Effects of dietary carbohydrate-to-lipid ratio on the growth performance and feed utilization of juvenile turbot (<Emphasis Type="Italic">Scophthalmus maximus</Emphasis>)

A 9-week feeding trial was conducted to investigate the effects of dietary carbohydrate to lipid ratio(CHO:LIP) on the growth performance and feed utilization of juvenile turbot Scophthalmus maximus(initial body weight 8.75 g ± 0.04 g). Four isonitrogenous and isoenergetic low protein level(39%) diets were formulated with increasing ratios of dietary carbohydrate to lipid(2:18, 6:18, 18:12 and 28:6). A high protein level(50%) diet with the 2:12 ratio of carbohydrate to lipid was used as the control. Results showed that the survival rate, contents of moisture, crude protein and ash in muscle were not significantly affected by dietary treatments. With the dietary CHO:LIP ratio increased from 2:18 to 18:12, weight gain rate significantly increased(P < 0.05). Higher dietary CHO:LIP ratio(28:6) resulted in the significantly decreased weight gain rate(P < 0.05). Meanwhile, this treatment also resulted in the highest daily feed intake and liver glycogen content, as well as the lowest feed efficiency(P < 0.05). Muscle glycogen content in fish fed the diet with 2:12 or 2:18 CHO:LIP ratio was significantly lower than those fed with the other three diets(P < 0.05). The present results confirmed that the juvenile turbot can utilize carbohydrate. Furthermore, the appropriate ratio of dietary carbohydrate to lipid was important to the growth and feed utilization of turbot. The proper CHO:LIP ratio based on the growth performance in the present study was determined to be 18:12 when the dietary protein level was 39%.     

Inbreeding depression on growth and survival of full-sib family of Manila clam (<Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>)

In present study, the inbreeding depression (ID) of growth and survival of Manila clam (Ruditapes philippinarum) was investigated at larval and juvenile stages. Nine inbred families (A ₂, B ₂, C ₂, D ₂, E ₂, F ₂, G ₂, H ₂ and I ₂) were established by mating within nine full-sib families with expected inbreeding coefficient of 0.25. Inbred families showed significant differences in shell length and hatching rate of D-larvae (straight-hinged larvae). The larvae of the nine inbred families grew slower than those of control group (CG), and their ID value ranged from 0.81% ± 6.09% to 16.10% ± 1.49%. The ID value of larval survival rate varied between 27.47% ± 9.36% and 70.50% ± 13.66%. The ID was also detected for juvenile growth in A ₂, B ₂, C ₂, and D ₂, which ranged from 4.60 ± 2.21 to 17.71 ± 7.73. The A ₂ family maintained the highest juvenile survival rate, whereas the other inbred families exhibited ID values varying between 62.79% ± 4.54% and 96.14% ± 0.87%. The linear relationship of estimated ID between growth and survival was negatively correlated (R = ?0.434, P < 0.05). The results of this study suggested that the ID of growth was common at the larval stage but was less prevalent at juvenile stage. In contrast, the ID of survival increased from larval to juvenile stage. A better understanding of the effect of inbreeding may aid to selective breeding of Manila clam.     

Establishment and characterization of a testicular Sertoli cell line from olive flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

The culture of Sertoli cells has become an indispensable resource in studying spermatogenesis. A new Sertoli cell line (POSC) that consisted predominantly of fibroblast-like cells was derived from the testis of the olive flounder Paralichthys olivaceus and sub-cultured for 48 passages. Analysis of the mtDNA COI gene partial sequence confirmed that the cell line was from P. olivaceus. Cells were optimally maintained at 25°C in DMEM/F12 medium supplemented with fetal bovine serum, basic fibroblast growth factor, and epidermal growth factor. The growth curve of POSC showed a typical “S” shape. Chromosome analysis revealed that the cell line possessed the normal P. olivaceus diploid karyotype of 2n=48t. POSC expressed dmrt1 but not vasa, which was detected using RT-PCR and sequencing. Immunocytochemistry revealed that the cells exhibited the testicular Sertoli cell marker FasL. Therefore, POSC appeared to consist of testicular Sertoli cells. Bright fluorescent signals were observed after the cells were transfected with pEGFP-N3 plasmid, with the transfection efficiency reaching 10%. This research not only offers an ideal model for further gene expression and regulation studies on P. olivaceus, but also serves as valuable material in studying fish spermatogenesis, Sertoli cell-germ cell interactions, and the mechanism of growth and development of testis.     

Effects of small peptides,probiotics, prebiotics,and synbiotics on growth performance,digestive enzymes,and oxidative stress in orange-spotted grouper, <Emphasis Type="Italic">Epinephelus coioides</Emphasis>, juveniles reared in artificial seawater

Aquaculture production efficiency may increase by using feed additives. This study investigated the effects of different dietary additives [w/w: 2% small peptides, 0.01% probiotics (Bacillus licheniformis) and 0.2% prebiotics (inulin)] on growth performance, digestive enzyme activities, and oxidative stress in juvenile Epinephelus coioides reared in artificial seawater of two salt concentrations (13.5 vs. 28.5). Weight gain rate was significantly higher in fish fed the diet supplemented with small peptides, B. licheniformis, inulin, or synbiotics than that in fish fed the basal diet; the greatest weight gain rate was found in fish fed the small peptide treatment [56.0% higher than basal diet]. Higher feed efficiency was detected in fish fed the diet supplemented with small peptides than that of fish in the other dietary treatments. Total protease activity in the stomach and intestines was highest in fish fed the small peptide-treated diet, whereas lipase activity was highest in those fed synbiotics (combination of Bacillus licheniformis and inulin) than that in fish fed the other treatments. Antioxidant enzyme (total superoxide dismutase and catalase) activities and hepatic malondialdehyde content were higher in fish receiving the dietary supplements and maintained in artificial seawater containing 13.5 salinity compared with those in the control (28.5). Hepatic catalase activity in grouper fed the diets with small peptides or synbiotics decreased significantly compared with that in control fish. Overall, the three types of additives improved growth rate of juvenile grouper and digestive enzymes activities to varying degrees but did not effectively improve antioxidant capacity under low-salinity stress conditions.     

Effects of dietary genistein on GH/IGF-I axis of Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

There is considerable concern that isoflavones, such as genistein in fish feed composed of soybean protein, aff ects somatic growth in fish. Our previous works demonstrated that 30 and 300 μg/g dietary genistein had no significant eff ect on growth performance in Nile tilapia (Oreochromis niloticus), but the higher level of genistein (3 000 μg/g) significantly depressed growth. This study was conducted to further examine the eff ects of dietary genistein on the endocrine disruption on growth hormone/insulin-like growth factor-I (GH/IGF-I) axis in Nile tilapia (O. niloticus). Juvenile fish were fed by hand twice daily to satiation with one of four isonitrogenous and isoenergetic diets, each containing either 0, 30, 300 or 3 000 μg/g genistein. Following an 8-week feeding period, plasma GH and IGF-I levels were investigated by radioimmunoassay and gene expression levels of gh, ghrelin, gnrhs, ghr, npy, npyrs, pacap, ghrs, igf-I, igf-Ir, and igfbp3 were examined by real-time PCR. The results show that no significant change in plasma GH and IGF-I levels in fish fed with diets containing 30 μg/g and 300 μg/g genistein. mRNA expression of genes along the GH/IGF-I axis remained unaff ected, except for igf-Ir, which was stimulated by the 300 μg/g genistein diet. While in fish fed the 3 000 μg/g genistein diet, the plasma GH and IGF-I levels decreased, and mRNA expression of gh, ghr2, npyr1, igf-I, and igf-Ir were also significantly depressed. In contrast, npy and igfbp3 mRNA expression were enhanced. This study provides convincing evidence for growth impediment by genistein by disturbing the GH/IGF-I axis in Nile tilapia O. niloticus.     

Carbon budget of bastard halibut<Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> in relation to body weight and temperature

The effects of body weight and temperature on the carbon budget of the juvenile bastard halibut ,Paralichthys olivaceus ,were studied at temperature 13.5,18,21.5 and 24℃,respectively.The carbon intake,faecal and growth carbon were measured ,and the carbon respiration was calculated using the carbon budget equation (Cc=Gc Fc Rc),The combined relationship between different components of the carbon budgent,body weight and temperature could be described by regression equations:Cc=1.0206 W^0.8126E^0.1483T;Gc=0.0042w^1.4096(-5.11 T^3 285.90T^2-5173.72T 30314.03);Fc=0.0485W^0.7711e^0.1624T;Uc=1.4333W^0.6715e^0.1487t,Body weight had no significant effect on the carbon absorption efficiency and the conversion efficiency.     

Low temperature stress on the hematological parameters and <Emphasis Type="Italic">HSP</Emphasis> gene expression in the turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis>

To study the effect of low temperature stress on hematological parameters and HSP gene expression in the turbot (Scophthalmus maximus), water temperature was lowered rapidly from 18 to 1°C. During the cooling process, three individuals were removed from culture tanks at 18, 13, 8, 5, 3, and 1°C. Blood samples and tissues were taken from each individual, hematological indices and HSP gene expression in tissues were measured. The red blood cell count, white blood cell count, and hemoglobin concentration decreased significantly (P < 0.05) as temperature decreased. Enzyme activities of plasma alanine transaminase and creatine kinase increased as temperature decreased, whereas aspartic transaminase and γ-glutamyl transpeptidase activities displayed no obvious changes above 1°C and lactate dehydrogenase activity increased first and then decreased. Blood urea nitrogen and uric acid levels were highest at 8°C, and creatinine concentration was highest at 3°C. The concentrations of plasma cortisol, cholesterol, and triglyceride all increased significantly (P < 0.05) as temperature decreased. The serum glucose concentration increased first and then decreased to the initial level. The HSP70 mRNA expression showed various patterns in different tissues, whereas HSP90 mRNA expression showed the same tendency in all tissues. Overall, these results indicate that temperature decreases in the range of 8 to 5°C may induce a stress response in S. maximus and that temperature should be kept above 8°C in the aquaculture setting to avoid damage to the fish.     

Interspecific competition and allelopathic interaction between <Emphasis Type="Italic">Karenia mikimotoi</Emphasis> and <Emphasis Type="Italic">Dunaliella salina</Emphasis> in laboratory culture

Algal allelopathy is a manifold ecological/physiological phenomenon that is focused on chemical interactions and autotoxicity. We investigated the allelopathic interactions between Karenia mikimotoi and Dunaliella salina in laboratory cultures based on diff erent temperature (15°C, 20°C, and 25°C) and lighting (40, 80, and 160 μmol/(m²·s)) conditions. The growth of D. salina in bi-algae culture (1:1 size/density) was significantly restrained. The results of cell-free filtrate culture indicate that direct cell-tocell contact was not necessary in interspecific competition. Further experimental results demonstrated that allelochemicals released from K. mikimotoi were markedly influenced by both temperature (P =0.013) and irradiance (P =0.003), resulting in diff erent growth characteristics of D. salina in filtrate mediums. Compared with the plateau period, K. mikimotoi exudates in the exponential phase had a stronger short-term inhibition effect on D. salina in normal conditions. A clear concentration-dependent relationship was observed in the effect of allelochemicals released from K. mikimotoi with low-promoting and high-repressing effects on D. Salina in a short time-scale. In addition, allelopathic substances remain stable and effective under high temperature and pressure stress. Many flocculent sediments adhering with D. salina cells were observed in all filtrate mediums, while the quantity and color depended on the original culture conditions.     

Feeding broodstocks different starfish diets affect growth and survival of larvae of trumpet shell (<Emphasis Type="Italic">Charonia lampas sauliae</Emphasis> Reeve 1844)

Trumpet shell (Charonia lampas sauliae) (Mollusca, Heterogastropoda, Cymatidae) has extensive economic value. Studies on the artificial larval development of C. lampas sauliae for aquaculture utilization have become especially important due to the finite natural resources. In the present study, the growth and survival rate of the larvae of C. lampas sauliae broodstocks fed three types of starfish diets, Asterina pectinifera Müller & Troschel 1842, A. amurensis Lütken 1871 and their mixture were compared. The larval size increased gradually between day 10 and day 20 after hatching at 15°C and 20°C. No difference was found in body size and specific growth rate (SGR) (two-way ANOVA; P > 0.05). However, during transition from trochophore to veliger stage 20 days after hatching, significant increases in larval survival and growth rates were observed. The maximum survival rate was observed on day 10. The mean survival rate was 0.463, 0.730 and 0.515 at 15°C, and 0.369, 0.713 and 0.444 at 20°C when A. pectinifera, A. amurensis and their mixture were fed, respectively. The SGR and survival rate of the larvae were definitely influenced by the diets (P < 0.05), and the effect of A. amurensis alone was higher than that of A. pectinifera alone and their mixture.     

Effects of starving and re-feeding strategies on the growth performance and physiological characteristics of the juvenile tongue sole (<Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis>)

Starving and re-feeding trials were conducted to evaluate the effects of starvation duration and recovery times on the growth performance and physiological characteristics of juvenile tongue sole Cynoglossus semilaevis, which included growth, body composition, intestinal morphology and digestive enzymes activities. The fish starved for one (D1), two (D2) and four (D4) days, respectively, were recovered for three (R3, R6, R12), seven (R7, R14, R28) and eleven (R11, R22, R44) folds of starvation days while the control fish were fed constantly. The experiment was conducted for 96 days. The fish individuals in D1R11, D2R14 and D2R22 caught up in weight with the control, indicating that complete compensatory growth existed in these fish individuals. The highest body weight gain and total specific growth rates were observed in D2R22. Food intake increased significantly in most treatments in comparison with that of the control except for D1R7 and D2R22 (P < 0.05). Food conversion efficiency and apparent digest coefficient in D2R22 were significantly promoted, being higher than that of the control (P < 0.05). The activities of trypsin and lipase were found to be closely related to the growth performance. The activities of liver and hindgut trypsin, also midgut and hindgut lipase in D2R22 were significantly higher than those of the control (P < 0.05). The fold height (HF) of foregut and midgut increased significantly in D2R22, and HF decreased significantly in D1R3, D2R6 and D4R12 (P < 0.05). On our findings, we may conclude that the optimum starving and re-feeding strategy is starving for 2 days and re-feeding for 22 days. And in this starving and re-feeding strategy, the compensatory growth could be mostly attributed to the promotion of food conversion efficiency and digestibility coefficiency.     

Effect of an escape vent in accordion-shaped traps on the catch and size of Asian paddle crabs <Emphasis Type="Italic">Charybdis japonica</Emphasis> in an artificial reef area

Accordion-shaped traps are widely used in China to catch the Asian paddle crab Charybdis japonica but traps of conventional design often catch juvenile crabs. A new type of accordion-shaped trap with an escape vent (L×W=4.3 cm×3.0 cm) was designed and a comparative study between the newly designed and conventional traps was performed in the artificial reef area of Zhuwang, Laizhou Bay, China from June to August 2012. The mean catch per unit effort (CPUE) of undersized crabs was significantly lower in the vented traps than in the conventional traps (paired t -test, n=30, P<0.001), while the CPUE of marketable crabs was significantly higher in the vented traps (paired t-test, n=30, P<0.001). The mean size of crabs (carapace length) caught in the vented traps was significantly larger than in conventional traps (paired t-test, n=29, P<0.001). The ratio of undersized crabs was 35.05±2.57% in conventional traps and 12.53±0.69% in vented traps (significantly lower, paired t-test, n=29, P<0.001). Therefore, a 4.3 cm×3.0 cm escape vent was considered appropriate for C. japonica fishing in the artificial reef area. This finding will assist the development of more sustainable and efficient crab fishing methods using accordion-shaped traps.     

Immunogenicity and protective role of antigenic regions from five outer membrane proteins of <Emphasis Type="Italic">Flavobacterium columnare</Emphasis> in grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

Flavobacterium columnare causes columnaris disease in freshwater fish. In the present study, the antigenic regions of five outer membrane proteins (OMPs), including zinc metalloprotease, prolyl oligopeptidase, thermolysin, collagenase and chondroitin AC lyase, were bioinformatically analyzed, fused together, and then expressed as a recombinant fusion protein in Escherichia coli. The expressed protein of 95.6 kDa, as estimated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was consistent with the molecular weight deduced from the amino acid sequence. The purified recombinant protein was used to vaccinate the grass carp, Ctenopharyngodon idella. Following vaccination of the fish their IgM antibody levels were examined, as was the expression of IgM, IgD and IgZ immunoglobulin genes and other genes such as MHC Iα and MHC IIβ, which are also involved in adaptive immunity. Interleukin genes (IL), including IL-1β, IL-8 and IL-10, and type I and type II interferon (IFN) genes were also examined. At 3 and 4 weeks post-vaccination (wpv), significant increases in IgM antibody levels were observed in the fish vaccinated with the recombinant fusion protein, and an increase in the expression levels of IgM, IgD and IgZ genes was also detected following the vaccinations, thus indicating that an adaptive immune response was induced by the vaccinations. Early increases in the expression levels of IL and IFN genes were also observed in the vaccinated fish. At four wpv, the fish were challenged with F. columnare, and the vaccinated fish showed a good level of protection against this pathogen, with 39% relative percent survival (RPS) compared with the control group. It can be concluded, therefore, that the five OMPs, in the form of a recombinant fusion protein vaccine, induced an immune response in fish and protection against F. columnare.     

Impact of <Emphasis Type="Italic">Vibrio parahaemolyticus</Emphasis> and white spot syndrome virus (WSSV) co-infection on survival of penaeid shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

White spot syndrome virus (WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect Litopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus (WSSV+Vp), or we first infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection (Vp+WSSV). The eff ect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of Vibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction (PCR) method. LvMyD88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the diff erent mortality rates. Our results show that (1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection; (2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed significant increases in the numbers of Vibrio (P<0.05); (3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 10⁵ to 10⁷ /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.     

Effect of dietary cottonseed meal on growth performance,physiological response,and gossypol accumulation in pre-adult grass carp, <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>

Cottonseed meal (CM) was used at up to 36.95% content in the diet (replacing 60% of dietary fish meal protein) without any negative eff ects on growth performance of pre-adult grass carp (initial body weight, 761 g) under outdoor conditions. A culture trial was conducted in net cages installed in a large concrete pond. Seven isonitrogenous and isoenergetic diets containing a gradient of CM concentrations (0, 12.2%, 24.4%, 36.6%, 48.8%, 54.8%, and 61.0%) as replacement for dietary fish meal protein (0, 20%, 40%, 60%, 80%, 90%, and 100%) were formulated. Dietary non-resistant starch (from maize) was inverse to dietary CM. Growth performance and feed utilization of fish fed the diets containing CM replacing 0–40% fishmeal protein were not aff ected after the 6-week feeding trial. Accumulation of hepatopancreatic total gossypol in the hepatopancreas was significantly correlated with free gossypol content in the diets (HTG=88.6+1.5×DFG, R ²=0.89, P<0.05). Intestinal α-amylase and γ-glutamyl transpeptidase activities rose along with increasing dietary CM level. The structure of the mid-intestinal tissues and the ultrastructure of the enterocyte microvilli were normal when dietary CM was <36.6% (60% protein replacement). Increasing dietary CM content increased serum alanine aminotransferase levels but decreased serum alkaline phosphatase, cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, and albumin (P<0.05).     

Global water vapor content decreases from 2003 to 2012: An analysis based on MODIS data

Water vapor in the earth′s upper atmosphere plays a crucial role in the radiative balance, hydrological process, and climate change. Based on the latest moderate-resolution imaging spectroradiometer(MODIS) data, this study probes the spatio-temporal variations of global water vapor content in the past decade. It is found that overall the global water vapor content declined from 2003 to 2012(slope b = –0.0149, R = 0.893, P = 0.0005). The decreasing trend over the ocean surface(b = –0.0170, R = 0.908, P = 0.0003) is more explicit than that over terrestrial surface(b = –0.0100, R = 0.782, P = 0.0070), more significant over the Northern Hemisphere(b = –0.0175, R = 0.923, P = 0.0001) than that over the Southern Hemisphere(b = –0.0123, R = 0.826, P = 0.0030). In addition, the analytical results indicate that water vapor content are decreasing obviously between latitude of 36°N and 36°S(b = 0.0224, R = 0.892, P = 0.0005), especially between latitude of 0°N and 36°N(b = 0.0263, R = 0.931, P = 0.0001), while the water vapor concentrations are increasing slightly in the Arctic regions(b = 0.0028, R = 0.612, P = 0.0590). The decreasing and spatial variation of water vapor content regulates the effects of carbon dioxide which is the main reason of the trend in global surface temperatures becoming nearly flat since the late 1990 s. The spatio-temporal variations of water vapor content also affect the growth and spatial distribution of global vegetation which also regulates the global surface temperature change, and the climate change is mainly caused by the earth's orbit position in the solar and galaxy system. A big data model based on gravitational-magmatic change with the solar or the galactic system is proposed to be built for analyzing how the earth's orbit position in the solar and galaxy system affects spatio-temporal variations of global water vapor content, vegetation and temperature at large spatio-temporal scale. This comprehensive examination of water vapor changes promises a holistic understanding of the global climate change and potential underlying mechanisms.     

Significance of different microalgal species for growth of moon jellyfish ephyrae, <Emphasis Type="Italic">Aurelia</Emphasis> sp.1

The scyphozoan Aurelia aurita (Linnaeus) sp. l., is a cosmopolitan species-complex which blooms seasonally in a variety of coastal and shelf sea environments around the world. The effects of different microalgal species on the growth of newly-released Aurelia sp.1 ephyrae were studied under laboratory conditions. We fed ephyrae with four different microalgal species (diatom, autotrophic dinoflagellate, heterotrophic dinoflagellate, and chlorophyta) plus Artemia nauplii for 12-24 d at 18°C. Results showed that the growth rate diverged significantly for Artemia nauplii compared to other food types. In addition, there was no significant variation between the growth rates for Skeletonema costatum and Prorocentrum donghaiense, and no significant variation was found in the growth rates for N. scintillans and P. subcordiformis. Artemia nauplii could support the energy requirement for the newly-released ephyrae to develop to meduase, and the ephyrae with Artemia nauplii showed a significant average growth rate of 25.85% d^?1. Newly-released ephyrae could grow slightly with some species of microalgae in the earliest development stage. Chain diatom Skeletonema costatum and autotrophic dinoflagellate Prorocentrum donghaiense, could not support the growth of the ephyrae, while heterotrophic dinoflagellate Noctiluca scintillans and chlorophyta Platymonas subcordiformis could support the growth of the ephyrae. However, none of the ephyrae fed with the tested phytoplankton could mature to medusae.     

The relationship between fat content and biological parameters of bigeye tuna (<Emphasis Type="Italic">Thunnus obseus</Emphasis>) in the Western Central Indian Ocean

Comparing fat content with physiological status can throw light on the reproduction and feeding behavior of the fish. The biological data of 586 bigeye tuna were collected from the longline fishery in the waters of Western Central Indian Ocean from November, 2012 to March, 2013. The spatial or temporal distribution of the fat content, and the relationships of fat content with gender, round weight, gonadal maturity and fork length were analyzed. A generalized additive model (GAM) was used to analyze the relationships between fat content and fork length (FL), gonadosomatic index (GSI), condition factor (K), and somatic index (SI). Results showed that: 1) the fat content of bigeye tuna was in the range of 3.1%–27.1% with the average 12.8%; 2) there were no significant geographical differences of average fat content (P > 0.05) among 1° squares in general; 3) there were no significant differences (P > 0.05) of the fat content for different genders, months, or gonad maturity stages; 4) there was an extremely significant correlation (P = 0.000) between fat content and FL and GSI. There was no significant correlation (P = 0.051) between fat content and K. There was a significant correlation (P = 0.003) between fat content and SI. The results of this study suggest that the fat content of the matured fish was relatively stable. The survey area was in a spawning region, and the survey period was the spawning season for bigeye tuna.     

Early embryo and larval development of inviable intergeneric hybrids derived from <Emphasis Type="Italic">Crassostrea angulata</Emphasis> and <Emphasis Type="Italic">Saccostrea cucullata</Emphasis>

To detect the intergeneric hybridization between the oyster Crassostrea angulata and Saccostrea cucullata coexisting along the southern coast of China, reciprocal crosses were conducted between the two species. Barriers for sperm recognizing, binding, penetrating the egg, and forming the pronucleus were detected by fluorescence staining. From the results, although fertilization success was observed in hybrid crosses, the overall fertilization rate was lower than that of intraspecific crosses. A large number of hybrid larvae died at 6–8 d after hatching, and those survived could not complete metamorphosis. C. angulata ♀× S. cucullata ♂ larvae had a growth rate similar to that of the maternal species, whereas S. cucullata ♀ × C. angulata ♂ larvae grew the slowest among all crosses. Molecular genetics analysis revealed that hybrid progeny were amphimixis hybrids. This study demonstrated that hybrid embryos generated by crossing C. angulata and S. cucullata could develop normally to the larval state, but could not complete metamorphosis and then develop to the spat stage. Thus, there is a post-reproductive isolation between C. angulata and S. cucullata.     

Optimization of protectant,salinity and freezing condition for freeze-drying preservation of <Emphasis Type="Italic">Edwardsiella tarda</Emphasis>

Novel preservation condition without ultra-low temperature is needed for the study of pathogen in marine fishes. Freeze-drying is such a method usually used for preservation of terrigenous bacteria. However, studies using freeze-drying method to preserving marine microorganisms remain very limited. In this study, we optimized the composition of protectants during the freeze-drying of Edwardsiella tarda, a fish pathogen that causes systemic infection in marine fishes. We found that the optimal composition of protectant mixture contained trehalose (8.0%), skim milk (12.0%), sodium citrate (2.0%), serum (12.0%) and PVP (2.0%). Orthogonal and interaction analyses demonstrated the interaction between serum and skim milk or sodium citrate. The highest survival rate of E. tarda was observed when the concentration of NaCl was 10.0, 30.0 and between 5.0 and 10.0 g L?1 for preparing TSB medium, E. tarda suspension and protectant mixture, respectively. When E. tarda was frozen at ?80°C or ?40°C for 6 h, its survival rate was higher than that under other tested conditions. Under the optimized conditions, when the protectant mixture was used during freeze-drying process, the survival rate (79.63%–82.30%) of E. tarda was significantly higher than that obtained using single protectant. Scanning electron microscopy (SEM) image indicated that E. tarda was embedded in thick matrix with detectable aggregation. In sum, the protectant mixture may be used as a novel cryoprotective additive for E. tarda.     

Diel variation in metabolism and ammonia excretion of <Emphasis Type="Italic">Marphysa sanguinea</Emphasis> (Polychaeta: Eunicidae)

Polychaetes provide an excellent food resource for fish and represent the dominant zoobenthos in marine ecosystems. Diel variation in the rates of metabolism and ammonia-N excretion of Marphysa sanguinea were studied. The worms were grouped according to their wet body weight into small (S; 1.24±0.06 g), medium (M; 4.00±0.30 g), and large (L; 8.54±1.08 g) categories. Their weight-specific metabolic rates, based on aerobic respiration (R), were measured at 16°C (±0.2°C) and classed as either routine (R _R) or standard (R _S) rates. Both respiration types decreased with increasing body weight. Respiration was described by R = a W ^b, where b was -0.400 9 and -0.532 0 for R _R and R _S, respectively. Diurnal changes in R _S for each group was relatively flat, with a slightly increasing trend with time, but was relatively stable as a whole. R _R of the diurnal variation of worms was higher than R S, but both had similar overall trends. The peak values of specific dynamic action (SDA) (R _SDA) in the S, M, and L groups were 2.704, 1.149, and 0.682 mg/(g?h), respectively. The durations of SDA were 13, 6, and 6 h, respectively and the energy expenditures of SDA were 377.98, 117.34, and 74.94 J/g, respectively. These data indicate that the metabolic rates were higher in smaller individuals, which is advantageous for their rapid growth.