Abstract:Follistatin (FST) could antagonize the TGF-β superfamily members and inhibit the activity of endo-myostatin via direct protein-protein interaction, thereby restoring muscle growth. Here, according to the splicing results of the high-throughput transcriptome sequencing data for Cyprinus carpio, we cloned the open reading frame (ORF) sequence of the Fst1 gene, which was 1 260 bp in length and encodes a protein of 320 amino acids. We then constructed a pTgf2-Mylz2-ccfst1 donor plasmid carrying both the left (220 bp) and right (185 bp) arms of goldfish Tgf2 transposon, the promoter of zebrafish Mylz2 and the ORF of carp Fst1. Transgenic carps were obtained via co-microinjection of the donor and the capped mRNA of the Tgf2 transposase synthesized in vitro. The average integration rate of the exogenous gene in transgenic carp was 44.7% as measured by PCR. The amplified products of four positive transgenic carp were recovered, cloned and sequenced to confirm that all of them contained the target fragment. Our results demonstrated the high transgenic efficiency of the Tgf2 transposon in C. carpio, which laid the foundation for further studies on the function of FST during carp muscle development.

Abstract:To understand the prevalence of white spot syndrome virus (WSSV) of cultured Penaeus vannamei (P. vannamei) in Guangxi Province and to evaluate the genetic diversity of deletion region ORF23/24 and the genetic evolution lineage among WSSV isolates of Guangxi Province and other places, we collected 306 diseased P. vannamei samples between May 2010 and September 2013 from Beihai, Qinzhou and Fangchenggang City in Guangxi Province, and the detection of WSSV among these samples was carried out using the nested polymerase chain reaction (nested PCR) recommended by Office International Des Epizooties (OIE). We selected 53 positive samples found by first step PCR for amplification of the deletion regions ORF23/24 of the WSSV genome, and 12 PCR amplifications were used to clone and sequence. We then conducted a pairwise and multiple alignment analysis to evaluate the degree of genetic divergence among different strains. The results were as follows: 82 of 306 samples were positive for WSSV, and the positive rate was 26.8%; 12 Guangxi strains were 2096 bp in length and carried a 10970 bp deletion in the ORF23/24 region relative to WSSV-TW, which has the longest nucleotide sequence in this region. By comparison of nucleotide sequences of ORF23/24 region, we found that 12 Guangxi strains were above 99.6% homology, only existed nucleotide substitution but no deletion and insert, and BH-3, QZ-1, FCG-1 and FCG-2 strains were complete homology. In the phylogenetic tree of WSSV deletion region ORF23/24, the Guangxi strains were near in genetics and formed a separate branch with India strain IN-05-Ⅰ, while far from WSSV-TW, WSSV-CN and WSSV-Korea. In conclusion, there was some extent of prevalence of WSSV infection in cultured P. vannamei of Guangxi; the results showed that the Guangxi WSSV strains had large deletion in middle region of the ORF23/24 gene, and there were no differences in spatio-temporal and they were closer in genetics to India strain IN-05-Ⅰ than other strains in GenBank.

Abstract:Based on the terminal inverted repeats of Tc1-like superfamily transposons from fish, a single primer was designed to amplify similar transposon in genome of Salmo trutta fario Linnaeus. Two Tc1-like transposons sequences were amplified. The sequencing results showed that the length of the obtained fragments were 1 607 bp and 1 473 bp, named Tbt1 and Tbt2. Sequence analysis showed that there was a 196 nt and a 225 nt inverted terminal repeats (ITR) at the ends of Salmo trutta fario Tbt1, respectively. Two 12 nt Sub-terminal inverted repeats (SIR) were found among the left and right ITR. In contrast, Salmo trutta fario Tbt2 transposon had a 32 nt and 31 nt short ITRs, and the left and right ITRs each contained only one 12 nt of SIR. During evolutionary process, 4 and 9 stop codon mutations had been accumulated among transposase coding region in Salmo trutta fario Tbt1 and Tbt2, respectively, which led both transposons to lose their capacity to produce transposases. The Salmo trutta fario Tbt2 had a sequence similarity less than 30% to other salmon Tc1-like transposons, while there had a sequence identity as high as 98% to goldfish Tca2 transposon. This indicates that this transposon is very likely originated in the horizontal gene transfer mode.

Abstract:The complete mitochondrial genome of Acrossocheilus wenchowensis was obtained by PCR based on 8 primers, which were designed on the basis of published fish mitochondrial genome (GenBank accession number: KC_495074). The complete mitochondrial genome was 16 591 bp in length, including 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and a non-coding control region. The organization and location of genes in the mitochondrial genome of A.wenchowensis were consistent with Cyprinidae fishes published in GenBank. Of 37 genes, 1 protein-coding gene(ND6) and 8 tRNA genes(tRNA^Gln, tRNA^Ala, tRNA^Asn, tRNA^Cys, tRNA^Tyr, tRNA^Ser(UCN), tRNA^Glu and tRNA^Pro) were encoded on the L strand, the remainders on the H strand. The nucleotide composition was 30.93% (A), 24.86% (T), 16.41% (G) and 27.81% (C). Most tRNA genes could form typical secondary structures except tRNA^Ser(AGN). The COⅠ gene used GTG as an initiation codon, with the remaining 12 using ATG of total 13 protein genes, 10 genes had complete stop codons,and COⅡ, ND4 and Cytb had an incomplete stop codon T. The phylogenetic analysis of A.wenchowensis on Cyprinidae fishes was made from the level of mitochondrial genome based on amino acids sequence of all protein-coding genes in 18 species of 17 genera, and the complete mitochondrial genome of A.wenchowensis will help to provide basic data for the classification of Acrossocheilu and Cyprinidae fishes.

Abstract:A starvation trial was conducted to determine compensatory growth of the juvenile golden pompano Trachinotus ovatus which were deprived of food for 1,2,5 and 7 days respectively. The results of tests in outdoor cement pools and net pens showed that, in the first 15 d, 30 d, the growth of 1 d, 2 'd-deprived groups was probably same with the control group, which accorded with fully compensatory description.However, the weight of 5 d,7 d-deprived fingerlings were lower than that of the control group, which accorded with the description of the patial compensatory growth. As the starvation prolonged, feed conversion efficiency of the starved groups increased and was higher than the control group.Meanwhile, feeding rate also increased, but the fish of 1 d,2 d-deprived groups was lower than that of the control group, and feeding quantity was reduced and lower than the control group. It suggested that the mechanism of compensatory growth was mainly due to improving feed conversion efficiency. Fish biochemical composition was analyzed: the water content and ash of fish sample increased due to starvation, while the lipid and protein decreased. The loss of lipid was higher than that of the protein, and the biochemical composition of fish in each group was restored to the control level by the end of the experiment. It suggested that Trachinotus ovatus may mainly consume lipid during the period of starvation.

Abstract:For the research of an economically closed recirculating aquaculture system, this study designed a new system integrating greenhouse, new water purification equipment and aerobic machines such as wave aerator. A running test farming California perch (Micropterus salmonides) with the period of 4 months was conducted for evaluating the performance of the system. During the period of culture, the water quality, fish growth and market prospect of the system were analyzed. The obtained results showed that after 4 months of farming, the fish density increased from 2.12 kg/m³ to 5.86 kg/m³, and the survival rate was 95.1%. Results from the water quality monitoring test showed that the average concentrations of ammonia nitrogen, nitrite nitrogen and dissolved oxygen (DO) were 0.66±0.35, 0.19±0.089 and 6.64±0.25 mg/L, respetively; the water temperature and pH were maintained at 27.34-28.00℃ and 6.73-7.34. Economic analysis showed that the annual profit was up to 174.2 thousand Yuan per 667 m², the payback period was 2.75 years, implying the high economic value of the new system. This study demonstrated that the constructed greenhouse pond recirculating aquaculture system is an economically feasible system with the advantages of high efficiency, energy saving and emission reduction.

Abstract:A basal diet without adding maggot meal(MGM) and six test diets containing 1.0%, 1.8%, 2.6%, 3.4%, 4.2%, 5.0% of the MGM were prepared to feed seven groups of black carp(Mylopharyngodon piceus) in triplicate with an initial average body weight of (127.42±4.83) g respectively for eight weeks. The results showed the Black carp fed the diet containing 5.0% MGM had higher final body weight(FBW), body weight gain(BWG), specific growth rate(SGR) and lower feed conversion ratio(FCR) than the others. The HIS and VSI decreased significantly when adding 5.0% MGM compared with control group (P<0.05). Body composition analysis showed that whole crude protein content increased significantly with the increasing of MGM levels(P<0.05). Compared with the control group, the crude protein content of fishes muscle increased significantly when adding 3.4%, 4.2%, 5.0% MGM(P<0.05). Intestine and hepatopancreas protease, intestine amylase activities of fish fed diet with 4.2% MGM were higher than those of fish fed the basal diet (P<0.05), hepatopancreas and intestine lipase activity increased with the increasing level of MGM supplement up to 5.0%. In conclusion, under the condition of this experiment, adding 4.2%-5.0% MGM to black carp diet could improve fish growth, digestive enzymes activities, whole fish crude protein content and muscle quality.

Abstract:The aim of the present study is to investigate the effects of high CO₂ in the ocean on the physiology of Aristotle's Lantern muscle. After 140-day long-term high CO₂ exposure, the muscle respiration in 3000 μL/L group decreased nearly 40% compared with control group. The ATP content was not significantly influenced by CO₂. Acetylcholinesterase content in high CO₂ groups decreased, but without significance. And the muscle contractile of high CO₂ groups was lower than that of control group. These experiments showed that CO₂ plays an important role in the muscle physiology.

Abstract:To further understand the development process of intermuscular bones in teleosts, a modified bone clearing and staining experiment was conducted to investigate morphological development of intermuscular bone in Cyprinus carpio var. color. It is showed that intermuscular bone has different shapes including I type, one-end-unequal-bifork type, one-end-equal-bifork type, one-end-multifork type, two-end-biofork type, two-end-multifork type and tree-branch type. Generally, the morphological complexity of most intermuscular bones in anterior body was greater than that in posterior body. The initial ossification of intermuscular bone was observed in the posterior part of the body at 32 dpf, and then each of the intermuscular bones is ossified anteriorly, and the morphological characters at 53 dpf are the same as those found in adult fish. The primary morphology of intermuscular bone is I type, and more complicated morphology of ossified intermuscular bones are gradually derived from I type during ossification. These results suggest similar ossification pattern of intermuscular bones existing in Cyprinidae, which will also be helpful for us to further understand the molecular mechanism underlying intermuscular bone development.

Abstract:In this study, we analyzed morphological characteristics for 178 individuals of Scatophagus argus collected from 2011 to 2013. Based on the analysis of the morphological index system and R-cluster analysis, the results indicated that S.argus could be classified by "body size"and "head feature". On the basis of this analysis, 11 characteristics were normalized and further used for stepwise discrimintion. Five characteristics were selected: snout length/head length, eye distance/head length, body breadth/body length, body depth/body length and head depth/body length.Thus the sex discriminant equation was built for classifying 178 samples and the accuracy rate was 85.96%. T-test showed that three standardized characteristics between male and female populations were significantly different (P<0.05), except for eye distance/head length and body breadth/body length. Therefore, we concluded that female S.argus were superior to males in the respects of body depth, head and snout length.

Abstract:This study collected 709 X. microlepis and 295 X. davidi specimens, and compared their age, growth and reproduction from September 2010 to August 2012 in the Qiandao Lake. The result showed that age range of X. microlepis and X. davidi were 1-5 years. The advantage age of 84% X. microlepis and 76% X. davidi were 2-3 years. The growth characteristics of length and weight about X. microlepis could be described as W=0.0223L^2.8785(R²=0.9439). The growth of X. microlepis could be described with von Bertalanffy equation as L_t = 41.99×[1-e^{-0.2264(t+0.9746)}],W_t=980.22×[1-e^{-0.2264(t+0.9746)}]^2.7172. The inflection point of X. microlepis growth age was 3.05 years, which body length was 25.33 cm and body weight was 244.78 g; The X. microlepi's sex ratio was 0.956 8：1, The individual absolute fecundity varied was 24 893-132 521 eggs(71 283 on average). The growth characteristics of length and weight about X. davidi could be described as W= 0.0381L^2.7172 (R²=0.9439). The growth of X. davidi could be described with von-Bertalanffy equation as L_t= 41.99×[1-e^{-0.2264(t+0.9746)}],W_t = 980.22×[1-e^{-0.2264(t+0.9746)}]^2.7172. The inflection point of X. davidi growth age was 3.45 years and the corresponding body length was 26.54 cm and body weight was 281.88 g; The X. davidi's sex ratio was 1.4513:1. The individual absolute fecundity was 39 813-228 541 eggs(109 141 on average).

Abstract:This study used Restriction Fragment Length Polymorphism (RFLP) and Internal Transcribed Spacer (ITS) PCR sequencing technology to identify 60 isolates of suspected pathogenic Saprolegnia strains collected from 7 provinces including Shanghai, Jiangsu, Jiangxi, Sichuan, Hebei, Zhejiang and Hubei. Taxonomical analysis was also performed with strains of Saprolegnia reported in recent years. Comparative studies of physiological and molecular characteristics of pathogenic strains isolated from both infected fishes and eggs were carried out. The results showed that the 60 isolates could be classified into 10 special genetypes and all the isolates belonged to the clade of S. ferax, S. australis and S. parasitia by ITS-RFLP. Based on the phylogenetic tree, S. ferax and S. parasitia were the main Chinese pathogenic Saprolegnia strains. In addition, no significant difference existed in morphology of both asexual and sexual organs between the two S.ferax isolates from infected fishes and eggs. The same isolate could infect amphibian eggs and fishes. Thus, S.ferax and S.parasitia were identified as major pathogenic strains for both fish and eggs. The results could serve as a reference for the control of Saprolegniasis.

Abstract:Florfenicol(FFC),a synthetic broad-spectrum antibiotic,with a structural analogue of thiamphenicol is active against many thiamphenicol-resistant strains. Due to its broad antibacterial spectrum and with no side effects of aplastic anemia, FFC has been widely used in livestock,poultry,and is also legally permissive to be used in aquaculture. To study the absorption characteristics of florfenicol in GIFT Nile tilapia, which was the biggest quantity export farming fish, the 10 hour blood samples were detected using high-performance liquid chromatography with fluorescence detection method after feed administration of 0,5,12,20,40,80,100 mg/kg of florfenicol at 30℃.Results showed:the concentration was (Mean±Standard Deviation,n≥6) 0,1.49±0.46,2.66±0.62,5.08±0.75,10.60±2.34, 13.74±2.87 and 14.42±0.49 μg/mL respectively. The concentration increased with the increase of dosage at the same time in blood,while the increase amplitude decreased, which indicated the decreased utilization of drug to the increasing dosage. In the same experiment condition, 0 (control), 12,20,40,100 mg/kg body weight of florfenicol a day were given to GIFT in feed for 7 consecutive days to study the histological toxicology of florfenicol.There were no mortalities or clinically observable changes found at any of the dose levels tested. Dose dependent ultrastructural damages were found in cells of kidney and liver at the dosages of 40 and 100 mg/kg,the damage could not be repaired after 7 days at the end of the test, while no obvious pathological changes were found at dosages under 20 mg/kg. We recommend the dosage of no more than 20 mg/kg, otherwise it may damage the function of liver and kidney,lower utiliztion and cause environmental pollution, etc.

Abstract:Based on the annual monitoring on the dissolved heavy metal and petroleum hydrocarbon in 2012, the present pollution situation and features were analyzed to clarify the environmental impact caused by the construction and operation of the Yangshan Port. The results indicated that the dominant heavy metal in Yangshan Port was Hg, followed by Cu, Pb and Zn. It can be classified into two stages throughout the year of 2012, that is, the first stage was from January to May and the second stage was from June to December. Contaminant levels were relatively high in the first stage which were mainly caused by construction and drainage based on the time distribution and cluster analysis of pollutants. The results of spatial distribution showed that areas of high concentration appear at the edge of Xiaoyangshan and the southeast region of Dayangshan Island. For the Yangshan Port was in the special stage that construction and operation is simultaneously underway, it was necessary to control the pollution of harbor.It is proposed to take the concept of development coefficient by using the modified GM(1,1) model in order to evaluate the variation tendency and pattern of contamination deeply. The results indicated that, of all heavy metals, Hg, Cr and Cd are difficult to be eliminated in the investigated waters. Measures need to be strengthened to prevent and control these pollutants.

Abstract:To investigate the influence of phosphorus form and pH on phosphorus release, SMT (Standard Measurements and Testing) method was applied to analyze the phosphorus chemical form in sewage sludge from different wastewater treatment plants. The release behavior of phosphorus from sewage sludge under seawater with different pH values was also investigated in lab. The results showed that: total phosphorus (TP) content of these sludge samples varied. The percentage of organic phosphorus(OP) was low, while the inorganic phosphorus (IP), accounting for 64.42% to 83.01%,was the predominant fraction of total phosphorus(TP).And the most abundant portion of inorganic phosphorus in these samples was the non-apatite inorganic phosphorus (NAIP) fraction. We have found from experiments that the phosphorus in sludge could release under acidic, neutral and alkaline conditions. Compared with neutral condition, acidic and alkaline conditions were more favorable to the release of phosphorus. The distribution of phosphorus chemical forms influenced phosphorus releasing, but does not depend on the content of total phosphorus and inorganic phosphorus simply.

Abstract:To understand the spatial and temporal distribution pattern of the fish communities in the south of northern Jiangsu shoal, seasonal bottom trawl surveys were conducted in May (spring) and September (autumn) of 2010 in the area. The dominant fish species, the impacts of the environment on the distribution of the fish communities, spawning and feeding habitat of dominant fish were also discussed. The result indicated that numerically the average density was 1.33×10³ ind/km² in spring and 2.02×10³ ind/km² in autumn, respectively. The average weight density was 76.35 kg/km² in spring and 35.12 kg/km² in autumn, respectively. The spatial distribution pattern was the same in spring and autumn, i.e. the density was higher in the near shore than that in the neritic waters. Generally, the species number was higher in autumn than that in spring. The distribution patterns of fish species and fish diversities are also similar to the density, and the value was higher in the near shore than in the neritic. Stepwise regression showed that the weight density or individual density was significantly related to the water depth, the fishes were abundant in near shore than in the neritic waters. In spring, numerically Larimichthy spolyactis was predominant species, whereas Miichthy smiiuy was most important dominant species by weight. In autumn, Collichthys lucidus was the predominant species both in number and in weight. Compared to the other coastal waters, the south of northern Jiangsu shoal was mainly composed of economically important fishes. Our study indicated that the northern Jiangsu shoal and its outer waters were important spawning and nursery ground for many economically important fishes. The northern Jiangsu shoal plays a role in completing the life histories for many economically important fishes of the Lvsi fishing ground.

Abstract:By using stable isotope techniques, the food sources and trophic levels of catches from gillnets were analyzed from September to November, 2011 in Qiandao Lake. Results showed that δ¹³C values of each food sources varied between -29.7 ‰——23.9 ‰, among which, the highest δ¹³C values was seen in phytoplankton, with the lowest in particulate organic matter (POM). The food sources of δ¹⁵N value range is small, with the highest phytoplankton (8.5 ‰), the lowest particulate organic matter the lowest (5.1 ‰). In contrast to isotope value distribution trend of food sources,consumers of δ¹³C values range was relatively small (-29.7‰——21.6‰), and then δ¹⁵N range was relatively large (5.1 ‰-14.8 ‰). Consumers of trophic level from 1.9 to 3.6, belong to the three nutrition levels, the trophic level of fishes all distributed between the third to the fourth trophic level having good consistency with the feeding characteristics. But as an invasive species, herbivores fish Megalobrama amblycephala and omnivorous fish Ietalurus punetaus trophic level as high as 3.4 and 3.0, which inicated that feeding habits changed greatly in Qiandao Lake. Isotope tracer method research results also showed that the carbon source primarily is phytoplankton in Qiandao Lake water ecosystem.

Abstract:CPUE (catch per unit effort) standardization is an essential task in fisheries stock assessment and GLM (generalized linear model) which has been used as a standardized method in the CPUE standardization. Before using GLM, the error structure, independent variables, and interaction between variables in the model had to be assigned and it would cause a great error if the assumption was wrong. Moreover, GLM could not be used to handle missing values automatically and to detect and extract complex interactions from the CPUE data. Outliers also had a great impact on the results estimated by using GLM. In contrast to GLM, regression trees may do a great job to deal with the above situations. In this paper, based on simulation data and chub mackerel (Scomber japonicus) catch and effort data from Chinese lighting-purse seine fishery in the East China Sea and Yellow Sea, we compared the performance of the regression tree and GLM in the CPUE standardization and the results showed that both models could do a good job if there were no outliers in the data and nonlinear relationships between nominal CPUE and abundance. Because the regression tree was characterized by a step function, the GLM was better in standardizing CPUE in this situation. However, if there were outliers and nonlinear relationships, the regression tree would harvest less root mean square errors and explain more deviations with fewer variables than GLM. The regression tree also could detect the complex relationships between independent variables and response variables by visualization which was ideally suited to explore and analyze the catch and effort data from fisheries.

Abstract:Based on the data collected in Jiaojiang inshore waters in April,July and November of 2010,the relationship between horizontal distribution and seasonal changes of zooplankton was discussed in this paper. The results showed that horizontal distribution of zooplankton had an obvious seasonal variation and zooplankton abundance peaked in spring (1 700.07 ind/m³), and it was the second highest in summer (258.65 ind/m³), the lowest in autumn (72.90 ind/m³). Jiaojiang runoff and Zhejiang coastal current play an important role in zooplankton abundance as a result of the relevantly weak upwelling in spring and autumn. Obvious variation of the three seasonal dominant species in investigated waters can be observed, Calanus sinicus in spring, Calanus sinicus,Acartia pacifica,Euchaeta concinna and Sagitta nagae in summer and Sagitta enflata and Diphyes chamissonis in autumn are the main species that result in the seasonal changes of phytoplankton abundance. Survey results were compared with the same period of phytoplankton findings of zooplankton, and phytoplankton and zooplankton abundance high value area is not consistent with the nutrient salt and water masses of different seasons of investigation waters changes.

Abstract:Squid is a marine animal with extremely ferocious feeding. The foraging behavior of captive Japanese common squid, Todarodes pacificus, was filmed in a circular tank and analyzed. Three shoal sizes (3, 7-12, and 44 individuals) were examined. Squid attacking on prey fish were observed, and the angle ranges for prey detection and attack were found wide and narrow, respectively. The mean mantle length of the squid was 20 cm, and most attacks on prey began when the squid were 1 to 6 mantle lengths away. Squids displayed dark patterns during the attack phases, which could have been a sign of intra-specific communication. In the large shoal size, squid chased bigger prey, and several squid sometimes fed together on a single prey. This might be an alternative strategy when food was scarce.

Abstract:Based on the data of the sinking depth, gear operating and the ocean environmental data collected on light falling-net fishing in the South China Sea during March to April 2013, the relationship between sinking depth(sinking speed) and time was studied, and the relationship between the maximum sinking depth and the relating was analyzed using multiple regression statistics. The results showed that the maximum sinking depths of the both ends of the lead line were deeper than that in the middle; the maximum sinking depths of the both ends of the lead line were consistent; the sinking speed of the both ends of the lead line was faster than the middle; the sinking speed of the both ends of the lead line achieved the maximum at depth of 10 m, and the middle achieved the maximum at 5 m; the relationship between the sinking depth and time could be described using the formula: H=-0.0008t²+0.4766t+1.2063(R²=0.9985); and the relationship between the sinking speed and time could be described using the formula: V=-1E-07t²-0.0018t+0.5269(R²=0.8813);the results of the multiple regression statistics showed that the shooting duration, the direction angle between current and the wind, pursing time had a significant effect on sinking depth(P<0.001), and the current speed and wind speed had no significant effect on the sinking depth(P<0.1); there was no multicollinearity existed among the relating factors(κ=2.396<100). Our observations provide reference for ways to improve sinking characteristics in light falling-net fishing and a guide for fishing operations.

Abstract:Cephalopods play the key roles in marine ecosystem as important invertebrate in the ocean. However, little information about its predation and dietary status has been known so far. As a novel applied technology, stable isotope has multiple utility functions with quick inspection and accurate result, so this method is applied in many researches including marine ecosystem. A large proportion of carbon, nitrogen and oxygen are accumulated in the body of cephalopods, which are both predators and food of top predators. The hard structures are the good carrier of the elements for stable isotope. Therefore by means of stable isotope tracing, the content and distribution of stable isotope in every section of hard issue is analyzed to understand the trophic level of cephalopods among the marine organism. In this paper we will give a brief introduction of the application of stable isotope in hard issues of cephalopods (such as statoliths, beaks and inner shell), give a comprehensive understanding of trophic ecology on cephalopods and have a looking in the future.