Analysis of bacterial communities in <Emphasis Type="Italic">Nannochloropsis</Emphasis> sp. cultures used for larval fish production

ABSTRACT:   Phytoplankton used in fish hatcheries is mass-cultured in the open air and usually contains large numbers of bacteria. In commercial fish production, the phytoplankton cultures are usually added into the larval rearing tanks; however, the numbers and types of bacteria introduced into the rearing tanks simultaneously are unknown. In this study, the bacterial community structures in Nannochloropsis sp. cultures were analyzed by using fluorescence in situ hybridization (FISH). A direct viable count (DVC)-FISH analysis was also performed as DVC is useful for the detection of actively growing cells. Total numbers of bacteria in Nannochloropsis sp. cultures ranged from 7.72 × 10⁵−2.39 × 10⁶ cells/mL. High proportions of the total bacteria (31.6–53.6%) in the Nannochloropsis sp. cultures showed growth potential. DVC-FISH analysis revealed that α-proteobacteria and the Cytophaga – Flavobacterium cluster were abundant in the bacterial community of actively growing cells. Thus, the high growth potentials of the distinct bacterial communities in Nannochloropsis sp. culture must influence the bacterial communities in larval rearing tanks.     

Fortification of <Emphasis Type="Italic">niboshi</Emphasis> with histidine prepared from <Emphasis Type="Italic">katsuobushi</Emphasis> protein digest

ABSTRACT:   Free and total histidine contents in the muscle of various fishes and invertebrates caught in the Seto Inland Sea were estimated. These contents were higher in the muscles from skipjack, yellowfin tuna, yellowtail and mackerel than in those from lizard fish, sea bream and hairtail. Invertebrate muscles, except squid, contained low free and total histidine. Boiled and dried anchovy ( niboshi ) contained lower free histidine than that of dried anchovy (suboshi). To prepare free histidine-rich extracts, the waste residue of soup stock from smoke-dried and shaved skipjack ( katsuobushi ) was digested with proteases. The extract of the enzyme digest contained high free histidine and was used to fortify niboshi with free histidine. If taken, the fortified niboshi may suppress food intake, and prevent obesity.     

Chemical composition of dried eyeballs from <Emphasis Type="Italic">Euphausia superba</Emphasis> and <Emphasis Type="Italic">Euphausia pacifica</Emphasis>

ABSTRACT:   Dried krill eyeballs were obtained from Euphausia superba and Euphausia pacifica by using a patented industry method and their chemical compositions were examined. Crude protein content was 77.7% and 80.8% of the dry matter of E. superba and E. pacifica , respectively. The dominating amino acids in both krill were glutamic acid, aspartic acid, glycine, arginine, leucine and tyrosine. Crude fat content was 10.9% and 5.4% of dry matter of E. superba and E. pacifica , respectively. The main lipid class of the extracted lipids was phospholipids at 88.5% in E. superba and 96.4% in E. pacifica . The dominating fatty acids in both krill were 22:6 ( n- 3), 20:5 ( n- 3), 16:0 and 18:1 ( n- 9). Astaxanthin (3, 3'-dihydroxy-β, β-carotene-4, 4'-dione) content of E. superba and E. pacifica was 566 mg/100 g and 252 mg/100 g of dry matter, respectively. Retinol of E. superba and E. pacifica was 153.0 mg/100 g (510 000 IU/100 g) and 57.6 mg/100 g (192 000 IU/100 g) of dry matter, respectively. The lipophilic extract of E. superba by using n -hexane contained 1923 mg/100 g of astaxanthin at approximately four times higher than the dried eyeballs.     

Suspension feeding and growth of ark shell <Emphasis Type="Italic">Anadara granosa</Emphasis>: comparison with ubiquitous species <Emphasis Type="Italic">Scapharca subcrenata</Emphasis>

The ark shell Anadara granosa is a species peculiar to the Ariake Sound. To determine why distribution of A. granosa in Japan is largely confined to this area, we examined feeding and growth of A. granosa as functions of environmental and biological variables. The results were compared with those of another ark shell Scapharca subcrenata, which is ubiquitous in Japan. Feeding experiments indicated that A. granosa is eurythermal and euryhaline, as is S. subcrenata, but is adapted to temperature slightly higher than S. subcrenata. Weight-specific clearance rate (CR) of A. granosa as a function of soft-body dry-weight (w) followed the power function of w (CR=2.7×w ^−0.37), with coefficient and exponent very close to those for S. subcrenata. Growth rate of A. granosa increased linearly with daily ration, similar to S. subcrenata. Thus, feeding and growth characteristics of A. granosa were comparable to those of S. subcrenata and no ‘peculiarities’ of the former were detected. Therefore, the factors that make A. granosa a species restricted to the sound are probably not directly related to feeding or growth characteristics.     

Sterols from bivalves <Emphasis Type="Italic">Calyptogena soyoae</Emphasis> and <Emphasis Type="Italic">Bathymodiolus septemdierum</Emphasis> living in deep sea

From the two species of bivalves, Calyptogena soyoae around a cold seep and Bathymodiolus septemdierum near hydrothermal vents in the sea, sterols were isolated using high-pressure liquid chromatography analysis of the lipid fraction guided by characteristic ¹H NMR signals of the sterol skeleton. The minor sterol composition of C. soyoae included 24-methylenecycloartanal, cycloeucalenol, and obutusifoliol, which are known phytosterols. From B. Septemdierum, lathosterol and cholesterol as main sterols together with more diverse sterols were obtained. The difference between these species and their sterol contents is most likely because of feeding modes and metabolism of nutrients from their habitat.     

Masculinization of Convict Cichlid (<Emphasis Type="Italic">Cichlasoma nigrofasciatum</Emphasis>) by immersion in <Emphasis Type="Italic">Tribulus terrestris</Emphasis> extract

We have examined the effects of Tribulus terrestris (TT), a non-toxic herb, on sex reversal in the Convict Cichlid Cichlasoma nigrofasciatum with the aim of introducing a new environmentally friendly method for masculinization in C. nigrofasciatum. TT is a natural plant product that elevates the testosterone levels in humans and animals. Different concentrations (0.0, 0.10, 0.20 and 0.30 g/l) of TT extract were tested for their effect on sex reversal in C. nigrofasciatum by immersing newly hatched offspring once weekly for 2 months in TT extract. Of the dosages used in the present study, 0.30 g/l TT was the most effective in terms of masculinization, resulting in a maximum male ratio of 87.23% (P < 0.001). Sex ratios of 79 and 85% at 0.10 and 0.20 g/l TT, respectively, were also significantly different from the expected 1:1 ratio (P < 0.001). Histological examinations revealed that the testes of fish treated with TT extract contained all stages of spermatogenesis, clearly demonstrating that the administration of TT extract to C. nigrofasciatum stimulated spermatogenesis. Total survival rates in all treatments and the control were uniformly high, ranging from 88.57 to 90% (P > 0.05). We concluded that TT had no negative effect on the survival rate of C. nigrofasciatum. In addition, all groups of TT-treated fish exhibited successful growth acceleration compared to the control group, but only the 0.30 g/l TT treatment significantly improved the growth rate of C. nigrofasciatum. (P < 0.01). Sex reversal in C. nigrofasciatum demonstrated that TT-treated 0-day-old larvae showed successful sex reversal, spermatogenesis and a better growth rate than untreated progenies.     

Optimum temperature and salinity conditions for growth of green algae <Emphasis Type="Italic">Chlorella ellipsoidea</Emphasis> and <Emphasis Type="Italic">Nannochloris oculata</Emphasis>

ABSTRACT:   The effects of temperature and salinity on growth of green algae Chlorella ellipsoidea and Nannochloris oculata were determined to compare the optimum culture conditions. A four-temperature (15, 20, 25, and 30°C) × three-salinity (10, 20, and 30) factorial design with triplicates was applied. Specific growth rate (SGR), maximum density, and duration to reach maximum density of C. ellipsoidea were significantly affected by both temperature and salinity. The highest SGR was observed in C. ellipsoidea at 25°C and salinity 10, but the maximum density was very low. The highest maximum density was achieved in C. ellipsoidea at 15°C and 10. The slope constant of the linear relationship between semilogarithmic growth of C. ellipsoidea and day of culture was highest at 15°C and 10. The SGR and duration to reach maximum density of N. oculata were significantly affected by both temperature and salinity. However, maximum density of N. oculata was significantly affected by temperature, but not salinity. The highest maximum density was achieved in N. oculata at 25°C and 30, but SGR was significantly lower than that of N. oculata at 25°C and 10. The slope constant of the linear relationship between semilogarithmic growth of N. oculata and day of culture was highest at 25°C and 30. Based on these results, the condition of 15°C and salinity 10 seemed to be optimal for maximum density of C. ellipsoidea , and the condition of 25°C and 10 and 30 for SGR and maximum density for N. oculata , respectively.     

Annual variation of potential predation impacts on larval and juvenile marbled sole <Emphasis Type="Italic">Pseudopleuronectes yokohamae</Emphasis> by sand shrimp <Emphasis Type="Italic">Crangon uritai</Emphasis> in Hakodate Bay,Hokkaido

An investigation was conducted to evaluate the annual variation in potential predation impact (PPI) on larval and juvenile Pseudopleuronectes yokohamae by Crangon uritai in Hakodate Bay using predator-prey size relationships. Laboratory experiments were designed to estimate the favorable prey size of C. uritai through back-calculation of body length (BL) of P. yokohamae from sagittal otolith diameter observed in the stomachs of C. uritai. The most favorable prey-predator size ratio (BL of P. yokohamae-total length (TL) of C. uritai) class was 0.15–0.19, and ranged from 0.12–0.31. There was a significant positive correlation between the BL increase of P. yokohamae and the bottom water temperature in the field, such that BLs stagnated during the cold year of 1999 from April to June, and increased during the warm year of 2002. In contrast, no significant correlation was found between the TL increase of C. uritai and the bottom water temperature. Moreover, there were no significant differences in the mean TL of C. uritai between months (April–June) or years (1998–2002). Variation of PPI depended on the relationships between the growth rates of P. yokohamae and bottom water temperature. Therefore, the warm year of 2002 resulted in higher potential predation impact (PPI≥20), and it was at least 20 days shorter than that of the cold year of 1999. These results suggest that increased bottom water temperature in the nursery area was one of the most important factors for cumulative predation loss.     

Identification of a telomere sequence type in three sponge species (Porifera) by fluorescence <Emphasis Type="Italic">in situ</Emphasis> hybridization analysis

The telomere sequence type (TTAGGG)_n is known to be distributed in various phyla in the Animalia and in Mastigophora (Protista). However, the telomere type of Porifera (sponges), a phylum comprising the lowest multicellular animals, has not been reported. In this study, we examined the three sponge species Leucetta chagosensis, Halichondria japonica, and Halichondria panicea for the presence of the telomere type (TTAGGG)_n by fluorescence in situ hybridization (FISH). The oligonucleotide probe (TTAGGG)₇ clearly displayed signals on the interphase nuclei of all three sponges. In contrast, the (TTAGG)₇ probe, which has one base fewer than (TTAGGG)₇, did not display the signals. These results suggest that the telomeres of the three sponges consist of (TTAGGG)_n, which is identical to the sequence type found in many higher multicellular animals and in Mastigophora. Additionally, this is the first study to reveal a telomere sequence type for Porifera. Moreover, these results suggest that Porifera are phylogenetically related to Mastigophora, and supports the general theory that Porifera evolved from Mastigophora. Further, this study strongly suggests that the origin of the (TTAGGG)_n telomere sequence is to be found in a common ancestor of either the Bilateria and Porifera, or the Protista.     

Induction of (<Emphasis Type="Italic">Z</Emphasis>)-8-heptadecene in marine green alga <Emphasis Type="Italic">Bryopsis maxima</Emphasis> by mechanical wounding

ABSTRACT:   After mechanical wounding of the marine green alga Bryopsis maxima in a phosphate buffer, one main organic component was released to the solution. The organic compound was characterized by a molecular ion at 238  m/z by gas chromatography–mass spectrometry (GC-MS) and was deduced as 8-heptadecene by the fragment pattern. To identify the stereochemistry of 8-heptadecene, synthetic ( Z / E )-8-heptadecene was coinjected to the solution of wounded B. maxima . Comparing retention times and mass spectra with the synthetic sample, the target hydrocarbon obtained from wounded B. maxima was coincident with ( Z )-8-heptadecene.     

Growth of wild pearl oysters <Emphasis Type="Italic">Pinctada fucata,Pinctada margaritifera</Emphasis> and <Emphasis Type="Italic">Pinctada sugillata</Emphasis> (Bivalvia: Pteriidae) in Taiwan

In order to understand growth features of pearl oysters in the genus Pinctada, i.e. Pinctada fucata, Pinctada margaritifera, and Pinctada sugillata in Taiwan, a total of 3062 wild individuals of these species from juvenile to adult were collected monthyly from March 2001 to April 2002 in Jukeng, Pingtung County, south-west Taiwan. Quantitative measurements of live oysters were conducted for shell height (SH), shell length (SL), shell width (SW), hinge length (HL), and wet weight (WW). Different cohorts were identified through multiple length frequency analysis on SH of P. fucata and P. margaritifera, and growth curves with seasonal variation were estimated for these species. Pinctada fucata in Taiwan had a different seasonal growth pattern from the Japanese population, but had similar growth rates during the high growth period. The growth rate of P. margaritifera in Taiwan was slower than in French Polynesia, the Solomon Islands, and the Red Sea. Comparisons of morphological growth features among the three species show large differences in the SW-related features. Pinctada fucata in Taiwan had larger SW than in Japan and Korea. The differences in growth rates and morphological features suggested that the wild Taiwanese oysters may retain genetically pristine characteristics, thus genetic conservation might be urgently needed.     

Effects of fish CYP inducers on difloxacin <Emphasis Type="Italic">N</Emphasis>-demethylation in kidney cell of Chinese idle (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>)

A drug–drug interaction occurs when the effect of one drug is altered by the presence of another drug which is generally associated with the induction of cytochrome P450s (CYPs) activity. Thus, unexpected treatment failures often happen resulting from inappropriate coadministration in fisheries. However, little information is available about CYP induction in fish. The reaction of difloxacin (DIF) biotransformation to sarafloxacin (SAR) belongs to N-demethylation catalyzed mainly by CYP(s). In order to supply useful information on CYP induction, the present study assessed the effects of fish-specific CYP inducers on DIF N-demethylation and enzyme kinetics in kidney cell of Chinese idle (CIK; grass carp (Ctenopharyngodon idellus)) by RP-HPLC. Results demonstrated that the amounts of SAR formation and enzymatic parameters Clint and Vmax were significantly increased due to β-naphthoflavone (BNF) pretreatment. Therefore, we suggest that CYP1A may be involved in DIF N-demethylation in CIK. This study provides instructive information to ensure treatment success via avoiding CYP induction in fisheries.     

Somatic growth and otolith microstructure of larval and juvenile pacific cod <Emphasis Type="Italic">Gadus macrocephalus</Emphasis>

ABSTRACT:   Microstructures of sagittae and lapilli were examined in relation to somatic growth for reared larvae and juveniles of Pacific cod. The Laird–Gompertz model was fitted to the daily age and somatic growth relationship. Growth increments were deposited on a daily basis in both kinds of otoliths, with a check formed at hatching. Two subsequent checks and an accessory primordia (AP) occurred in the sagittae. The lapillus was adequate for increment width measurement through the early life stages. Sagittal and frontal plane of sagitta was adequate for measurement in the pre-AP and post-AP formation stages, respectively. The shift of desirable plane was caused by changes in otolith and increment shapes with AP formation. Back-calculated total lengths using the biological intercept method did not significantly differ with certain body lengths ( P  > 0.05), suggesting validity of back-calculation in this species. Using the back-calculated total length, morphological and ecologic changes that seemed to affect checks and AP formations are discussed.     

Effects of capture,loading density and transport stress on the mortality,physiological responses,bacterial density and growth of rohu <Emphasis Type="Italic">Labeo</Emphasis><Emphasis Type="Italic"> rohita</Emphasis> fingerlings

This study evaluated the effects of three levels of loading density (200, 300 and 400 g L⁻¹) and four levels of duration (1, 3, 6 and 9 h) on the mortality, plasma cortisol and chloride levels and growth of rohu Labeo rohita fingerlings in a transport simulation. While total immediate mortality was low (4–12%), delayed mortality was high (27–49%). Initial capture and handling were found to constitute the most stressful phase as indicated by elevated plasma cortisol. The levels of cortisol continued to rise at 6 and 9 h after transport, with the highest level observed at 9 h after transport. The level of plasma chloride fell after transport across all density levels and treatment durations. Specific growth rate (SGR, %/day) and total bacterial density (cfu/g) were not different between and within treatments, and between treatments and the controls. This study demonstrates that transport has a profound effect on the stress responses of rohu fingerlings. Although the level of immediate mortality was low, the higher level of delayed mortality has serious implications in terms of production inefficiency for grow-out farmers.     

Effects of heavy metals in river waters in Japan on immobility and mortality of <Emphasis Type="Italic">Daphnia magna</Emphasis> and <Emphasis Type="Italic">Oryzias latipes</Emphasis> larvae

ABSTRACT:   Samples of river waters containing high concentrations of zinc and other heavy metals but low concentrations of other anthropogenic contaminants were collected to investigate the relationship between toxicity of heavy metals and naturally present organic matters or hardness, as well as the effects of heavy metals on aquatic organisms. Acute toxicity tests were conducted for the water samples using Daphnia magna and medaka Oryzias latipes . Almost all the D. magna died in river waters containing high concentrations of zinc, but O. latipes in the same waters were hardly affected. Since the test organisms were not only exposed to zinc but also other heavy metals in the river waters, we examined the toxicity using toxic units composed of zinc, copper, lead, and cadmium. The results of a bioassay with the river waters showed that the mortality of D. magna did not depend solely on the total number of toxic units of heavy metals. The organic matters and the hardness of the river waters could decrease the acute toxicity of zinc and other heavy metals to D. magna .     

Improvement of larval rearing technique for mass seed production of snow crab <Emphasis Type="Italic">Chionoecetes opilio</Emphasis>

Larval rearing experiments were conducted to examine the potential for mass seed production of the snow crab Chionoecetes opilio using a total of eight 20-kL tanks. Tanks were equipped with agitators, which move the water using a rectangular blade to prevent the zoeas sinking to the bottom of the tank. Larval rearing water was treated with sodium nifurstyrenate once a week to reduce the chance of larval infection by pathogenic bacteria. Zo eas were fed with rotifers and Artemia nauplii. A total of 122 830 megalops and 16 660 first-stage crabs were produced. Thus, the potential for mass seed production of snow crab was determined. Survival rates up to the megalopal stage were high in tanks with a feeding regime that fed rotifers to larvae through an entire zoeal stage. This study also describes the fatty acid composition of snow crab larvae. It revealed that the first zoeas had a high DHA content and DHA/EPA ratio, but these values significantly decreased in the second stage zoeas and megalops. Improving the DHA content and/or DHA/EPA ratio of larvae should be important in studies on mass seed production technology of the snow crab.