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Table of Content

    25 June 2020, Volume 42 Issue 3
    High-throughput sequencing analysis of soil microbial diversity from Antarctic Fildes Peninsula
    2020, 42(3):  195-204. 
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    The study of microbiomes using metagenomic high-throughput sequencing enables the analysis of uncultivated microbial populations that may play important roles in the environment. The Antarctic is an extreme environment where pole-end temperature, radiation, and desiccation limit most of life activities. Through long term adaptation and evolution, the microbial populations form special biodiversity and genetic backgrounds, develop various metabolic pathways that produce enzymes and active substances with special functions. These precious bioresources can be used as biofuels, medicine or for environmental protection and other fields widely. In order to discover the biodiversity and community structure of the microorganisms in the Antarctic soil, we collected 3 soil samples from Antarctic Fildes Peninsula as the study objectives. Initially, we obtained the diversity information through metagenomic high-throughput sequencing. MetaPhlAn2 and GraPhlAn were applied to analyze the sequencing data and visualize the microbial composition results, respectively. The microbes identified from the three soil samples belonged to 8 phyla, 14 classes, 21 orders, 32 families, 48 genera and 62 species. Among them, Pseudomonas was the dominant bacteria and Candida albicans was the dominant fungi. This discovery was consistent with the results from Liu Chunying, who has previously established some studies of Antarctic soil microorganisms. The bacterial abundance of S61 was the highest among the 3 samples and the dominant genus was Pseudomonas. Compared with bacterial species identified from S59 and S62, Janthinobacterium lividum, Pseudomonas veronii and Ralstonia were exclusive in S61. The relative abundance of bacteria and fungi in S59 was accounted for 59.83% and 39.92%, respectively. In which, Actinoplanes exclusively existed and Pseudomonas mandelii was the dominant bacterial species, while C. albicans was the dominant fungi species. The fungi abundance in S62 was slightly higher, which was about 58.13%. In this sample the dominant fungi was the same with S59 and the dominant bacteria was Pseudomonas. These results suggested us that the Antarctic had abundant microbial diversity and the species were distributed differently in different regions. The study of environmental microbial communities through metagenomic high-throughput sequencing provided supports for further investigations of metabolic potentials of microbiomes, in addition to their taxonomic composition. This greatly improved the ability to interpret and predict functional interactions and population dynamics of microbiomes.
    Morphological variation study among nine wild populations of Ptychidio jordani on the main stream of Xijiang River in Guangxi
    2020, 42(3):  205-215. 
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    According to the traditional morphological measurement of 333 Pthchidio jordani collected from upstream (Tian’e, Donglan, Dahua, Du’an and Heshan), midstream (Wuxuan, Guiping and Pingnan) and downstream (Wuzhou) on the main stream of Xijiang River in Guangxi, the morphological difference among nine geographical populations of P. jordani were explored by principal component analysis, one-way analysis of variance (One-way ANOVA), discriminant analysis and cluster analysis. Thereinto, Principal component analysis extracted a total of four principal components (PC1~PC4) which could explain 81.429% of morphological difference. The contribution rates of PC1 and PC2 were 61.830% and 7.863%,respectivel, and mainly reflected morphologic changes on the dorsal-ventral axis and tail of the fish. By making a scatter plot on the scores of PC1 and PC2, the morphological differences of most populations were easier to distinguish on the PC1 than PC2, of which Guiping, Pingnan and Wuzhou had deviated significantly from other populations. Further one-way ANOVA revealed that there were significance differences (P<0.05) in the characters of dorsal-ventral axis (D3-6 and D5-8) of these three populations with most populations. In the discriminant analysis, the comprehensive discriminant accuracy of each geographical population was 96.36% and it reached to 91.30% after cross validation. Cluster analysis found that Wuxuan population was most similar to the five populations from upstream with morphological characters, which first clustered into a single clade, then in turn clustered with far morphological distance of Wuzhou, Guiping and Pingnan populations together. In conclusion, there were significant difference about morphological of the nine wild populations of P. jordani on the main stream of Xijiang River in Guangxi. This study would provide reference for the protection management and rational utilization of the resources of P. jordani.
    Observation and correlation analysis of body size and body weight traits of bivalve, Cultellum scalprum in Funing Bay of Fujian Province
    2020, 42(3):  216-222. 
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    In this paper, the measured body weight traits (body weight, fresh and dry weight of soft tissue, fresh and dry weight of the shell), body size traits (shell length, shell height and shell width) and condition index of 1a~2a bivalve of Cultellum scalprum from Funing Bay in Fujian Province were described. The correlation between the traits, growth types of the traits, and the effect of body size traits on body weight traits of the bivalve were analyzed with correlation,regression and path analysis, respectively. The results showed that variable coefficient (CV) of weight traits was 28.09%~43.10%, CV of body size traits was ≤ 15%, CV of weight traits was larger than that of body size traits of bivalves. There were significant correlation coefficients between body size and weight traits of the bivalve (P<0.01), and the regression relationship between the traits could be expressed by power function equation. Compared with body size traits (shell length, shell height and shell width), the body weight of the bivalve showed positive abnormal growth (b>1.0, P<0.01); shell width and shell length showed isokinetic growth (b=1.0, P<0.01); shell height and shell length, as well as shell width and shell height showed negative allometric growth respectively (b<1.0, P<0.01). The multiple regression equation was obtained to estimate body weight of the bivalve: Y=-2.535 1+0.046 0X1+0.060 4X2+0.295 8X3 (shell length: X1, shell height: X2 and shell width: X3). Based on path analysis, the direct effects of shell width on weight traits (body weight, the fresh weight and dry weight of the soft tissue) were the largest; and the direct effects of shell length on body weight, shell height on fresh and the dry weight of the soft tissue were the second; the indirect effects of shell height through the shell width on body weight, fresh weight and the dry weight of the soft tissue were the largest, respectively.
    Primary research on the impacts of marine environment ecology from the offshore wind farm opreation——a case study of the experimental offshore wind farm in Fuqing Xinghua Bay
    2020, 42(3):  223-233. 
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    For the project of the experimental offshore wind farm in Fuqing Xinghua Bay, based on the marine environment monitoring between pre-construction and operation period, we analysed and discussed the impacts of environmental factors (e.g. water quality, sediment quality, marine ecology, and fishery resources) during the project implement. We found that the marine environmental ecology impacts of the experimental offshore wind farm operation were in forecast range, and the influence level of some indices were better than prediction.
    Primary study on DNA damage in hepatopancreas cells of Ruditapes philippinarum exposed to bifenthrin
    2020, 42(3):  234-240. 
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    To evaluate the genotoxicity of bifenthrin, the present study investigated the damage effects of different concentrations of bifenthrin on the DNA of hepatopancreas cells in Ruditapes philippinarum. After exposure to 0.05,0.10,0.15,0.20 mg/L bifenthrin for 25 days, hepatopancreas cells of Ruditapes philippinarum were extracted and assayed by using the single cell gel-electrophoresis (SCGE).The percentage of comet tail DNA (TDNA%), comet tail length (TL), olive tail moment (OTM)and Tail rate (TR) were used as the indicators of DNA damage. The results showed that, all indicators in all treatment groups increased significantly compared with those in the control group (P<0.05). All indicators increased gradually according to the increased concentrations of bifenthrin. The correlation coefficient of r was greater than 0.99, which indicated that the concentration-effect relationship was observed in the experimental concentration range. TDNA% and OTM were the better experimental indices in the assessment of comet assay. The multiple-dimensional regression equation had significant statistic meaning (P<0.01), which might be an effective way to estimate the exposure concentration of bifenthrin.
    Extraction conditions of saponins from sea cucumber
    2020, 42(3):  241-247. 
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    Sea cucumbers have long been used as a kind of food and traditional medicine in Asian countries. Stichopus hermanni, Thelenota ananas, Thelenota anax, Holothuria fuccogilva, and Actinopyga mauritiana are the main highly-valued species. Sea cucumber is rich in active substance such as triterpene glycosides, polysaccharide collagens and chondroitin sulfates. It is well known that holothurians are of high nutrition and pharmic values. Saponins are the main secondary metabolite of sea cucumber, and they are also the chemical basis for their defense. Sea cucumber saponins are composed of aglycone and sugar chain through β-glycosidic bond, mainly of holostane-type. Sea cucumber saponins are more polar, soluble in water, hot methanol, and ethanol. Moreover, there are many phospholipids in sea cucumbers, so alcohol extraction is generally used to remove fatty acids and save costs. At present, the extraction methods of sea cucumber saponins are mostly reflux extraction, alcohol-alcohol extraction, water extraction, and cold soak extraction. The separation and purification method mainly use a series of chromatographic techniques, such as macroporous resin column chromatography, dextran gel column chromatography, normal phase silica gel column chromatography, reversed phase silica gel column chromatography and HPLC. Pharmacological experiments showed that most of the sea cucumber saponins had strong physiological activities, including anti-tumor, anti-fungus, anti-radiation, hemolytic and anti-choline effects. Sea cucumber saponins was the main active substance of sea cucumber, which had great development value and potential application. In recent years, more and more researches on sea cucumber saponins have been carried out, and a variety of sea cucumber saponins have been gradually discovered. At present, at least 147 kinds of sea cucumber saponins have been isolated. But the content of sea cucumber saponins was low in sea cucumber. The metabolism process of sea cucumber is complex, the secondary metabolites in the body are abundant, and the sea cucumber saponins are more isomers, which bring great interference to the separation and purification and structural identification of saponin monomers, difficult to fully extract and analyze. In order to reduce the interference of polysaccharides and proteins in the preparation of sea cucumber saponins and simplify the subsequent separation process, the effects of ethanol concentration, temperature and solid-liquid ratio on the extraction of sea cucumber saponins were investigated. The results showed that the yield of sea cucumber saponins was higher when the concentration of ethanol was 0% and 80%, and the polysaccharide content in the extract was less when the concentration of ethanol was 0% and 80%, which was more conducive to subsequent separation; the saponins were more stable when extracted at 20 and 30℃, and less impurities when extracted at 30℃; and the content of sea cucumber saponins was higher when the ratio of material to liquid was 1:15, and the extraction time was controlled Impurities are produced and more saponins can be obtained. The results showed that when extracting sea cucumber saponin, it was extracted with 0% ethanol (pure water), the ratio of material to liquid was 1∶15, 20 °C, and the extraction time was controlled at 8 h, which could effectively increase the yield of sea cucumber saponin, control the generation of impurities, save extraction costs and simplify the separation and purification steps.
    Auxiliary protection of oyster extracts on chemical liver injury in mice
    2020, 42(3):  248-253. 
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    This study investigated the protective effect of oyster extract on acute liver injury in mice. A model of alcohol-induced acute liver injury in mice was established to monitor the body weights of mice, the contents of MDA (Malondialdehyde), GSH (glutathione) and TG (triglycerides) in liver tissues and the effects of liver histopathology. The results showed that there was no significant difference in the weight of mice between all dose groups of oyster extract and model group and normal group (P>0.05). MDA and TG levels in high-dose group were significantly lower than those in model group, and GSH levels in high-dose group were significantly higher than those in model group (P<0.05). The degree of hepatic steatosis in high-dose group was significantly lower than that in model group, with no significant difference (P<0.05). Therefore, it could be revealed that oyster extract had an auxiliary protective function on acute chemical liver injury in mice.
    A comparative study on the low temperature resistance of larvae from different breeding populations of Litopenaeus vannamei
    2020, 42(3):  254-261. 
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    In order to explore the breeding techniques and methods of Litopenaeus vannamei, the comparative experiment on the low temperature resistance was conducted based on the larvae from four breeding populations of L. vannamei (A, B, C and D), including inbred progenies and hybrid progenies according to different mating combinations. In this experiment, the growth period of larvae was P11~P12; the body length was 6.09~7.54 mm; the water temperature was 13.5~19.0℃; and the experimental period was 48 h. According to the survival rates at the time of 24 h, 36 h and 48 h, the low temperature resistance of larvae from different provenances was evaluated comprehensively in all groups by combining the factor of body length. The results showed that there was a significant difference in the survival rate among all groups from the low temperature test of 24 h. At the time of 48 h during the experiment, the mating combination in which 4 groups had the highest survival rate and were related to seed shrimps of Population A (unrelated to those of Population D) had an average survival rate of 10.35% in these 16 mating combinations. An average survival rate of 0 was found in the mating combination in which 4 groups had the lowest survival rate and were not related to seed shrimps of Population A and also in the mating combination in which 3 groups were related to Population D. There was an extremely significant difference in the low temperature resistance of larvae from different provenances.
    Analysis of water environment variations for industrialized culture system of Litopenaeus vannamei
    2020, 42(3):  262-268. 
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    The features of water environment variations were analyzed in an industrialized culture system of Litopenaeus vannamei. The results showed that NO3- was the major form of DIN, which had a proportion of 55%~85%. NH4+ fluctuated in a low concentration range of 0.005~0.060 mg/L. NO2- accumulated during the culture process in the 5# ponds, which had the same trend as NO3-. PO43- ranged from 0.012 mg/L to 0.400 mg/L with an overall upward trend. Contents of Chl-a in the ponds increased at the beginning and then decreased at the end of the culture, with a highest value in September. The number of total heterotrophic bacteria in cultural water was 2.80×104~2.40×105 CFU/mL, and 8.00×102~1.00×104 CFU/mL for Vibrio. Proportions of Vibrio in total heterotrophic bacteria were below 10% all the time.
    Investigation of the main diseases on cultured Rana spinosa in Fujian Province
    2020, 42(3):  269-276. 
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    Through the establishment of fixed investigation points to carry out regular investigation, supplemented by door-to-door visits and irregular itinerant investigation, from 2015 to 2017, the types, epidemic situation and harm degree of some diseases in the breeding area of Rana spinosa farm in Fujian Province were investigated and analyzed. The results showed that there were 17 kinds of diseases occurring on cultured R. spinosa in Fujian Province, among which 10 were in tadpole-period and 7 were in frog-period. The most serious diseases in tadpole-period were Hexamita disease and Suctorida disease, while the average annual incidence of which was 22.5% and 25.2%, and the annual average mortality rate was 13.1% and 12.4%, respectively. The most serious disease in frog-period was head-crooked disease, and the annual average morbidity and annual average mortality rate were 16.5% and 8.6%, respectively. The occurrence and death of diseases were mainly concentrated in the high temperature season of summer and autumn, while the incidence and mortality of diseases were low in winter and spring. By the investigation, the occurrence of diseases on cultured R. spinosa in Fujian Province were preliminarily found out, which provided scientific basis for the prevention and control of diseases in breeding production.
    Rainbow trout (Oncorhynchus mykiss) culturing experiment with natural flow-through aquaculture system
    Fang-Ping CHEN
    2020, 42(3):  277-281. 
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    The present study determined the hatching rate, survival rate, and net weight of rainbow trout, as well as water temperature, dissolved oxygen level, and ammonia-nitrogen content in natural flow-through aquaculture system (NFTAS) based on a culture experiment. The results showed that the hatching rate of egg in NFTAS was 89.2%, the survival rate was 92.1%. The total fish harvest after 12-months-feeding was 13 262.72 kg, the mean body weight was 1.36 kg per fish, and the mean fish harvest per mu was 6 270 kg. The coefficient of feeds was 1.07. The water temperature, dissolved oxygen content and ammonia nitrogen content in NFTAS were maintained at normal levels which were close to the suitable growth temperature of the rainbow trout, indicating that the flow of natural stream was sufficient to keep the normal growth of the rainbow trout. Heaters, equipment for heating or oxygen were no necessary in NFTAS. This culture experiment provided the practical basis for NFTAS promoting.
    Application of DNA molecular marker technology in Pyropia haitanensis
    2020, 42(3):  282-288. 
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    Pyropia haitanensis, an important economic cultured species in China, has attracted more and more attention from scholars. With the rise of molecular markers development, they have been widely used in the study of algae, providing efficient and valuable information for P. haitanensis. This paper introduced the main molecular marker technologies, including restriction fragment length polymorphism (RFLP), randomly amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), sequence characterized amplified regions (SCAR), sequence related amplified polymorphism (SRAP), simple sequence repeats (SSR), inter-simple sequence repeat (ISSR), and their application in genetic diversity, germplasm identification, fingerprint mapping construction, population and genetic relationship of P. haitanensis. Furthermore, the existing problems of molecular marker in the research field of P. haitanensis were expounded. The limitations of molecular marker technology and particular properties of P. haitanensis and the prospects for future development were given.
    3D cell culture technology and its application in aquaculture
    2020, 42(3):  289-294. 
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    Cell culture technology has made important contributions since 19th century, in various disciplines such as genetics, immunology, experimental medicine, and oncology. Cell culture of aquatic animals began in the 1960s. It was originally used for the isolation and purification of viruses and is of great significance for the prevention and treatment of diseases in aquatic animals. Three-dimensional (3D) culture of cells is a new culture method developed on traditional cell culture technology, which can better simulate the growth environment and condition of cells in living organisms, and has paid more and more attention by researchers. Based on the summary of the characteristics and development of 3D cell culture technology, this paper briefly introduced and prospected the application of 3D cell culture technology in aquaculture.