Abstract:Separation and purification of transgenic cyanobacteria has became an indispensable step in the research of its physiology, biochemistry, molecular biology, nutritional value, etc. However, has been were no research on the separation and purification of transgenic cyanobacteria under the circumstance of cross contamination during the production of transgenic product. The method used in this article separated and purified Anabaena at three levels:species, cell and gene. In this study, the separation and purification between wild type, transgenic type and empty carrier type began at biochemical level via different culture solutions with antibiotics. Then, RT-qPCR and Western blot were applied to distinguish transgenic type and empty carrier type at molecular level. Our study not only can separate three substrain, but also obtain a result of product increase and higher growth rate. This study provides a solution to the cross contamination during the production of transgenic product and can be positive to the development of transgenic microalgae industry.

Abstract:Located at the confluence of the East China Sea and the Yellow Sea, the Yangtze River Estuary is the area of overlapping distribution for two subspecies of the narrow-ridged finless porpoise (Neophocaena asiaeorientalis).In recent years, finless porpoise deaths occur frequently in this area, which has aroused widespread concern of the public.In this study, the genetic diversity and population dynamics were investigated using 16 polymorphic microsatellite loci in 36 samples of the East Asian finless porpoise (N. asiaeorientalis sunameri) from the Yangtze River Estuary. The results showed that 14 microsatellite loci did not deviate from Hardy-Weinberg equilibrium except for 2 loci. A total of 129 alleles were obtained in all of 16 loci with an average number 8.1. The average of observed heterozygosity (Ho) was 0.733; the average of expected heterozygosity (He) was 0.758, and the value of the polymorphism information content (PIC) was above 0.5. The level of genetic diversity is similar to marine mammals such as the Indian-Pacific finless porpoise(N. phocaenoides) and the seal (Phoca largha), but significantly higher than valuable terrestrial mammals such as the giant panda (Ailuropoda melanoleuca) and the Amur tiger (Panthera tigris altaica). In addition, the heterozygosity excess test and Mode-shift analysis showed that the population has not experienced genetic bottleneck effects during the recent historical period. Meanwhile, 4 simulations on Msvar software exhibited virtually no detectable effective population size change in their recent past, and estimated the recent effective population size about 5 623 individuals, which revealed that the population of N.asiaeorientalis sunameri is large and stable in the Yangtze River Estuary.

Abstract:The aim of this study was to explore the genetic variation characteristics and the level of genetic variation of Sinipercine fishes in the National Aquatic Germplasm Resources Protection Area of Jinjiang River. Based on PCR amplification and sequencing technologies, the mitochondrial control region (D-loop) sequences of Siniperca kneri, Siniperca scherzeri, Coreoperca whiteheadi(3 groups, 97 fish)were compared and analyzed. The results showed that:the sequence length of Siniperca kneri was 843 bp with no variation sites and 1 haplotype was identified; The Coreoperca whiteheadi length sequence had two types of 852 bp and 856 bp, and 28 polymorphic loci and 11 unique haplotypes were identified; The length sequences of Siniperca scherzeri were 845 bp and 846 bp with 5 polymorphic loci and 5 haplotypes were identified. The genetic distance were 0.000 0, 0.002 0±0.000 9 and 0.008 7±0.002 0 in the three groups; The haplotype diversity and nucleotide polymorphism were 0.000 0, 0.859 9, 0.7367 and 0.000 0, 0.008 7, 0.002 0, respectively. The experiment and analysis indicated that there were differences among the termination sequence regions and the conserved sequence regions. For the three populations, Coreoperca whiteheadi was a stable population with great genetic variation and rich diversity. Due to bottleneck effect, although the Siniperca scherzeri had rich haplotype diversity, but had low nucleotide diversity; Siniperca kneri had single hereditary composition, and diversity was very poor. Presumably, there may be a invasive breeding population. Jinjiang, as the biggest river of Fanjing Mountain National Nature Reserve area, once with rich resources of the Sinipercine, but had been badly damaged. It is necessary to strengthen the research and protection of the genetic resources.

Abstract:To study the effect of different diets on digestive tract index and amylase activity of largemouth bass, digestive tract index and amylase activity of largemouth bass fed the formulated feed(feed group)and the ice fish (chilled group) were analyzed comparatively. The results indicated that the intestinal weight/body weight, hepatopancreas weight/body weight, splanchnic weight/body weight of feed group were significantly higher than those of the chilled group (P<0.05). The amylase activities in different organs were sequenced as follows:hepatopancreas > pyloric caeca > intestine > stomach in two groups. The amylase activities in hepatopancreas were significantly higher than those in other organs (P<0.05). For the amylase activities of chilled group and feed group, the optimum pH were 7.2 and 6.8,6.4 and 6.8,6.4 and 6.8,6.8 and 6.8 in stomach, hepatopancreas, pyloric caeca and intestine, respectively. The optimum temperature were 30℃ and 30℃,35℃ and 30℃,50℃ and 55℃,60℃ and 60℃, respectively. The concentration of starch were 8% and 8%,0.5% and 2%,8% and 8%,8% and 8%, respectively. The amylase activity of digestive organs in the feed group was significantly higher than that of the chilled group at same temperature and concentration of starch (P<0.05). The results would provide some reference for adding enzyme preparations and optimizing artificial feed formulation in the process of artificial feed preparation.

Abstract:To explore the impact of light on the growth and animal welfare of Japanese eel(Anguilla japonica), the eel were cultured at different light intensity and light photoperiod in recirculating aquaculture system (RAS). This study was designed with the light intensity (10 lx, 100 lx, 300 lx) combining with the photoperiod (12L:12D, 18L:6D, 24L:0D) according to double-equation irrepetitive testing method. Growth indexes, routine blood physiochemical and cortisol levels were measured. The results showed that the final weight gain and daily gain of the 10 lx group were significantly lower than those of 100 lx and 300 lx (P <0.05). There was no significant difference in the survival rates of all groups at different intensity. The specific growth rate of 10 lx group was significantly lower than that of the 300 lx group (P <0.05). While the growth efficiency and feed conversion ratio of 300 lx group was significantly better than those of 10 lx group and 100 lx group (P <0.05). The final weight gain and daily gain of the 12L:12D and 18L:6D groups were significantly lower than those of the 24L:0D group (P <0.05), while the specific growth rate of 18L:6D group was significantly lower than those of 12L:12D and 24L:0D groups (P <0.05). The activities of glutamic oxalacetic transaminase of the 24L:0D and 12L:12D groups were significantly lower than 18L:6D group (P <0.05), and the activities of glutamic-pyruvic transaminase of 10 lx group conditions were significantly higher than that of 100 lx group (P <0.05). Triglyceride of 24L:0D group was significantly higher than the 12L:12D group (P <0.05). The SOD activity of 18L:6D group was significantly higher than 12L:12D group at 10lx and 300 lx (P <0.05), while the opposite of those at 100 lx. Light intensity and photoperiod have a significant difference (P <0.05) for serum cortisol of Japanese eel, 12L:12D was significantly lower than 18L:6D and 24L:0D groups at 10 lx and 300 lx. While the 12L:12L and 18L:6D group was lower than 24L:0D at 100 lx (P <0.01). The 10 lx group was significantly lower than 100 lx group in 12L:12D and 24L:0D, while the opposite of those in 18L:6D. So in RAS, the best welfare conditions for the growth of Japanese eel were 300 lx and 12L:12D.These conditions could reduce the stress of fish, and promote the development of eel industry.

Abstract:To compare the budget of nitrogen and phosphorus in Megalobrama amblycephala, which was cultivated in separated pond and sequencing pond,two kinds of ponds were selected for sample analysis, from August to November 2016, with the traditional pond as the control. The results showed that feed was the main source of nitrogen and phosphorus of Megalobrama amblycephala in ponds. The feed input proportion of nitrogen was traditional pond(68.53%) < separated pond(72.03%) < sequencing pond(76.22%). The feed input proportion of phosphorus was traditional ponds (42.87%) < separated pond (53.37%) < sequencing pond(56.64%). In these three kinds of ponds, the most important expenditure of nitrogen was sedimentation and water discharge. Among them, sedimentation and water discharge of nitrogen in the sequencing pond was the lowest, followed by the separated pond, and the traditional pond was the largest. The most important expenditure of phosphorus was aquaculture outputs. As for the proportion of phosphorus in aquaculture output to total phosphorus output, the highest is separated pond, up to 54.55%, the traditional pond is 52.20% and the sequencing pond is 43.38%. The results showed that the proportion of nitrogen and phosphorus sedimentation and water discharge in separated pond and sequencing pond was lower than that of traditional pond. It can reduce the nitrogen and phosphorus sedimentation and water discharge, increase the utilization rate of nitrogen and phosphorus, and improve the efficiency of feed utilization by constructing separated pond and sequencing pond. In particular, sequencing pond was used to raise the efficiency of material utilization in Megalobrama amblycephala farming.

Abstract:In view of precision feeding machine developed by Shanghai Ocean University, with CFX simulation analysis, combined with single factor experiments and orthogonal experiments, the influence of measurement accuracy of the feeder feeding tube cone angle, length and feed tube in tube access port velocity is studied. The single factor test found that the feed mass flow that is measured by the weight device is determined by the dip angle α of the feeding tube, the tube length L and the feed speed v₀ at the access port of the feeding tube; the best parameter of accurate feeder feeding machine based on the design of orthogonal test should be as followed. The feeding tube dig angle is 38°. The length is 410 mm. The tube access port velocity is 0.32 m/s. The best measurement accuracy under these parameters is got; the primary and secondary order of measurement accuracy is obtained by range analysis. The feed speedat the tube access port is the main factor affected the accuracy of measurement, and dip angle is the second, tube length is last.

Abstract:The concentrations and distributions of heavy metals Cu, Zn, Pb, Cd, Hg, and As in surface sediments were investigated with the samples collected from the Qiantang River. Then we analyzed the potential ecological risk of heavy metal with the Hakanson ecological risk index and searched for its source based on Pearson correlation coefficient and principal component analysis(PCA). Results showed that the concentrations of Cu, Zn, Pb, Cd, Hg and As in surface sediments of Qiantang River were rather high, the overall potential ecological risk level was extremely high. Cd and Hg were the prominent factors for ecological risks in surface sediments, showing the intensity of potential risk as extremely strong and strong, respectively. There was a certain distribution difference with different sections of the river, among which the concentrations and potential ecological risks in S3 section were both higher than others. It was likely that the sources of heavy metals in surface sediments were similar; industrial pollution, agricultural non-point source pollution, traffic pollution and domestic wastewater were the probable sources by the method of PCA. Besides, we determined the effects of heavy metal with surface sediment on fish in Qiantang River. Results showed that the biota-surface sediment accumulation factor(BSAF) of Hg was 0.861, which presented a bioaccumulation to fishes. However, the average levels of heavy metals in surface sediments presented low correlations with fish on overlying water samples. It may be caused by the fact that the water in Qiantang River flowed fast and the majority of fish investigated were from proliferation of releasing of young age.

Abstract:By Miseq high-throughput sequencing technology, soil microbial community dynamics and the associated soil physiochemical characteristics were studied. The reclaimed Scirpus mariqueter wetland in Shanghai Nanhui Dongtan coastal area was the study site, which is just undergoing large scale wetlands reclamation impacts. Results showed the dense area and the light beach of S. mariqueter soil microbial communities were significantly different (P<0.001). The dense area of S. mariqueter is higher than the light beach area. The seasonal variation of microbial diversity and abundance in the same area was significant (P<0.001), and the spring and summer microbial diversity and abundance was lower than that in the autumn and winter. Proteobacteria, Bacteroidetes, Acidobacteria, Chloroflexi, Planctomycetes, Actinobacteria, Cyanobacteria, Gemmatimonadetes, and Nitrospirae were relatively abundant, and the seasonal dominant microbial community was different. Redundancy analysis showed that microbial community structure was significantly correlated with factors including soil salinity, dry wet ratio and nitrate nitrogen. This study is helpful to understand the micromechanism during S. mariqueter wetlands spatial sprawling and promote the fast remediation of S. mariqueter wetlands.

Abstract:Dishui Lake(DSL) has received a great deal of attention since it built, there are many researches on the quality of water in it, but the research on the bacterial diversity overall was insufficient. Therefore, this study combined 16S rRNA gene denaturing gradient gel electrophoresis (DGGE), 16S rRNA gene metagenome and bioinformatic analysis to understand bacterial diversity and community structure in surface water of DSL, including water fecal pollution indicator bacteria, water eutrophication typical bacteria, pathogens and conditional pathogens. The results show that:bacterial diversity in these 5 water samples can represent it in surface water of DSL at sampling time. 428 OTUs have been clustered from 5 DSL water samples and belong to 17 phyla, 39 classes, 66 orders, 130 families, 191 genera and 282 species. Actinobacteria, Cyanobacteria, Proteobacteria, Bacteroidetes, Verrucomicrobia and Planctomycetes are six dominant phyla in these 5 samples and account for 97% of total bacterial sequences in these samples. Except for a few sequences of Shigella (0.1%), sequences of water fecal pollution indicator bacteria, Pathogens and conditional pathogens were not found in metagenome. In conclusion, the characteristics of bacterial diversity in surface waters of DSL are similar to those of typical freshwater lakes and there are no water-contaminating bacteria detected in the lake, and there is no tendency that blue algae will cause blooms.

Abstract:Here we report the synthesis and characterization of magnetic Fe₃O₄ loaded chitosan microparticles for low cost, biocompatible heavy metal removal. Magnetic Fe₃O₄ loaded chitosan microparticles are prepared based on ionic gelation and characterized by Transmission electron microscope (TEM), Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) respectively, which confirm that magnetic Fe₃O₄ nanoparticles have been incorporated into chitosan microparticles successfully and the particle size falls into the range of 164.05-768.69 nm. Their adsorption efficiencies for Cu (Ⅱ), Cd (Ⅱ) and Zn (Ⅱ) have been investigated, in which small size magnetic Fe₃O₄ loaded chitosan microparticles present higher adsorption efficiency than that of other ones. And their maximum adsorption capacity for Cu (Ⅱ), Cd (Ⅱ) and Zn (Ⅱ) could reach 51.66, 97.86 and 82.24% respectively. Moreover, the adsorbing mechanism can be well explained by pseudo-second-order model (R²=0.998-0.999). Notably, magnetic Fe₃O₄ loaded chitosan microparticles also exhibited excellent biocompatibility when co-cultured with MCF-7 cells with nearly 100% viability. This study demonstrates that small size magnetic Fe₃O₄ loaded chitosan microparticles is an effective, low cost and bio-compatible heavy metal adsorbent and could potentially be widely used in many different fields, for example, wastewater treatment in food and agriculture.

Abstract:The urgency demanding to reuse the aquaculture water is more and more strengthen with the shortage of the water resources and the constraint of the environment pollution. How to correctly evaluate whether the treated aquatic water can be used ofr aquaculture again is the basis for avoiding insuffient or excessive treatment and improving treatment efficiency. Few reports about evaluating the reusing possibility was found until now. The reuse avaluation of aquaculture water can draw lessons from the evaluation criteria of aquaculture headwaters, but there should be differences. The current paper proposed the evaluating parameter used to of the reuse aquaculture water based on the effect of the aquaculture activity on the water quality. Furthermore, variation criteria and fixed criteria are proposed according to these parameters. For established production system and aquaculture animals, the recording and analysis of existing production behavior is the basis for obtaining information that can guide the actual aquaculture activities.

Abstract:By regularly monitoring and analyzing the water chemical indexes and the content variation trend of S^2-, S⁰ and TS in sediments of the outdoor ponds of Litopenaeus vannamei, their relationships with DO, pH, TP, TN and TOC were investigated for guiding the healthy aquaculture of Litopenaeus vannamei. The results indicate that S^2- content in sediments of the outdoor ponds and surrounding rivers were respectively 0.31~5.86mg/kg and 18.13-232.56 mg/kg, which were all lower than the safety low-limit of sulfide content in fishery sedimentary environment; S⁰ and TS contents in sediments of the outdoor ponds were 0.39~3.06 mg/kg and 221-4421 mg/kg, respectively. S^2- and TS contents showed wave-like increase trend with aquaculture time, and S⁰ content varied within a cube-type range. S^2- and S⁰ proportions of TS in sediments of outdoor ponds were respectively 0.12%-1.76% and 0.12%-1.04%, and those in sediments of water source were respectively 4.30%-17.85% and 0.09%-0.37%; The main water chemical indexes met the Litopenaeus vannamei aquaculture demand. TOC and TN contents showed wave-like increase trend versus aquaculture time but the variation trend of TP content was not obvious. Additionally, it was found that S^2- and TS had a significant positive correlation with TOC and TN but a negative correlation with DO and pH. The results demonstrate that accumulation of organic matters including residual feeds, metabolites and biological carcasses in shrimp pond could result in higher oxygen consumption and the sulfide content increase under low-oxygen condition. Hence, it was suggested that the key solutions to reducing the sulfide and organic matter contents in shrimp pond sediments were the rational feeding, the maintenance of abundant dissolved oxygen in shrimp ponds and the thorough desilting.

Abstract:Crustaceans hard structures, which can record biological information, disappear periodically after molting.Thus, studyon the age and growth of crustaceans has been a tricky problem. In this paper,Procambarus clarkii were collected from the river channel of NanhuiNew City, PudongNew District, Shanghai Municipality. The eyestalks microstructure of the species wasanalyzed, and the relationship between growth incrementsinthe eyestalks and the length and weight of Procambarus clarkii were studied. Clear microstructures were observed both in cross and vertical section of the eyestalks, which are composed of four parts (ie. epicuticle, exocuticle, endocuticle and membranous layer).Each growth increments consist of one light and one dark increment.The number of growth increments of sampled specimens are form8 to 31.The width of growth increments become narrow fromthe junction of exocuticle and endocuticle to the junction of endocuticle and membranous layer.The number of growth increments in eyestalks and the length and weight of Procambarus clarkiiare both suit for Logistic relationships, and their functions are L=(96.66)/(1+e^{-0.25(t-2.54)}) and W=(22.48)/(1+e^{-0.41(t-11.78)}) respectively.

Abstract:The study carried out the fishing tests of 3 kinds of LED lamps and 1 kind of traditional metal halide lamp to analyze if the LED lamp was suitable for inshore falling-net fishery of South China Sea. The results show that:(1) there are two peaks in the relative spectral power distribution of white light LED used in the fishing test, and two peaks are in blue bands, green and yellow bands respectively; green light LED has one peak and the peak is in green bands; blue light LED has one peak and the peak is in blue bands; metal halide lamp has multiple peaks and the peak wavelength is mainly in red bands; (2) The catch rates of 3 kinds of LED lamps are all higher than metal halide lamp, and white light LED has obvious advantages; (3)Loligo chinensis is the common dominant specie among 4 kinds of lamps, and the catch rate of white light LED is significantly higher than the other 3 kinds of lamps; (4) one-way analysis of variance shows that there is no significant difference in the mantle length and weight of Loligo chinensis between individuals caught by white light LED and metal halide lamp. The study suggests that LED lamp, especially white light LED is suitable for inshore falling-net fishery.

Abstract:Based on the basic data of fishery resources by set-nets in Northern Hangzhou Bay from May to August, 2017, a preliminary analysis of biological characteristics of reproductive stocks of Collichthys lucidus was carried out. The results indicated that body length of reproductive stocks of Collichthys lucidus ranged from 87 to 150 mm, and average body length was (116.4±9.9) mm, while body weight of reproductive stocks of Collichthys lucidus ranged from 12.6 to 53.3 g, and average body weight was (27.4±7.2) g. There was an increasing trend of average body length and body weight, and independent T-test displayed a significant difference in average body length and body weight between May and August. Sex ratio (female:male) of Collichthys lucidus was 2.53:1, the number of female was significantly higher than the number of male. According to chi-square test, sex ratio was not 1:1. The gonad maturity stages of female was mainly from Ⅳ to Ⅴ, accounting for 78.2%, and the gonad maturity stages of male was mainly from Ⅲ to Ⅳ, accounting for 97.9%, and body length of individuals whose gonad maturity stages had achieved Ⅳ or above mainly ranged from 100 to 130 mm. The feeding levels of reproductive stocks of Collichthys lucidus was mainly 0 and 1, accounting for 72.8%, and the empty stomach ratio was 39.8%. The research showed that Northern Hangzhou Bay was the feeding ground of Collichthys lucidus. Collichthys lucidus has become smaller, and management for protection of reproductive stocks of Collichthys lucidus in Northern Hangzhou Bay should be enhanced.

Abstract:As an important method for surimi preservation, frozen storage can cause degeneration of myofibrillar protein, resulting in the decrease of surimi quality. Addition of cryoprotectants is considered to be one of the most effective ways to prevent denaturation of aquatic protein. In this paper, the regularity of protein freeze denaturation, physicochemical change and mechanism of cryoprotectants were discussed. On this basis, the progress of research and application of trehalose, algae extract, polyphenols, protein hydrolysates and other green antifreeze agents were reviewed. Finally, the existing problems of cryoprotectant were analyzed, and its development trend was prospected, aiming to provide reference for the frozen surimi industry.

Abstract:Ultrasonic-assisted extraction of polysaccharides from Gelidium pacifium Okam. was optimized by response surface methodology based on the single factor experiment, The experiment results showed that the optimal extraction conditions were ultrasonic temperature 85℃, extraction time 50 min, and material-to-liquid ratio of 36 mL/g. Under these extraction conditions, the extraction yield of polysaccharides was 31.21%. The antioxidant activities of the extracted polysaccharides were evaluated by measuring the scavenging ratio of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical,2,2'-zainobis (3-ethylbenzothi azoline-6-sulfonic acid) ammonium salt radical (ABTS⁺) and superoxide anion radical. The results of antioxidation experiment indicated that the polysaccharides from Gelidium pacifium Okam. removed DPPH radical, ABTS radical and superoxide anion radical, showing a dose-effect relationship. The antioxidant activity of polysaccharides obtained by ultrasonic-assisted extraction was higher than that obtained by aqueous extraction.