.0 and 9.0 and lost 100?% of its activity at 70?°C. It recognized the carbohydrates d-galactose, 1-O-methyl-d-galactopyranoside, and d-fucose, showing antibacterial activity for Gram-negative bacteria. Its activity showed significant differences between the summer and winter (p?<?0.05, Tukey test), thereby corroborating observations that tambaqui becomes more susceptible to mortality from diseases caused by bacteria and fungi during the winter. With this information, new tools may be developed for gaining a better understanding of the role of these proteins in the immune system of the tambaqui, ultimately resulting in the improved management of this fish by pisciculturists.     

Dynamics of free amino acids in the hemolymph of Pacific whiteleg shrimp Litopenaeus vannamei exposed to different types of stress

We analyzed the dynamics of amino acids, ammonia-N, and carbohydrates in hemolymph when Pacific whiteleg shrimp Litopenaeus vannamei was subjected to air-exposure and low-salinity stresses. Glycine, l-arginine, and d- and l-alanine levels in hemolymph were increased under both stress conditions. Ammonia-N, a product of amino acid catabolism, also increased in level. These results suggest that the above-mentioned amino acids are used as energy sources. Levels of total carbohydrates, the sum of glucose and other carbohydrates, in the hemolymph showed different dynamics between the two types of stress and were not always high. This suggests that other energy sources, such as amino acids, are important when animals are subjected to stress. It is possible that particular amino acids act as energy sources under various stress conditions.     

Purification and characterization of trypsin from the pyloric ceca of orange-spotted grouper, Epinephelus coioides

Trypsin from the pyloric ceca of orange-spotted grouper, Epinephelus coioides, was purified by fractionation with ammonium sulfate, ionic exchange, and affinity chromatography. The protein was purified 161.85-fold with a yield of 4%. Purified trypsin had an apparent molecular weight of 24 kDa according to an SDS-PAGE analysis. Optimal profiles of temperature and pH of the enzyme were 50°C and 8–10, respectively, using Nα-benzoyl-l-arginine ethyl ester as the substrate. The results of thermal and pH stability assays showed that the enzyme was stable at temperatures of up to 50°C and in the pH range of 6–8. Trypsin activity decreased with an increasing NaCl concentration (0–0.6 M). The activity of purified trypsin was effectively inhibited by a soybean trypsin inhibitor and N-p-tosyl-l-lysine chloromethyl ketone, and was slightly inhibited by iodoacetic acid, ethylenediaminetetraacetic acid, 1-(l-trans-epoxysuccinyl-leucylamino)-4-guanidinobutane, and pepstatin A. Protein identification of the purified protease showed that the sequences of two peptides, LGEHNI and NLDNDIML, were highly homologous to other fish trypsins. The measurement of trypsin activity in different tissues showed that the highest activity was detected in pyloric ceca, followed by anterior intestine, middle intestine, hind intestine and spleen, but very low activities were found in other tissues. An inverse relationship between the trypsin activity in four tissues of pyloric ceca, anterior intestine, middle intestine and hind intestine and fish body weight as a result of increased pepsin in stomach indicated grouper growth status was increased.     

ATP depletion inhibits retraction of chromatophores induced by GABA in cephalopod Todarodes pacificus

Color changes in cephalopods are generated by the expansion or retraction of chromatophores located under the dermis. The behavior of the chromatophores is regulated by neurotransmitters; l-glutamate (l-Glu) is an excitatory transmitter that causes the chromatophores to expand. To date, serotonin [5-hydroxytryptamine (5-HT)] is the only neurotransmitter known to stimulate retraction of chromatophores. We found that the chromatophores in the Japanese squid Todarodes pacificus were regulated by γ-aminobutyric acid (GABA), an inhibitory neurotransmitter, and that GABA caused expanded chromatophores to retract. We also found that adenosine 5′-triphosphate (ATP) concentrations in skin samples remained stable at their initial values for more than 24 h after the death of each squid; therefore, the chromatophores could respond to both l-Glu and GABA during that period. Furthermore, we attempted to reduce the levels of ATP by storing skin sample in sodium azide solution. The chromatophores in sodium azide-treated skin samples were induced to expand by l-Glu, but these expanded chromatophores could not be induced to retract by GABA. Based on these observations, we conclude that ATP is essential for retraction, but not expansion, of chromatophores.     

The influence of \upbeta-hydroxy- \upbeta-methylbutyrate (HMB) on cell-mediated immunity in tench Tinca tinca (L.): in vitro and in vivo study

$\upbeta$ -Hydroxy- $\upbeta$ -methyl butyrate (HMB) has been shown to counteract many of the negative effects of intensive fish production methods and results in increased growth and protection against diseases. In the present study, the in vitro and in vivo effect of HMB on the immunocompetence cell activity in tench Tinca tinca (L.) was examined. In the in vitro study pronephric phagocytes and lymphocytes were isolated from the fish and grown in culture medium (RPMI-1640) containing 0, 0.1, 1, 5, 10, 25, 50 or $100\,\upmu$ g HMB cm^?3 of medium. The effects of HMB on the respiratory burst activity (RBA), the potential killing activity (PKA) and lymphocyte proliferation stimulated by concanavalin A (ConA) or lipopolysaccharide (LPS) were examined. The in vitro study showed that HMB increased the RBA and PKA of the phagocytes, compared to the medium over that of cells grown without HMB. Lymphocyte proliferation stimulated by ConA and LPS was also increased approximately when HMB was added to the culture medium at concentrations between 10 and $100\,\upmu$ g HMB cm^?3. In the in vivo study fish were fed daily with pellets containing HMB at doses 0, 10, 25 and 50 mg kg^?1 body weight day^?1. The study showed that HMB statistically increased the RBA and PKA and highest activity at 50 mg kg^?1 body weight day^?1 were observed. Also lymphocyte proliferation stimulated by ConA and LPS were maximally stimulated at dose 50 mg kg^?1 body weight day^?1. In conclusion, the current study shows that HMB could potentially improve immunocompetence cell activity in tench through increased cell proliferation and functionality.     

Embryonic development and influence of egg density on early veliger larvae and effects of dietary microalgae on growth of brown mussel Perna perna (L. 1758) larvae under laboratory conditions

The effect of egg density on embryonic development and larval quality as well as the lipid and fatty acid contents (eicosapentaenoic acid, EPA; docosahexaenoic acid, DHA) of cultured microalgae fed to Perna perna larvae was studied under controlled conditions to provide information needed for development of an experimental hatchery. Embryonic development followed the common sequence exhibited by other bivalves. d-larva stage was attained 40–44 h post-fertilisation at 21 ± 1 °C. The umbo-stage was reached in 11 days, and pediveliger larvae were observed 26 days post-fertilisation. Low egg density (range 20–100 eggs cm^?2) produced high proportions of normal d-larvae. Larval development showed two growth phases: 1st—the mixotrophic stage and 2nd—the exotrophic stage where the composition of diets had significant effects on larval growth with higher rates in larvae fed with the mixed microalgae (Isochrysis galbana + Chaetoceros calcitrans, I. galbana + Phaeodactylum tricornutum and I. galbana + Skeletonema costatum) in comparison with the monospecific diet (I. galbana). Fatty acid analysis showed that larval growth and survival were strongly influenced by proportions of dietary DHA and EPA. These results indicate that DHA and EPA are the key factors in determining larval performance, considerably more than the total amount of other fatty acids.     

Inactivation kinetics of formaldehyde on N-acetyl-β-d-glucosaminidase from Nile tilapia (Oreochromis niloticus)

Formaldehyde is a widely used sanitizer in aquaculture in China, while the appropriate concentration is not available to be used effectively and without damage to tilapia much less to its reproductive function. N-acetyl-β-d-glucosaminidase (EC 3.2.1.52, NAGase), hydrolyzing the oligomers of N-acetyl-β-d-glucosamine into monomer, is proved to be correlated with reproduction of male animals. In this paper, NAGase from spermary of tilapia was chosen as the material to study the effects of formaldehyde on its activity in order to further investigate the effects of formaldehyde use on tilapia reproduction. The results showed the relationship between the residual enzyme activity and the concentration of formaldehyde was concentration dependent, and the IC₅₀ value was estimated to be 3.2 ± 0.1 %. Appropriate concentration of formaldehyde leaded to competitive reversible inhibition on tilapia NAGase. Moreover, formaldehyde could reduce the thermal and pH stability of the enzyme. The inactivation kinetics of formaldehyde on the enzyme was studied using the kinetic method of substrate reaction. The inactivation model was setup, and the rate constants were determined. The results showed that the inactivation of formaldehyde on tilapia NAGase was a slow, reversible reaction with partially residual activity. The results will give some basis to determine the concentration of formaldehyde used in tilapia culture.     

Culture-independent analysis of the bacterial community during fermentation of pa-som,a traditional fermented fish product in Laos

Pa-som is a traditional salty fermented food made from varieties of freshwater fish with garlic and rice in Laos. A similar product, plaa-som, is made in Thailand. To investigate the fermentation process of pa-som, lactic acid production and pH were monitored during laboratory-scale production of pa-som. Furthermore, the variety of bacteria involved and their succession during fermentation were examined using culture-independent polymerase chain reaction (PCR) denaturing gel gradient electrophoresis targeting the 16S ribosomal RNA (rRNA) gene. d- and l-lactic acid production and a decrease in pH were observed in the samples after 1 day of fermentation. The lactic acid content and pH continued to increase and decrease, respectively, until 4 days of fermentation. Overall, six lactic acid bacteria and eight other indigenous bacteria species were detected during pa-som fermentation. Among the lactic acid bacteria detected, two Lactococcus and one Weissella species were detected in samples after 1–4 days of fermentation, suggesting that these three species play major roles in pa-som fermentation from the initial phase. This is the first report to investigate the fermentation process in pa-som at the microbial level. This research approach should be applicable to a wide variety of pa-som and related fermented fishery products in Laos to help understand their microbial diversity and to identify beneficial bacterial species for improving quality.     

Role of catecholamines and nitric oxide on pigment displacement of the chromatophores of freshwater snakehead teleost fish,Channa punctatus

We are reporting for the first time that the catecholamines (adrenaline and noradrenaline) inhibit the effect of nitric oxide (NO) on melanosome dispersion in freshly isolated scales of the freshwater snakehead fish, Channa punctatus. We studied the effect of NO and catecholamines on the pigment displacement by observing the changes in the melanophore index. The scales when treated with solution containing NO donor sodium nitroprusside (SNP) showed dispersion of melanosomes, whereas NO synthase blocker N-omega-Nitro-l-arginine suppresses this action of SNP. Treatment with adrenaline and noradrenaline on the isolated scales caused aggregation of melanosomes. Scales treated with solution containing catecholamines and SNP resulted in aggregation of melanosomes suggesting that catecholamines mask the effect of SNP. These results suggest that the catecholamines are inhibiting the effect of NO and causing the aggregation of the melanosomes may be via surface receptors.     

Novel method for improving the antioxidative properties of fish meat by direct injection of sodium l-ascorbate into the blood vessels of live fish

The color of the red muscle of pelagic fish is used as an indicator of freshness, and sustaining a bright red color is important for maintaining the commercial value of pelagic fish. Feeding fish with antioxidant compounds can delay metmyoglobin (metMb) formation, but the process requires long-term feeding. Live cultured Japanese horse mackerel Trachurus japonicus were used in this study. After anesthetization, 2 ml of blood was drawn from blood vessels, and a parenteral solution including an antioxidant compound was injected. Browning, metMb formation, and lipid oxidation of dark muscle during chilled storage were delayed by injecting sodium l-ascorbate as an antioxidant compound in the blood vessels of live fish.     

Digestive enzyme activity in juvenile Nile tilapia (Oreochromis niloticus, L) submitted to different dietary levels of shrimp protein hydrolysate

The effect of different dietary concentrations of shrimp protein hydrolysate (SPH) on digestive enzyme activity of Nile tilapia juveniles was evaluated. SPH concentrations in diets were 0, 15, 30 and 60 g kg^?1 (treatments SPH0, SPH15, SPH30 and SPH60, respectively). Hemoglobin, azocasein, BApNA (Nα-benzoyl-dl-arginine-p-nitroanilide), SApNA (Suc-Ala-Ala-Pro-Phe p-nitroanilide), aminoacyl of β-naphthylamide and starch were used as substrates for enzyme activity determinations. The activity of total alkaline protease was significantly higher (P < 0.05) in fish under SPH15 and SPH60 treatments than in the control (SPH0). However, the effect was not dose-dependent. Substrate-SDS-PAGE was also performed to evaluate changes in the profile of Nile tilapia digestive proteases caused by SPH. Substrate-SDS-PAGE revealed 12 active proteolytic bands, eight of which responded to SPH dietary incorporation. Inhibition substrate-SDS-PAGE indicated a decrease in the activity of three enzymes, with trypsin activity decreasing with the increase of SPH concentration, whereas the opposite occurred for two aminopeptidases. Distinct protease profiles were also found for each treatment, suggesting adaptability of digestive proteases from Nile tilapia to the different diets.     

The optimal dietary level of ascorbic acid for juvenile Pacific bluefin tuna, Thunnus orientalis

A feeding experiment was conducted to evaluate the efficacy of, and determine the requirements for dietary AsA (l-ascorbic acid) using its popular derivative l-ascorbyl-2-monophosphate-Mg (APM) for juvenile Pacific bluefin tuna (PBT). The five test diets (with APM-0, -400, -800, -1200 and -1600) were prepared by adding 0, 400, 800, 1200 and 1600 mg of APM kg diet^?1, respectively. These diets, together with chopped sand lance (the control diet), were administered, in duplicate to 0.27 g PBT at 25 days after hatching (300 juveniles per 15 m³ tank). The PBT were fed to apparent satiety 6 times a day for 2 weeks. While final body weight, feed conversion and survival showed no differences among diets APM 400–1600, PBT fed APM-0 indicated lower growth performance together with anorexia, dark pigmentation and ataxia from 5 days and reached 80 % mortality at 12 days after the start of the feeding trial. In addition, liver and brain ascorbic acid (AsA) concentrations increased with increasing dietary APM levels, the values were similar in PBT fed diets with APM-1200 and APM-1600, and liver and brain concentrations of AsA were significantly higher in these fish than in the PBT fed the other diets (P < 0.05). The PBT responded to increasing dietary APM, and the fish fed APM-1200 had the highest weight gain, protein and lipid content. The data indicate that PBT can utilize APM as an AsA source and suggest that dietary supplementation with APM-1200 (454 mg AsA kg^?1 diet) would result in satisfactory growth.     

Microbiological characteristics of Vibrio scophthalmi isolates from diseased olive flounder Paralichthys olivaceus

In 2005, massive mortality occurred in olive flounder Paralichthys olivaceus farms in Korea, and five isolates were collected from diseased fish. In this study, microbiological and pathogenic characteristics of these isolates were studied. The isolates gave negative results in lysine and ornithine decarboxylase, ortho-nitrophenyl-??-galactoside, and citrate tests, and positive results in urease, esculinase, and nitrate reduction tests. The isolates produced acid from adipate, fructose, d-glucose, and maltose, and gave positive results in alkaline phosphatase, esterase lipase, leucine arylamidase, and naphthol-AS-BI-phosphohydrolase. According to genetic analysis, 16S rRNA gene sequences showed 98?C100?% identity with both Vibrio scophthalmi and V. ichthyoenteri. The dnaJ gene sequences presented a higher identity with V. scophthalmi than with V. ichthyoenteri. Thus, the isolates were identified as V. scophthalmi. Pathogenicity of the five isolates in olive flounder was different and LD₅₀ values were from 10⁶ to 10⁸?CFU/g fish. Symptoms included darkening of skin, hemorrhage of liver and intestine, ascites, and distended abdomen. Histopathological changes included hemopoiesis dilatation and epithelial hyaline droplets in kidney, macrophage infiltration and ellipsoid dilatation in spleen, vascular dilatation, submucosal edema, and serosa inflammation of intestine. Cumulative mortality was 25?% for fish singly infected by isolate A19008 or Streptococcus parauberis, and increased to 87.5?% in super-infection group with these two pathogens.     

Sperm characteristics and androgens in Acipenser ruthenus after induction of spermiation by carp pituitary extract or GnRHa implants

Spermiation and changes in androgen (testosterone, T and 11-ketotestosterone, 11-KT) levels were studied in sterlet (Acipenser ruthenus) treated with GnRH agonist implants (dAla⁶-Pro⁹-LHRHa) at 25 and 75?μg?kg^?1 b.w. and compared with those males treated with 4?mg?kg^?1 b.w. of carp pituitary extract (CPE) and 3 pellets of Ovopel kg^?1 b.w., which contains dAla⁶-Pro⁹NEt-mGnRH and metoclopramide. Sperm quality (sperm mass, spermatozoa concentration and sperm motility and velocity) was evaluated 24, 48 and 72?h after hormonal treatments. Males did not release sperm in the control group injected with physiological solution, while sperm could not be collected 7?days after treatments in all hormonally treated groups. Spermiation rates were 100?% in the CPE and Ovopel groups and 25–50?% in the GnRHa-treated groups. Sperm production was significantly lower in the GnRHa-treated groups than in the CPE and Ovopel groups and decreased 72?h after hormonal treatment. Sperm motility and velocity were higher in the Ovopel and GnRHa (75?μg) groups compared to the CPE and GnRHa (25?μg) groups and decreased 72?h after hormonal treatment. Androgens were only affected in spermiating males and changed in the Ovopel and GnRHa (75?μg) after hormonal treatment. Significant correlations were observed between sperm production, sperm motility and sperm velocity, but not androgens. The present study suggests involvement of dopamine in sturgeon spawning. Additionally, better sperm quality observed in the Ovopel group and particularly sperm motility in the GnRHa (75?μg) suggests enhancement of sperm quality in sturgeon treated with GnRHa. Therefore, further study is needed to induce fully spermiation using GnRHa implants in combination with a dopamine inhibitor.     

In vitro effect of levamisole on the cell viability,phagocytosis and respiratory burst of Barbel chub (Squaliobarbus curriculus) macrophages

The present study was to determine in vitro effect of levamisole on the immune functions of Barbel chub (Squaliobarbus curriculus), head–kidney‐derived (HKM), spleen‐derived (SM) and peripheral blood monocyte‐derived macrophage (PBM). Macrophages were incubated with levamisole 10³, 10¹, 10⁻¹ and 10⁻³ ng mL⁻¹ to assay the cell viability, respiratory burst and phagocytosis. The results showed that macrophages treated with levamisole 10⁻³ ng mL⁻¹ gave a maximum respiratory burst response, whereas levamisole 10³ ng mL⁻¹ had no effect. Phagocytosis activity of the macrophages treated with levamisole enhanced significantly when compared to control, maximum response being at 10⁻³ ng mL⁻¹. While using the methylthiazoletetrazolium method to measure the cell activity for 12, 24, 36, 48, 60 h, there was no significant role on proliferation and the cell viability began to decline after 24 h. In conclusion, levamisole is a potent enhancer of Barbel chub macrophage activity with low concentration.     

Purification and characterization of chymotrypsin from viscera of vermiculated sailfin catfish, Pterygoplichthys disjunctivus, Weber, 1991

Pterygoplichthys disjunctivus viscera chymotrypsin was purified by fractionation with ammonium sulfate (30–70 % saturation), gel filtration, affinity, and ion exchange chromatography. Chymotrypsin molecular weight was approximately 29 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), shown a single band in zymogram. Electrofocusing study suggested being an anionic enzyme (pI ≈ 3.9), exhibiting maximal activity at pH 9 and 50 °C, using Suc-Ala-Ala-Pro-Phe-p-nitroanilide (SAAPNA) as substrate. Enzyme was effectively inhibited by phenyl methyl sulfonyl fluoride (PMSF) (99 %), and N-tosyl-l-phenylalanine chloromethyl ketone (TPCK) (94 %). Enzyme activity was affected by the following ions in decreasing order: Hg²⁺, Fe²⁺, Cu²⁺, Li¹⁺, Mg²⁺, K¹⁺, Mn²⁺, while Ca²⁺ had no effect. Chymotrypsin activity decreased continuously as NaCl concentration increased (from 0 to 30 %). K _m and V _max values were 0.72 ± 1.4 mM and 1.15 ± 0.06 μmol/min/mg of protein, respectively (SAAPNA as substrate). Results suggest the enzyme has a potential application where low processing temperatures are needed, such as in fish sauce production.     

The production of a lymphokine (macrophage activating factor) by rainbow trout, Oncorhynchus mykiss (Walbaum), leucocytes stimulated with the extracellular products of Mycobacterium sp.

The production of macrophage activation factor (MAF) by rainbow trout, Oncorhynchus mykiss (Walbaum) , head kidney leucocytes was examined after culturing in vitro with extracellular products (ECP) collected from Mycobacterium sp. Cultures of leucocytes were prepared from naive fish, or fish previously vaccinated with either the ECP or with formalin killed whole cell preparations (WC) of the bacterium. The cells were then incubated with the ECP in vitro and the ability of their supernatants to activate macrophages assessed. Macrophages from control fish were incubated with the supernatants, and their ability to reduce nitroblue tetrazolium (NBT) measured as an indicator of macrophage activation. Incubation of head kidney macrophages from naive fish directly with 1, 10 or 100 μg mL^–1 of ECP for 48 h significantly enhanced macrophage activation compared with control macrophages. Vaccination of fish with either ECP or WC had no significant effect on the respiratory burst of control macrophages 4 weeks post-vaccination. By the eighth week, however, absorbance levels of respiratory burst reflecting both the primary (cells from vaccinated fish cultured in vitro with PBS) and the secondary (cells cultured in vitro with ECP) MAF responses of fish vaccinated with ECP and WC, had peaked and these were significantly different from the non-vaccinated controls. This activity had fallen to levels similar to control fish by week 12 for fish vaccinated with WC.     

Age, growth, and reproductive characteristics of dolphinfish Coryphaena hippurus in the waters off west Kyushu, northern East China Sea

The growth and reproductive characteristics of dolphinfish Coryphaena hippurus collected in the waters off western Kyushu from May 2008 to April 2011 were determined based on scale and otolith readings and gonad histological examinations, respectively. Based on annual increments in scales and daily increments in sagittal otoliths, the von Bertalanffy growth curves in male and females were determined as $ FL_{t} = 1049[1 - \exp \{ - 0.835(t + 6.975 \times 10^{ - 14} )\} ] $ and $ FL_{t} = 938[1 - \exp \{ - 1.029(t + 6.975 \times 10^{ - 14} )\} ] $ , respectively, where FL _t is the mean fork length (mm) at age t. The spawning period was found to last from June to August for dolphinfish, based on an examination of the monthly changes in the gonadosomatic index and histological observations. Therefore, based on the relationship between the fork length and the developmental stage of the testes or ovaries, male and female dolphinfish were found to reach sexual maturity by the following spawning season after hatching in the northern East China Sea.     

Dietary l-tryptophan alters aggression in juvenile matrinxã Brycon amazonicus

This study evaluated the effect of dietary supplementation with l-tryptophan (L-TRP), a serotonin precursor, on the aggressiveness of juvenile matrinxã Brycon amazonicus. Fish were kept in individual aquaria for 7 days receiving the diets: D1 (control: 0.47% of TRP), D2 (0.94% of TRP), D3 (1.88% of TRP), and D4 (3.76% of TRP). After this, they were grouped with an intruder fish to establish a resident–intruder relationship during periods of 20 min. Blood cortisol, glucose, chloride, sodium and calcium; hemoglobin, hematocrit, red blood cell count and volume; liver glycogen and lipids were measured. Territoriality had significant effect on the aggressiveness of matrinxã (the residents were more aggressive than intruders, P < 0.001) and tryptophan significantly affected their behavior. Fish fed with the D2 diet presented a longer latency until the first attack (P = 0.0069) and bit the intruder fewer times (P = 0.0136) during the period of observation, compared to the control group. The frequency of bites and chases after the first attack was not affected by the dietary supplementation of TRP. Physiological variables were not significantly affected by the diet, except for a moderate increase in cortisol level in fish fed with D2 diet after the fight, indicating slight activation of the hypothalamus–pituitary–interrenal axis. The results show that juvenile matrinxã have aggressive and territorial behavior and that a diet containing 9.4 g TRP kg^?1 alter their aggressiveness, without affecting the stress-related physiological parameters.