Effects of lectin in the scleractinian coral Ctenactis echinata on symbiotic zooxanthellae

We report herein the presence of a lectin in the scleractinian coral Ctenactis (Fungia) echinata. The lectin bound preferentially to lactose, melibiose, and d-galactose. The purified lectin CecL was composed of several isolectins, and it was found to have a molecular mass of 67.4 kDa via gel filtration. Glycopeptidase F-treated CecL showed a single band at 32.5 kDa. The mass/charge ratios of the reduced CecL peaks were equivalent to half those of the native peaks. These results suggest that CecL is composed of two glycosylated polypeptides linked by interchain disulfide bonds. In a biological activity test using a zooxanthellal culture (Dinoflagellate Symbiodinium) clonally isolated from Fungia cf. fungites, CecL transformed the flagellated motile form of Symbiodinium into the nonmotile coccoid form, a form equivalent to the symbiotic stage. The activity of CecL on Symbiodinium cells was concentration dependent, and 100 μg/ml CecL arrested Symbiodinium cells in the coccoid form for 5 days. CecL also suppressed the growth of Symbiodinium cells, unlike the octocoral lectin derived from Sinularia lochmodes, which arrests Symbiodinium cells in the coccoid form but does not affect the growth of the coccoid. This result provides further evidence that coral lectins play a role in symbiont engagement and maintenance in zooxanthellae–coral symbiosis.     

Genetic variations within Symbiodinium clade C among zooxanthellate corals (Scleractinia) in the temperate zone of Japan

In a recent study most of the temperate-living corals were found to harbor stress-sensitive Symbiodinium clade C, which is common in the tropical zone. In this study, we investigated the genetic variations within Symbiodinium clade C among corals living in temperate waters of Japan (17 genera; 26 species) using rDNA internal transcribed spacers (ITSs) to clarify the genetic differences between climatic zones. We also focused on two widespread coral species, Acropora hyacinthus and Pocillopora damicornis, to compare the genetic differences of Symbiodinium clade C between subtropical and temperate regions. We found multiple haplotypes of Symbiodinium ITSs in clade C, which formed nine subclades. Our data indicate that five genera (Acropora, Cyphastrea, Stylocoeniella, Porites, Pocillopora) contained host-specific Symbiodinium, while members of the family Faviidae and others contained several Symbiodinium ITS haplotypes from two or three subclades. A comparative analysis using Acropora and Pocillopora also revealed that the temperate zone coral population formed a single subclade of Symbiodinium clade C that was genetically isolated from those of subtropical populations. In summary, the majority of corals living in temperate waters of Japan, such as faviid corals, have several ITS haplotypes which possibly are utilized as an opportunistic survival strategy under severe environmental conditions, whereas Acropora and Pocillopora at the very least harbor temperate zone-specific Symbiodinium.     

Establishment of a model for chemoattraction of <Emphasis Type="Italic">Symbiodinium</Emphasis> and characterization of chemotactic compounds in <Emphasis Type="Italic">Acropora tenuis</Emphasis>

Corals harbor symbiotic dinoflagellates, Symbiodinium spp., acquired from surrounding environments. Because Symbiodinium are present at low densities in the water column, corals may attract these symbionts using chemotactic compounds. To examine whether corals contain chemotactic compounds, we established an assay to measure the chemotactic activity for Symbiodinium using an extract of the coral Acropora tenuis, a major reef-building coral in Japan. Our assay revealed that Symbiodinium strain NBRC102920 (clade A), which is taken up by juvenile A. tenuis polyps, is attracted to crude A. tenuis extracts. We found that the chemotactic compounds are water-soluble, heat-labile macromolecules and that the chemotactic activity was inhibited by N-acetyl-d-glucosamine (GlcNAc). We separated the GlcNAc-binding fraction (Fr-ActL) and identified it as the most plausible candidate for the chemoattractant, since the chemotactic activity of the crude A. tenuis extract appeared to be mainly attributable to the activity of Fr-ActL and was also inhibited by the addition of GlcNAc. These results indicate that chemoattraction is mediated via the binding of Symbiodinium to Fr-ActL.     

Molecular characterization of coral sulfate transporter homolog that is up-regulated by the presence of symbiotic algae

Reef-building corals are in symbiosis with the dinoflagellates Symbiodinium spp. In our previous study, the expression of two mRNAs (AtSym-01 and 02) was up-regulated by the presence of Symbiodinium cells (strain PL-TS-1) in juveniles of the reef-building coral Acropora tenuis. In this study, the AtSym-01 mRNA was found to encode a cnidarian ortholog of the vertebrate SLC26A11 sulfate transporter. The AtSym-01 and human SLC26A11 proteins exhibited 46% identity over 542 amino acids. Real-time polymerase chain reaction analysis showed that the expression level of the AtSym-01 mRNA was also increased by the presence of Symbiodinium strain CCMP2467 cells. Immunohistochemical analysis was performed using polyclonal antibodies against the AtSym-01 protein, in order to study the distribution of the protein in A. tenuis tissues. Cell-specific immunoreaction was observed in diverse tissues in juvenile and adult specimens. Notable immunoreaction was observed in mucocytes (mucus cells) in the outer epithelium of juveniles, and gastrodermal cells located between the coelenteron and skeleton of the adult colony. These observations suggest the possibility that the AtSym-01 protein is involved in uptake of sulfate ion for synthesis of sulfated macromolecules that are contained in the mucus and organic matrix of the calcified skeleton.     

Recruitment patterns and early growth of acroporid corals in Manado, Indonesia

A coral settlement device was used to examine recruitment patterns of Acropora at two stations (st.; A and B) in Bunaken Island, Manado. The recruitment timing of Acropora was not determined because of mixing with Isopora. A marine block (MB) was used to observe the growth of settled acroporid corals at st. A. Within a year of settlement (February–May of the following year), the corals which had settled on the MB had grown to a size of 13.4 ± 5.86 mm. They were dominated by Isopora, with a small proportion of Acropora. A supplementary experiment again using the MB was conducted at three other sites (st. C, D, E) where Isopora was not abundant. The size of the corals which settled on the MB was nearly the same as that of the corals at st. A. Juvenile Acropora corals (≤3 cm, encrusting form) were measured on the Manado coast (st. F) between February and May. At this site, Acropora corals were dominant and no Isopora corals were observed; the size of Acropora was 18.5 ± 5.01 mm. Acroporid corals in February and March at st. A appeared smaller than those at st. F and grew to nearly the same size in early April.     

基于线粒体COⅠ基因和形态学方法的3种梭子蟹科溞状幼体物种鉴定

为调查黄海蟹类资源情况,并识别梭子蟹科溞状幼体各阶段形态,利用2018年5月下旬至8月上旬在黄海海州湾及邻近水域的鱼类浮游生物大面综合调查获得的海洋蟹类幼体样品,基于线粒体COⅠ基因和形态学方法对梭子蟹科溞状幼体种类进行了鉴定。结果显示,春夏季调查水域共出现3种梭子蟹科幼体:三疣梭子蟹(Portunus trituberculatus)、日本蟳(Charybdis japonica)和双斑蟳(Charybdis bimaculata);线粒体COⅠ基因测序可准确识别3个物种。对3种梭子蟹溞状幼体的形态学观察结果表明,背棘与额棘形态、尾叉外缘小刺的有无可作为区分上述3种梭子蟹溞状幼体的主要形态学指标。根据3个物种幼体形态特征建立了梭子蟹科溞状幼体分类检索表,基于检索表对幼体进行分类鉴定,并选择8月上旬航次形态学鉴定的80个三疣梭子蟹溞状幼体、24个日本蟳溞状幼体、70个双斑蟳溞状幼体样品进行分子测序验证。结果表明,上述3种蟹类溞状幼体的形态学鉴定准确率分别达到93.75%、100%和97.14%,误判皆发生于溞状幼体Ⅰ期梭子蟹科3个物种之间。研究初步建立了黄海中部梭子蟹科溞状幼体形态鉴定...     

Effect of p-Nonylphenol on sperm motility in Japanese medaka (Oryzias latipes)

This study examined the effects of exposure to p-Nonylphenol (NP) on the motility of the spermatozoa of Japanese medaka (Oryzias latipes). Spermatozoa collected from adult males reared for 2 weeks in aqueous solutions of 20 and 100 μg/L NP showed similar swimming speeds but significantly lower percentages of motile spermatozoa than those of untreated controls. These results suggest that even a short exposure to NP might reduce significantly spermatozoan viability in fish.     

Hatchery selection may depress the number of motile sperm but intensify selection for their swimming velocity in the Arctic charr

The ability of captive breeding programs to maintain genetic diversity and fitness has often been questioned. Recent studies suggest that fitness loss can be extremely rapid in various traits, but it is poorly known how captive breeding affects sperm quality and thus male fertility. We studied the potential effects of hatchery-induced selection on traits indicative of semen quality, in four generations of captive bred Arctic charr Salvelinus alpinus L. We found that the number of motile sperm cells decreased, but that the swimming velocity of the sperm increased over generations. The independent effects of inbreeding and hatchery selection on semen traits could not be separated, but since in small captive broodstocks, both of them often act together, the present results should indicate real changes of semen traits in such situations. Taken together, the present data suggest that the fitness loss in some semen traits (number of motile sperm) can be extremely rapid, but selection on other, closely related traits (swimming velocity) may delay or counteract the overall deterioration of male fertilizing ability during captivity.     

Visualization of musculature of the larval greater amberjack Seriola dumerili (Risso) using whole‐mount immunostaining with special reference to cranial muscles

Muscles play important roles in feeding, respiration and swimming during not only adult stage but also larval stage in fish. However, there is no information of the muscle development in larval greater amberjack, Seriola dumerili (Risso). Here, we investigated muscle development in the greater amberjack, focusing primarily on the cranial muscles from 0 days post hatch (dph) to 12 dph using a modified whole‐mount immunohistochemical staining method. We found that the muscles required for feeding develop by 3 dph, when the larvae begin to feed. Subsequently, muscle composition in the dorsal branchial arches changes to the adult form between 5 and 8 dph. At 8 dph, all the muscles required for feeding and respiration appear, whereas the dorsal, pelvic and caudal fin muscles required for swimming develop later. This report provides fundamental information on larval greater amberjack muscle development, which will enable the detection of abnormal larval muscles and improve larval rearing techniques by modifying environmental conditions.     

Reciprocal hybrids of tench Tinca tinca (L.) × bream Abramis brama (L.), and tench × carp Cyprinus carpio L., and some characteristics of their early development

Reciprocal hybrids between tench Tinca tinca (L.) and carp Cyprinus carpio L., and tench and bream Abramis brama (L.) were produced artificially. The survival of all these hybrids during embryogenesis was quite high. The highest survival rate (over 60%) at the eyed stage was observed for tench and bream hybrids (both sex combinations). The hatching rates of these hybrids were also over 60%. The number of larvae with some abnormalities (i.e. deformed body) was low. In contrast, the hatching rates of tench and carp hybrids were very low (0.2%). From over 1000 fertilized eggs, only three specimens started swimming, and only one specimen survived to juvenile stage. Embryos of hybrids and their parental species differ in morphological features. These differences were also visible in the juveniles. Body parameters of juvenile hybrids produced from three species had intermediate values in comparison to parental fish.     

Effects of a research trawl on a hard-bottom assemblage of sponges and corals

The effects of a research trawl on several sponge and coral species was assessed in a shallow-water, hard-bottom area located southeast of Savannah, Georgia. The study entailed a census of the numerically dominant species in replicate 25-m² quadrats located along five transects established across a trawling alley. The density of undamaged sponges and corals was assessed in trawled and non-trawled (control) portions of each transect immediately before, immediately after, and 12 months after a 40/54 roller-rigged trawl was dragged through the alley once. Some damage to individuals of all target species was observed immediately after trawling, but only the density of barrel sponges (Cliona spp.) was significantly reduced. The extent of damage to the other sponges (Ircinia campana, Haliclona oculata), octocorals (Leptogorgia virgulata, Lophogorgia hebes, Titanideum frauenfeldii) and hard corals (Oculina varicosa) varied depending on the species, but changes in density were not statistically significant. Twelve months after trawling, the abundance of specimens counted in the trawled quadrats had increased to pre-trawl densities or greater, and damage to the sponges and corals could no longer be detected due to healing and growth. Trawl damage observed in this study was less severe than the damage reported for a similar habitat in a previous study. Differences between the two studies are attributed to (1) differences in the rollerrig design of the trawls used, and (2) differences in the number of times the same bottom was trawled.     

Fatigue analysis of the jack mackerel <Emphasis Type="Italic">Trachurus japonicus</Emphasis> by electrocardiographic monitoring during prolonged swimming

The effect of fatigue on swimming performance was examined by measuring the swimming endurance time and heart rate of the jack mackerel Trachurus japonicus [15.7 ± 0.8 cm fork length (FL), n = 15] during forced exercise in a flume tank at fixed swimming speeds of 4, 5 and 6 FL/s. Electrocardiographic (ECG) monitoring during the experimental process from control (0.8 FL/s) to exercise phase revealed a rapid cardiac response of T. japonicus to the elevation of swimming speed. The heart rate of T. japonicus significantly increased from the control level of 52.9 beats/min at a slow flow speed of 0.8 FL/s to 148.2 beats/min at 4 FL/s, 168.6 beats/min at 5 FL/s and 183.2 beats/min at 6 FL/s. During the fixed speed test, the heart rate of each individual fish was stabilized without any recognizable increase or decrease until the fish failed to swim because of fatigue. Fatigue analysis on endurance time demonstrated that prior swimming experience at prolonged speeds would impair the endurance performance during subsequent swimming exercise. Recovery time of the heart rate after the fish was fully exhausted by prolonged fast exercise increased with increasing swimming endurance time.     

Evaluation of the pigmentation stages and body proportions from the glass eel to yellow eel in Anguilla japonica

The pigmentation stages of Anguilla japonica were classified from the glass eel to yellow eel stage using 412 wild-caught eels from Hamana Lake and detailed developmental observations of 10 laboratory-reared eels. The sequential appearance of pigment was similar to that in A. anguilla, except for delayed pigment formation on the nerve cord of A. japonica. The general classification of pigmentation stages (V_A–VI_B) in A. anguilla was applicable to A. japonica, but the V_B stage could be separated into two stages (V_B1, V_B2) in A. japonica. The completion of guanine deposition on the intra-abdominal membrane may be a trait to discriminate the yellow eel stage (VII) from the VI_B stage. In natural conditions, wild-caught 0-age eels (n = 3,298) did not increase in size and retained a slender body form until VI_A4, and then became thicker during VI_B. The transition between body forms and growth may correspond to their ecological change from using passive transport in glass eels with tidal flow into rivers, to their settlement and further movements of elvers upstream. Considering this behavioral transition and pigmentation progression, it is proposed that the terms "glass eel" and "elver" could be defined as stages V_A–VI_A4 and VI_B, respectively.     

Early development of Lamprey Lampetra japonica (Martens, 1868)

Studies of lamprey early development have produced major insights into primitive developmental patterns and the evolution of vertebrates. In this study, the effectiveness of the morphological and histological characteristics of Lampetra japonica during embryological, yolk‐sac larval and ammocoete stages were examined. Based on the external and histological morphology, the embryological phase was divided into eight stages, including the zygote, cleavage, blastula, gastrula, neural plate and groove, head protrusion, prehatching and hatching stages. A set of key morphological and histological transformations occurred through the yolk‐sac larval stage, reflecting the larva‐ammocoete transition. In the ammocoete stage, all organ systems were differentiated. Except for the melanophores, the lamprey did not develop any further morphological complexity beyond the ammocoete stage. The total length and body weight of larva and ammocoete generally increased over time. We hope that this study might form the basis for future studies of the phylogenetic development of this interesting group of animals.     

Analysis of germ cell proliferation, apoptosis, and androgenesis in the Lake Van fish (Chalcalburnus tarichi) during testicular development

In the present study, the testis histology, gonadosomatic index (GSI), germ cell proliferation and apoptosis, and the plasma 11-ketotestosterone (11-KT) and testosterone (T) levels of male Chalcalburnus tarichi were analyzed. According to the histological examinations of the specimens that were caught between February 2009 and January 2010, three testicular stages were determined. Those stages were as follows: (1) recrudescence or prespawning (July–April), (2) spawning (May–June), and (3) postspawning (July). It was observed that the GSI increased gradually, starting from the recrudescence stage, and it reached peak values at the spawning stage, while the lowest values were in the postspawning. Germ cell proliferation in the testis was detected using a proliferating cell nuclear antigen (PCNA), and germ cell apoptosis was detected by transferase dUTP nick end labeling staining. The germ cell PCNA and apoptosis index values were calculated. It was indicated that germ cell proliferation was observed in all of the testicular stages. The highest germ cell PCNA index (PI) levels were detected in July, August, and September, which then dropped in October and stabilized between February and April. The lowest PI values were detected in the spawning stage (May–June). Germ cell apoptosis was observed in all of the months, and the highest apoptotic index values were detected in August, September, October, May, and June. Plasma 11-KT and T levels were at their highest levels in May and June, and it was detected as stabile in the other months. There was a correlation between GSI, PI, and plasma androgen levels. In conclusion, the present data illustrate testicular development stages for C. tarichi and show changes in the level of GSI and sex steroid biosynthesis through spermatogenesis.     

条石鲷仔鱼饥饿试验及不可逆点的确定

在盐度29、水温22~24℃的条件下,进行了条石鲷仔鱼饥饿试验和不可逆点(PNR)的确定;观察了饥饿条件下条石鲷初孵仔鱼、3、6、9、12、15日龄的存活、生长、卵黄囊与油球利用和游泳行为的变化。结果表明:饥饿会延缓初孵仔鱼对油球的利用。随着饥饿时间的延长,仔鱼的生长与正常条件下仔鱼差异显著(P<0.05)。条石鲷仔鱼从初次摄食到PNR期为2.5~3 d,这个耐受饥饿的时间临界点在孵化后的5~5.5 d。在饥饿的条件下初孵仔鱼、3、6、9、12、15日龄仔鱼的全部死亡时间分别为6.5、6、5、4、65、d;后5个日龄仔鱼半数死亡时间分别为4、3.5、3.5、3.5、3.5 d。说明9日龄是条石鲷早期发育中最为敏感的阶段。饥饿仔鱼体长较短,头大且体瘦,长期饥饿后脑后部下陷。饥饿仔鱼表层集群游动觅食,缓慢游动反应迟钝,静伏底。     

Development of ceramic settlement devices for coral reef restoration using <Emphasis Type="Italic">in situ</Emphasis> sexual reproduction of corals

A ceramic coral settlement device (CSD) was developed to raise juvenile corals in situ in preparation for transplantation. The CSD is designed to be deployed in the water before mass spawning of corals, allowing larvae to settle and grow on the CSD before it is fixed to the reef structure. CSDs were aggregated and fixed on 0.2-m² frames (either 648 or 720 CSDs per frame) and deployed in the Sekisei Lagoon around the full moons of April and May in 2002. Seven frames were collected in August 2002 and settled corals were counted in the laboratory. The maximum number of coral colonies settled on CSDs in one frame was 617, with 94% belonging to the genus Acropora. The aggregated CSDs successfully functioned as a coral settlement substratum, with more appropriate arrangement of CSDs on the frame, enabling greater exposure to sunlight and currents, which is expected to increase the amount of settlement.     

The effect of K+, Ca2+, and Mg2+ on sperm motility in the perch,Perca fluviatilis

This study investigated the effect of 0.25–5 mM K⁺, Ca²⁺, and Mg²⁺ on sperm motility in the perch, Perca fluviatilis. In 75 mM NaCl, the used motility-activating solution, motility rate, and swimming velocity decreased within the first 4 min after activation, and the rate of locally motile sperm increased. Thereafter, the motility parameters remained constant for periods >20 min. Based on the decrease in sperm motility, two types of semen samples could be distinguished. Semen samples of type I retained a high motility rate of >65 % after 20 min, and the rate of locally motile sperm was <20 %. In semen samples of type II, the motility rate decreased to values <30 % after 20 min, and the rate of locally motile sperm exceeded >50 %. Ca²⁺ and Mg²⁺ concentrations of 0.25–0.5 mM had no effect on the sperm motility parameters 10 s after activation, while 0.25 mM K⁺ increased the swimming velocity. K⁺, Ca²⁺, and Mg²⁺ concentrations ≥1.5 mM had suppressive effects on the sperm motility 10 s after activation. No differences were found between the two semen types. Twenty minutes after activation, type I semen was not affected by the tested cations. On the contrary, 0.25–2.5 mM K⁺, 0.25 mM Mg²⁺, and 0.25–2.5 mM Ca²⁺ significantly increased the sperm motility rate and/or sperm velocity of type II semen. Therefore, supplementation of saline solution with cations might stabilize the motility of perch sperm, which can be a benefit for experimental purposes and for specific handling procedures in aquaculture.     

三疣梭子蟹CYP302a1基因克隆及其表达分析

细胞色素P450(CYP)302a1是昆虫蜕皮激素合成通路中的关键酶,为了研究其在三疣梭子蟹蜕皮过程中的调控作用,实验采用反转录PCR(RT-PCR)和cDNA末端快速扩增(RACE)技术,克隆得到了三疣梭子蟹CYP302a1全长cDNA序列(GenBank登录号:KM596851).该序列全长为3 171 bp,包含一个长度为1 626 bp的开放阅读框,编码541个氨基酸;序列分析显示该氨基酸含helix-C、helix-K、helix-I、PERF及heme-binding共5个P450特征保守区域;系统进化树分析发现三疣梭子蟹CYP302a1与日本剑水蚤CYP302a1聚为一小支,再与其他物种CYP302a1聚为一支.采用实时荧光定量PCR(qRT-PCR)技术分析了CYP302a1在不同组织和蜕皮过程中的表达水平变化.结果显示,CYP302a1的表达量在Y器中最高,而在其他组织中均极低(P＜0.05);蜕皮过程中,CYP302a1在蜕皮后期(A、B期)表达量最低,从蜕皮间期(C期)开始上升,至蜕皮前期的D1亚期达到最大值,随后下降.结果表明CYP302a1在三疣梭子蟹蜕皮调控中可能起着重要作用.     

Morphological characterization of larval stages and first juvenile of the freshwater prawn Cryphiops caementarius (Molina, 1782) (Decapoda: Palaemonidae) under laboratory conditions

Research has been in progress for several years on various aspects of the biology and ecology of the freshwater prawn Cryphiops caementarius, an inhabitant of rivers in northern Chile. The commercial value of this prawn fomented the accomplishment of studies on its reproduction and development with the aim of producing juveniles under controlled conditions, to be followed by growout to commercial size in managed culture systems. The present study describes larval culture of this species from eggs of gravid females obtained in the field, from the first developmental stage (Zoea I) through the first juvenile stage. The larvae were cultured at 25 °C in UV sterilized water at variable salinities based on the requirements of the developmental stages. Larvae were fed with Nannochloris, Isochrysis and Artemia nauplii as required. This report describes in detail the 18 larval stages of this prawn, as well as its first juvenile form.