Age,growth and mortality estimates for populations of red snappers <Emphasis Type="Italic">Lutjanus erythropterus</Emphasis> and <Emphasis Type="Italic">L. malabaricus</Emphasis> from northern Australia and eastern Indonesia

Lutjanus erythropterus and L. malabaricus were examined for life-history differences among northern Australian and eastern Indonesian populations. Formation of opaque growth increments in otoliths began during April to September for northern Australian fishes and September to April for eastern Indonesian fishes. Maximum observed ages were greater than previously reported: 42 and 48 years for L. erythropterus and L. malabaricus, respectively. Eastern Indonesian red snappers grew faster than northern Australian fish. Growth was similar for northern Australian L. erythropterus populations. However, Kupang and Sape populations of L. erythropterus in eastern Indonesian showed different growth from the Arafura North population, which was similar to northern Australian fish. There were growth differences among northern Australian populations of L. malabaricus. Arafura South and Timor populations were similar, but differed from Groote and Weipa populations. There were no significant differences in growth among populations of L. malabaricus in eastern Indonesia. Total mortality estimates were similar between northern Australian and eastern Indonesian fish: 0.09 and 0.16 year⁻¹ for L. erythropterus and 0.11 and 0.14 year⁻¹ for L. malabaricus, respectively. Life-history characteristics of these red snappers are typical of other tropical snappers: slow-growing and long-lived with low mortality. However, population growth differences suggest that their management should be based on biological information from distinct stocks.     

PGF<Subscript>2α</Subscript> and gonadal steroid plasma levels of successful and unsuccessful spawning <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> (Teleostei,Characiformes) females

Gonadal steroid and prostaglandin F₂α (PGF_2α) plasma levels were evaluated in successfully (SP) and unsuccessfully ovulated (UN) female Piaractus mesopotamicus. Forty-one females were injected with crude carp pituitary extract (0.6 and 5.4 mg kg^?1 with a 24-h interval between the doses) and sampled to determine the plasma concentration of 17β-estradiol (E₂), 17α-hydroxyprogesterone (17α-OHP), 17α,20β-dihydroxy-4-pregnen-3-one (DHP), PGF_2α, and testosterone (T) after each injection (first—A1 and second—A2), and at the time of ovulation for SP and UN. Two clusters were obtained using multivariate analysis: 1—composed of all A1, all A2, and some UN; and 2—composed of all SP and some UN. Median values of E₂ plasma levels were similar between clusters; however, plasma levels of T, 17α-OHP, DHP, and PGF_2α of cluster 2 (predominantly formed by SP) were higher than those of cluster 1. Since cluster 2 contained all SP and females of this cluster presented higher levels of PGF_2α, T, 17α-OHP, and DHP, here we evidently shown in an unprecedented manner that concomitant increased levels of these substances were associated with successful ovulation in this species, but such an increase was not determinant for successful ovulation due to the presence of some UN females in the same cluster 2. These findings highlight the unexplored potential of PGF_2α to be used as an accessory tool for inducing successful ovulation for fish farming purposes.     

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.     

Reproductive dynamics and nursery habitat preferences of two commercially important Indo-Pacific red snappers <Emphasis Type="Italic">Lutjanus erythropterus</Emphasis> and <Emphasis Type="Italic">L. malabaricus</Emphasis>

Red snappers were examined for reproductive biology and age-0 habitat preferences. Spawning in red snappers occurred throughout the year in northern Australia and eastern Indonesia; at least 10–30% of females and 40–80% of males were in ripe or spawning condition in most months. Northern Australian populations showed a spawning peak from July to December (L. erythropterus) and September to March (L. malabaricus). Eastern Indonesian L. malabaricus had a less defined pattern with two peaks: January–March and October. Size at first maturity was 240 mm for males and 250–300 mm for females. L ₅₀ estimates were similar between species in northern Australia: 270–280 mm (males) and 350–370 mm (females). Maximum batch fecundity was 676,100 oocytes for L. erythropterus and 997,000 oocytes for L. malabaricus. Higher mean abundances of age-0 L. erythropterus were found in silty and coarse sand/rubble estuarine habitats of northern Australia (456 ± 119 fish/km²) compared with sandy coastal habitats (5 ± 3 fish/km²). Most age-0 snapper caught at Sape (eastern Indonesia) were L. malabaricus (91%) with mean abundances of 312 ± 14 fish/km². The similarities in the reproductive characteristics of red snappers suggest that successful management approaches adopted in northern Australia should be considered in eastern Indonesia.     

Effect of trawling with traditional and ‘T90’ trawl codends on fish size and on different quality parameters of cod <Emphasis Type="Italic">Gadus morhua</Emphasis> and haddock <Emphasis Type="Italic">Melanogrammus aeglefinus</Emphasis>

The effect of trawling on fish size and on different quality parameters of cod (Gadus morhua) and haddock (Melanogrammus aeglefinus) was evaluated after conducting 16 valid hauls using two trawls in a double rig fitted with a traditional and a novel ‘T90’ codend, respectively. The total catch volume during the fishing period was 47.6 metric tons, with an average catch per codend of 1.5 (range 0.5–2.9) tons. The mean haul duration was 5 h. The catch was assessed according to fish size, mortality, external damage, initial white muscle pH and development of rigor mortis. Fillet quality (colour, blood spots, gaping) was assessed after 1 week of freeze-storage. Our results showed there was no difference between the two types of nets in terms of catch volume, but significantly slightly bigger fish were caught with T90 than with the traditional trawl net (p < 0.05). Haddock caught with the traditional trawl net had more external injuries related to the trawl gear than haddock caught with the T90 gear (p < 0.05). The gaping frequency for cod caught with the traditional trawl net tended to be higher than cod caught with the T90 gear, but the difference was not significant (p = 0.07). No other differences in fish quality between fish caught in the trawl nets were observed.     

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.     

cDNA cloning and primary structure comparison of tauropine dehydrogenase and β-alanopine dehydrogenase from the limpet <Emphasis Type="Italic">Cellana grata</Emphasis>

The primary structures of β-alanopine dehydrogenase (β-AlDH) and tauropine dehydrogenase (TaDH) from the limpet Cellana grata were determined by amino acid sequence analysis and complementary DNA (cDNA) cloning. β-AlDH and TaDH cDNAs comprised 1,479 nucleotides and 1,444 nucleotides, respectively, and both included an open reading frame of 1,206 nucleotides corresponding to 402 amino acids. The enzymes showed very high homology, with 96% amino acid identity. These enzymes were homologous to other marine invertebrate opine dehydrogenases (OpDHs), except TaDH of the marine sponge Halichondria japonica. The highest homologies were to alanopine dehydrogenase from Fusitriton oregonensis, being 57% for both enzymes. A phylogenetic tree constructed on the basis of marine invertebrate OpDHs and developed using a sequence distance method and neighbor-joining algorithm showed a tendency for the classification of animals from taxonomically derived evolutionary trees. Additionally, Cellana grata OpDHs belong to the same group as the Gastropoda OpDHs. This represents the first report concerning the primary structure of marine invertebrate β-AlDH, and primary structure comparisons of clearly different enzymes from the same species.     

Genetic and morphological evidence of hybridization between <Emphasis Type="Italic">Nematalosa japonica</Emphasis> and <Emphasis Type="Italic">N. come</Emphasis> (Clupeiformes: Clupeidae) off Okinawa Island,Ryukyu Archipelago,Japan

Two morphologically similar species of gizzard shad, Nematalosa japonica Regan and N. come (Richardson), sympatrically distributed off Okinawa Island, Japan, were examined using an allozyme locus (SOD*) and two nuclear polymerase chain reaction (PCR)-based DNA markers (ITS-1 and CaM), which provided diagnostic identification of each species. In addition, a multiplex PCR-based mitochondrial DNA (mtDNA) marker (16S) was used to characterize the distribution of mtDNA haplotypes among specimens. The species composition of sympatric and allopatric population samples from Tungkang, southern Taiwan, to Okinawa and the Shikoku Islands, Japan, were also examined. Gizzard shad with hybrid genotypes were detected in three populations from Okinawa Island, with hybrid frequency ranging from 1 to 67%. A backcross level of 2% was detected in the dominant hybrid frequencies of one population sample only. Morphological examination of hybrids showed intermediate forms, with hybrid indices of three meristic characters falling between those of the parental species (range 39–53; mean 45). Although principal component analysis showed differences between N. japonica and N. come based on the first principal component scores, hybrids were difficult to identify. Accordingly, satisfactory identification of species and hybrids could be achieved only using genetic tools. We also discuss the cause of hybridization and its relationship with recently conducted reclamation on Okinawa Island.     

Experimental infection of <Emphasis Type="Italic">Betanodavirus</Emphasis> in freshwater fish <Emphasis Type="Italic">Gambusia affinis</Emphasis> (Baird and Girard, 1853)—a potential infection model for viral encephalopathy and retinopathy

Betanodaviruses are the causative agents of the disease known as viral nervous necrosis (VNN) or viral encephalopathy and retinopathy (VER) in a variety of marine and freshwater fish species. The aim of this study was to demonstrate experimental infection of an isolate of betanodavirus (RGNNV genotype) in freshwater fish, Gambusia affinis, for elucidation of transmission mechanism and potential use as a laboratory model. Morbidity and mortality rate was significantly higher by injection route of infection as compared to immersion by bath and resembled the natural infection of juvenile marine fish. The fish in disease affected group showed severe neurological disorders accompanied by extensive vacuolar degeneration and mild to moderate neuronal necrosis of the brain in comparison to control. Amplification of ~?427 bp product in the variable region of the coat protein gene of betanodavirus was achieved by RT-PCR with 100% sequence homology to RGNNV genotype.     

Hematologic and serum biochemical values for jundiá (<Emphasis Type="Italic">Rhamdia quelen</Emphasis>)

The determination of hematologic parameters for jundiá Rhamdia quelencultivated in ponds is an easy and useful tool for the control of their sanitary and nutritional conditions. In the present work hematologic indices (RBC, WBC, Ht, Hb, MCV, MCH, MCHC), electrolytes (Cl, Na, K, Ca, Mg, P), metabolic products (creatinine, urea, ammonia, glucose, total protein, triglycerides, cholesterol, bilirubin) and enzymes (alanine and aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase) were measured in a population of jundiá. The mean ± SEM and range were established.     

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.     

Dietary β-glucans and mannanoligosaccharides improve growth performance and intestinal morphology of juvenile pacu <Emphasis Type="Italic">Piaractus mesopotamicus</Emphasis> (Holmberg, 1887)

The objective of this study was to evaluate the effect of increased levels (0.0, 0.1, 0.2, 0.4, and 0.8%) of a combination of β-glucans and mannanoligosaccharides (MOS) in isonitrogenous (23% of digestible protein) and isoenergetic (13.38 MJ of digestible energy kg^-1) pacu diets, corresponding to five treatments and four replicates. A 30-day feeding trial was conducted to assess the effects on growth performance, hematological parameters, and intestinal morphology. Fish (n = 160, 30.92 ± 0.46 g) were distributed randomly in 20 aquaria (300 L) with a recirculating water system with controlled temperature (26.20 ± 0.32 °C) and fed four times a day until apparent satiation. A quadratic effect (P < 0.05) was observed for weight gain (WG), feed conversion ratio (FCR), and protein efficiency ratio (PER). The hepatosomatic and viscerosomatic indexes, hematocrit, hemoglobin, mean corpuscular volume (MCV), and mean corpuscular hemoglobin concentration (MCHC) did not show differences (P > 0.05) among treatments. Pacu fed β-glucans and MOS at 0.1 and 0.2% resulted in the greatest (P < 0.05) villus height and perimeter. The diet containing 0.2% β-glucans and MOS promoted the best growth response, feed efficiency, and intestinal morphology, without detrimental effects on the hematological parameters for juvenile pacu.     

Leptin and its receptor in turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis>: cloning,characterization and expression response to ratios of dietary carbohydrate–lipid

In the present study, the full-length cDNA sequences of leptin (LEP) and its receptor (LEPR) from turbot Scophthalmus maximus were cloned. The cDNA of tLEP was 1126 bp in length encoding 157 amino acids. The amino acid sequence shared low identity with human LEP (18.8 %), but the three-dimensional structures of these two LEPs were strongly conserved. The deduced 1173-amino acid sequence of tLEPR was 28 % identical to human LEPR, and 82 % too range-spotted grouper LEPR, containing all functionally important domains conserved in vertebrate LEPR. Tissue distribution analysis showed that tLEP was abundantly expressed in brain, eyes and liver. The highest level of tLEPR mRNA was found in liver and kidney. After a 9-week feeding trial using diets with different ratios of carbohydrate–lipid (1:6, 1:2, 2:1 and 14:1), it was found that the increase in dietary carbohydrate-to-lipid ratios from 1:6 to 2:1 did not significantly influence tLEP and tLEPR expression in turbot liver (P > 0.05). The hepatic tLEP expression was significantly elevated in treatment with 14:1 dietary carbohydrate-to-lipid ratio (P < 0.05). The hepatic tLEPR mRNA level in group with 14:1 dietary carbohydrate-to-lipid ratio was significantly lower than that in 1:6 group (P < 0.05), but had no significant difference with the other two groups (P > 0.05). These results revealed the important relationship between dietary carbohydrate-to-lipid ratio and LEP expression in turbot.     

Estimation of the spawning grounds of chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> and spotted mackerel <Emphasis Type="Italic">Scomber australasicus</Emphasis> in the East China Sea based on catch statistics and biometric data

The spawning grounds of the chub mackerel (Scomber japonicus) and spotted mackerel (Scomber australasicus) in the East China Sea were estimated based on catch statistics of the Japanese large- and medium-type purse seine fishery from 1992 to 2006. Biometric data were obtained from specimens caught by purse seiners in the East China Sea from 1998 to 2006. Gonadosomatic index (GSI) at 50% sexual maturity of chub mackerel and spotted mackerel females was 2.5 and 2.6, respectively. Using this criterion for GSI, chub mackerel larger than 275 mm and spotted mackerel larger than 310 mm in fork length were considered to be mature. Mature chub mackerel was observed in the area of 15–22°C sea surface temperature (SST), and mature spotted mackerel was observed in the area of 17–25°C SST. The spawning period of chub mackerel ranged from February to June, and that of spotted mackerel ranged from February to May in the East China Sea. The spawning grounds were estimated from the distributions of catch per unit effort (CPUE) of spawners and SST. As a result, the spawning ground of chub mackerel was estimated to be in the central and southern part of the East China Sea and the area west of Kyushu in February, March, and April, and in the central part of the East China Sea, the area west of Kyushu and Tsushima Straight in May, and in Tsushima Straight and western part of the Sea of Japan in June. The spawning ground of spotted mackerel was estimated to be in the central and southern part of the East China Sea and southern coastal area of Kyushu in February, March, and April, and the central and southern part of the East China Sea and the area west of Kyushu in May.     

Nucleotide sequence and mRNA expression analysis of <Emphasis Type="Italic">β</Emphasis>-actin gene in the orange-spotted grouper <Emphasis Type="Italic">Epinephelus coioides</Emphasis>

The full-length cDNA, encoding the orange-spotted grouper β-actin and spanning 1920 bp including a poly (A) tail, was cloned from its brain cDNA library. The open reading frame encodes a protein of 375 amino acids. Sequence analysis indicated that it contained the typical structural features of cytoplasmic actins, and showed higher homology with other vertebrate β-actin than any other members of the actin family. The partial genomic sequence indicated that the organization of the β-actin gene in the orange-spotted grouper might also be conserved. Northern blot analysis indicated that it was expressed at high levels in the brain, spleen, adipose tissue, ovary, and liver, but at low levels in the gill filament and heart, and at a very low level in the kidney. The expression of β-actin gene in the skeletal muscle was barely detectable. These results indicated that the expression of the orange-spotted grouper β-actin gene showed significant variation in different tissues. Therefore, caution should be taken when using β-actin gene as an internal control in the normalization of gene expression among tissues. Whereas, semi-quantitative RT-PCR analysis indicated that treatment with 17α–methyltestosterone (MT) had little effect on the mRNA expression of β-actin gene in the in vitro incubated hypothalamus, pituitary, and ovary fragments of the orange-spotted grouper, suggesting β-actin can be used as an internal control for RT-PCR analysis of MT effects on gene expression in these tissues.