Existence of a 400 kDa glycoprotein in the dermis of sea cucumber Apostichopus armata: partial purification and characterization

A large size (400 kDa) non-collagenous protein was detected as a major component in the extract, with neutral salt solution, from the dermis of sea cucumber Apostichopus armata. On SDS-PAGE analysis, the 400 K component shifted to a lower molecular weight component (about 200 K) by reduction with 2-mercaptoethanol, and they were both reactive for periodic acid-Schiff (PAS) reaction staining. From these results, this protein was suggested to be a glycoprotein consisting of disulfide-bonded two subunits with almost equal molecular weight (200 K). In addition to relatively high contents (>100/1,000 residues) of aspartic and glutamic acids, cysteine was also detected (6.1/1,000 residues) in amino acid analyses of this protein partially purified by anion-exchange column chromatography. These combined results suggest the structural similarity of the 400 K component to fibronectins from other vertebrate and invertebrate animals.     

An analysis of the effect of diet and genotype on protein and energy utilization by the black tiger shrimp,Penaeus monodon – why do genetically selected shrimp grow faster?

Selected (G8) and wild‐type (W) genotypes of black tiger shrimp (Penaeus monodon) juveniles (initial weight G8 = 9.14 ± 0.36 g per animal and W = 8.44 ± 0.10 g per animal) were fed either of two diet types in a clear‐water tank trial to examine the effects of diet type and genetics on growth and feed utilization parameters. Animals were fed twice daily at one of the five ration levels from starvation to apparent satiety. All uneaten feed was accounted for and moults removed. Starved animals were measured after 3 weeks; those fed were measured at both three and 6 weeks. Diet type varied by protein content, raw material choice and the presence [high‐specification diet (HSD)] or absence [low‐specification diet (LSD)] of bioactive substances. At the end of the study, faecal samples were also collected to determine the digestible protein and energy content of each diet by each genotype. Whole animal protein and energy content were also assessed from samples from the initial populations and those from each tank. Growth after 6 weeks of those animals fed to satiety showed that the G8 animals fed the HSD diet had grown at a rate of 2.56 g week^?1, significantly faster than any other treatment. Those G8 animals fed the LSD diet (1.81 g week^?1) had grown significantly faster than the W animals fed the HSD diet (1.25 g week^?1), while those W animals fed the LSD diet (0.61 g week^?1) grew the slowest. Using the data from the varying ration levels, we were able to define that the growth gains of the G8 animals were achieved not only by a greater appetite, but also through lower maintenance energy costs (29 versus 57 kJ kg^?0.8 day^?1) and a more efficient energy conversion (19.5% versus 11.6% when fed the HSD diet). Use of a low‐specification diet with the G8 and W shrimps limited their growth and impaired their potential as demonstrated by a curvilinear response of growth to intake. By comparison, those shrimp fed the HSD diet had a relatively linear growth response to intake.     

The effects of bioturbation by the Venus clam Cyclina sinensis on the fluxes of nutrients across the sediment–water interface in aquaculture ponds

A mesocosm experiment was conducted to study the effects of bioturbation by the marine bivalve Cyclina sinensis in an aquatic system and to examine how the bioactivities of this clam alter the rate at which nutrients are exchanged across the sediment–water interface. A dark incubation experiment was performed to determine the fluxes of inorganic nutrients (NH₄ ⁺, NO₃ ^? + NO₂ ^? and PO₄ ^3?), benthic chlorophyll a (Chl a), organic matter and sediment oxygen consumption (SOC) on days 1, 10, 20 and 30. C. sinensis destroyed the initial sediment surface and partially enhanced oxygen penetration into the sediment. The clam also increased the water content and volume of the oxic sediment. The analysis showed that increases in the clam density and time period produced higher fluxes of all the nutrients (NH₄ ⁺, NO₃ ^? + NO₂ ^? and PO₄ ^3?) in the bioturbated chambers compared with the control (non-bioturbated chambers). The concentration of Chl a in the sediment increased significantly at the end of the experiment (day 30) (p < 0.05). The SOC rate increased significantly with the increase in density and with the time period from day 10 to day 30 (p < 0.05). As a result, the fluxes of nutrients increased with increased SOC, producing oxygen-induced sediment mineralization. Our findings suggest that the activities of the clams could positively influence SOC and cause an increase in the fluxes of inorganic nutrients. These effects may substantially improve the primary productivity and water quality of earthen pond ecosystems.     

PCR cloning and identification of the β-haemolysin gene of Aeromonas hydrophila from freshwater fishes in China

In this study, β-haemolysin gene (AHTPS30HEM) of Aeromonas hydrophila was cloned from diseased fish in mainland China. AHTPS30HEM gene (AB021152) resulted in a 1589 bp fragment which covers the open reading frame (ORF) in region 5–1486 coding for 493 amino acids. Multiple alignment of AHTPS30HEM with other β-haemolysin amino acid sequences showed 18 amino acid substitutions between AHTPS30HEM and A. hydrophila β-haemolysin. Although the ORF sizes between AHTPS30HEM and Aeromonas species are different, four cysteins and four potential N-glycosylation sites were conserved. To identify the β-haemolysin-producing virulent or pathogenic A. hydrophila, a specific PCR to amplify 208 bp target DNA of β-haemolysin gene was established. Twenty strains containing pathogenic A. hydrophila, Aeromonas sobria, Vibrio anguillarum, Cytophaga columnaris, Pseudomonas fluorescens, and Yersinia yuckeri were investigated by PCR. Based on the cloned β-haemolysin sequences, the specific PCR method for identification of the β-haemolysin gene of A. hydrophila was established, and surveyed on those samples. The results indicated that β-haemolysin-specific PCR might be useful in the detection of pathogenic A. hydrophila.     

Origin identification of dried seaweed product “nori” by PCR–RFLP analysis of Pyropia yezoensis in the internal transcribed spacer ITS-1 region

Nucleotide sequences in internal transcribed spacer (ITS)-1 region derived from dried nori products produced in Japan, China, and the Republic of Korea were compared. Thalli contained in the Japanese products were genetically homogenous, and their nucleotide sequences in ITS-1 were identical to those of the reference strains of Pyropia yezoensis f. narawaensis. In Chinese products, the thalli were related to P. yezoensis strain Minomiasakusa. In contrast, the thalli in the Korean products were genetically heterogeneous, and several different P. yezoensis strains and other Pyropia spp. were used for dried nori products. In some thalli produced in both China and Korea, the DNA sequences of the ITS-1 region were identical with that of Japan, suggesting that the cultivar strains might have been transplanted from Japan to China in recent years. The 432-bp-long nucleotide sequences in the ITS-1 region of thalli derived from Japanese origin were cleaved to two restriction fragments at 154 and 278 bp by cleavage of PCR-amplified products using MspI. Conversely, almost all of the corresponding sequences derived from China and Korea were lacking MspI or other restriction patterns, except for nori products from some areas that cultivate a closely related strain to the Japanese cultivar.     

Growth and fatness of 1975–2002 year classes of Japanese sardine in the Pacific waters around northern Japan

We examined individual growth and fatness in the 1975–2002 year classes of Japanese sardine. Samples were collected at the feeding grounds in the Pacific waters off northern Japan during drastic fluctuations in the population in the 1970s to 2000s. Growth rates for ages 1–3 of the 1979–1988 year classes, which included low-recruitment year classes subsisting during the high population levels of the 1980s, were apparently slower than for other year classes. There was no obvious trend when comparing year classes, growth during the first year of life (age 0), and maximum body length (BL) at age ≥5. The condition factors (CF, indicating fatness) for adult sardines of BL ≥19 cm in the 1979–1983 year classes during the maximum population level of the mid-1980s were significantly lower than for other year classes. However, there were no apparent trends in CF variations for small sardines of BL <19 cm. The apparent decreases in growth rate and fatness were strongly related to the cumulative sum of population abundance that each year class experienced. It is thought that insufficient food owing to the density-dependent effect of an abundant population at feeding grounds resulted in a decrease in the growth rate for small-bodied sardines that are investing their energy intake in body growth, and a decrease in fatness for large-bodied adults that are accumulating fat for the next reproduction.     

Study of food web structure and trophic level in the sea ponds of an optimized culture model (jellyfish–shellfish–fish–prawn)

The food sources of the main economic animals and trophic levels of biotic communities from an optimized culture model (Rhopilema–Sinonovacula constricta–Paralichthys olivaceus–Penaeus chinensis) in a sea water pond in Donggang, Liaoning province were studied using the stable carbon and nitrogen isotopes technique. The results indicated that the values of δ¹³C range from (?27.28 ± 0.35) ‰ to (?16.65 ± 0.20) ‰ and the values of δ¹⁵N range from (3.68 ± 0.23) ‰ to (13.91 ± 0.26) ‰, both of which exhibited significant fluctuations. The δ¹³C values of P. chinensis, Macrobrachium mipponensis and P. olivaceus were comparatively higher than those of other aquaculture animals, and the δ¹⁵N values of P. olivaceus was also comparatively higher. The contribution to the food sources of aquaculture animals (Rhopilema, S. constricta, P. olivaceus and P. chinensis) was analyzed by using the IsoSource software. The results indicated that fish meat had the greatest contribution to Rhopilema, S. constricta and P. chinensis; P. chinensis had the greatest contribution to P. olivaceus, followed by the M. mipponensis. The trophic level of the biotic communities under that optimized culture model in a sea water pond was 3.54, in which P. olivaceus was in the fourth level (3.54); P. chinensis, Synechogobius hasta, Ablennes anastomella, M. mipponensis, Ditrema temmincki Bleeker, Chelon haematocheilus were in the third trophic levels; Rhopilema, Engraulis japonius, S. constricta and zooplankton (1.00) in the second trophic levels, suspended matters (0.15) including phytoplankton, bacteria, humus, etc., were in the primary trophic levels.     

CYP1A1 immunolocalization in the olfactory organ of rainbow trout and its possible induction by β-naphthoflavone: analysis in adults and embryos around hatching

Cytochrome P4501A1 (CYP1A1) isoform, which is known as being of major toxicological significance, has been well-studied in the mammalian olfactory mucosa. Only few studies have dealt with this biotransformation system in the fish olfactory organ which is particularly vulnerable to waterborne xenobiotics since sensory neurons are in direct contact with the aquatic environment. The present immunocytochemical study describes the cellular and subcellular distributions of CYP1A1 in the olfactory organ of rainbow trout in both adults and embryos around hatching. The enzyme inducibility in response to a 4-day exposure to waterborne -naphthoflavone (0.1 mg l^–1), a model inducer of CYP1A1, was also examined. In untreated adult fish, CYP1A1 was almost exclusively expressed in the nonsensory epithelium which covers the edges and the tip of the lamellae. Both goblet and ciliated nonsensory cells appeared immunoreactive. In -naphthoflavone-treated fish, in addition to a strong labeling in the nonsensory epithelium, ciliated nonsensory cells in the olfactory epithelium appeared well-labeled. Four days before hatching, only a few cells were weakly stained in the placodal epithelium of some embryos. By 7 days post-hatching, the enzyme expression was increased in the olfactory pit and it was restricted to ciliated nonsensory cells. No evident CYP1A1 induction was detected in either embryos or alevins. Results suggest the presence of a two-line CYP1A1 biotransformation system in the adult fish olfactory organ: a basal level of enzyme expression insured by the nonsensory epithelium and an additional line in which the sensory epithelium is activated in response to CYP1A1 inducers. This system might take place during development in parallel with the onset of the nonsensory epithelium.