2-methylisoborneol implicated as a cause of off-flavour in channel catfish, Ictalums punctatus (Rafinesque), from commercial culture ponds in Mississippi

Abstract. Samples of water, sediment and fish were collected from six commercial channel catfish, Ictalurus punctalus (Rafinesque), ponds from May to October and analysed for muddy-odour producing compounds. Three episodes of off-flavour in the fish occurred in late summer. The episodes were of 2–3 months' duration and correlated with the levels of 2-methylisoborneol in water, sediment and fish. Oscillatoria agardliii was the dominant phytoplankton in ponds with off-flavour fish. Other known muddy odour-producing compounds such as geosmin, 2-isopropyl-3-mcthoxypyrazinc, 2-isobutyl-3-methoxypyrazine and 2-isobutyl-6-methoxypyrazinc were not detected in the water sediment or at concentrations greater than 5·0 ng/l.     

Efficacy of Netminder® silicone release coating for biofouling reduction in bay scallop grow‐out and comparative effects on scallop survival,growth and reproduction

Netminder^®, a water‐based silicone barrier release coating, was highly effective in reducing biofouling of lantern nets during grow‐out/overwintering of bay scallops, Argopecten irradians irradians, in a large‐scale restoration programme in New York, USA. Compared with untreated nets, scallops held in Netminder^®‐coated nets experienced reduced survival after 8 months, probably due to higher loads of epibionts on their shells. However, this was countered by higher reproductive and overall condition of scallops in treated vs. untreated nets; there were no consistent differences in shell growth in different net treatments. Although further testing is recommended under a wider range of conditions, we conclude that for the purposes of our scallop restoration work, coating nets with Netminder^® would eliminate the need for a gear change in spring, thus reducing labour costs; the coating process should be refined so that Velcro closures of nets do not become sealed shut.     

Microsatellite marker isolation and cultured strain identification in <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>

Microsatellites have become the preferred molecular markers for strain selection and genetic breeding in fish. In this study a total of 105 microsatellites were isolated and identified in gibel carp (Carassius auratus gibelio) by microsatellite sequence searches in GenBank and other databases and by screening and sequencing of positive clones from the genomic library enriched for AG and GATA repeats. Moreover, nineteen microsatellites were randomly selected to design locus-specific primer pairs, and these were successfully used to identify and discriminate different cultured strains of gibel carp including strains A, D, L, and F. Three different types of microsatellite pattern were distinguished by the number and length of fragments amplified from the 19 primer pairs, and some microsatellite primer pairs were found to produce different microsatellite patterns among strains and strain-specific fragments. In addition, some duplicated alleles were also detected in two microsatellite patterns. Therefore, the current study provides direct molecular markers to discriminate among different cultured strains for selective breeding and aquaculture practice of gibel carp.     

Morphological and histological changes inthe genital ducts ofthe male Atlantic stingray, Dasyatis sabina, during the seasonal reproductive cycle

Changes inthe morphology and histologyofthe epididymis and seminal vesicle in male Atlantic stingrays (Dasyatis sabina) were examined in relation tothe seasonal reproductive cycle. Observations onthe size and structureofthese organs were accompanied by analysisof cell proliferation in genital duct epithelia using proliferating cell nuclear antigen (PCNA) as a marker for mitotic activity. No signsof reproductive tract growth or histological alteration were apparent duringthe initial stagesof spermatogenesis. However, increased PCNA expression inthe seminal vesicle epithelium was observed during this period, suggesting that this organ begins to undergo preparatory changes at an early stage inthe reproductive cycle. During late spermatogenesis, heightened expressionof PCNA in epithelial cellsofthe epididymis and seminal vesicle was observed in conjunction with dramatic increases in size and semen contentofthese organs. Extensive changes inthe histological architectureofthe genital ducts also were evident at this time, including regressionofthe stroma and an increase in size and secretory activityofthe epithelium. Althoughthe epididymis regressed in size and structure shortly after sperm production was completed,the seminal vesicle retained its appearance and activity forthe durationofthe 7-month copulatory period. Afterwards, immune cell content increased considerably in both ducts, likely reflecting clearanceof residual spermatozoa in preparation forthe subsequent reproductive season.the present study provides a basis for future reports onthe physiological regulationof reproductive tract function in elasmobranchs, and establishes PCNA immunocytochemistry as a novel tool for such investigations.     

Validation,use, and disadvantages of enzyme-linked immunosorbent assay kits for detection of cortisol in channel catfish,largemouth bass,red pacu,and golden shiners

Measurement of cortisol response is an important tool to asses stress in fisheries research. Radioimmunoassay (RIA) is a common method for the measure of cortisol in fish. Use of enzyme-linked immunosorbent assays (ELISA) to detect cortisol would eliminate health hazards, costs of handling radioisotopes, and the short stability time associated with RIA. Enzyme-linked immunosorbent assays have been used for the determination of cortisol in several fish species. However, the ELISA method of cortisol determination in fish lacks proper validation testing. We conducted validation procedures for multiple commercial cortisol ELISA kits and compared the results to RIA. The assays were tested for four species: (1) channel catfish Ictalurus punctatus, (2) largemouth bass Micropterus salmoides, (3) red pacu Piaractus brachypomus, and (4) golden shiners Notemigonus crysoleucas. We evaluated the ELISA methods against RIA, and determined that at least one kit is suitable (accuracy: mean recovery of spiked samples, 102.8%; reproducibility: interassay coefficient of variation < 10.5% for all species; precision: intra-assay coefficient of variation < 16.8% for all species; linearity: R ² > 0.96 for all species) for the measurement of cortisol response in fish and comparative determination of stress. All of the ELISA assay results varied by more than 10% from the cortisol concentrations detected by the RIA. The high variability of the kit results indicates that commercial ELISA kits could be utilized for qualitative determination of cortisol in fish, but should be fully validated in each laboratory for each species before being used for research.     

Constitutive nitric oxide synthase (NOS) activities in big-head carp (Aristichthys nobilis)

Nitric oxide synthase (NOS) is an enzyme that catalyzes the formation of nitric oxide (NO), an important biological messenger from L-arginine. There are considerable evidence showing the expression of NOS in mammalian tissues. Information on distribution of NOS activities in various organs and tissues of fish is rare. Non-functional NOS activities were documented in fish semi-quantitatively either by an indirect nicotine-adenine-dinucleotide-phosphate diaphorase (NADPH-d) activity histochemical staining method or by an immunohistochemical method using a cross-reacting antibody to brain NOS. Report on the functional levels of NOS activities in fish is lacking. This report represent the first attempt to document the functional NOS levels in various fish organs and tissues. Constitutive NOS (cNOS) activities in various organs of big-head carp (Aristichthys nobilis) was measured by a chemiluminescence method with a detection limit as low as 10 mol of NO produced. It was found that constitutive NOS activity was highest in the brain, followed by the intestine, stomach, retina, olfactory lobe, swim bladder, skeletal muscle, heart, kidney, ovary and liver. NOS activity could not be detected in the gill filaments. Omission of NADPH in the reaction mixture caused a 57–100% decrease in cNOS activities. However, omission of arginine in the mixture only caused a 56–87% drop in cNOS activities. When compared with cNOS activities documented from other species, a similar pattern of cNOS activities in the various organs and tissues of big-head carp could be seen.     

Relationship between zooplankton abundance and the early marine life history of juvenile chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis> in eastern Hokkaido,Japan

Interannual variations in abundance, timing of outmigration from rivers, growth rate and condition of juvenile chum salmon (Oncorhynchus keta) were studied in the Nemuro Strait (eastern Hokkaido, Japan) during 1999–2002 to establish a possible relationship to zooplankton abundance. The otolith microstructure of juveniles was examined each year in late June to determine their time and size at sea entry (i.e., outmigration), and to estimate the early marine growth rates. Salmon outmigration peaked in mid- or late May, which coincided, in three of the four study years, with the peak release of juveniles into rivers within the study area. Abundance, growth rate and condition of fish were higher in 2001, when—compared to other years—smaller fish experienced higher growth rates, coinciding with greater zooplankton abundance for that year. Our results suggest that high zooplankton abundance positively influenced juvenile chum salmon growth and the condition of the fish during their early marine life despite their small size at sea entry.     

Development of an in vitro culture system for producing eel larvae from immature ovarian follicles in Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

To standardize conditions during the final maturation and ovulation of ovarian follicles from Japanese eel, we have developed a culture system for the production of fertilizable eggs from post-vitellogenic ovarian follicles in vitro. Post-vitellogenic ovarian follicles were incubated in culture medium supplemented with 17α,20β-dihydroxy-4-pregnen-3-one (DHP) with or without bovine serum albumin (BSA) to assess the effects of protein concentration. Eggs that ovulated during incubation were fertilized, and the remaining follicles were incubated in prostaglandin F_2α (PGF_2α) for a further 3 or 6 h before fertilization. Male eels were injected repeatedly with human chorionic gonadotropin. The quality of eggs obtained under the different culture conditions was evaluated after artificial fertilization in terms of hatching success. Hatching rates tended to decrease with increasing concentrations of BSA in the incubation medium in a dose-dependent manner. The addition of PGF_2α drastically increased the number of eggs that ovulated, but the rate of hatching was greatly decreased compared with eggs obtained earlier by DHP incubation alone. The larvae obtained from artificially fertilized eggs produced in vitro survived for 14 days without feeding. We conclude that in vitro culture systems thus have a great potential for the acquisition of good quality eggs under tightly controlled artificial conditions, culminating in the production of eel larvae.     

Analysing the growth of turbot (<Emphasis Type="Italic">Psetta maxima</Emphasis>) in a commercial recirculation system with the use of three different growth models

The growth data of a commercial aquaculture recirculation system were analysed to investigate the growth performance of reared turbot (Psetta maxima). Three common growth models (von Bertalanffy, Gompertz and Schnute) were fitted to the growth data documented over a time period of 6 years. To determine the most suitable model, three different criteria were used: (1) the Akaike index criterion, (2) the sum of squared residuals and (3) the average daily deviation between the estimated final weight and the observed final weight. The evaluation of the growth models showed that the Schnute model had the lowest Akaike index, the lowest sum of squared residuals and the lowest daily deviation between estimated and real weight of all tested growth models. The Schnute model produced sigmoid growth curves. The estimated growth coefficients were the most realistic ones in regard to biological interpretation. In contrast, the von Bertalanffy growth model and the Gompertz model estimated inaccurate exponential growth curves and are therefore unable to simulate the growth data as well as the Schnute model. The results indicate that the von Bertalanffy growth model is not the optimal model to simulate the present growth data and that the growth potential of reared turbot has probably not yet been fully exploited in the aquaculture system(s) examined (so far).