Distribution patterns of settlement-stage juveniles of <Emphasis Type="Italic">Girella punctata</Emphasis> and <Emphasis Type="Italic">Girella leonina</Emphasis> on the rocky coast of the Kanto–Izu region,Japan

The early life history of girellid fishes in Japanese waters is unclear, and little is known about their species-specific reproductive strategies. We examined seasonal changes of distribution patterns for settlement-stage juveniles of Girella punctata and Girella leonina on the rocky shore in the regions of Kanto and Izu, Japan, to infer the influence of the Kuroshio Current on their reproduction. We collected 813 settlement-stage juveniles mainly in Sagami Bay and genetically identified the species. The juveniles of G. punctata were collected on the rocky shore in Sagami Bay during April to August, with the abundant catch in May and June. Thus, we infer that juvenile G. punctata ubiquitously inhabit the rocky shore in the area in spring and summer. By contrast, juveniles of G. leonina were rarely collected in Sagami Bay, with a total catch of only 66. Notably, no juveniles were collected during the wintertime in Sagami Bay, although an abundant catch of G. leonina had been previously reported for Sagami Nada off Sagami Bay during January to March. This clear-cut difference between the areas likely reflects the difference in proximity to the path of the Kuroshio Current. We expect that the Kuroshio Current strongly influences the reproductive success of G. leonina.     

Effects of dietary riboflavin levels on antioxidant defense of the juvenile grouper <Emphasis Type="Italic">Epinephelus coioides</Emphasis>

This study was conducted to evaluate the effects of dietary riboflavin on antioxidant defense in the juvenile grouper Epinephelus coioides. Graded levels of riboflavin (0.9, 1.6, 4.4, 6.7, 12.9 and 19.4 mg kg⁻¹ dry diet) were fed to grouper juveniles (mean weight: 14.90 ± 0.46 g) for 12 weeks. Higher levels of liver thiobarbituric acid reactive substances (TBARS) content were observed in grouper fed low doses (0.9 and 1.6 mg kg⁻¹ diet) of riboflavin. Both liver glutathione reductase (GR) activity and its activation coefficient (GR-AC) poorly responded to riboflavin deficiency. In addition, other indices of the glutathione-dependent defense system, including the activities of glutathione peroxidase (GSH-PX) and glutathione-S-transferase (GST), and the content of glutathione (GSH), were also non-significantly affected by dietary riboflavin levels. However, the activities of liver superoxide dismutase (SOD) and catalase (CAT) were significantly lower in fish fed 0.9 mg kg⁻¹ diet, with a positive correlation between the different groups. In conclusion, the present study indicated that the juvenile grouper fed the riboflavin-unsupplemented diet was susceptible to lipid peroxidation (LPO), with lower SOD and CAT activities in the liver. However, the glutathione-dependent defense system of grouper was not affected by dietary riboflavin levels.     

The effects of hairtail protein hydrolysate–Fe<Superscript>2+</Superscript> complexes on growth and non-specific immune response of red swamp crayfish (<Emphasis Type="Italic">Procambarus clarkii</Emphasis>)

The experiment was conducted to investigate the effects of prepared hairtail protein hydrolysate–Fe²⁺ (PH–Fe²⁺) complexes on growth and non-specific immune responses of crayfish (Procambarus clarkii). The diets with five different levels of PH–Fe²⁺ (200, 400, 600, 800, and 1000 mg/kg) were fed to crayfish for 60 days. The results indicated that the survival rate, weight gain percentage, specific growth rate, and muscle index of crayfish with 400–1000 mg/kg of PH–Fe²⁺ feeding were significantly higher, 9.94–10.10, 8.55–8.72, 0.24–0.29 %/day, and 2.39–3.05 %, respectively, than the control values after 60 days of feeding. Additionally, after 30 days of feeding, 200–400 mg/kg of PH–Fe²⁺ showed no significant (P > 0.05) improvement effect on activities of superoxide dismutase (SOD), phenoloxidase (PO), lysozyme (LSZ), and acid phosphatase (ACP) in serum or hepatopancreas of crayfish. However, significant improvement effects were observed in 800–1000 mg/kg groups. After 60 days of feeding, 400–600 mg/kg of PH–Fe²⁺ significantly (P < 0.05) improved the activity of SOD and LSZ, but did not affect the activity of PO and ACP significantly (P > 0.05). Moreover, the activities of SOD, PO, LSZ, and ACP in serum and hepatopancreas were all significantly enhanced upon treatment with 800–1000 mg/kg of PH–Fe²⁺. For these reasons, PH–Fe²⁺ can be recommended as a supplement in crayfish feed to increase the growth and immunity.     

Ontogenetic development of digestive enzymes and effect of starvation in miiuy croaker <Emphasis Type="Italic">Miichthys miiuy</Emphasis> larvae

The ontogenetic development of the digestive enzymes amylase, lipase, trypsin, and alkaline phosphatase and the effect of starvation in miiuy croaker Miichthys miiuy larvae were studied. The activities of these enzymes were detected prior to exogenous feeding, but their developmental patterns differed remarkably. Trypsin activity continuously increased from 2 days after hatching (dah), peaked on 20 dah, and decreased to 25 dah at weaning. Alkaline phosphatase activity oscillated at low levels within a small range after the first feeding on 3 dah. In contrast, amylase and lipase activities followed the general developmental pattern that has been characterized in fish larvae, with a succession of increases or decreases. Amylase, lipase, and trypsin activities generally started to increase or decrease at transitions from endogenous to exogenous feeding or diet changes, suggesting that these enzymatic activities can be modulated by feeding modes. The activities of all the enzymes remained stable from 25 dah onwards, coinciding with the formation of gastric glands and pyloric caecum. These results imply that specific activities of these enzymes underwent changes due to morphological and physiological modifications or diet shift during larval development but that they became stable after the development of the digestive organs and associated glands was fully completed and the organs/glands functioned. Trypsin and alkaline phosphatase were more sensitive to starvation than amylase and lipase because delayed feeding up to 2 days after mouth opening was able to adversely affect their activities. Enzyme activities did not significantly differ among feeding groups during endogenous feeding; however, all activities were remarkably reduced when delayed feeding was within 3 days after mouth opening. Initiation of larvae feeding should occur within 2 days after mouth opening so that good growth and survival can be obtained in the culture.     

Effect of nitrite exposure on oxygen-carrying capacity and gene expression of <Emphasis Type="Italic">NF-κB</Emphasis>/<Emphasis Type="Italic">HIF-1α</Emphasis> pathway in gill of bighead carp (<Emphasis Type="Italic">Aristichthys nobilis</Emphasis>)

Nitrite (NO₂^?) contamination of water can severely impact the health of aquatic life and is a major concern for commercial aquaculture. In order to study the effect of nitrite on Aristichthys nobilis, we investigated the oxygen-carrying capacity, NF-κB/HIF-1α pathway, and the gill tissue structure under nitrite stress. In the current study, bighead carp (initial weight 180.05?±?0.092 g) were exposed to nitrite (48.634 mg/L) for 96 h and then for 96 h recovery test. After nitrite exposure for 6 h, hypoxia-inducible factor-1α (HIF-1α) and nuclear factor-κB (NF-κB) mRNA expression increased significantly in the gill of bighead carp (P?<?0.05). After nitrite exposure for 12 h, hemoglobin (Hb) and methemoglobin reductase (MHBR) content in blood decreased significantly (P?<?0.05); TLR4 mRNA expression increased significantly (P?<?0.05). After nitrite exposure for 24 h, methemoglobin (MetHb) content increased significantly (P?<?0.05). After recovery test, all the indicators except TLR4 mRNA expression level recovered to initial level. In conclusion, nitrite exposure can affect hemoglobin dynamics, as oxidization of nitrite by hemoglobin results in the reduction of Hb to MetHb leading to hypoxia and nitrite exposure can also result into gill tissue damage. In the face of nitrite exposure, NF-κB and HIF-1α mRNA expression level increased immediately to protect the body from oxidative damage and eased hypoxic condition caused by nitrite. It was also observed that nitrite damage is recoverable in Aristichthys nobilis, but it may be need more than 96 h.     

Determination of poly-β-hydroxybutyrate production by <Emphasis Type="Italic">Bacillus</Emphasis> spp. isolated from the intestines of various fishes

In this study, 30 strains of the genus Bacillus were isolated from various fresh fishes obtained from fish markets in Ankara, Turkey. They were identified as Bacillus pasteurii, Bacillus badius, Bacillus circulans, Bacillus licheniformis, Bacillus megaterium, Bacillus thuringiensis, Bacillus brevis, Bacillus cereus, Bacillus sphaericus, Bacillus subtilis, Bacillus coagulans, Bacillus lentus, and Bacillus pumilus. When poly-β-hydroxybutyrate (PHB) production by these strains was determined by spectrophotometry, it was found that PHB production ranged from 0.81–23.38% (w/v). PHB production and productivity were highest for B. pasteurii P2 (20.58% w/v) and B. lentus P18 (23.38% w/v). Furthermore, B. pasteurii P2 and B. lentus P18 were examined for PHB production using different carbon and nitrogen sources and in combined medium containing both carbon and nitrogen sources. The amounts of PHB produced by the strains in these media were more different than the amounts of PHB produced in nutrient broth medium.     

The impact of net-isolated polyculture of tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>) on plankton community in saline–alkaline pond of shrimp (<Emphasis Type="Italic">Penaeus vannamei</Emphasis>)

We used 12 land-based experimental enclosures (6 m × 5 m) in a saline–alkaline pond of shrimp (Penaeus vannamei) to determine the impact of net-isolated polyculture of tilapia (Oreochromis niloticus) on plankton communities for 40 days. Tilapias were stocked in net cages suspended in enclosures, in polyculture ponds including tilapia and shrimp. Four tilapia biomass were tested: 0, 39, 115 and 227 g m⁻². Shrimp stocking biomass were 0.7 g m⁻² in all treatments. There were three replicates in each treatment. Our results showed that the presence of tilapia significantly reduced phytoplankton biomass directly through predation and indirectly through top-down effect. The stocking of tilapia reduced zooplankton biomass, particularly rotifer biomass. However, copepod biomass was not been significantly affected. So, net-isolated polyculture of tilapia can thus have a strong impact on phytoplankton allowing the co-existence of large numbers of copepods with planktivorous fish and improving the water quality of shrimp ponds.     

Effects of <Emphasis Type="Italic">Euphorbia hirta</Emphasis> plant leaf extract on growth performance,hematological and organosomatic indices of hybrid catfish, <Emphasis Type="Italic">Clarias macrocephalus </Emphasis>× <Emphasis Type="Italic">C. gariepinus</Emphasis>

This study was conducted to evaluate the effects of Euphorbia hirta leaf extract on the growth performance, hematological and organosomatic indices of hybrid catfish, Clarias macrocephalus?×?C. gariepinus. The fish were treated with 0 (control), 300, 500 and 800 mg/kg (ppm) for 90 days. The weight gain, average daily growth rate, and specific growth rate were at significantly higher levels in fish from all the treatment groups on days 75 and 90, while the feed efficiency and protein efficiency ratio were consistently higher in fish from all the treatment groups from day 60 up until day 90. The feed conversion rate significantly decreased from day 60 until day 90 in all treatment groups when compared with the control group, and the survival rate was significantly different from day 30 until day 90; a consistently higher rate was observed in fish fed 800 mg/kg. The highest viscerosomatic index and intraperitoneal fat were observed in the group fed 500 mg/kg (p?<?0.05). The hepatosomatic index was significantly increased alongside increasing levels of E. hirta extract. The total white blood cell count in the control group was significantly higher on day 30, but on day 90 all the treatment groups showed higher levels. Hematocrit percentage was significantly different on days 30 and 90. Lymphocyte, eosinophil and thrombocyte levels were shown to be significantly different (p?<?0.05) when different groups of fish were compared. In conclusion, 300 mg/kg of E. hirta leaf extract could improve growth performance, hematological and some organosomatic indices in hybrid catfish.     

Development of digestive enzyme activity in larvae of spotted sand bass <Emphasis Type="Italic">Paralabrax maculatofasciatus</Emphasis> II: Electrophoretic analysis

The activities of several digestive enzymes during larval development of the spotted sand bass (Paralabrax maculatofasciatus) were evaluated using electrophoretic techniques. The results show the presence of three isoforms of alkaline protease from day 2 after hatching (ah) and the early appearance of one pepsin-like band from day 12 ah onwards. In addition, two lipase bands first appeared on day 2 ah, and there was a change in the molecular weight of one band from day 15 ah onwards. Several α-amylase isoforms were observed from hatching up to day 5 ah. These results indicate that the important digestive enzymes develop rapidly in these larvae, supporting the possibility of early weaning at day 12 ah using artificial diets.     

Effects of light and temperature conditions on the expression of GnRH and GtH genes and levels of plasma steroids in <Emphasis Type="Italic">Odontesthes bonariensis</Emphasis> females

In this study we examined the endocrine mediation between environmental factors (temperature and photoperiod) and the brain–pituitary–gonadal axis in females of pejerrey Odontesthes bonariensis. Changes in the expression of brain gonadotropin-releasing hormones (GnRHs) and gonadotropin (GtH) subunit [follicle stimulating-β (FSH-β), luteinizing hormone-β (LH-β), glycoprotein hormone-α (GPH-α)] genes, plasma gonadal steroids [estradiol (E₂) and testosterone (T)], gonadal histology, and gonadosomatic index (GSI) in adult females exposed to combinations of short-day (8 h) or long-day (16 h) photoperiods and low (12°C) or high (20°C) temperatures after winter conditions (8 h light, 12°C) were analyzed. Pejerrey females kept under the short photoperiod had low GSIs, and their ovaries contained only previtellogenic oocytes regardless of the experimental temperature. In contrast, females exposed to the long photoperiod had high GSIs and ovaries with vitellogenic oocytes at both temperatures. These fish also showed a significantly higher expression of sGnRH, pjGnRH, cGnRH-II (the three different GnRH variants found to date in the pejerrey brain), FSH-β, LH-β and GPH-α genes and plasma E₂ levels than those at the shorter photoperiod. No significant changes were observed in plasma T levels. Based on these results, we concluded that the increase in day length but not that of temperature triggers the maturation of pejerrey females after the winter period of gonadal rest and that this occurs by an integrated stimulation of the various components of the brain–pituitary–gonad axis.     

A comparison of growth and biochemical composition of <Emphasis Type="Italic">Mytilus galloprovincialis</Emphasis> (Lmk.) and <Emphasis Type="Italic">Mytilus edulis</Emphasis> (L.) on the West coast of Cotentin,Normandy, France 1999–2000

The blue mussel, Mytilus edulis has been reared along the Normandy coast line since the 1960s. The gonadal cycle of this mussel species shows a sharp decrease in meat quality during the winter period after spawning. This decline in meat quality is so severe that sales have to be suspended from December to July. Another species of mussel, Mytilus galloprovincialis, which is reared in the lagoons along the French shore of the Mediterranean Sea has a different spawning cycle. An experiment was undertaken to study the meat quality of M. Galloprovincialis throughout the year after the spat were transferred from the Mediterranean Sea to the Normandy coast. This species showed an immediate adaptation to the gonadal cycle of M. edulis. Despite suggestions from researchers, no interest was expressed to consider such transfers of M. galloprovincialis in the future.     

Use of biomarkers to assess the welfare of the edible clam, <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>: may it be a tool for proving areas of origin?

The use of biomarkers as early-warning signals to assess the impact of pollutants and other environmental stressors in coastal habitats has been internationally recognized as a useful tool in marine coastal monitoring. We propose a similar approach to verify the welfare of edible shellfish, Ruditapes philippinarum, thus exploiting its possible application as an effective tool to verify the origin of fishing/aquaculture products. A selected battery of biomarkers was chosen and applied to organisms from two different habitats in the Lagoon of Venice (a farming site and a natural area where fishing is currently banned, because of pollution problems) and from a local market. Biological responses were evaluated physiologically (survival in air test and condition index), behaviourally (reburrowing rate), and histochemically (quantification of neutral lipids and lipofuscin). Micropollutants (heavy metals, PCBs, and PAHs) were also determined in the soft tissues. Results indicated clams from the farming site were in the best condition with regard to both chemical and biological measurements. In contrast, the market sample contained the highest concentrations of PAHs. PCBs and heavy metals varied slightly and quite irregularly among the three samples. The worst values of the biomarkers were observed for clams collected at the polluted site, where, in particular, the lowest physiological and behavioural indexes and the highest accumulation of lipofuscin in the digestive tissue were recorded.     

Purification and characterization of α<Subscript>1</Subscript>-proteinase inhibitor and antithrombin III: major serpins of rainbow trout (<Emphasis Type="Italic">Oncorhynchuss mykiss</Emphasis>) and carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis>) blood plasma

The main serine proteinase inhibitors of rainbow trout (Oncorhynchuss mykiss) and common carp (Cyprinus carpio) blood plasma were isolated and purified. The investigated inhibitors, α₁-proteinase inhibitor (α₁-PI) and antithrombin III (AT III), act by forming stable complexes with target proteinases. The association rate constants k _on for the interaction of fish plasma inhibitors with several serine proteinases have been determined: k _on for both carp and rainbow trout α₁-PI were >10⁷ M⁻¹ s⁻¹ for human neutrophil elastase, and in the case of bovine trypsin and chymotrypsin k _on values were 2.0–5.2 × 10⁶ M⁻¹ s⁻¹. Association rate constants k _on for the interaction of carp and rainbow trout AT III with bovine trypsin and thrombin were about 1.3 × 10⁴–7.9 × 10⁵ M⁻¹ s⁻¹ without and >10⁷ M⁻¹ s⁻¹ in presence of heparin; so antithrombins require the presence of heparin to become effective proteinase inhibitors. The high degree of homology of the estimated amino acid sequences of fish inhibitors reactive site loops confirms their similarity with other proteinase inhibitors from the serpin family.     

Effect of dietary nutrient composition on the growth of olive flounder (<Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>) with different feeding regimes

We have examined the effect of dietary nutrient composition on the growth, body composition and blood chemistry of juvenile olive flounder (Paralichthys olivaceus) maintained on two different feeding regimes. Three experimental diets [HP (high protein), HE (high energy) and IPE (intermediate protein and energy)] were crossed in a two-factor design with the two feeding regimes, which consisted of 2WS + 2WF (2-week starvation and then 2-week feeding) and 4WS + 4WF (4-week starvation and then 4-week feeding)], for 8 weeks. The total days of feeding in all treatments were the same. The final weight of fish was affected by feeding regime, but not by dietary nutrient composition. Long-term starving and feeding (the 4WS + 4WF treatments) yielded higher growth than the alternating short-term starving and feeding regime (2WS + 2WF treatments). The feed consumption of fish was affected by dietary nutrient composition, but not by feeding regime, while the feed efficiency of fish was affected by feeding regime, but not by dietary nutrient composition. Moisture, crude protein and crude lipid contents of the liver of the fish were affected by dietary nutrient composition, but not by feeding regime.     

Influence of cell volume changes on protein synthesis in isolated hepatocytes of air-breathing walking catfish (<Emphasis Type="Italic">Clarias batrachus</Emphasis>)

The present study aimed at determining the effect of cell volume changes on protein synthesis, measured as the incorporation of [³H]leucine into acid-precipitable protein, in isolated hepatocytes of air-breathing walking catfish (Clarias batrachus). The rate of protein synthesis, which was recorded to be 10.02 ± 0.10 (n = 25) nmoles mg⁻¹ cell protein h⁻¹ in isotonic incubation conditions, increased/decreased significantly by 18 and 48%, respectively, following hypo- (−80 mOsmol l⁻¹)/hypertonic (+80 mOsmol l⁻¹) incubation conditions (adjusted with NaCl), with an accompanying increase/decrease of hepatic cell volume by 12 and 20%, respectively. Similar cell volume-sensitive changes of protein synthesis were also observed when the anisotonicity of incubation medium was adjusted with mannitol. Increase of hepatic cell volume by 9%, due to addition of glutamine plus glycine (5 mM each) to the isotonic control incubation medium, led to a significant increase of protein synthesis by 14%. Decrease of hepatic cell volume by 15 and 18%, due to addition of dibutyl-cAMP and adenosine in isotonic control incubation medium, led to a significant decrease of protein synthesis by 30 and 34%, respectively. Thus, it appears that the increase/decrease of hepatic cell volume, caused either by changing the extracellular osmolarity or by the presence of amino acids or certain other metabolites, leads to increase/decrease of protein synthesis, respectively, and shows a direct correction (r = 0.99) between the hepatic cell volume and protein synthesis in walking catfish. These cell volume-sensitive changes of protein synthesis probably help this walking catfish in fine tuning the different metabolic pathways for better adaptation during cell volume changes and also to avoid the adverse affects of osmotic stress. This is the first report of cell volume-sensitive changes of protein synthesis in hepatic cells of any teleosts.     

Biochemical properties of the sensitivity to GABA<Subscript>A</Subscript>ergic ligands,Cl<Superscript>−</Superscript>/HCO<Subscript>3</Subscript><Superscript>−</Superscript>-ATPase isolated from fish (<Emphasis Type="Italic">Cyprinus carpio</Emphasis>) olfactory mucosa and brain

This paper presents a comparative study of the roles of Cl^? and HCO₃ ^? in the functioning of the GABA_AR-associated Cl^?/HCO₃ ^?-ATPase of the plasma membranes of the olfactory sensory neurons (OSNs) and mature brain neurons (MBNs) of fish. The ATPase activity of OSNs and its dephosphorylation were increased twofold by Cl^?(15–30 mmol l^?1), whereas the enzyme from MBNs was not significantly affected by Cl^?. By contrast, HCO₃ ^?(15–30 mmol l^?1) significantly activated the MBN enzyme and its dephosphorylation, but had no effect on the OSN ATPase. The maximum ATPase activity and protein dephosphorylation was observed in the presence of both Cl^?(15 mmol l^?1)/HCO₃ ^?(27 mmol l^?1) and these activities were inhibited in the presence of picrotoxin (100 μmol l^?1), bumetanide (150 μmol l^?1), and DIDS (1000 μmol l^?1). SDS-PAGE revealed that ATPases purified from the neuronal membrane have a subunit with molecular mass of ~?56 kDa that binds [³H]muscimol and [³H]flunitrazepam. Direct phosphorylation of the enzymes in the presence of ATP-γ-³²P and Mg²⁺, as well as Cl^?/HCO₃ ^? sensitive dephosphorylation, is also associated with this 56 kDa peptide. Both preparations also showed one subunit with molecular mass 56 kDa that was immunoreactive with GABA_AR β3 subunit. The use of a fluorescent dye for Cl^? demonstrated that HCO₃ ^?(27 mmol l^?1) causes a twofold increase in Cl^? influx into proteoliposomes containing reconstituted ATPases from MBNs, but HCO₃ ^? had no effect on the reconstituted enzyme from OSNs. These data are the first to demonstrate a differential effect of Cl^? and HCO₃ ^? in the regulation of the Cl^?/HCO₃ ^?-ATPases functioning in neurons with different specializations.