Differences in oxygen and carbon stable isotope ratios between hatchery and wild pink salmon fry

We examined differences in otolith oxygen (δ¹⁸O_otolith) and carbon (δ¹³C_otolith) stable isotope ratios between hatchery and wild pink salmon fry. The δ¹⁸O_otolith values of hatchery and wild fry were ?7.7 ± 0.2 ‰ and ?8.3 ± 0.3 ‰ (1σ), respectively. This difference reflected differences in temperature conditions experienced by each fry. The δ¹³C_otolith values of hatchery and wild fry were ?19.2 ± 0.3 ‰ and ?11.1 ± 1.8 ‰ (1σ), respectively. The lower δ¹³C_otolith values of hatchery fry were probably related to their intake of artificial diets. Discriminant analysis of δ¹⁸O_otolith and δ¹³C_otolith values demonstrated a highly significant difference between hatchery and wild fry with 95.8 % classification accuracy. Therefore, analysis of δ¹⁸O_otolith and δ¹³C_otolith precipitated in the fry stage may be useful for discriminating the origin of returning adult pink salmon. The discrimination method for returning adult fish would provide important information for evaluating the effect of hatchery release and their impact on the wild population.     

Identification of ommastrephid squid paralarvae collected in northern Hawaiian waters and phylogenetic implications for the family Ommastrephidae using mtDNA analysis

ABSTRACT:   A total of 110 adult individuals from four ommastrephid (family Ommastrephidae) squid species ( Ommastrephes bartramii, Sthenoteuthis oualaniensis, Eucleoteuthis luminosa, and Hyaloteuthis pelagica ) were used to obtain diagnostic DNA markers for species identification. Restriction fragment length polymorphism (RFLP) analysis of a partial segment (855 bp) of the mitochondrial DNA cytochrome oxidase I (COI) gene amplified by polymerase chain reaction (PCR) revealed that the restriction profiles of two endonucleases ( Alu  I and Tsp 509 I) were diagnostic for species identification. The restriction assay partially supplemented with nucleotide sequence analysis successfully assigned 69 damaged and morphologically equivocal ommastrephid paralarvae collected in northern Hawaiian waters, identifying 60 O. bartramii , eight S. oualaniensis , and one E. luminosa . The family Ommastrephidae appears to be monophyletic. Although the phylogenetic relationships among genera were not resolved well due to apparent homoplasy and large genetic divergence between species, COI sequence data without transitions provided support for subfamily level relationships.     

Genetic variability and stock structure of red tilefish Branchiostegus japonicus inferred from mtDNA sequence analysis

Sequence analysis based on the anterior part of the mitochondrial DNA control region was carried out to reveal the genetic diversity, stock structure, and historical demography of the red tilefish. Nucleotide sequences of 388 bp in length were determined for 280 individuals taken from eight localities. The molecular diversity, haplotype diversity, and nucleotide diversity were relatively high (average h = 0.929 ± 0.011; average π = 0.008 ± 0.005). The mismatch distribution was not significantly different from the expected distribution for a rapidly expanding population (P = 0.453). The minimum spanning network connecting with full-sequence haplotypes contained star-like topologies derived from multiple ancient lineages, supporting the mismatch distribution analysis. No significant genetic differentiation was observed among eight localities (maximum pairwise F _ST = 0.023, reduced-sequence data set). Our results suggest a large panmictic population of the red tilefish along the coast of Honshu to the East China Sea.     

Lethal effects caused by suspended particles and sediment load on zoospores and gametophytes of the brown alga <Emphasis Type="Italic">Eisenia bicyclis</Emphasis>

ABSTRACT:   The effects caused by suspended particles in seawater and sediment cover of the substrate on the attachment to the substrate of zoospores, and on the subsequent growth and survival of gametophytes of the brown alga Eisenia bicyclis were examined in the laboratory. The attachment rate to the substrate of the zoospores was remarkably reduced by the sediment cover on the substrate, and it decreased to 3.8% of the no-sediment control by a slight sediment cover of 3.0 mg/cm² (0.048 mm in thickness). The growth of gametophytes was not inhibited at 10 mg/cm² or less of sediment cover, though it was stopped at 30 mg/cm². The survival rate of gametophytes became 39.4% at 5 mg/cm² on the substrate, and gametophytes were all blighted at 30 mg/cm². The ratio of male to female of the surviving gametophytes became 81.5:18.5, when the sediment cover was 10 mg/cm². The rate of total loss ( TL [%]) of zoospores and gametophytes of Eisenia bicyclis was obtained from the following equation: TL  = 100 (1 − exp[−0.0339  C ] exp[−1.24  Q ]), where, C (mg/L) and Q (mg/cm²) indicate the concentration of suspended particles in seawater and the sediment on the substrate, respectively.     

Effect of Artificial Diets on Growth,Survival and Condition of Adult Cuttlefish, <Emphasis Type="Italic">Sepia officinalis</Emphasis> Linnaeus, 1758

The effects of artificial diets on growth and body condition of adult cuttlefish, Sepia officinalis were tested in two experiments. Supplemented prepared diets (fish myofibrillar protein concentrate) were fed during a 30-day and a 21-day experiments. Growth, feeding rate and food conversion of group-reared cuttlefish were analyzed. The first of these experiments tested four artificial diets, made with increasing levels of lysine, on adult cuttlefish. According to the chemical analysis, diets 1–3 had limiting concentrations of lysine and other essential amino acids (compared to mantle composition of the cuttlefish), while diet 4 was the only one where almost all essential amino acids were present in concentrations similar or higher than the ones present in cuttlefish mantle. A second experiment was conducted by isolating 16 adult cuttlefish individually, and feeding them the same four artificial diets, in order to obtain individual data. During Experiment 1, only the diet with the best chemical score (diet 4) produced growth (p < 0.05), with a mean instantaneous growth rate (MIGR) of 0.30% wet body weight (BW) d⁻¹. Similarly, individually reared cuttlefish fed diet 4 produced the highest IGR’s (0.26, 0.38 and 0.48% BW d⁻¹) and grew larger (p < 0.01). Comparison of cuttlefish fed the artificial diets vs. thawed shrimp and unfed cuttlefish indicated that cuttlefish fed the artificial diets were in an intermediate state. Growth rates obtained with the artificial diets (<0.4% BW d⁻¹) were considerably lower compared to natural prey, live or frozen, reported by other authors.     

Molecular characterization of microalgae used in aquaculture with biotechnology potential

Microalgae are the main component of first tropic level in aquatic food chain; it is for this reason that they are used as food in aquaculture. Also due to its biotechnological potential properties, they are used in the production of diverse components, dyes, antioxidants, enzymes, emulsifiers, etc. The extended ways of microalgae applications require physiologically and genetically stable cultures as well as correctly identified organisms to guarantee reproducibility and reliability. But the variety of species and the morphological similarity between some of them make difficult the identification of some microalgae. The use of molecular markers has supplied a very useful tool for identification of microalgae in fast mode, such as in classification. The present study has worked on the molecular characterization of main species of microalgae used in aquaculture in base of the molecular markers 18S rRNA and 16S rRNA. Microalgae DNA has been amplified and sequenced, and the resultant sequences were analyzed and reflected in phylogenetic trees. The phylogenetic analyses obtained reflect as both molecular markers allow to differentiate the main genus used in aquaculture.     

Expression of HSP70 in response to heat-shock and its cDNA cloning from Mediterranean blue mussel

Expression of HSP70 in response to heat-shock was investigated at the protein and mRNA levels in Mediterranean blue mussel. Western and Northern blot analyses revealed that HSP70 was expressed following heat-shock in the mantle at both protein and mRNA levels, suggesting that gene expression of HSP70 is implicated in the cellular response to heat-shock stress in mussel. It was then attempted to clone HSP70 cDNA in order to determine the primary structure of mussel HSP70. As a result, two full-length cDNA encoding HSP70 were isolated from a cDNA library prepared from the heat-shocked mantle. The isolated cDNA consist of single open reading frames of 2067 bp and 1911 bp which encode proteins of 689 amino acids and 637 amino acids, respectively. Both HSP70 cDNA encode an ATPase do main, and a substrate-binding do main in addition to a Glu-Glu-Val-Asp (EEVD) peptide motif that is specific for cytosolic HSP70. These findings suggest that the cDNA clones obtained in the present study encode cytosolic HSP70.     

Age estimation of the wild population of Japanese mantis shrimp <Emphasis Type="Italic">Oratosquilla oratoria</Emphasis> (Crustacea: Stomatopoda) in Tokyo Bay,Japan, using lipofuscin as an age marker

ABSTRACT:   In this study, the age composition of the Japanese mantis shrimp, Oratosquilla oratoria , in Tokyo Bay, central Japan, was investigated using lipofuscin, an autofluorescent pigment, as an age marker. Lipofuscin in a histological section of protocerebral bridge cell mass (PBCM) in the brain was identified by confocal microscopy, and its concentration was quantified by image analysis. Modal analysis of the lipofuscin concentration showed four regularly-spaced modes that could each be regarded as a distinct age group. This implied a constant lipofuscin accumulation in PBCM at a 6.5 × 10⁻²% volume fraction per year; it also implied the existence of individuals that are at least 4 years old. The lipofuscin concentration was found to be a more appropriate index than body length for estimating the age of O. oratoria , because the modal analysis on the body–length histogram failed to detect apparent age groups. This was probably a result of the declining growth rate with age and the individual variations in growth through molting leading to considerable overlap in the size between different age groups. The lipofuscin analysis suggested that fast-growing individuals in each cohort are recruited to the fishery, and most individuals attain the exploitable size between 2 and 3 years of age.     

The effects of vitamin E on non-specific immune response of the juvenile soft-shelled turtle <Emphasis Type="Italic">Pelodiscus sinensis</Emphasis>

ABSTRACT:   This study was conducted to determine the effect of dietary vitamin E supplements on non-specific immune responses in juvenile soft-shelled turtles. Turtles were fed diets supplemented with vitamin E at 0 (control), 50, 250, 500, 1000 and 5000 mg/kg, respectively, for 4 weeks. Results showed that blood cell phagocytosis and serum bactericidal activity were significantly improved in turtles from two diets (250 and 500 mg/kg vitamin E supplementation), while had no significant improvement in turtles from the three remaining diets (50, 1000 and 5000 mg/kg vitamin E supplementation) when compared to turtles from the control group. Serum bacteriolytic activity in turtles from diets with 50, 250, 500 and 1000 mg/kg vitamin E supplementation were higher than activity levels quantified for the control group, but no significant difference was observed between the 5000 mg/kg vitamin E supplementation diet and the control diet. The results suggest that vitamin E has an upper and lower threshold for improving non-specific immune function in soft-shelled turtles, and the optimal supplementation may be between 250 and 500 mg/kg.     

Measurements of auditory sensitivity in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> by the auditory brainstem response technique and cardiac conditioning method

ABSTRACT:   Recently, the non-invasive auditory brainstem response (ABR) technique has been applied to the determination of hearing sensitivity in fish. The technique has some advantages over, and methodological differences from, the classical techniques. The auditory thresholds of common carp were measured by the ABR technique. The classical method was applied to measure cardiac response by electrocardiogram (ECG) after conditioning with electric shock. The most sensitive frequency obtained by ABR was 505 Hz, and by ECG, 1000 Hz. The shapes of the audiogram obtained by ABR and ECG were similar, though threshold levels in both audiograms differed from each other. Furthermore, audiograms obtained by using behavioral techniques were compared with those taken by the ABR and ECG techniques to assess the validity of the ABR technique for testing fish hearing sensitivity. The measured threshold levels increased in the order of behavioral, ABR, and ECG techniques, in the range below 1000 Hz.