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.     

Dietary value of benthic diatoms for post-larval abalone <Emphasis Type="Italic">Haliotis diversicolor</Emphasis> associated with feeding transitions

ABSTRACT:   The feeding behavior and growth of post-larval Haliotis diversicolor with initial shell lengths (SL) of approximately 500 μm (Exp. 1-1 and 1-2), 800 μm (Exp. 2), and 1200 μm (Exp. 3) were studied in a laboratory setting while they fed on four species of benthic diatom Achnanthes longipes , Cocconeis sublittoralis , Cylindrotheca closterium , and Navicula ramosissima . Exp. 1-1 and 1-2 revealed no marked differences in post-larval growth rates (mean 24–39 μm SL/day) among the diatom species. However, marked differences in growth rates among the species were revealed in Exp. 2 and 3. Three species, A. longipes , Co. sublittoralis, and Cy. closterium , produced faster growth (Exp. 2 mean 29–51 μm/day, Exp. 3 mean 36–44 μm/day) than N. ramosissima (Exp. 2 mean 18 μm/day, Exp. 3 mean 23 μm/day). Post-larvae fed N. ramosissima had lower digestion efficiency (42.8%) than those fed other diatom species (90.7–100%). Diatom extracellular substances appeared to be principally used from post-settlement to 800 μm SL, and diatom cell contents were required to produce rapid growth of larger post-larvae (>800 μm SL). It is likely that the availability of each diatom for post-larvae was affected by diatom morphology, attachment strength, frustule strength, and post-larval size.     

Purification and characterization of the amylase from a small abalone <Emphasis Type="Italic">Haliotis sieboldii</Emphasis>

ABSTRACT:   Amylase, with MW of 59 kDa, was purified from small abalone Haliotis sieboldii by ammonium sulfate fractionation, CM Sepharose Fast Flow and Sephacryl S-100 HR chromatographies. The optimal pH and temperature of purified amylase were 6.0 and 37°C, respectively. The purified enzyme was stable at pH 6.0–8.0 and low temperatures. It was activated by Ba²⁺, Mg²⁺, Ca²⁺, Co²⁺, Ni²⁺, Mn²⁺, K⁺, Ag⁺, Na⁺ and Li⁺, but completely or partially inhibited by Al³⁺, Cu²⁺, Cd²⁺, Hg²⁺ and Zn²⁺. EDTA could completely inhibit, while iodoacetamide, N-ethylmaleimide and urea partially inhibit the purified amylase. According to the digestion mode of various polysaccharides, the purified enzyme was considered to be an α-amylase.     

Ontogenetic habitat shift in abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis>: a review

Ecological literature on abalone Haliotis discus hannai populations is reviewed to identify processes and factors driving ontogenetic shifts in habitat. Abalone recruitment is related to the algal community type, with abalone shifting with growth from deeper crustose coralline algal (CCA) habitats to shallower kelp bed habitats via algal turfs. Timing of the habitat shifts is linked to ontogenetic changes in diet, from benthic diatoms to a diet dominated by macroalgae. Susceptibility to predation appears to change, concomitant with the shift from cryptic microhabitats during early-life stages to more exposed habitats during latter-life stages. Interstitial spaces between cobbles and boulders in the CCA habitat are considered to be important for reducing predation risk. The loss of CCA nursery habitat through sedimentation and macroalgal overgrowth likely negatively affects abalone recruitment. Preservation of diverse algal communities, including suitable habitats for the different abalone growth stages, is critical for successful abalone recruitment.     

Complete nuclear ribosomal DNA sequence analysis of Pacific abalone <Emphasis Type="Italic">Haliotis discus hannai</Emphasis>

Pacific abalone Haliotis discus hannai (Haliotidae, Gastropoda) is an economically important shellfish species in northern China. The complete nuclear ribosomal DNA (nrDNA) of Pacific abalone was amplified, sequenced, and analyzed. The length of the nrDNA was determined to be around 10.7 kb, and to contain, in order, small subunit ribosomal RNA (nrSSU) genes (1871 bp), internal transcribed spacer (ITS, 759–762 bp), large subunit ribosomal RNA (nrLSU) gene (3411 bp), and an intergenic spacer (IGS, 4624–4654 bp). The SSU and LSU regions were almost identical in different individuals, and show little variation from those of other abalone species. The two different variations of the ITS2 region were presented, and this phenomenon also existed in other species. A phylogeny tree was constructed, based on ITS region sequence datasets, to determine the evolutionary relationships of abalones. Abalones have two major subclades, mainly distributed in the North Pacific, Europe and Australia. The IGS region of the nrDNA was sequenced and analyzed for the first time. Several repeat fragments were present upstream of the sequence, and were significantly different between individuals (93.86% sequence identity). The complete nrDNA sequence will be useful for the classification, identification, phylogeny, germplasm management, and breeding of this shellfish.     

Genetic difference between Ezo-awabi <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> and Kuro-awabi <Emphasis Type="Italic">H. discus discus</Emphasis> populations: Microsatellite-based population analysis in Japanese abalone

ABSTRACT:   This study documents the genetic population relationships between Ezo-awabi Haliotis discus hannai and Kuro-awabi H. discus discus by means of the microsatellite technique with an emphasis on the extent of genetic difference between Ezo- and Kuro-awabi populations. Eight markers were employed to screen five populations each collected from Ezo- and Kuro-awabi habitats. All eight loci showed polymorphisms in all populations (number of alleles per locus, 11.1–14.1; averaged expected heterozygosity, 0.64–0.70). The number of loci that yielded significant genetic heterogeneities (allele frequency distribution and/or pairwise θ ) between Ezo- and Kuro-awabi populations was larger than that generated in comparisons between populations within Ezo- and Kuro-awabi habitats. According to the bootstrap neighbor-joining trees constructed on the basis of two genetic distance measures ( D _A and D _ST), the 10 populations fell into two clusters of populations (Ezo-awabi and Kuro-awabi habitat groups), and the results of hierarchical amova testing also supported the neighbor-joining clustering. The outcomes presented here suggest that the microsatellite markers used in this study are potentially an efficient means to differentiate Ezo- and Kuro-awabi populations.     

Identification of juvenile abalone <Emphasis Type="Italic">Haliotis diversicolor</Emphasis> based on number of open and sealed respiratory pores

ABSTRACT:   Changes in the number of respiratory pores (open pores) and their imprints (sealed pores) in post-larvae and juveniles were observed and compared for four abalone species Haliotis diversicolor , H. discus discus , H. madaka , and H. gigantea . The first open pore was evident at a shell length (SL) of 1.5 mm in H. diversicolor , 1.9–2.0 mm SL in H. discus discus , and 2.3–2.4 mm SL in H. madaka and H. gigantea . The number of open pores in H. diversicolor gradually increased with growth, with four to five pores at 2.5–18.0 mm SL and five to six pores at 18.0–27.0 mm SL. The other three species maintained four to five open pores after they reached 3.4–4.5 mm SL. The total number of open and sealed pores (TNP) was greater in H. diversicolor than in the other species at the same SL. Juvenile H. diversicolor were identified among field-caught abalone by the difference in the relationship between SL and TNP (SL–TNP relationship) and also by the monoclonal antibody reaction method. The results of the two methods were in perfect agreement, indicating that our method using the SL–TNP relationship is reliable for the identification of H. diversicolor .     

Anti-allergic effects of the brown alga <Emphasis Type="Italic">Eisenia arborea</Emphasis> on Brown Norway rats

To investigate the anti-allergic effects of the brown alga Eisenia arborea. A strain of Brown Norway rats know to strongly respond to immunoglobulin E (IgE) were used as an allergy model animal. The rats were immunized with ovalbumin by oral administration. The levels of serum IgE and histamine were suppressed in the rats fed a diet supplemented with dried E. arborea powder. As for the cytokine pattern, the interferon-γ production in the spleens and mesenteric lymph nodes (MLN) was enhanced, and the interleukin-4 (IL-4) production in the spleens and/or IL-10 production in the spleens and MLN were suppressed. These results, together with the change in the Th1/Th2 balance, indicate that the rats fed with E. arborea became more anti-allergic, suggesting that E. arborea might possess anti-allergic effects.     

Dietary supplementation of curcumin augments heat stress tolerance through upregulation of <Emphasis Type="Italic">nrf-2</Emphasis>-mediated antioxidative enzymes and <Emphasis Type="Italic">hsps</Emphasis> in <Emphasis Type="Italic">Puntius sophore</Emphasis>

Heat stress is one of the major environmental concerns in global warming regime and rising temperature has resulted in mass mortalities of animals including fishes. Therefore, strategies for high temperature stress tolerance and ameliorating the effects of heat stress are being looked for. In an earlier study, we reported that Nrf-2 (nuclear factor E2-related factor 2) mediated upregulation of antioxidative enzymes and heat shock proteins (Hsps) provide survivability to fish under heat stress. In this study, we have evaluated the ameliorative potential of dietary curcumin, a potential Nrf-2 inducer in heat stressed cyprinid Puntius sophore. Fishes were fed with diet supplemented with 0.5, 1.0, and 1.5% curcumin at the rate 2% of body weight daily in three separate groups (n = 40 in each group) for 60 days. Fishes fed with basal diet (without curcumin) served as the control (n = 40). Critical thermal maxima (CTmax) was determined for all the groups (n = 10, in duplicates) after the feeding trial. Significant increase in the CTmax was observed in the group fed with 1.5% curcumin- supplemented fishes whereas it remained similar in groups fed with 0.5%, and 1% curcumin-supplemented diet, as compared to control. To understand the molecular mechanism of elevated thermotolerance in the 1.5% curcumin supplemented group, fishes were given a sub-lethal heat shock treatment (36 °C) for 6 h and expression analysis of nrf-2, keap-1, sod, catalase, gpx, and hsp27, hsp60, hsp70, hsp90, and hsp110 was carried out using RT-PCR. In the gill, expression of nrf-2, sod, catalase, gpx, and hsp60, hsp70, hsp90, and hsp110 was found to be elevated in the 1.5% curcumin-fed heat-shocked group compared to control and the basal diet-fed, heat-shocked fishes. Similarly, in the liver, upregulation in expression of nrf-2, sod, catalase, and hsp70 and hsp110 was observed in 1.5% curcumin supplemented and heat shocked group. Thus, this study showed that supplementation of curcumin augments tolerance to high temperature stress in P. sophore that could be attributed to nrf-2-induced upregulation of antioxidative enzymes sod, catalase, gpx, and the hsps.     

Spatial learning-induced <Emphasis Type="Italic">egr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> expression in telencephalon of gold fish <Emphasis Type="Italic">Carassius auratus</Emphasis>

The immediate-early gene (egr-1) expression was used to examine the neuron’s response in telencephalon of goldfish during spatial learning in small space. Fishes were pre-exposed in the experimental apparatus and trained to pick food from the tray in a rectangular-shaped arena. The apparatus was divided into identical compartments comprising three gates to provide different spatial tasks. After the fish learned to pass through the gate one, two more gates were introduced one by one. Fish made more number of attempts and took longer time (P < 0.05) to pass through the first gate than the gate two or three. This active learning induces the expression of egr-1 in telencephalon as established by western blot analysis. Subsequently, the fish learn quickly to cross the similar type of second and third gate and make fewer errors with a corresponding decline in the level of egr-1 expression. As the fish learned to pass through all the three gates, third gate was replaced by modified gate three. Interestingly, the level of egr-1 expression increased again, when the fish exhibit a high exploratory behavior to cross the modified gate three. The present study shows that egr-1 expression is induced in the telencephalon of goldfish while intensively acquiring geometric spatial information to pass through the gates.     

Dietary total phosphorus requirement of juvenile yellow catfish <Emphasis Type="Italic">Pelteobagrus fulvidraco</Emphasis>

The present experiment was conducted to determine the dietary total phosphorus requirement of juvenile yellow catfish Pelteobagrus fulvidraco. Six diets with increasing dietary phosphorus concentration (0.43, 0.55, 0.78, 0.90, 1.05 and 1.18% of dry matter, respectively) were fed to triplicate groups of 20 fish each (mean initial body weight, 2.68 ± 0.08 g, mean ± SD) to apparent satiation for 7 weeks. Weight gain and specific growth rate (SGR) increased with increasing dietary phosphorus level from 0.43 to 0.90% and then declined over dietary phosphorus level of 0.90% (P < 0.05). Phosphorus retention increased with increasing dietary phosphorus level from 0.43 to 0.55% and then declined over dietary phosphorus level of 0.55% (P < 0.05). Dietary phosphorus levels significantly influenced whole body crude protein and ash contents (P < 0.05), but not whole body lipid content (P > 0.05). Vertebrae phosphorus content increased with dietary phosphorus level from 0.43 to 0.78% (P < 0.05) and then plateau over the level of 0.78% (P > 0.05). Dietary phosphorus level significantly influenced condition factor, viscerosomatic index and hepatosomatic index (P < 0.05). The relationship between SGR and whole body ash content against dietary phosphorus levels could be expressed as a second-order polynomial equation and the points of 0.89 and 0.85% were considered to be the optimal dietary total phosphorus level, respectively. Based on broken-line analysis of vertebrae phosphorus content, the minimal dietary total phosphorus requirements for maintaining maximum phosphorus storages were estimated to be 0.76% phosphorus.     

Differences in longitudinal distribution patterns along a Honshu stream of brown trout <Emphasis Type="Italic">Salmo trutta</Emphasis>, white-spotted charr <Emphasis Type="Italic">Salvelinus leucomaenis</Emphasis> and masu salmon <Emphasis Type="Italic">Oncorhynchus masou</Emphasis>

Brown trout Salmo trutta were first introduced into Japan in 1892, and they currently naturally reproduce in several rivers in Honshu and Hokkaido, Japan. Although negative impacts of brown trout introductions on native salmonid fishes have been documented in some Hokkaido rivers, studies of ecological interactions between brown trout and native salmonid fishes on Honshu are limited. In this study, we describe the longitudinal distribution patterns of introduced brown trout, white-spotted charr Salvelinus leucomaenis and masu salmon Oncorhynchus masou in a 4 km stretch of a stream in central Honshu. Underwater observations were conducted in all pools within upstream, middle and downstream sections (190–400 m in length) of this stretch in order to estimate the densities of these species. Only white-spotted charr was observed in the upstream section, while brown trout and masu salmon were observed in the middle and downstream sections. Masu salmon densities, however, were much lower than brown trout densities. In the downstream section, white-spotted charr was absent. These results are consistent with results from previous studies of Hokkaido rivers, where it was found that white-spotted charr in low-gradient areas tend to be displaced by brown trout.     

Use of thraustochytrid <Emphasis Type="Italic">Schizochytrium</Emphasis> sp. as source of lipid and fatty acid in a formulated diet for abalone <Emphasis Type="Italic">Haliotis asinina</Emphasis> (Linnaeus) juveniles

The effects of using thraustochytrid Schizochytrium sp. as source of lipid and fatty acids in a formulated diet on growth, survival, body composition, and salinity tolerance of juvenile donkey’s ear abalone, Haliotis asinina, were investigated. Treatments consisted of diets either containing a 1:1 ratio of cod liver oil (CLO) and soybean oil (SBO) (Diet 1) or thraustochytrid (Diet 2) as source of lipid and fatty acids at 2 % level. Natural diet Gracilariopsis heteroclada (Diet 3) served as the control. No significant difference in growth was observed in abalone fed Diet 3 (SGR: 5.3 % BW day^?1; DISL: 265 μm day^?1) and Diet 2 (SGR: 5.2 % BW day^?1; DISL: 255 μm day^?1). Survival ranged from 78 to 85 % for all treatments and was not significantly different from each other. A 96-h salinity stress test showed highest survival of 84 % in abalone fed Diet 2 compared with those fed diets 1 and 3 (42 %). The high growth rate of abalone fed Diet 2 and high tolerance to low salinity could be attributed to its high DHA content (8.9 %), which resulted to its high DHA/EPA ratio of 10.5 %. These fatty acids play a significant role in abalone nutrition. The fatty acid profile of abalone meat is a reflective of the fatty acid profile of the oil sources in the diet. The present study suggests that the use of Schizochytrium oil in lieu of CLO and SBO can support good growth of abalone which is comparable with abalone fed the natural seaweeds diet.     

<Emphasis Type="Italic">Neoheterobothrium hirame</Emphasis> (Monogenea) alters the feeding behavior of juvenile olive flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

We monitored feeding behavior and survival of starved juvenile olive flounder experimentally infected with the gill monogenean Neoheterobothrium hirame. Infected flounder increased amount of the time spent in the water column by 117% when trying to capture live mysids, Neomysis sp. They also showed different feeding patterns from those of uninfected fish and made fewer attacks towards prey during one feeding attempt. Although the average numbers of mysids captured by individuals were similar between infected and uninfected fish, heavily infected fish tended to catch less prey. These results indicate that N. hirame reduces the feeding efficiency of the host for capturing live prey and possibly makes them more vulnerable to predation during feeding. We could not detect any obvious difference in survival rates between uninfected, lightly and heavily infected fish during 3 months of starvation. There was no evidence that starvation makes fish more susceptible to N. hirame. The present study provides first experimental evidence that N. hirame affects feeding behavior of juvenile olive flounder and supports the idea that this parasite indirectly reducing the host’s survival and may be responsible for the recent reduction of the flounder population in Japan.     

Characterization of <Emphasis Type="Italic">kiss2</Emphasis> and <Emphasis Type="Italic">kissr2</Emphasis> genes and the regulation of kisspeptin on the HPG axis in <Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>

Reproduction allows organisms to produce offspring. Animals shift from immature juveniles into mature adults and become capable of sexual reproduction during puberty, which culminates in the first spermiation and sperm hydration or ovulation. Reproduction is closely related to the precise control of the hypothalamic–pituitary–gonadal (HPG) axis. Kisspeptin peptides are considered as the important regulator of HPG axis in mammalian. However, the current understanding of kisspeptin in flatfish is not comprehensive. In this study, we cloned and analyzed the kiss2 and kissr2 genes in Cynoglossus semilaevis. Interesting alternative splicing in the 5′-untranslated regions (UTR) of the Cskissr2 gene was found. The expression profiles of Cskiss2 and Cskissr2 showed relative high messenger RNA (mRNA) levels at the late gastrula stage during embryonic development, at total length = 40 mm during early gonadal differentiation, and in the brains and gonads of all investigated tissues. These results suggested that the kisspeptin system participated in embryogenesis and in the regulation of gonadal differentiation and development. Considering that the control and regulatory mechanisms of kisspeptin in the central reproductive axis are still unclear, we documented that the intramuscular injection of kisspeptin caused different sGnRH and cGnRH mRNA levels in a dose- and tissue-dependent manner. The mRNA expressions of FSH and LH were stimulated in the ovary and were inhibited in the testis under the kisspeptin treatments. These results provided foundations for understanding the roles of kisspeptin in the neuroendocrine system in fish. The manipulation of the kisspeptin system may provide new opportunities to control the gonadal development and even reproduction in fish.     

Low-salinity tolerance of juvenile fugu <Emphasis Type="Italic">Takifugu rubripes</Emphasis>

ABSTRACT:   To study the adaptability of juvenile fugu Takifugu rubripes to low-salinity environments, fish were transferred from full-strength seawater (100% SW) to freshwater (FW) and 25, 50, 75 and 100% SW, and checked for mortality over 3 days. No mortality was observed in 25–100% SW, whereas all fish died in FW. In fish transferred to 25–100% SW, blood osmolality was maintained within a physiological range. To further explore the lower limit of salinity that fugu could tolerate, fish were transferred from 100% SW to FW and 1, 5, 10, 15 and 25% SW. All fish survived in 5–25% SW, but fish died in FW and 1% SW. In fish surviving transfer to FW and 1 and 5% SW, blood osmolality was decreased to a near sublethal level of approximately 300 mOsm/kg·H₂O. Therefore, the lower limit of salinity tolerance is estimated to lie between 5 and 10% SW. Preacclimation in 25% SW for 7 days did not essentially affect the survival salinity range. Although survival rates and blood osmolality were slightly improved by preacclimation in 25% SW, blood osmolality was markedly decreased in salinities less than 10% SW, as was seen in the direct transfer. Neither chloride cell morphology nor sodium-potassium adenosinetriphosphatase activity in the gills showed a significant change following transfer to low salinities. These findings indicate that fugu can be adapted to hypoosmotic environments to some extent, exerting hyperosmoregulatory ability, although chloride cells are less likely to absorb ions in hypoosmotic environments.     

A dynamic simulation analysis of Japanese abalone (<Emphasis Type="Italic">Haliotis discus hannai</Emphasis>) production in Chile

In this paper the author develops a dynamic simulation model and uses it to analyze the economics of pond cultivation of Japanese abalone in Chile. The analysis is done in continuous time by defining discounted free cash flows on the basis of explicit relationships between several biological, technical, and economic functions. In a culture batch starting with 600,000 seedlings when abalone has an average length of 19 mm, and the optimal harvest time is day 763 from stocking, harvesting at that optimum time implies a per-unit cost of production of approximately CH$320 per abalone. This includes consumption of 443,644 kg algae per generation, providing approximately 1.46 kg algae per abalone, and an electric energy requirement of about 433,200 kWh. Even at the optimum time of harvest, the profits are rather low. This explains why Japanese abalone culture has been substantially reduced in the north of Chile. To obtain abalones with an average weight of 100 g, harvest should be on day 1,460, when operating net present value is negative. In addition, the effect of changes of some key parameters on the results is analyzed via a sensitivity analysis.