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SUPPLEMENTAL DESCRIPTION OF MYXOBOLUS HAICHENGENSIS CHEN, 1958 (MYXOZOA: MYXOSPOREA) INFECTING THE GILLS OF ABBOTTINA RIVULARIS BASILEWAKY: MORPHOLOGICAL AND MOLECULAR DATA
Peng LI, Xin ZHAO, Bing-Wen XI, Jun XIE
 Available online  
[Abstract](0) [PDF 995KB](0)
Abstract:
Myxobolus haichengensis Chen, 1958 forms numerous small plasmodia on the gill filaments of wild cyprinid Abbottina rivularis Basilewaky. The species described originally was lacking important characters, which made the accurate identification difficult. Here, we supplemented its characteristics with morphological and molecular data. Plasmodia of M. haichengensis are oval. Mature spores are ellipsoidal-shaped in frontal view and fusiform-shaped in lateral view, measuring (10.8±0.7) μm (10.1—11.5 μm) long, (8.1±0.5) μm (7.5—9.0 μm) wide, and (5.7±0.4) μm (5.2—9.0 μm) thick; two unequal polar capsule are pyriform with tapering anterior, large polar capsule averaging (4.7±0.5) μm (4.8—6.7 μm) long and (2.5±0.2) μm (3.2—4.3 μm) wide; small polar capsule averaging (4.4±0.2) μm (4.1—4.8 μm) long and (2.2±0.1) μm (2.0—2.5 μm) wide; polar filaments coil with four to five turns. The nuclear 18S rDNA sequence was obtained and deposited in GenBank (KY965936), and sequences alignment analyses revealed that M. haichengensis was most similar with the actinosporean Hexactinomyxon type 2 (AY162272, 97%) released from the freshwater tublificid oligochaete Limnodrilus hoffmeisteri.
INVESTIGATING THE STOICHIOMETRIC COUPLING BETWEEN MESOZOOPLANKTON ASSEMBLAGE AND SESTON IN FRESHWATERS
Hong GENG, Feng CHENG, Song-Bo WANG, Hong DU, Lai-Yan WU
 Available online   , doi: 10.7541/2018.106
[Abstract](122) [FullText HTML](20) [PDF 597KB](18)
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Food quality can significantly affect the phosphorus content, growth, reproduction, as well as physiological behavior of zooplankton, leading to regulate the energy transfer efficiency at the primary producer-consumer interface. However, the elemental coupling between zooplankton and food is scarcely observed at the community level. In this study, the elemental coupling and the elemental imbalances between seston and mesozooplankton assemblage in addition to the ecological impacts of such imbalances on zooplankton-phytoplankton interaction were detailed by analyzing the compositions of carbon, nitrogen, and phosphorus in seston and mesozooplankton assemblage derived from 13 samples containing water bodies in the Hubei province, China. The achieved results showed that both C﹕N and C﹕P ratios in mesozooplankton were positively correlated with the corresponding ratios in seston. Besides, the elemental imbalances between seston and mesozooplankton were positively correlated with the corresponding seston ratio (C﹕N,r=0.97, P<0.001; C﹕P,r=0.81, P=0.001). Further analysis also revealed that the energy transfer efficiency between zooplankton and phytoplankton declined with the increase of elemental imbalance in C﹕P ratio (r= –0.58, P=0.037). Moreover, a similar decline in zooplankton biomass was found as well as increase of imbalance in C﹕P ratio (r= –0.59, P=0.033). These results indicated that seston stoichiometry could remarkably impact the elemental stoichiometry of mesozooplankton assemblage, and the elemental imbalances tended to be greater with the increase of seston stoichiometry; consequently, the energy transferred to mesozooplankton was decreased as well.
THE EUTROPHICATION AND ITS REGIONAL HETEROGENEITY IN TYPICAL LAKES OF CHINA
Na LI, Jia-Xi LI, Guo-Wen LI, Ye LI, Bei-Dou XI, Yi-Wen WU, Cao-Le LI, Wei LI, Lie-Yu ZHANG
 Available online   , doi: 10.7541/2018.105
[Abstract](87) [FullText HTML](15) [PDF 1456KB](14)
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In order to comprehensively evaluate the eutrophication status in China lakes, this study scientifically analyzed the eutrophication and its regional heterogeneity in 22 representative lakes of China, and the quantitative relationship between TP and eutrophication was also studied. The results showed that 59.1% of the surviving lakes are in different eutrophication status from 2010 to 2011. Among them, Yunnan-Guizhou Lake area has the highest eutrophication level, but Menggu-Xinjiang Lake area has a bipolar levels. The lakes at Northeast Mountain and Plain area, and also the lakes at eastern area generally have a level between middle nutrition to mild eutrophication. The lowest eutrophication level is at the area of Qinghai-Tibet Plateau. Based on the relationship between sunshine amount, frost-free period, temperature, water depth, altitude, precipitation and lake nutrition status, it reveals that the geographical location of lakes is the fundamental factor affecting lake eutrophication and its regional distribution. The results of comprehensive analysis showed that there is a significant correlation between chlorophyll a and total phosphorus concentrations in lakes nationwide. Cubic curve regression equation fits the relation between chlorophyll a and total phosphorus concentrations at the areas of eastern plain, northeastern plain - mountain, Qinghai - Tibet Plateau and Yun - Gui Plateau. S-curve equation fits the relation between concentrations of chlorophyll a and total phosphorus in the Menggu-Xinjiang Lake area. With the increase of total phosphorus concentration, there was a minimum value of chlorophyll concentration in the lake areas of eastern plain, northeastern plain - mountain, Qinghai -Tibet Plateau at the beginning, and then followed by a maximum value. The minimum concentrations of total phosphorus in the three lake areas were 0.054 mg/L, 0.089 mg/L and 0.072 mg/L, respectively. This study can provide information for the control of lake eutrophication in future.
EFFECTS OF LIGHT INTENSITY ON GROWTH AND ANTIOXIDANT ENZYME ACTIVITY OF POTAMOGETON CRISPUS
Yun CAO, Shan-Shan LUO, Bin-Xiang CHEN
 Available online   , doi: 10.7541/2018.104
[Abstract](57) [FullText HTML](12) [PDF 740KB](9)
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Submerged macrophytes have an important effect on the restoration of the degraded aquatic ecosystem, and the light intensity is the main limiting factor for its growth and development. This study investigated 7 different light intensities (100%, 60%, 40%, 20%, 10%, 5% and 1% of natural light) on the morphological index (plant height, leaf number) and physiological index (chlorophyll, root activity, antioxidant enzyme activity and malondialdehyde), of Potamogeton crispus to provide the theoretical basis for the construction of eutrophic water plant community. The results showed: (1) With the increase of light intensity, the plant heights, leaf numbers and biomass growth rate increased significantly, but the growth was inhibited in high light intensity (CK group) and the low intensity (1% group). (2) The chlorophyll content of P. crispus increased first and then decreased with the increase of light intensity. The contents of Chl. a, Chl. b, Chl. a + b and Car in 1% and CK groups were lower than those of the others. (3) With the extension of the test time, the root activity increased first and then decreased with higher root activity in the group of 20%, 40% and 60%. (4) The activity of SOD, POD and CAT increased first and then decreased during the whole experiment, and in the later stage of experiment, 1% and CK groups had higher content of SOD but lower content of POD and CAT. The content of MDA of leaf increased during the experiment associated with reduced growth of P. crispus. The best growth conditions of P. crispus were the 20%—60% of natural light, and the sustained weak light stress would cause the metabolism disorders of active oxygen, cells damage, and ultimate death.
EFFECTS OF IAA ON THE PHYSIOLOGICAL AND TOXIN-PRODUCING CHARACTERISTICS OF MICROCYSTIS AERUGINOSA
Chun QING, Hui-Ling ZHANG, Yi-Qing LIN, Jun LI, Ting HU, Ji-Hai SHAO
 Available online   , doi: 10.7541/2018.102
[Abstract](47) [FullText HTML](9) [PDF 713KB](3)
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In order to investigate the effects of indole-3-acetic acid (IAA) on physiological characteristics of Microcystis aeruginosa (M. aeruginosa), the growth, photosynthetic pigment content, chlorophyll fluorescence, lipid oxidation, and microcystin biosynthesis characteristics were studied. The achieved results showed that there was no significant variation in cell density, chlorophyll content, chlorophyll fluorescence characteristics, and microcystin contents at low concentrations of IAA (e.g., 0.04 and 0.2 mg/L). However the contents of phycocyanin, allophycocyanin, and malondialdehyde (MDA) decreased in comparison with the control group. High concentrations of IAA (0.04 and 0.2 mg/L) accelerate the algal growth and increased chlorophyll content, while that inhibited the contents of phycocyanin and allophycocyanin and reduced the peroxidation of membrane lipid peroxidation as well as synthetization of intracellular algal toxins. The findings also demonstrated that low concentrations of IAA have no clear influence on the growth and photosynthesis of M. aeruginosa CHAB-6301. However, high concentrations of IAA can promote the growth and photosynthesis of this alga, which may increase the possibility of formation of Microcystis-based water blooms.
THALASSIOSIRA ALLENII VAR. STRIATA AS A NEW VARIETY IN THE GENUS THALASSIOSIRA CLEVE
Xiao-Hui GUO, Ya-Qiong GUO, Yang LI
 Available online   , doi: 10.7541/2018.101
[Abstract](37) [FullText HTML](9) [PDF 662KB](2)
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Thalassiosira Cleve is one of the most speciose marine diatom genera, that has been globally distributed. To assess the diversity of the genus Thalassiosira species, monoclonal strains were isolated and established from Chinese coast. The morphology was observed by using a light microscope (LM) and a scanning electron microscope (SEM). Sequences of the large subunit (LSU) and small subunit (SSU) of the nuclear rRNA-encoding genes (rDNA) were obtained to form a phylogenetic relationship. Among 14 established Thalassiosira allenii (T. allenii) Takano strains, morphological variation was found involving some with and without marginal ribs on the valve edge. After comparing with the original description of T. allenii, the strains without marginal ribs were assigned as a type of T. allenii, and those with marginal ribs were proposed as a new variety, T. allenii var. striata. Morphological divergence was supported by molecular data as well. On the phylogenetic trees inferred from both LSU and SSU of the nuclear rDNA, accordingly, T. allenii was clustered with var. striata, and formed as sister groups (Bayesian posterior probability (BPP) > 0.90), demonstrating their firm relationship. SSU sequences are totally identical, without any different base pair, between these two varieties. However, 11 different base pairs among 556 LSU base pair, which used for analysis, could be found, and divergence of distance among them is only 0.01.
OPTIMIZATION OF HETEROTROPHIC GROWTH CONDITIONS FOR A SCENEDESMUS STRAIN WITH HIGH CONTENT OF α-LINOLENIC ACID
Xia MENG, Hong GAO, Ren-Qiu KONG, Xu-Dong XU
 Available online   , doi: 10.7541/2018.100
[Abstract](49) [FullText HTML](9) [PDF 582KB](9)
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HSJ296, a Scenedesmus strain isolated and purified by this lab, can grow heterotrophically in the dark and contains a relatively high amount of α-linolenic acid. Comparisons of heterotrophic growth at different temperatures and concentrations of glucose, or with different nitrogen sources, indicated that the optimal culture conditions included 30℃, 4 g/L urea and 20—40 g/L glucose. As shown with fatty acid composition analyses under different conditions, the fatty acids of HSJ296 were mainly composed of hexadecanoic acid (C16:0), oleic acid (C18:1), linoleic acid (C18:2) and α-linolenic acid (α-C18:3), and the α-linolenic acid content in total lipids was between 35%—45% in independent experiments. The fermentation product of Scenedesmus HSJ296 may be used as a fish dietary supplement to supply α-linolenic acid and other nutrients.
IDENTIFICATION AND LIPID ANALYSIS OF AN OIL-PRODUCINGCHLORELLA STRAIN FROM SOIL
Hang SU, Jun-Ping LÜ, Qi LIU, Shu-Lian XIE, Jia FENG
 Available online   , doi: 10.7541/2018.099
[Abstract](41) [FullText HTML](9) [PDF 1089KB](2)
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In this study, an oil-producing alga was isolated and identified from soil at Pangquangou National Nature Reserve (Shanxi province, China), and cultivated under axenic conditions. Based on morphological observations, it was similar to the genus Chlorella. The phylogenetic analysis according to ribulose-bisphosphate carboxylase gene (rbcL) and 18S ribosomal RNA (18S rDNA) showed that it has the highest homology with Ch. vulgaris, so the strain was identified as Ch. vulgaris PQG67. It produced a lipid content within 30% after continuous culture with different light intensities, as well as more than 40% after affecting by salt (NaCl) stress. Fourier-transform infrared spectroscopy (FTIR) of the produced oil showed lipid band intensity from spectra vCO at 1634/cm, and continued to increase in Ch. vulgaris PQG67 cells during experimental test. The alga possessed high biomass and oil-producing potentials for utilization in biodiesel production.
THE EFFECTIVE FECUNDITY IN MALE ILLEX ARGENTINUS IN SOUTHWEST ATLANTIC OCEAN
Si-Peng XUAN, Xin-Jun CHEN, Dong-Ming LIN, Zi-Mo CHEN
 Available online   , doi: 10.7541/2018.098
[Abstract](48) [FullText HTML](10) [PDF 1135KB](6)
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Illex argentinus is one of the most important ommastrephid squids, due to its annual landing volume, and its key role as transient ‘biological pumps’ in southwest Atlantic ecosystem. In order to understand the fecundity characteristics of male ommastrephids, specimens of male I. argentinus, collected from December 2012 to March 2013, were used to analyze the fecundity and its relationship to reproductive development with biological statistics analysis. The results showed that the minimum biological size was 207.29 mm for mantle length (ML) and 162.55 g for body weight (BW). The effective fecundity (EF), defined as the sum of normal spermatophores in spermatophoric complex and spermatophoric sac, was in an average of (374±280.33) spermatophores. The mean value of relative effective fecundity by mantle length was (179.18±117.66) spermatophores per millimeter. The length of spermatophores (SL) varied from 6.34 to 33.14 mm, and its mean value was (21.57±4.17) mm, which was (10.22±1.82)% of mantle length. Both EF and SL significantly increased with the onset of sexual maturation (P<0.05). EF attained the maximum value at functionally maturity stage (VI), with a mean value of (811±181.34) spermatophores. SL increased up to the mating stage (VII), whereby it attained a mean value of (23.89±3.87) mm. Meanwhile, both EF and SL showed significantly linear relation with mantle length (EF=–581.92+4.56ML, R2=0.17, P<0.05;SL=5.77+0.075ML, R2=0.23, P<0.05) and body weight (EF=16.55+1.77BW, R2=0.37, P<0.05;SL=17.96+0.017BW, R2=0.17, P<0.05), indicating that individual with bigger size has larger EF and longer SL. Furthermore, the compare datasets of fitted model analysis revealed that there was no significant difference between the linear relationship of EF-ML and that of EF-BW (P>0.05). Based on multiple regression analysis, however, the influence on EF from body weight was more significant than that from mantle length (P<0.05). The compare datasets of fitted model analysis for SL showed that there was significant difference between the linear relationship of SL-ML and that of SL-BW (P<0.001), although the multiple regression analysis suggested a comparative influence on SL from both mantle length and body weight (P<0.001). These findings indicated that the EF of maleI. argentinus was a strategy of adaptability to its short lifespan by producing and accumulating spermatophores along with reproductive development and body growth, and the length of spermatophores will also increase with time.
REPRODUCTION TRAITS OF CAPTIVE ASIAN GIANT SOFTSHELL TURTLES, PELOCHELYS CANTORII
Xiao-You HONG, Xin-Ping ZHU, Chen CHEN, Jiao ZHAO, Zhao-Yang YE, Quan-Bo QIU
 Available online   , doi: 10.7541/2018.097
[Abstract](42) [FullText HTML](12) [PDF 474KB](7)
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The Asian giant softshell turtle Pelochelys cantorii (Trionychidae) is seriously endangered and has designated as the first grade of protected animal in China in 1989. In 2000, it has been listed as endangered species by the International Union for Conservation of Nature (IUCN) Red List of Threatened Species, and put on par along with the Giant panda Ailuropoda melanoleuca and the white-flag dolphin Lipotes vexillifer. This animal typically suffers from gradual reduction in its total population because of inadequate safety, food, and health protections. Sparse data, extensively referred in the literature, emphasize on the reproductive for this species, in order to enhance individuals’ knowledge about the reproduction of P. cantorii. For this purpose, we attempted to study the nesting behavior, clutch size, egg size, incubation period, as well as other reproductive characteristics of 4 adults (2♂, 2♀) under captive conditions from 2015 to 2017. Oviposition season is commenced from May to August; the egg-laying behaviors are typically occurred at the night. Clutches are made up of eggs with spherical and rigid shells without parental care behaviors. All adult females laid 5—6 clutches [mean egg number=(42.6±5.3); (32—55); n=21] during the breeding season in 2015 and 2016. In 2016, the mean egg’s mass, egg’s diameter, and mass of the newly hatchings were (16.82±1.99) g (n=476), (3.10±0.18) cm (n=476), and (13.60±0.85) g (n=202), respectively. Analysis of reproductive data obtained by 2016 based on the size and mass of the eggs, which were positively correlated (r=0.916, P<0.01), showed that there was no significant difference in average size of clutch between these two females (Using analysis of variance (ANOVA),F1,9=0.442, P=0.283). The weight of female No. 1 was more than that of female No. 2, the mass of female No. 1’s egg and newly hatchings were both more than that of female No. 2 as well. The juveniles fed by live fish seedlings could be reared to (510.30±82.77) g in the greenhouse for one year. The achieved findings affiliated with reproduction traits in P. cantorii can shield some light, and those results might be used to improve the conservation of the species.
ISOLATION, IDENTIFICATION AND ANTIBIOTIC SENSITIVITY OF CHRYSEOBACTERIUM INDOLOGENES FROM TRIONYX SINENSIS
Yi-Bin YANG, Xiao-Hui AI, Yi SONG, Hai-Peng CAO, Xian-Le YANG, Jia-Yun YAO, Jin-Yu SHEN
 Available online   , doi: 10.7541/2018.096
[Abstract](54) [FullText HTML](9) [PDF 529KB](3)
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This study conducted isolation, identification and drugs sensitivity of pathogen from sick Trionyx sinensis to provide a reference for control and prevention disease for Trionyx sinensis. The pathogenic bacteria were isolated and purified from skin, liver, kidney and spleen of Trionyx sinensis. The identification of the isolated pathogen was analyzed by biochemical reactions and 16S rRNA gene sequence. The artificial infection test was performed with intraperitoneal injection, and drugs susceptibility test was conducted by K-B method and double broth dilution method. The results showed that strain J22 was the pathogen of Trionyx sinensis, and the LD50 was 3.30×104 CFU/g. According to morphological and biochemical characteristics as well as 16S rRNA gene sequence analysis, strain J22 was identified as Chryseobacterium indologenes. Strain J22 was susceptible to neomycin, gentamicin and amoxicillin and other 9 antibiotics but resistant to florfenicol and doxycycline, etc. The chlorine dioxide, bleaching powder and potassium ferrate had great antiseptic effect on the isolated pathogeb. These results indicate that Strain J22-induced disease of Trionyx sinensis can be treat by drugs such as neomycin, gentamicin and amoxicillin for oral administration, and that chlorine dioxide, bleaching powder and potassium ferrate are suitable for external use in farming.
ESTABLISHMENT AND APPLICATION OF AN ELISA TECHNIQUE FOR DETECTING CHINESE STURGEON VITELLOGENIN
Xiao-Qian LENG, Huan YE, Hao DU, Chuang-Ju LI, Qi-Wei WEI
 Available online   , doi: 10.7541/2018.095
[Abstract](48) [FullText HTML](10) [PDF 577KB](3)
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In this study, the cDNA sequence of Vitellogenin (Vtg) gene in Chinese sturgeon Acipenser sinensis was amplified with the rapid amplification of cDNA end (RACE) method, and its theoretical molecular mass is 196 kD. The antigenic site of the encoded amino acid sequence was cloned, which was used to construct recombinant plasmid and express recombinant protein. The polyclonal antiserum was obtained by immunizing rabbits with purified recombinant protein, and its specificity to Vtg was examined by western blotting. A competitive Enzyme-linked immunosorbent assay (ELISA) was established to detect Vtg in the serum of A. sinensis using antiserum of Vtg as antibody and the purified recombinant protein as antigen. The linear regression of the standard curve was y=–0.2916x+0.6794 (R2=0.9976). The sensitivity of this method was 4.12 μg/mL, and the lowest threshold for detection was 0.3 μg/mL. The inter- and intra-assay coefficients of variation was 2.52% (n=3) and 3.42% (n=3), respectively. The test results of blood sample for the female A. sinensis at different developmental stages showed that the method could be used to monitor the gonadal development of female Chinese sturgeon.
ANALYSIS ON NUTRIENT COMPOSITIONS IN THE MUSCLES OF FOUR STRAINS OF PELODISCUS SINENSIS
Jun ZHANG, Lu CHEN, Peng YU, Qing-Yan ZHOU, Xiao-Li YANG, Wei-Shang ZHOU, Bao-Ping SHEN, Shi-Ping SU, Quan WAN, Jian-Fang GUI
 Available online   , doi: 10.7541/2018.094
[Abstract](62) [FullText HTML](15) [PDF 436KB](9)
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Ink turtle is one stain of P. sinensis in south Anhui province, which is very popular in local market. In this study, the differences of basic nutrient compositions, including amino acid and fatty acid contents in the muscles, and collagen contents in the calipash of four strains of P.sinensis were compared, which are ink turtle, wild turtle, Huai River turtle, and T. japonicus. The results showed as follows: (1) the moisture contents in the muscles of four strains were 80.40%, 79.48%, 81.57% and 79.25%, respectively. The moisture contents in ink turtles and Huai River turtles were significantly higher than those in the wild turtle and T. japonicas (P<0.05). The crude protein contents in the four strains were 17.82%, 17.60%, 15.93% and 16.40%, respectively. The crude protein content in ink turtle was the highest, which was significantly higher than that in the Huai River turtle andT. japonicus (P<0.05). The crude fat contents were 0.42%, 1.43%, 0.65% and 1.06%, respectively. The crude fat content in ink turtles was significantly lower than that in other strains (P<0.05), while the ash showed no significant difference among each other (P>0.05). (2) The ink turtle had the highest contents of TAA and DAA in muscles. (3) The UFA in the four strains ofP. sinensis was 65.19%, 56.44%, 59.20% and 54.73%, respectively, which was significantly higher in the ink turtle than in other strains (P<0.05). The content of MUFA in ink turtle was significantly lower than in wild turtle, while the content of PUFA in ink turtle was significantly higher than in other strains (P<0.05). The contents of SFA in the four strains ofP. sinensis were 35.20%, 41.85%, 39.41% and 39.98%, respectively, which was significantly lower in ink turtle than that in other strains (P<0.05). (4) The contents of DHA and EPA in ink turtle were significantly higher than those in the Huai River turtle andT. japonicus , which had no significant difference with those in wild turtle. AA in muscles of ink turtle was significantly higher than that in other stains (P<0.05). (5) There was no obvious difference in the collagen content at calipash among the four strains ofP. sinensis (P>0.05). Therefore, the ink turtle is an indigenous strain with high nutritive value and delicious flavor.
AGE STRUCTURE AND GROWTH CHARACTERISTICS OF CULTER ALBURNUS FROM DIANSHAN LAKE
Da-Wei LÜ, Yan-Feng ZHOU, You GE, Chen-He WANG, Yang YOU
 Available online   , doi: 10.7541/2018.093
[Abstract](54) [FullText HTML](12) [PDF 1106KB](4)
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Age structure and growth characteristics of Culter alburnus provide a theoretical basis for the artificial reproduction, sexual maturity age, survival rate to optimize the structure of fish stocks, the scientific use of its germplasm resources and to provide reference resource utilization and conservation of the species. To study the relationship between age structure and growth characteristics, a total of 452 Culter alburnus individuals were collected in the Dianshan Lake from May 2016 to July 2017. The results showed that the body length of Culter alburnus ranged from 15.32 to 77.91 cm, and that body weight ranged from 43 to 5567 g. There were no significant differences in body length and body weight between female and male (P>0.05). The relationship between body weight and body length was described by the linear equation: W=0.00002L2.9211 (R2=0.9143, n=452), which is consistent with uniform growth characteristics. The growth pattern of Culter alburnus was described by the von Bertalanffy equation as Lt=99.65[1–e–0.1357(t+0.6287)], Wt=11874.27[1–e–0.1357(t+0.6287)]2.9211, selection of scale identification age, measurement scale radius. The age of the fish ranged from one to six years with dominated ones with three year-old (55.71%), which revealed a growth trend of youth and miniaturization. The inflection point of the growth curve occurred at 7.2711 years with a body length of 65.54 cm and weight of 3471.79 g.
THE PHENOTYPIC BASIS, ECOLOGICAL CONSEQUENCES AND FASTING RESPONSES TO ANGLING OF JUVENILE GIBEL CARP (CARASSIUS AURATUS GIBELIO)
Ling-Qing ZENG, Meng-Lu LI, Mao-Qin XIA, Fang-Hui GU, Shi-Jian FU
 Available online   , doi: 10.7541/2018.092
[Abstract](40) [FullText HTML](9) [PDF 626KB](2)
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According to previous studies on fish angling, a correlation has been established between vulnerability to angling and metabolic traits in fish. However, it still remains unknown that whether other phenotypes (i.e., swimming performance and personality) of fish are related to the vulnerability to angling. To examine the phenotypic basis, ecological consequences and fasting responses to angling in Cyprinids fish, juvenile gibel carp (Carassius auratus gibelio) was used as experimental model in this study. This study first measured the phenotypic traits (i.e. energy metabolism, swimming performance and personality) of the experimental fish, and then the fish were angled in the buckets (named as control group) under laboratory condition. After all measurements of phenotypes and angling having been completed, all fish were fasted for 7 days and angled again at the end of fasting period (named as fasting group), following by a 14-day period of continuous feeding (named as refeeding group). There were four parallel angling samples (n=40 individuals per sample) in each angling test, which was ceased when the 20th individual was successfully angled. The angled 20 individuals were considered as angling group (i.e. higher vulnerability to angling), and the other 20 individuals were considered as un-angling group (i.e. lower vulnerability to angling). The factorial aerobic scope of the angling group was smaller than that of the un-angling group. The results showed that no differences in morphology (body weight, body length and condition factor), energy metabolism (standard metabolic rate, SMR; maximum metabolic rate, MMR; and aerobic scope, AS), swimming performance (maximum accelerated swimming capacity, Ucat; and gait transition speed, Ugt), and personality (exploration, activity and boldness) between the angling group and the un-angling group. The fish had a higher percent latency in the exploration test than that in the boldness test, which led to shorter time spent in moving and lower frequency in passing through door in the exploration test than those in the boldness test. SMR was not related to Ucat and Ugt (P>0.05), whereas MMR and AS were positively correlated with bothUcat and Ugt (P<0.05). The energy metabolic parameters were related to some personality parameters. Fasting increased the total angling time, average individual angling time and coefficient variance of the individual angling time in juvenile gibel carp. The specific growth rate (SGR) of body mass was higher in the un-angling group than that in the angling group during the fasting period, but no difference in SGR was found between the un-angling group and the angling group during the refeeding period. With the exception of MMR and AS, SMR was negatively related to SGR during both the fasting period and the refeeding period (P<0.05), suggesting that higher SMR individuals decreased their body mass faster during fasting, and grew slower during refeeding. Our study suggested that the juvenile gibel carp may not have the phenotypic basis, and would decrease their vulnerability to angling due to fasting. There was difference in the ecological consequences between two phenotypes of vulnerability to angling, but this difference disappeared after the 14-day period of refeeding, indicating that the vulnerability to angling of the juvenile gibel carp is context-dependent to some extent.
THE EFFECT OF DHA FORTIFICATION ON BODY WEIGHT LOSS AND LIPID METABOLISM OF STARVED COMMON CARP (CYPRINUS CARPIOL.) IN TWO DIETARY LIPID LEVELS
Ji-Shu ZHOU, Yan-Zi CAO, Hong JI, Hai-Bo YU
 Available online   , doi: 10.7541/2018.091
[Abstract](42) [FullText HTML](9) [PDF 406KB](3)
Abstract:
Starvation is a special period when fish cannot effectively obtain food, resulting to some problems (i.e., lack of energy required). DHA (Docosahexaenoic acid), being highly unsaturated fatty acids retained in body during starvation, is postulated to contain special energy regulation effect on the starving fish body. To explore the effect of DHA fortification on body weight loss and lipid metabolism of starved common carp (Cyprinus carpio L.) in two dietary lipid levels, a feeding trail, which followed with starvation, was conducted. Firstly, 3% DHA product was added to 6% and 12% lipid levels diets to formulate four different kinds of diet respectively, (i.e., basic diet, basic-DHA diet, high lipid diet, and high lipid-DHA diet). A total of 360 common carp (14.81±0.13) g/tail were randomly divided into four groups (triplicate per each group), fed with four diets and reared in a recycling water rearing tanks, respectively. After 74 days of feeding, some fish per each tank were sampled and eventually 6 fish per each tank were left to endure 36 days of starvation. Then, all staved fish were sampled and weight of fish and the relative indexes were determined. Result showed that the body weight loss ratio of fish in DHA groups was significantly higher than that of the non-DHA groups in the same lipid level. Besides, the hepatocyte diameter and lipid content in muscle and intestine of starved fish in DHA groups were remarkably lower than that of the non-DHA groups (P<0.05). The lipid content in hepatosomatic and serum biochemical index of fish showed that there is no significant difference. The achieved result demonstrated that DHA fortification dietary groups had significantly lower body weight, hepatocyte diameter, as well as lipid content of muscle and intestine in starved common carp, which may reflect that DHA fortification had no great advantages in fostering common carp endure starvation.
EFFECTS OF DIETARY PROTEIN LEVELS ON GROWTH PERFORMANCE, BODY COMPOSITION, LIVER BIOCHEMICAL INDICES, AND DIGESTIVE ENZYME ACTIVITIES OF JUVENILE CHINESE CRUCIAN CARP
Yong-Ming SANG, Yao YANG, Hang YIN, Yu-Ru WU, Hai-Fu YIN
 Available online   , doi: 10.7541/2018.090
[Abstract](43) [FullText HTML](8) [PDF 539KB](5)
Abstract:
The study was conducted to evaluate the effects of dietary protein levels on evolution, body composition, liver biochemical indices, as well as digestive enzymes activities of juvenile Chinese crucian carp. Six isolipidic and isoenergetic diets were formulated containing 30.18%, 33.13%, 36.16%, 39.34%, 42.32%, and 45.46%, respectively. A total of 540 juvenile Chinese crucian carp with the average initial weight of (3.10 ± 0.01) g were randomly distributed into six groups with three replicates per group, containing 30 juvenile Chinese crucian carp per replicate, which were fed for eight weeks. The results indicated that with the increase of dietary protein level, the weight gain rate (WGR), specific growth rate (SGR), feed efficiency (FE), protein efficiency ratio (PER), the whole body protein content (WBPC), the hepatic glutamic-pyruvic transaminase (GPT), and intestinal trypsin (TPS) activities of fish in 36.16% group were significantly higher than those in other groups (P<0.05). The hepatic total cholesterol (HTC) and triglyceride (TG) content of fish in 30.18% group were remarkably higher than those in other groups except for 33.13% group. Based on the quadratic regression analysis of theSGR, PER, and dietary protein levels, it was revealed that the rate of optimal dietary protein for Chinese crucian carp ranges from 35.29% to 37.07% under the experimental condition.
EFFECTS OF DIETARY SUPPLEMENTATION WITH GLUTATHIONE ON TISSUE GSH LEVEL, IMMUNITY, AND ANTIOXIDANT CAPACITY OF JUVENILE YELLOW CATFISH (PELTEOBAGURS FULVIDRACO)
Yan-Ling ZHOU, Yu-Ping SUN, Yan-Hua HUANG, Guo-Xia WANG, Bin CHEN, Wei-Min WANG, Jun-Ming CAO, Chang-Jin YI
 Available online   , doi: 10.7541/2018.089
[Abstract](60) [FullText HTML](9) [PDF 470KB](2)
Abstract:
To assess the effects of dietary supplementation with glutathione (GSH) on tissue GSH levels, immunity, and antioxidant capacity of juvenile yellow catfish (Pelteobagrus fulvidraco), a total of 800 yellow catfish with a mean body weight of (1.32±0.01) g were considered and randomly divided into 5 groups with 4 replicates per each group and 40 fish per each replicate as well. During 56d, the fish were fed by a basal diet and four test diets, which supplemented with 100, 300, 500, and 700 mg/kg of reduced GSH, respectively. The results indicated that, except for 100 mg/kg group, supplementation with GSH significantly increased the GSH content in liver and serum of yellow catfish (P<0.05). When GSH level was 300 mg/kg or higher, GSH content in liver and serum was in a stable state. With the increase of glutathione level, the immunity and antioxidant indices were initially increased, and then decreased. The lysozyme and alkaline phosphatase activities in 300 and 500 mg/kg groups, immunoglobulin M (IgM) and complement component 4 (C4) contents in 300 mg/kg group, and acid phosphatase activity in 500 mg/kg group markedly increased, compared with the control group (P<0.05). There were no significant differences (P>0.05) in complement component 3 (C3) and nitric oxide contents among all groups. Compared with the control and 700 mg/kg groups, the liver superoxide dismutase, catalase and glutathione peroxidase activities, total antioxidant capacity and the serum superoxide dismutase, as well as glutathione peroxidase activities in 300 mg/kg group were significantly higher (P<0.05), while the serum malondialdehyde content in that group was considerably lower (P<0.05). Under ammonia-nitrogen stress for 96h, compared with the control group, the liver and serum superoxide dismutase, catalase, and glutathione peroxidase activities in 300 mg/kg group were significantly greater (P<0.05), however, the serum malondialdehyde content in the mentioned group was significantly lower (P<0.05). In conclusion, dietary supplementation with GSH could enhance the tissue GSH level, immunity and antioxidant capacity of juvenile yellow catfish, and the effective range of this enhancement was 300 to 500 mg/kg as well.
THE REPLACEMENT OF FISH MEAL AND SOYBEAN MEAL TO FERMENTED SOYBEAN MEAL AND ITS EFFECTS ON THE GROWTH PERFORMANCE, SERUM BIOCHEMICAL INDICES, AND IMMUNE GENE EXPRESSION IN GIANT RIVER PRAWN, MACROBRACHIUM ROSENBERGII
Jing-Feng YANG, Xue-Ming HUA, Zi-Hao GUO, Tao LIU, Chun KONG, Yue FENG, Gang WANG
 Available online   , doi: 10.7541/2018.088
[Abstract](47) [FullText HTML](9) [PDF 589KB](7)
Abstract:
An experiment was conducted to investigate the effects of replacing fish meal (FM) and soybean meal (SBM) by fermented soybean meal (FSBM) on growth performance, serum biochemical indices, and immune genes in giant river prawn, Macrobrachium rosenbergii, and to find a suitable proportion of FSBM to replace FM in diets. Five isonitrogenous and isoenergetic diets were formulated for giant river prawn, among which a basal diet (T0) containing 30% FM and 18% SBM was treated as the control group, and the FM and SBM (2﹕1) in the basal diets replaced by 2% (T2), 5% (T5), 8% (T8) and 15% (T15) of FSBM respectively were treated as experimental diets. Giant river prawns with initial average body weight of (0.17±0.02) g were cultured for 64 days in indoor net cages at a cement tank. Results showed that with increasing FSBM supplemental level, the growth rate and specific growth rate increased firstly and then decreased, and the highest value was found in group T8. Serum superoxide dismutase activity and malondialdehyde content firstly increased and then decreased. MDA of all experimental groups was higher than that of the control group. Serum aspartate transaminase, alanine aminotransferase activity and total protein content had decreasing trend followed by an increasing trend, and the values in all experimental groups were lower than the control group. Gills Toll receptor mRNA and NF-κB mRNA expression levels and hepatopancreas heat shock protein mRNA relative expression levels in group T15 were the highest among all groups. All these results indicated that the replacement of FM and SBM with FSBM would affect growth, antioxidant ability and immunity, and the optimum amount of FSBM was 8% in the diets in this experiment.
EFFECTS OF DIETARY PROTEIN LEVELS ON THE GROWTH PERFORMANCE, ACTIVITY OF NON-SPECIFIC IMMUNITY AND PROTEIN SYNTHESIS CAPACITY OF RHYNCHOCYPRIS LAGOWSKII DYBOWSKI
Lan YANG, Li-Fang WU, Zi-Hui QU, Kai ZHOU, Jing DUAN, Jing-Yao WANG, Xiu-Jie ZU
 Available online   , doi: 10.7541/2018.087
[Abstract](68) [FullText HTML](9) [PDF 698KB](11)
Abstract:
To evaluate the effects of different dietary protein levels on the growth performance, non-specific immunity and protein metabolism enzymes of Rhynchocypris lagowskii Dybowski, diets with 24.98%, 30.02%, 34.99%, 40.01% and 44.98% protein formulated with fish meal, soybean meal, rapeseed meal and cottonseed meal were fed to Rhynchocypris lagowskii Dybowski (6.98±0.01) g for 8 weeks in this study. After the feeding trial, a national standard method was used for determining muscle nutrients, and kits were used to measure serum alkaline acid enzyme (AKP), acid phosphatase (ACP), superoxide dismutase (SOD) and lysozyme (LZM) activity. The results showed that, with the increase in protein levels, final mean body weight, weight gain rate and specific growth rate firstly increased with increasing protein levels, and then decreased. When the protein levels were 34.99% and 40.01%, the final mean body weight, weight gain rate and specific growth rate were significantly higher than that in the groups with 24.98% level (P<0.05). With increasing protein levels in diets, feed and protein efficiencies ofRhynchocypris lagowskii Dybowski firstly increased, and then decreased. Feed and protein efficiencies in 40.01% groups were significantly higher than those in 24.98% groups (P<0.05), but there was no significant difference among groups with 30.02%, 34.99%, and 40.01% protein levels (P>0.05). According to the polynomial regression analysis, the specific growth rate ofRhynchocypris lagowskii Dybowski was the highest when protein level was at 35.89%, and the feed efficiency was the highest when the protein level was at 36.11%. The content of crude protein in muscles increased first, and then decreased with increasing protein levels in diets. Among them, the crude protein content in muscles of 40.01% group was significantly higher than that in the other four groups (P<0.05). However, the crude fat content in muscles decreased first, and then increased with increasing protein levels. The crude fat content in 40.01% group is significantly lower than that in the 24.98%, 30.02%, and 34.99% groups (P<0.05), but the 44.98% group showed no significant difference from other groups (P>0.05). The activities of alkaline phosphatase (AKP), acid phosphatase (ACP), superoxide dismutase (SOD) and lysozyme (LZM) in hepatopancreas increased first, and then decreased with increasing protein levels. The activities of AKP, ACP, SOD, and LZM in the 40.01% group were significantly higher than those in the other four groups (P<0.05). The RNA and RNA/DNA ratios in muscles increased firstly, and then decreased with increasing protein levels. Among them, the RNA content and RNA/DNA in 40.01% group were significantly higher than those in the other four groups (P<0.05). According to the polynomial regression analysis, the RNA content in white muscle was highest when the protein level was 36.10%, and the RNA/DNA ratio was highest in white muscle when the protein level was 35.91%. Therefore, in the formulated diets, the most suitable protein level is 34.99% to 40.01%.
EFFECTS OF DIETARY SUPPLEMENTATION WITH HAEMATOCOCCUS PLUVIALIS POWDER ON OVARIAN DEVELOPMENT, COLORATION, ANTIOXIDANT CAPACITY AND BIOCHEMICAL COMPOSITION OF ADULT FEMALE SWIMMING CRAB, PORTUNUSTRITUBERCULATUS
Ren-Fu WU, Xiao-Wen LONG, Wen-Jie HOU, Gui-Ping PAN, Jian-Guo LIU, Yong-Xu CHENG, Xu-Gan WU
 Available online   , doi: 10.7541/2018.086
[Abstract](55) [FullText HTML](8) [PDF 658KB](7)
Abstract:
Swimming crab (Portunus trituberculatus) is an important mariculture crab, and the light color of the pond-reared crabs may be related to low level of dietary astaxanthin. This study was designed to investigate the effects of dietary astaxanthin on ovarian development, coloration, antioxidant capacity, immune performance and biochemical composition of female P. trituberculatus. Haematococcus pluvialis powder was used as natural astaxanthin source to formulate four experimental diets with different astaxanthin levels (0, 26.60, 41.62, and 81.37 mg/kg, defined as Diet 1#—4#). The four experimental diets were fed to female P. trituberculatus for 45 days during the ovarian development; therefore, there were four treatments in this study. The results showed as follows: (1) Dietary astaxanthin had no significant effects on gonadosomatic index (GSI) and hepatosomatic index (HSI). (2) Total carotenoids content, redness (a*) and yellowness (b*) values of hepatopancreas, ovaries and carapace increased significantly with increasing dietary astaxanthin, while the lightness (L*) values of these tissues showed a decreasing trend (P<0.05). (3) For antioxidant indices, the activities of super oxide dismutase (SOD) and peroxidase (POD) in treatments (Diet 2#—4#) with supplementation of astaxanthin were significantly lower than those in Diet 1# treatment without supplementation of astaxanthin. The activities of total antioxidant capacity (T-AOC) and glutathione peroxidase (GSH-Px) in hemolymph as well as the SOD, T-AOC in hepatopancreas increased significantly with elevated dietary astaxanthin, while a decreasing trend was found for the malondialdehyde (MDA) content in the hemolymph and hepatopancreas. (4) For the immune indices, Diet 4# treatment had the highest activity of acid phosphatase (ACP) in hemolymph and hepatopancreas, while Diet 1# treatment had the significantly higher hemolymph hemocyanin (Hc) content than other treatments (P<0.05). (5) As for the biochemical composition, the increasing trend was found on the contents of total lipids and total carbohydrate in the muscle and hepatopancreas as well as ovarian lipids with increasing dietary astaxanthin, while Diet 2# treatment had the highest total carbohydrate content in the ovaries among the four treatments (P<0.05). In conclusion, dietary supplementation of astaxanthin had no significant effect on ovarian development ofP. trituberculatus, but could improve the coloration, antioxidant capacity and the contents of total lipids and total carbohydrate in edible tissues. These results suggest the appropriate level of astaxanthin in fattening diet was approximately 50 mg/kg for adult female P. trituberculatus.
THE CLONING AND BIOLOGICAL CHARACTERISTICS OF MYD88 IN LAMPREY AND THE EXPRESSION PATTERN OF ITS DOWNSTREAM PROTEINS
Chun-Hui ZHAO, Pei-Zhe ZHAO, Xin LIU, Qing-Wei LI
 Available online   , doi: 10.7541/2018.085
[Abstract](45) [FullText HTML](8) [PDF 543KB](6)
Abstract:
Myeloid differentiation factor 88 (MYD88) is a key adapter protein in Toll-like receptor (TLR) signaling, which plays significant role on the innate and adaptive immunity. In this study, a myd88 gene from lamprey (Lampetra japonica) was obtained. The ORF of myd88 is 852 bp in length, encoding a polypeptide of 283 amino acids. The theoretical molecular weight of lamprey Myd88 is 32.432 kD with a theoretical isoelectric point of 6.25. Lamprey Myd88 was predicted to have no signal peptide. Multiple protein sequence alignment revealed highly conserved death domain in N-terminal, and Box1, Box2, and Box3 in the C-terminal TIR domain, which indicated that lamprey Myd88 is homologous with the MYD88 in other species. Quantitative real-time PCR analysis showed lamprey myd88 gene was extensively expressed in all detected tissues. The highest expression level was observed in gill, which was followed by marrow and kidney. Under in vivo stimulation by lipopolysaccharide (LPS), the expression of myd88 significantly increased in leukocytes, followed by gill, which implicated its role in the defense of L. Japonica against bacteria. Furthermore, increasing expressions of downstream proteins in Myd88-dependent TLR signaling pathway, including Irak1, Traf6, Ikkβ, and Nfkb, were detected in the tested tissues stimulated with LPS. These results suggested that a conserved Myd88-dependent TLR signaling pathway was found in the lamprey, which play a fundamental role on the exploration of the origin and evolution of the signaling pathway in immune response in future.
MICROSATELLITE ANALYSIS OF GENETIC VARIATION IN THE FAST GROWTH FAMILIES OF THE 2nd GENERATION OF TEGILLARCA GRANOSAAND CORRELATION WITH GROWTH TRAITS
Shuang-Shuang TENG, Jun FANG, Yan-Qing SHAO, Xing-Guan LIN, Xue-Liang CHAI, Guo-Qiang XIAO
 Available online   , doi: 10.7541/2018.084
[Abstract](41) [FullText HTML](15) [PDF 609KB](2)
Abstract:
In this study, microsatellite markers were used to assess the genetic structure and genetic diversity of the second generation of Tegillarca granosa. In addition, the correlation of these markers with growth-related traits was assessed to explore their potential use for marker-assisted breeding. Genome-wide identification and structural analysis herein led to detect 59 alleles through 18 pairs of primers. Among these, the average values of alleles (Na) in the families of F19, F21, and F22 were 2.500, 2.722, and 2.722, respectively. The average observed heterozygosity (Ho) was 0.446, 0.510, and 0.628; the average expected heterozygosity (He) was 0.394, 0.433, and 0.464, respectively. In addition, the corresponding values of polymorphism information content (PIC) were 0.346, 0.379, and 0.403, respectively. Correlation analyses of these polymorphic markers and growth traits revealed significant association of three loci with shell height, shell length, shell width, and total weight. Specifically, genotype BB of Teg-30 in F19 was remarkably associated with shell height and total weight, while genotype BB of Teg-03 and genotype BC of Teg-20 in F21 showed a significant correlation with the shell height, shell length, shell width, and total weight. As a result, the growth-related markers, identified in the present study, may provide precious genetic information and novel insights for molecular marker-assisted breeding of T. granosa.
CLONE AND EXPRESSION ANALYSIS OF TRANSCRIPTION COFACTOR LBH-B in GIBEL CARP
Wen-Hua LI, Yang WANG, Zhi LI, Jian-Fang GUI, Li ZHOU
 Available online   , doi: 10.7541/2018.083
[Abstract](49) [FullText HTML](8) [PDF 619KB](5)
Abstract:
LBH (limb-bud and heart) is a novel high-conserved transcription cofactor in vertebrates involved in embryonic development and pathogenesis of some human disease. We screened the lbh-b gene that was abundantly expressed in the pituitary of gibel carp, and cloned the lbh-b (Cglbh-b) cDNA sequence of lbh gene family from gibel carp cDNA library by RACE-PCR. The full-length of Cglbh-b cDNA was 1526 bp with a 549 bp long open reading frame (ORF) coding a 182 amino acid protein. Bioinformatics analysis showed that CgLbh-b protein shared high homology (>68%) with other vertebrate LBH and is one of intrinsic disordered proteins.Cglbh-b was abundantly expressed in pituitary, telencephalon, ovary and eye of adult gibel carp. The expression of Cglbh-b in female pituitaries was 4.66 times higher than that in male pituitaries, and Cglbh-b in pituitary was up-regulated during the process of oocyte maturation. During the early embryonic development, maternal mRNA of Cglbh-b was detected in embryos from 4-cells stage to gastrula stage, and its transcripts were synthesized at bud stage. The results of whole mount in situ hybridization showed that Cglbh-b was distributed on the brain and eyes of embryos from 2 days post fertilization (dpf) to 3dpf. These results suggest that Cglbh-b may play important roles in brain and retina development, and reproductive regulation in oocyte maturation of gibel carp.
GENE CLONING OF GPR43 AND ITS DISTRIBUTION IN THE TISSUE OF BLUNT SNOUT BREAM AND THE EFFECT OF BERBERINE ON ITS MRNA EXPRESSION
Dan-Hong CHEN, Qing-Qing CHEN, Wen-Bin LIU, Ding-Dong ZHANG, Wei-Na XU
 Available online   , doi: 10.7541/2018.082
[Abstract](57) [FullText HTML](8) [PDF 740KB](5)
Abstract:
This study was conducted to explore the effects of berberine on the mRNA expression of G protein coupled receptors 43 (GPR43) and the lipid metabolism performance of blunt snout bream fed with high-fat diet. GPR43 in blunt snout bream was cloned and sequenced with a RACE assays. The effects of berberine on GPR43 mRNA expression in tissues were also investigated. The obtained full-length nucleotide sequence covers 2026 bp, which has an open reading frame of 981 bp, and translates a total of 326 amino acids. Moreover, GPR43 mRNAs were detected in different tissues of blunt snout bream. In blunt snout bream, the expression of GPR43 in intestine, hepatopancreas, gill and white muscle were the highest. 320 fish (80.00±0.90) g were randomly divided into four trial groups (each has four replicates) and fed with 4 diets (normal diet, normal diet with 50 mg/kg berberine, high-fat diet, and high-fat diet with 50 mg/kg berberine, among which normal diet has 5% lipid and high-fat diet has 10% lipid), respectively. The expression of GPR43 mRNA in the high-fat diet group decreased compared with control group, while it significantly increased (P<0.05) after adding in berberine. Compared with the control group, the contents of free fatty acids (FFA), triglycerides (TG), cholesterol (CHO), and mRNA expressions of Cytokinins protein kinase (p38), and Peroxidase value-added factor (PPARγ) all increased (P<0.05) in the high-fat group, but a dramatic decrease (P<0.05) happened after adding in berberine. The same result was found on the composition of Polyunsaturated fatty acids (PUFA) in hepatopancreas and muscle tissues. A completely inverse variation was presented on the mRNA expressions of Carnitine palm acyltransferaseⅠ (CPTⅠ), Peroxidase value-added factor (PPARα&β) and AMP dependent protein kinase (AMPK), as well as on the compositions of Saturated fatty acids (SFA) and Monounsaturated fatty acids (MUFA) in both tissues. Nevertheless, the difference between berberine supplemental normal diet group and control group was out of regular pattern. so further work is still needed to explore the role of berberine in normal circumstances. Overall, the results showed the supplement of berberine could slow down lipid deposition in hepatopancreas, and restore lipid homeostasis which has been damaged by continuous high-fat diet feeding.
GENETIC DIVERSITY AND REPRODUCTIVE CHARACTERISTICS OF CORBICULA FLUMINEA FROM THE LAODAO RIVER IN LIUYANG CITY
Jian-Ping WANG, De-Liang LI, Cong ZENG, Peng-Fei ZHU, Jiang-Fan ZHAN
 Available online  
[Abstract](51) [FullText HTML](14) [PDF 717KB](9)
Abstract:
This study attempted to theoretically support and comprehensively perceive genetic diversity and reproduction of Corbicula fluminea from the Laodao River in Liuyang city was investigated. Partial sequences of the cytochrome oxidase c subunit 1 (COⅠ) of mitochondrial genes were utilized to evaluate the genetic diversity, and reproductive characteristics were observed by gonadal histology and sperm morphology using light microscopy. Among examined 40 individuals, only 4 haplotypes and 17 variable sites were detected in the partial sequences of the COⅠ, with an average haplotype diversity, nucleotide diversity, and number of variable sites equal to 0.664±0.042, 0.014±0.006, and 8.595, respectively. Hermaphrodite was a predominant reproduction in the population studied, and the hybrid hermaphrodite-sex ratio was 6﹕3﹕1. Furthermore, there were two types of reproductive follicles in hermaphroditic individuals, including sperm and egg cells in the same and in different follicular cells. The observation of sperm morphology revealed that biflagellate spermatozoa were discovered in both hermaphrodite (n=23) and male (n=8) cases. The results also showed that Corbicula fluminea samples from the Laodao River of Liuyang city contained a relatively low genetic diversity, but with multiple reproduction modes, and the predominant mode was androgenesis. This study attempted to present significant biological reproduction information required for Corbicula fluminea protection in China.
ULTRASTRUCTURE of THE EMBRYONIC DEVELOPMENT AND METAMORPHOSIS OF RHOPILEMA ECULENTUM
Chun-Sheng LIU, Yi WAN, Fei GAO, Si-Qing CHEN, Ai-Ming WANG, Zhi-Feng GU
 Available online  
[Abstract](22) [FullText HTML](9) [PDF 1107KB](7)
Abstract:
This study investigated cell ultrastructure of Rhopilema esculentum during embryogesis and metamorphosis into a polyp using scanning and transmission electron microscopy combined with protargol staining method. Our study showed that (1) Cell cleavage was almost equal from fertilized egg to gastrula. All divided cells were similar in morphology, and cell junctions were observed among adjacent cells; (2) Vacuoles in cell were firstly observed at ectoderm of early planula, and became bigger as developing to 4-tentacle polyp, while the vacuoles of entoderm were firstly found at 4-tentacle polyp stage. The suspected apoptotic body were observed at scyphostoma and polyp stages; (3) Immature cnidocytes were firstly observed in ectodermal area closed to mesoglea ectoderm at early planula stage, moved to outside, located at outermost layer, and matured at polyp stage; (4) Cilium was observed at early planula stage, gradually degraded, and completely disappeared at polyp stage; (5) Lots of yolk bodies with dark stain were observed at all developmental stages, and the phagocytosis of ectodermal cell of planula was firstly observed. These phenomena suggest that planulae of R. esculentum were both planktotrophic and lecithotrophic, which would provide the scientific basis for illuminating the developmental pattern of embryonic and metamorphosis of cnidarians.
EFFECT OF ISOLEUCINE ON mTOR SIGNALING PATHWAY AND NITROGEN METABOLISM OF CHINESE PERCH (SINIPERCA CHUATSI)
Kang HUANG, Xu-Fang LIANG, Shan HE, Jiao LI, Shu-Lin TANG, Zhen ZHANG
 Available online  
[Abstract](21) [FullText HTML](9) [PDF 659KB](2)
Abstract:
In the present study, the effects of isoleucine on the mammalian target of rapamycin signaling (mTOR) pathway and nitrogen metabolism were investigated through Intraventricular (ICV) administration in Chinese perch (Siniperca chuatsi). In one hand, the ammonia excretion significantly increased, and mRNA levels of nitrogen metabolism genes involving glutamate dehydrogenase (GDH), glutamic oxaloacetic transaminase (GOT), and adenosine monophosphate deaminase (AMPD) remarkably increased after ICV injection of isoleucine as well (P<0.05). On the other hand, the results showed that blood glucose levels markedly decreased at 0.5 h post-injection. The expression of liver ribosomal protein S6 was also notably enhanced, demonstrating that mTOR signaling pathway was activated. The results also indicated that isoleucine could activate the mTOR signaling pathway, mediate amino acid metabolism, and enhance ammonia-N excretion via nitrogen metabolism genes in Chinese perch.
EFFECTS OF FERMENTED SOYBEAN MEAL ON THE GROWTH AND INTESTINAL HISTOLOGY AND MICROBIOTA OF LARGE YELLOW CROAKER LARIMICHTHYS CROCEA
Jiao-Jiao HE, Ping WANG, Jian FENG, Yu-Dong LOU
 Available online  
[Abstract](29) [FullText HTML](8) [PDF 1229KB](5)
Abstract:
The fermented soybean meal (FSM) was used as a replacement of fish meal (FM) in a practical diet with 45% protein and 10% lipid, to investigate its effects on the growth, intestinal histology, and the structure of intestinal dominant bacteria in juvenile large yellow croaker. FM was replaced by 0, 15%, 30%, 45%, 60% and 75% of FSM (FSM0, FSM15, FSM30, FSM45, FSM60 and FSM75), respectively. The fish of 10.49 ± 0.03 g was hand-fed daily to satiation twice for 56 days. Results showed that the survival rate (SR) was not significantly affected with all the diets (P>0.05), but in groups FSM60 and FSM75, theSR had a downward trend. In comparison with FM0, the weight gain rate (WGR) and specific growth rate (SGR) in FSM60 and FSM 75 decreased significantly (P<0.05), while the feed conversion rate (FCR) increased significantly (P<0.05). Feed intake (FI) increased significantly in FSM60 and FSM75 (P<0.05), compared with that in FSM0, FSM15, FSM30 and FSM45. The observed intestinal histology results showed that Mucosa thickness, Mucosafold height, Laminapropria width and Gobletcell quantity were not significantly affected in all the diets (P>0.05). Sequencing results of intestinal contents from Illumina-MiSeq high-throughput sequencing technology showed that the index of Chao1, Shannon, Simpson and Good coverage in FSM0 (TC the control), FSM45 (TB the best group in growth) and FSM75 (TW the worst group in growth) were not significantly affected (P>0.05).Firmicutes group totally dominated the bacterial communities in juvenile large yellow croaker. Paenibacillus genus was the most dominant bacteria in microbial ecosystem. Bacterial composition had certain effect on the growth of large yellow croaker: compared with TW group, the species richness of Paenibacillus and Alkaliphilus increased significantly in TB group and TC group (P<0.05). However, compared with TW group, the species richness ofEnhydrobacter in TB and Paracoccus in TC decreased significantly (P<0.05). These results suggested that the optimal dietary protein level was about 45% without negtive effects on the growth, intestinal histology, and the structure of intestinal dominant bacteria in juvenile large yellow croaker.
RESPONSE OF STREAM FISH ASSEMBLAGES TO THE SIZE AND FUNCTION OF LOW-HEAD DAMS: A CASE STUDY IN THE HEADWATER STREAMS OF THE WANNAN MOUNTAINS
Qiang LI, Dong ZHANG, Feng-Ying WAN, Yu-Ru LI, Ling CHU, Yun-Zhi YAN
 Available online  
[Abstract](30) [FullText HTML](8) [PDF 505KB](6)
Abstract:
It has been discovered that low-head dam could impact local habitat conditions, and alter fish assemblages in streams. However, how fish assemblages respond to the size and function of low-head dams is still not clear. Based on the data collected from 15 impounding segments upstream and 15 plunging areas downstream of low-head dams in the Wannan Mountains at July and November 2016, respectively, we examined how habitat factors and fish assemblages varied seasonally, and the differences in their response to dam size and function. Furtherly, after classifying the collected samples into two types (indigenous species and native-invasive species), we determined whether the indigenous fishes and native-invasive fishes were different in their responses to dam disturbance. A total of 24 fish species representing four orders and ten families were collected, including 11 indigenous species and 13 native-invasive species. Although fish assemblages in the plunging areas did not vary seasonally, the assemblages in the impounding areas was significantly different between July and November, where fishes were more abundant in November than in July. Only the assemblage structure of native-invasive fishes in the plunging areas was influenced by dam size. However, both native fish assemblages in the impounding areas and native-invasive fish assemblages in the plunging areas showed difference in their response to the between-dam-function, suggesting that dam function was more important in impacting fish assemblages than dam size. Compared to those associated with the impounding dams, the irrigating dams had less Zacco platypus but more Cobitis sinensis and Ctenogobius spp. in the impounding areas, and more Rhodeus ocellatus and Misgurnus anguillicaudatus in the plunging areas. In addition, the response of the whole fish assemblages (including both indigenous and native-invasive species) to dam disturbance was similar to the indigenous fish assemblages, instead of the native-invasive fish assemblages. Our results suggest that, in this study area, dam function was more important in influencing local fish assemblages than dam size. Due to their potential differences in response to dam disturbance for the indigenous species and native-invasive species, it is necessary for us to distinguish these two types of species when we assess how anthropogenic activities affect fish diversity in streams.
A REVIEW OF INTERNATIONAL FISHWAYS ADAPTIVE MANAGEMENT SYSTEMS AND MANAGEMENT PROSPECTS FOR CHINA
Wei-Wei LI, Jiang-Hui BAO, Chao-Shuo ZHANG, Li-Wei WANG, Ye QIAO, Jing-Ya WEN, Hong-Tao LI, Tang-Lin ZHANG, Jia-Shou LIU, Zhong-Jie LI, Ming DUAN
 Available online  
[Abstract](28) [FullText HTML](8) [PDF 417KB](4)
Abstract:
Many efforts have been devoted to developing, constructing and refining fishways or fish passage facilities to facilitate the target fish species to pass barriers in fluvial systems. Once dominated by an engineering-focused approach, fishways science nowadays involves a wide range of disciplines from fish behaviour to socioeconomics, and to complex modeling of passage prioritization options in river networks. Designing efficient fishways, with minimal passage delay and post-passage impacts, requires adaptive management and continued innovations. After the completion of fishways construction, corresponding management systems usually need to be improved as well. In the current study, we first reviewed international fishways adaptive management systems (e.g., aspects of development, construction, and management) from 1991 to 2017 reporting on " fishway”, " fish passage”, " fish” and " dam”. The functions of fishways gradually transferred from conservation of economic stocks to overall biodiversity conservation, from facilitating upwards to both upwards and downwards passages, from developing a single project to considering the entire river basin, from fish passing function to habitats restorations as well, and from pure engineering construction to considering adaptive management. Finally, we discussed adaptive management systems and management prospects of fishways for China.
FISH ASSEMBLAGE RESPONSES TO DIFFERENT SECONDARY CHANNEL DESIGNS IN THE LOWER MISSISSIPPI RIVER, U.S.A.: A TEMPLATE FOR RIVER RESTORATION
Joseph E. KAISER, Shannon C.F. SMITH, Jr. Harold L. SCHRAMM, Michael A. EGGLETON
 Available online  
[Abstract](27) [FullText HTML](8) [PDF 640KB](4)
Abstract:
The lower Mississippi River (LMR) has been heavily modified for multiple human purposes such as navigation, flood control, and bank stabilization. However, the LMR simultaneously supports a diverse fish fauna that includes recreational and commercial fisheries. Due to river training and diversion structures constructed during the past 80 years, the historic characteristics of the LMR have been drastically altered and have likely influenced fishes and fisheries in the system. One common restoration measure used throughout the LMR has been to " notch” wing-dike structures that close secondary (side) river channels. Dike notching allows year-round flows through secondary channels, which enhances habitat diversity and promotes biological productivity at the ecosystem scale. Although notching is presumed good for LMR fishes and other biota, few studies have examined its effects on fish assemblages. In this study, fish assemblages were sampled at seven LMR secondary channels spanning from river kilometer (rkm) 628 (Louisiana-Mississippi, U.S.A.) upstream to rkm 1504 (Missouri-Kentucky, U.S.A.). Four secondary channels were termed " permanent” (i.e., with notched dikes) while three secondary channels were termed " temporary” (i.e., without notched dikes). Fishes were sampled by boat-mounted electrofishing conducted during falling and low stages from 1995—1997. Fish assemblages differed between permanent and temporary secondary channels, and varied somewhat between falling and low stages. Gizzard shad (Dorosoma cepedianum), threadfin shad (D. petenense), and white bass (Morone chrysops) demonstrated consistent preferences for low-current conditions associated with temporary secondary channels. Conversely, blue catfish (Ictalurus furcatus), flathead catfish (Pylodictis olivaris), and freshwater drum (Aplodinotus grunniens) were more associated with permanent secondary channels. Future restoration strategies in the LMR should consider dike notching and resultant maintenance of permanent secondary channels in selected river reaches. However, temporary secondary channels also contain unique fish species, and also appear to be important sites of riverine primary production. Restoration strategies should consider a balance of both secondary channel types, which should support the greatest biodiversity for the LMR ecosystem.
ANESTHETIC EFFECTS OF MS-222 ON SCHIZOTHORAX O’CONNORI LLORD IN TWO SIZE RANGES
Yan-Chao LIU, Hai-Ping LIU, Shu-Yun LIU, Xi-Feng LI, Xiao-Guang SONG
 Available online  
[Abstract](24) [FullText HTML](9) [PDF 686KB](1)
Abstract:
This study examined anesthetic behaviors of Schizothorax o’connori Llord in two size ranges as induced by MS-222, to acquire useful information on anesthesia of said highland fish species in out-of-water activities (surgery, labeling, measurement, etc.). Test results indicated that for both large (25.0±1.5) cm and small-sized (14.8±2.3) cm specimens, respiration rates showed no significant change in anesthetic stages I to III, and only begun to significantly decline after entering stage IV. The effective concentrations of MS-222 were 150—180 mg/L and 150 mg/L for two sizes, respectively. The 150—180 mg/L caused large specimens to enter anesthetic stage IV within 5min, and recovered within 5min; the 150 mg/L caused small specimens to enter stage IV within 5min, and recovered within 7min; both doses ensured a 100% survival rate after immersion for 20min. For large specimens anesthetized at 180 mg/L for 5min and exposed to air for 0—15min, there was no significant difference in recovery times (P>0.05); for small specimens anesthetized at 150 mg/L for 5min and exposed to air for 0—15min, there were significant differences in recovery (P<0.05).
LENGTH-WEIGHT RELATIONSHIPS OF 18 FISH SPECIES FROM LAKE KUILEI, YANGTZE RIVER BASIN, CHINA
X. Cai, W. Li, Z. Li, T. Zhang, S. Ye, H. Fan, J. Liu
 Available online  
[Abstract](42) [FullText HTML](8) [PDF 293KB](2)
Abstract:
This study is based on monthly sampling (from April 2013 to April 2015) of 18 fish species representing three families. Samples were obtained using benthic fyke-nets, block nets, and multi-mesh gillnets in Lake Kuilei, a shallow lake of the Yangtze River basin, China (N 31º24', E 120º51'). Using ordinary least-squares linear regression, length-weight relationships of these fish species were estimated, with equations for 3 species reported for the first time. The maximum total length records of the 15 species exceeded those reported in FishBase. This paper provides new LWR equations for these species to be included in FishBase.
ECOLOGICAL RISK ASSESSMENT OF MICROCYSTIS BLOOMS IN ERHAI LAKE
Yi LIU, Shao YANG, Xiao-Feng LIU
 Available online  
[Abstract](29) [FullText HTML](8) [PDF 491KB](4)
Abstract:
The ecological risk of a Microcystis bloom was evaluated on the basis of the ecotoxicological risk assessment method. The effects of Microcystis bloom on benthic fauna, zooplankton, fish, and submerged macrophytes were assessed by using exposures of acute and chronic and findings reported by previous studiesThe algal cell densities were calculated using toxicity percentile rank (TPR) method, and aquatic ecological risk assessment for Microcystis bloom was performed using risk quotient approach. The criteria required for acute and chronic ecological risk assessment of Microcystis bloom were defined based on the algal cell density, algal bloom coverage in lake, as well as duration of algae blooms. The ecological risk of an acute was classified as low-, moderate-, and high- levels. A Microcystis bloom with cell density less than 3.4×106 cells/L was defined as low risk-level, with cell density at the range of 3.4×106—3.4×107 cells/L was denoted as moderate-risk level, and with cell density higher than 3.4×107 cells/L was defined as high-risk level. Similarly, an ecological risk of a chronic toxicity was also classified as low-, moderate-, and high-levels of risk. The Microcystis bloom with cell density less than 1.1×106 cells/L was classified as low risk, cell density between 1.1×106—1.1×107 cells/L was classified as moderate risk, the cell density greater than 1.1×107 cells/L was classified as high risk. In practical applications, cell density, algal bloom coverage, and duration of algae blooms are essential factors to formulate a robust ecological risk assessment strategy for Microcystis bloom in Erhai Lake. In presence of moderate-risk level, early warning of Microcystis bloom should be alarmed, and improvement of water quality or implementation of biological control should be carried out. Under the high risk level, emergent removal or control of algal bloom should be undertaken as well.
EFFECTS OF ARTIFICIAL SUBSTRATES ON THE CARP GROWTH PERFORMANCE AND WATER QUALITY IN PONDS EQUIPPED BY USING A POLY-CULTURE SYSTEM
Ji Shu ZHOU, Qing An LIAN, Er Meng YU, Jun XIE, Hong JI, Hai Bo YU
 Available online  
[Abstract](23) [FullText HTML](8) [PDF 598KB](2)
Abstract:
To investigate the influence of artificial substrates (AS) on the growth performance of fish in pond poly-culture systems, a feeding experiment was conducted in six individual earthen ponds, which were divided into two groups. The ponds with AS in the water were labeled as the " AS group” and that without the use of AS were labeled as the " control group” (three ponds per group). A total of 3867 common carp [Cyprinus carpio, (310±11) g], being artificial-feeds feeding fish, and 370 bighead carp (Aristichthys nobilis) and silver carp [Hypophthalmichthys molitrix, (810±15) g], being plankton filter feeding fish, were divided into six groups and cultured in the ponds respectively. The common carp in each pond were fed with commercial particle feeds three times a day and the feeding period was 62 days. Water quality, concentration of plankton and the bacterial community presenting in the water and sediment were monitored during the experiment. At the end of the experiment, all fish were harvested and the growth of the fish and feed efficiency ratio were determined. Results showed that the weight gain rate and feed efficiency ratio of common carp in AS group was significantly higher than that of the control group. Final body weights of bighead carp and silver carp in the AS group were significantly lower than that of the controls. Water transparency and diversity indices of the bacterial community in the AS group were significantly higher than that of the control (P<0.05) and the concentration of plankton in the AS group was lower than that of the control. Results showed that the application of AS in poly-culture ponds was beneficial for the growth of common carp, being artificial-feeds feeding fish, while it was not beneficial for the growth of bighead carp and silver carp, being plankton filter feeding fish.
Diagnosis of iridovirus disease in Sciaenop socellatus
Yun-Qing XI, Rui-Rong QI, Jia-Xin TIAN, Hui-Jun ZHANG, Yan LEI, Shao-Lin TANG, Gui-Qin WANG
 Available online  
[Abstract](25) [FullText HTML](8) [PDF 1595KB](2)
Abstract:
To examine the cause of the disease of Sciaenop socellatus of cultured pond in Zhuhai, Guangdong Province from June to October, the epidemiological investigation, the symptom observation, the pathogenic detection, and the pathology detection were used. These results showed that the disease had obvious infectious. The anatomy examination revealed the anemia of the gill filament, swollen spleen and kidney with hemorrhagic spots, and no observed large number of parasites by microscopy. Meanwhile, no pathogenic bacteria were isolated from the liver, spleen, kidney and intestine of the diseased fish. A large number of swelling cells were observed in pathological sections of liver, spleen and kidney of diseased fish. The 570 bp fragment was amplified by PCR using the specific primers of OIE. MCP sequence alignment showed that the amplification of the gene sequence had 99.2% of homology with gene sequence of RSIV. The comprehensive results showed that the cause of the disease of the Sciaenop socellatus is infected by the iridovirus of Megalocytivirus genus.
THREE NEW KINDS OF TOXIC PSEUDO-NITZSCHIA SPECIES DERIVED FROM CHINA’S COAST
Huan-Chang DONG, Chun-Xiu HUANG, Yang LI
 Available online  
[Abstract](51) [FullText HTML](6) [PDF 739KB](7)
Abstract:
In order to investigate the diversity of Pseudo-nitzschia species in China’s coast and their ability in producing neurotoxin domoic acid (DA), a total of fifteen monoclonal Pseudo-nitzschia strains were isolated and established. Using a liquid chromatography tandem-mass spectrometry (LC-MS/MS) technique, DA was accordingly detected from ten monoclonal strains. Based on the morphology observed by using a light microscope (LM), a transmission electron microscope (TEM), as well as molecular analysis inferred from an internal transcribed spacer region, these fifteen strains were precisely identified which belonged to three different species involving, P. cuspidata (Hasle) Hasle, P. pseudodelicatissima (Hasle) Hasle, and P. fraudulenta(Cleve) Hasle. In addition, three out of eleven established P. cuspidata strains were nontoxic, while other eight cases were toxic with the DA abundance rate of 0.4—5.5 fg/cell. Also, one P. pseudodelicatissima strain was nontoxic, while DA production rate of 1 fg/cell was detected in another strain. Besides, no DA was tested in the two P. fraudulenta strains. Several strains were selected and their DA productions were induced by the available Artemia salina. As a result, it was shown that the ability of DA production decreased in P. cuspidate (MC4049) and P. pseudodelicatissima (MC3015) strains, from 2 and 1 to 0.2 and 0.4 fg/cell, respectively. However, the ability of DA production in P. fraudulenta(MC4074) strain swiftly increased, from an undetected level to 17.5 fg/cell. The results achieved in this study enrich the diversity of toxic Pseudo-nitzschiaspecies in China’s coast, and may establish strong research basis for conducting further studies.
EFFECTS OF METSULFURON-METHYL ON THE GROWTH AND PHOTOSYNTHETIC SYSTEM OF MICROCYSTIS
Xiao-Jie MA, Yu-Liang WANG, Han-Yang LU, Yang LIU, Xiao-Yu LI, Ren-Hui LI
 Available online  
[Abstract](44) [FullText HTML](10) [PDF 1070KB](3)
Abstract:
Metsulfuron-methyl is one of the sulfonylurea herbicides, which have been widely used throughout the world because of its significant advantages and safety. However, long-term use of Metsulfuron-methyl will impose a threat to aquatic ecosystems. Algae are frequently the major primary producers in aquatic ecosystems, often as an indicator to study water pollutants. In this study, dominant species of cyanobacteria (Microcystis aeruginosa PCC7806) were selected to deeply perceive the effect of Metsulfuron-methyl on the growth and photosynthetic of Microcystis aeruginosa. The achieved results show that the growth of M. aeruginosa PCC7806 is significantly inhibited when the concentration of Metsulfuron-methyl exceeds 80 mg/L. The EC50 of the growth of M. aeruginosa PCC7806 is 81.998 mg/L by regression and Probit analysis. The results of cytochrome studies demonstrate that the content of chlorophyll a and carotenoids in unit cell of the treatment group on the 6th day is lower than that of in the control group, and when the concentration of Metsulfuron-methyl is 80 mg/L, the carotenoids content is remarkably lower than that of in the control group. The results of kinetics of chlorophyll fluorescence showed that the energy (ET0/RC) captured by the unit reaction center for electron transport (φE0) is used for quantum yield of photosynthetic of electron transport, which is extraordinary suppressed on the 6th day of the experiment. The results of chlorophyll fluorescence induction and cytochrome analysis show that Metsulfuron-methyl could negatively affect the electronic properties of the electron acceptor in the photosynthetic reaction center. In summary, Metsulfuron-methyl may act on the electron acceptor side of the photosynthetic response center, thereby likely affecting the photosynthetic system of Microcystis aeruginosa.

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GENETIC DIVERSITY AND STRUCTURE OF CRUCIAN CARP (CARASSIUS AURATUS) BASED ON G-SSR AND EST-SSR MARKERS
Bao-Jiang GAN, Mei-Xia PANG, Xiao-Mu YU, Jin-Gou TONG
2018, 42(3): 451-462   doi: 10.7541/2018.057
[Abstract](58) [FullText HTML](26) [PDF 1061KB](8)
Abstract:
This study investigated genetic diversity and genetic structure of six wild crucian carp populations collected from Heilongjiang River, Yangtze River, Fenghua River and Huaihe River by 8 genome microsatellites (G-SSR) and 8 transcriptome microsatellites (EST-SSR). The mean Na, Ne, Ho, He and PIC of the eight G-SSR loci were 22, 12.9, 0.769, 0.893 and 0.879, respectively. Fst values ranged from 0.008 to 0.085, indicating that there were different degrees of genetic differentiation among the six wild crucian carp populations, among which two populations from the Heilongjiang River system were different from all the others. There was no significant genetic differentiation among the four populations in the Yangtze River, Fenghua River and Huaihe River. The Nei’s genetic distances of six wild crucian carp populations were between 0.203—0.701. UPGMA cluster analysis and Bayesian analysis supported that six crucian carp populations could be divided into two large branches with two populations from Heilongjiang water system as one group while the others as another group. The average Na, Ne, Ho, He and PIC of the eight EST-SSR loci were 19, 9.5, 0.728, 0.870 and 0.855 respectively. The Fst values and Nei’s genetic distances among the six populations were between 0.005—0.084 and 0.117—0.683, respectively. Genetic differentiation analysis, UPGMA cluster analysis and Bayesian analysis based on EST-SSR markers also separated six populations of crucian carp into two major groups: the Heilongjiang water system group as well as the Yangtze River, Fenghua River and the Huaihe River system group. Based on G-SSR and EST-SSR markers, the average polymorphism information content (PIC) of the six crucian carp populations ranged between 0.786—0.864 and 0.761—0.833, respectively, indicating that all the six wild crucian carp populations had a high genetic diversity. Although the polymorphism of the EST-SSR markers was slightly lower than that of the G-SSR markers, the two types of SSR markers revealed similar genetic structure and pattern of differentiation for crucian carp in three major waters of China. This study provides basic and new information for the conservation of crucian carp germplasm resources and the evaluation of EST-SSR markers in genetic studies of fish populations.
CLONING AND TISSUE EXPRESSION OF FOUR WNT GENES IN YELLOW CATFISH PELTEOBAGRUS FULVIDRACO AND THEIR RESPONSE TO COPPER EXPOSURE
Li-Han ZHANG, Zhi LUO, Wen-Jing YOU, Dan-Dan LI, Kun WU, Yi-Huan XU
2018, 42(3): 463-472   doi: 10.7541/2018.058
[Abstract](36) [FullText HTML](22) [PDF 767KB](3)
Abstract:
Four full length Wnt cDNA sequences, designated Wnt5a, Wnt5b, Wnt7a and Wnt9b, were amplified by RT-PCR and RACE approaches from yellow catfish Pelteobagrus fulvidraco. The validated cDNAs of Wnt5a, Wnt5b, Wnt7a and Wnt9b were 1984, 2905, 2158 and 1622 bp in length, encoding peptides of 375, 375, 350 and 358 amino acid residues, respectively. The amino acid alignment and phylogenetic analysis revealed that Wnt sequences were relatively conserved among different vertebrates. All Wnt mRNAs were expressed in tested tissues (brain, spleen, kidney, gill, heart, muscle, mesenteric fat, liver and ovary) but with variable levels. Cu exposure for 28 days highly induced Wnt7a mRNA levels only at 30 μg Cu/L group. Cu exposure had no impact on mRNA levels of Wnt5a, Wnt5b and Wnt9b at day 28. On 56-day Cu exposure, mRNA levels of Wnt5b were not lowest at 60 μg Cu/L group and Wnt9b were the highest at 30 μg Cu/L group. Wnt5a and Wnt7a showed no significant differences among three groups. The present study indicated that functional differentiation exists in members of Wnt family, and some members may mediate the effect of Cu exposure on ovary development of yellow catfish.
CLONING AND FUNCTIONAL STUDY OF PGRP-S IN CHINESE GIANT SALAMANDER ANDRIAS DAVIDIANUS
Shi-Si REN, Zhi-Tao QI, Ming-Xian CHANG
2018, 42(3): 473-479   doi: 10.7541/2018.059
[Abstract](28) [FullText HTML](24) [PDF 661KB](4)
Abstract:
Peptidoglycan recognition proteins (PGRPs) are a class of pattern recognition receptors that are highly conserved from invertebrates to mammals. So far, PGRP is rarely reported in vertebrate species except for bony fish and mammals. This study constructed the eukaryotic expression plasmid of PGRP-S, a short peptidoglycan recognition protein in Chinese giant salamander, and explored its functins. Sequence analysis showed that the N-terminal of PGRP-S had no signal peptide, but had two closely spaced cysteine residues and two Zn2+ binding sites. Moreover, PGRP-S could be secreted to the exterior and also be retained within the cells. In vitro antibacterial experiments revealed that overexpression of PGRP-S significantly inhibited the proliferation of Edwardsiella tarda in HEK293T cells intracellularly and extracellularly. In addition, overexpression of PGRP-S enhanced NF-κB promoter activity and bound Lys-type and Dap-type peptidoglycan but without degradation ability.These results indicate that the Chinese giant salamander PGRP-S is functionally similar to mammalian PGRP-S.
ISOLATION AND IDENTIFICATION OF T-LYMPHOCYTES FROM PERIPHERAL BLOOD AND IMMUNE RESPONSE ANALYSIS OF TCRβ CHAIN GENE IN CYNOGLOSSUS SEMILAEVIS
Lin-Qing WANG, Hang WANG, Ya-Dong CHEN, Guang YANG, Li BAI, Lu-Ming SUN, Zhen-Xia SHA
2018, 42(3): 480-487   doi: 10.7541/2018.060
[Abstract](22) [FullText HTML](40) [PDF 1078KB](1)
Abstract:
The density gradient method was used to isolate lymphocytes from peripheral blood of Cynoglossus semilaevis, and the isolated lymphocytes were cultured in complete DMEM medium (with 0.3 μg/mL PHA) for 3—4 days. Peripheral blood lymphocytes were separated to non-adherent lymphocytes and adherent lymphocytes using self-made nylon hair column. Non-adherent lymphocytes specifically bind with mouse anti-human FTIC-CD3 monoclonal antibody, while adherent lymphocytes specifically bind to anti-human FTIC-CD19 monoclonal antibody.TCRβ specifically expressed on non-adherent lymphocytes. Taken together, non-adherent lymphocytes were confirmed as T lymphocytes. The results of qRT-PCR showed that TCRβ was expressed in all detected 13 tissues with high expression level in the intestine, stomach, spleen, head kidney and the lowest level in muscle. The expression of TCRβ gene was significantly induced in the liver, spleen and gill with peak level at 72—96h afterV. Anguillarum infection. These results indicate that TCRβ gene plays an important role in adaptive cellular immune response.
CLONING AND EXPRESSION ANALYSIS OF SRBI GENE IN DIFFERENT SHELL-COLOR STRAINS OF MERETRIX MERETRIX
Bao-Yue CUI, Ying-Hui DONG, Jia-Xi ZHAO, Ling-Wei HU, Xiao-Ying LI, Han-Han YAO, Zhi-Hua LIN
2018, 42(3): 488-493   doi: 10.7541/2018.061
[Abstract](21) [FullText HTML](17) [PDF 579KB](1)
Abstract:
To explore the molecular structure and biological function of SRBI in the process of carotenoids accumulation in Meretrix meretrix, we cloned cDNA of Mm-SRBI by SMART RACE technique and conducted the bioinformatics, expression analysis and intron amplification. The full length cDNA of Mm-SRBI is 1723 bp that contains a complete 1515 bp ORF encoding 504 amino acids with one functional domain of Mm-SRBI protein (CD36). Mm-SRBI is highly homologous with Mimachlamys nobilis and shares 55% similarity in amino acid sequence. It shares 34%—40% similarity with vertebrates. We also amplified 12 introns of Mm-SRBI, which followed the principle of GT-AG. Mm-SRBI gene expressed in all six tested tissues with the highest level in mantle (P<0.01), suggesting thatMm-SRBI gene maybe relate to high carotenoid in mantle. Moreover, the expression of Mm-SRBI gene in red shell strain was significantly higher than that of white shell strain (P<0.05). In conclusion, this study provides helpful foundation to explore the shell colors on this species.
GENETIC VARIATION AND RELATIONSHIP ANALYSIS OF SCALLOPS BASED ON THE SEQUENCES OF ITS1 AND 5S rDNA
De-Jie LIAO, Shan-Mao CAO, Jin-Gou TONG, Ying ZHOU, Xiao-Mu YU, Yang LIU, Xiao WANG
2018, 42(3): 494-502   doi: 10.7541/2018.062
[Abstract](20) [FullText HTML](16) [PDF 535KB](1)
Abstract:
Rock scallop Crassadoma gigantea was introduced to China in 2012, and was successfully bred in 2015. It is in the stage of popularizing culture now. To understand the genetic relationships and the genetic variations among C. gigantea, Argopecten irradians, Chlamys farreri, and Patinopecten yessoensis, the whole sequences of ITS1 and 5S rDNA were sequenced after the amplification by PCR. The sequence characterization, genetic diversity, genetic distance, and phylogeny were determined by using a suite of biology softwares. Based on the sequences of ITS1, the numbers of haplotype and polymorphic sites, haplotype diversities, nucleotide diversities, and average nucleotide differences were varied from 2 to 6, 1 to 12, 0.425 to 0.800, 0.00081 to 0.00486 and 0.366 to 2.44, respectively. In addition, based on the sequences of 5S rDNA, their same genetic variation parameters varied from 2 to 5, 1 to 14, 0.533 to 0.841, 0.00108 to 0.1058 and 0.533 to 5.726, respectively. The result showed that all these four species had a low diversity level (π<0.01). Based on the sequences ofITS1 and 5S rDNA, the genetic distance were varied from 0.040 to 0.124, and 0.043 to 0.457, respectively, which demonstrated that C. gigantea appeared to be closely-related to P. yessoensis. Meanwhile, phylogeny trees of the genetic distances confirmed the genetic relatedness between C. gigantea and P. yessoensis. This work would provide basic information for future studies on germplasm resources management, as well as on the genetics and breeding strategies of C. gigantea.
THE FUNCTION AND MECHANISM OF BURSICON GENE IN CARIDINA
Li-Qi REN, Jie-Yang WENG, Xin WANG, Jin-Sheng SUN, Ran LI
2018, 42(3): 503-511   doi: 10.7541/2018.063
[Abstract](92) [FullText HTML](27) [PDF 739KB](7)
Abstract:
To study the function of bursicon gene in Caridina during molting and cuticle formation, its open reading frame (ORF) containing two subunits genes were amplified by PCR respectively. The ORF of bursicon-α is 441 bp in length, encoding a protein with 146 amino acid residues. The ORF of bursicon-β is 411 bp in length, encoding a protein with 136 amino acid residues. The dynamic changes in the expression of both bursicon subunit genes during the molt cycle in Caridina have been analyzed by the real-time quantitative PCR. Relative expression of both bursicon-α and bursicon-β had different levels in different stages of the molt cycle, which increased through premolt stage (D stage) and reached their peak level at the D3 stage. The mRNA accumulation decreased to its lowest level at the molt stage (E stage). RNA interference-mediated knockdown of bursicon-α and bursicon-β retard the molting process and ecdysis behavior of Caridina. The cuticle of dsRNA-treated Caridina was thinner than that in the control group. These findings demonstrate that bursicon is involved in the thickening and hardening of cuticle in newly formed exoskeleton. Moreover, bursicon plays an important role in molt cycle.
MOLECULAR PHYLOGENY OF ONYCHOSTOMA (CYPRINIDAE) BASED ON MITOCHONDRIAL GENOMES
Chen ZHANG, Qi CHENG, Hong GENG, Ai-Hua LIN, Hong-Ying WANG
2018, 42(3): 512-516   doi: 10.7541/2018.064
[Abstract](24) [FullText HTML](13) [PDF 394KB](5)
Abstract:
The Onychostoma belongs to cyprinid group and contains 19 species from essentially morphological classification. However, the molecular phylogenetic relationship of Onychostoma, was not very clear. Here, we presented molecular phylogenetic analysis of 8 species in Onychostoma based on the whole mitochondrial genome sequence with Danio rerio and Spinibarbus denticulatus as outgroups. The results indicated that Onychostoma did not appear monophyletic which included 3 groups: Onychostoma macrolepis, Onychostoma lini andOnychostoma barbatum. O. barbatulum in clade 1, Onychostoma gerlachi and Onychostoma simum in clade 2, Onychostoma rara and Onychostoma alticorpus in clade 3. The result was roughly the same as the traditional morphological classification, which divides Onychostoma into three branches. The mitochondrial whole genome was firstly used in Phylogenetic research of Onychostoma and filled the shortage of Phylogenetic studies of this genus.
EFFECTS OF EXOGENOUS FATTY ACID ON THE PROLIFERATION AND DIFFERENTIATION OF NILE TILAPIA PREADIPOCYTES
Ya-Wen WANG, Fang QIAO, Mei-Ling ZHANG, Yong-Yi JIA, Zhen-Yu DU
2018, 42(3): 517-524   doi: 10.7541/2018.065
[Abstract](18) [FullText HTML](14) [PDF 1624KB](1)
Abstract:
To investigate the effects of fatty acids on the proliferation and differentiation of preadipocytes from Nile tilapia (Oreochromis niloticus), 100 μmol/L Palmitic acid (PA), Oleic acid (OA), Linoleic acid (LA) and α-Linolenic acid (LNA) were used. SRB (Sulforhodamine B) staining and oil red O staining were used to detect cell proliferation and preadipocyte differentiation. The expression level of genes related to proliferation and differentiation was detected by Real-time qPCR. The results showed that exogenous fatty acids promoted the preadipocyte proliferation in 8 days, and significantly increased the expression level of proliferation related genes (c-fos and c-myc), lipolysis related genes (ATGL) and adipogenesis related genes (PPAR, CD36 and FAS) (P<0.05). In addition, fatty acid inhibited preadipocyte differentiation into adipocyte by decreasing lipid droplets area and increasing the lipid droplets number. During differentiation, expression level of β-oxidation related gene (CPT-1a) significantly increased (P<0.05), while the expression level of lipolysis related genes (ATG) and adipogenesis related genes (PPARγ and FAS) declined. Our results showed that exogenous fatty acid promoted the proliferation and inhibited preadipocyte differentiation. In the process of proliferation, excess fatty acids stored in the preadipocyte that could be metabolized by lipolysis and β-oxidation during adipogenesis to help cells adapt to high concentrations of fatty acids in the environment. While during the differentiation, the exogenous fatty acids can inhibit adipogenesis and lipolysis of adipocyte and promote β-oxidation to reduce adipocyte differentiation.
EFFECTS OF α-KETOGLUTARATE SUPPLEMENTATION IN LOW-PHOSPHOROUS DIETS ON GROWTH PERFORMANCE, BODY COMPOSITION AND SERUM BIOCHEMICAL INDEXES OF SONGPU MIRROR CARP
Feng AI, Lian-Sheng WANG, Jin-Nan LI, Chang-An WANG, Zhi-Gang ZHAO, Liang LUO, Xue DU, Qi-You XU
2018, 42(3): 525-532   doi: 10.7541/2018.066
[Abstract](26) [FullText HTML](20) [PDF 472KB](5)
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To study the effects of α-ketoglutarate (AKG) supplementation in low-phosphorous diets on growth performance, body composition and serum biochemical indexes of Songpu mirror carp, triplicate groups of Songpu mirror carp (6.67±0.84) g were fed with practical diets containing available P (AP) 0.73% (normal P) and 0.47% (low-P) with graded levels of 0, 0.4%, 0.8%, 1.2%, 1.6% and 2.0% of AKG to satiation for 8 weeks, respectively. The results showed that the feed coefficient (FCR) of the low phosphorus group was significantly higher than that of the normal phosphorus group (P<0.05), but the weight gain rate (WGR), specific growth rate (SGR), protein efficiency (PER), VSI, and CF were similar between normal P and Low-P groups (P<0.05). Compared with low P group,FCR of 0.4% AKG group and the levels of phosphorus and albumin (ALB) in 1.6% AKG group in normal P group were significantly increased (P<0.05), respectively. Compared with normal P group, ALT activity of the 0.8% AKG and AST activity of the 1.6% AKG in the low P group were significantly increased, respectively (P<0.05). AKG had no effect on serum other indexes (P>0.05). Compared with the normal phosphorus group, the content of crude ash and the deposition rate of calcium and phosphorus in the low phosphorus group decreased significantly (P<0.05). Compared with the low phosphorus group, the phosphorus deposition rate and bone calcium content of 0.4% AKG and bone phosphorus content of 0.8% AKG in normal P group increased significantly, respectively (P<0.05). Adding AKG in low phosphorus diet had no significant effect on other body composition indexes (P>0.05). Thus, the suitable supplementation of AKG in the low phosphorus diets can reduce theFCR and improve the content of calcium and phosphorus in bone of Songpu mirror carp.
EFFECTS OF DIFFERENT LIPID SOURCES ON THE GROWTH, LIPID METABOLISM AND ANTIOXIDANT ABILITY OF BRACHYMYSTAX LENOK
Yang LIU, Jian GAO, Xiao-Juan CAO, Jia-Sheng YIN, Hong-Bai LIU
2018, 42(3): 533-541   doi: 10.7541/2018.067
[Abstract](23) [FullText HTML](17) [PDF 464KB](6)
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A 52-day growth trial was conducted to evaluate the effects of different dietary lipids on the growth performance, lipid metabolism, and antioxidant ability of Brachymystax lenok. Fish with average initial weight of (22.19±1.10) g were fed with four experimental diets that contained fish oil (FO), colza oil (CO), palm oil (PaO), and soybean oil (SO), respectively. The lowest weight gain, specific growth rate, and the feed conversion ratio were observed with CO treatment, while the highest values of these parameters occurred at the experiments of FO and SO, with no significant differences between these two conditions (P>0.05). Crude protein and lipid in the muscle ofBrachymystax lenok were not affected by lipid sources (P>0.05). However, the DHA level ofBrachymystax lenok fed with SO increased, which indicated that Brachymystax lenok had the ability to desaturate and elongate from 18:3n-3 to 22:6n-3. Compared to which fed with FO, an increase of TG and decrease of TC were observed in the serum of Brachymystax lenok fed with plant oils. In addition, the mRNA expression of Δ6 Fad in the liver of those fed with plant oils also increased, but the mRNA expression of ACC1 and FAS decreased. The values of ROS, MDA, GPx and TBARS in liver were significantly affected by different lipid sources (P<0.05). The results suggest that FO and SO could be used in the diet for juvenileBrachymystax lenok, but CO and PaO are unsuitable as single lipid sources for juvenile Brachymystax lenok.
NUTRITIONAL COMPOSITION OF MUSCLE IN CULTURED SINIBOTIA REEVESAE POPULATION AT DIFFERENT AGES
Yong-Ming WANG, Jin-Rong SHI, Bi-Wen XIE, Rui TANG, Yu WANG, Ze-Min QI
2018, 42(3): 542-549   doi: 10.7541/2018.068
[Abstract](20) [FullText HTML](15) [PDF 463KB](4)
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Sinibotia reevesae, an endemic fish, live only in the upper reaches of the Yangtze River. To study nutritional composition in muscle during different growth phases, the main nutrient components in the muscles of age 1, 2 and 3 farmed (fed formulated feed) S. reevesae were analyzed, and the nutritional value and product flavor were evaluated. The general nutrient compositions were determined by routine methods, the amino acid compositions were measured with an amino acid analyzer and the fatty acid compositions by gas chromatography-mass spectrometry (GC-MS). The results showed that the contents of crude protein and crude fat in S. reevesae muscles increased with fish growth. With the increase of age, the moisture content decreased, whereas the contents of crude protein and crude fat increased. The crude protein in muscles of age-group 3 was significantly higher than that of the other two groups (P<0.05), while no significant difference was found in crude protein content between age-group 2 and 3 (P>0.05). The contents of different amino acids in muscles ofS. reevesae among 3 groups were stable. Glutamic acid, aspartic acid and lysine were the most abundant among 16 kinds of amino acids detected in muscles of each group. The essential amino acid index (EAAI) and the ratio of branched chain amino acid to aromatic amino acid (F value) of S. reevesae were very high. According to nutrition evaluation in amino acid score (AAS) and chemical score (CS), the first limiting amino acid for muscles of S. reevesae during different growth phases was valine acid. Totally 7 saturated fatty acids (SFA), 6 mono-unsaturated fatty acids (MUFA), and 7 poly-unsaturated fatty acids (PUFA) were found in dry muscles of S. reevesae from each group. The content of SFA was significantly lower than that of unsaturated fatty acids (UFA) (P<0.05), and the total content of MUFA, especially C18:1n9c, was significantly higher among UFA. PUFA in age-group 2 was significantly higher than that of the other two groups (P<0.05). The total content of EPA and DHA was lower than other Cobitidae fishes with the tendency to descend with the increasing age. These results indicate thatS. reevesae is a kind of high nutritional value, delicious, and high quality fish, and that S. reevesae in age 2 and 3 age are great for consume because of higher contents of crude protein and crude fat, better amino acid and fatty acid compositions in muscle.
THE VALIDITY OF SUBSPECIES OF BRACHYMYSTAX LENOK TSINLINGENSIS LI BASED ON MORPHOLOGICAL DIFFERENCE ANALYSIS
Yan-Xiao MENG, Gui-Hua WANG, Dong-Mei XIONG, Hai-Xia LIU, Jian-Lu ZHANG, Ji-Long WANG, Li-Xin WANG, Xiao-Lin LIU
2018, 42(3): 550-560   doi: 10.7541/2018.069
[Abstract](21) [FullText HTML](18) [PDF 713KB](5)
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The taxonomic status of Brachymystax lenok tsinlingensis Li, distributing in Qinling area of China, is controversial since it was found. In this study, the morphological differences among three Brachymystax groups (B. lenok lenok (Pallas, 1773) and B. tumensis (Mori, 1930) from Amur river basin and B. lenok tsinlingensis (Li, 1966) from the Qinling area) were investigated, including five meristic characters and thirty-four standardized morphometric characters through multivariate morphometrics methods. Results of one-way ANOVA showed that five meristic characters and thirty-two standardized morphometric characters were significantly different (P<0.01) among three groups. Principal component analysis indicated that the cumulative contribution of the top three principal components was 92.779%. Effective linear discriminant formulas were obtained from fourteen standardized morphometric characters, with the discriminating accuracy of 99.4% in the three groups. The tree diagram of these groups based on the squared Euclidean distance showed thatB. lenok tsinlingensis had a close distance with B. lenok lenok, but it is distant from B. tumensis. The coefficient of difference (CD) for the outer gill raker number in the first gill arch between B. lenok tsinlingensis and B. lenok lenok is larger than the threshold value of 1.28, and the CD for the thirty-one standardized morphometric characters and two meristic characters in B. lenok tsinlingensis and B. tumensis was also greater than 1.28. According to the results listing above, and the geographical isolation of B. lenok tsinlingensis together with its morphological differences from B. lenok lenok and B. tumensis, it is deduced that the morphological difference between B. lenok tsinlingensis in Qinling and B. lenok lenok in Amur river basin has at least reached subspecies level.
INDIVIDUAL VARIATION AND PHENOTYPIC CORRELATIONS IN SWIMMING PERFORMANCE OF JUVENILE CRUCIAN CARP
Ling-Qing ZENG, Cheng FU, Shi-Jian FU
2018, 42(3): 561-570   doi: 10.7541/2018.070
[Abstract](18) [FullText HTML](14) [PDF 627KB](3)
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The swimming ability is a fundamental organismal trait to influence growth, survival and even the Darwinian fitness of fish. Energy metabolism involves oxygen uptake, locomotor capacity includes aerobic swimming performance and anaerobic swimming performance, which exhibits great variations in these two physiological functional traits. Trade-off may occur when these two traits can not be optimized simultaneously since aerobic and anaerobic swimming performance pose conflicting demands on the same morphological feature of design. The present study examined the intra-individual variations and phenotypic trait correlations of swimming ability of the cyprinid fish, and investigated the effects of different acceleration speed on the constant acceleration swimming capacity in the cyprinid fish. Swimming performance of the juvenile crucian carp (Carassius auratus) including critical swimming speed (Ucrit), and gait transition speed (Ugt) was performed in a Ucrit test. The metabolic rates including resting metabolic rate (RMR), maximum metabolic rate (MMR), aerobic scope (AS=MMR–RMR) as well as factorial aerobic scope (FAS=MMR/RMR) were measured by both of theUcrit test and the test of excess post-exercise oxygen consumption (EPOC). A constant acceleration test (Ucat) was employed to examine the Ucat and Ugt in the juvenile crucian carp at four acceleration rates (0.083, 0.167, 0.250 and 0.333 cm/s2). Our study found that Ucrit were correlated with MMR and AS with the exception of RMR while Ugt was not related to any parameters of metabolic rates (MMR, AS and RMR) in the juvenile crucian carp. There were no significant difference in parameter average of metabolic traits (MMR, AS and FAS) between two induced exercise methods, and high repeatability was detected in intra-individual level between the two methods. Distinct correlations were found among traits of metabolism. The Ucrit and Ugt conducted by Ucrit test were smaller than those conducted by Ucat test. The acceleration rate had a determinable influence on Ucat but not Ugt. Ugt correlated positively with either Ucrit or Ucat and the constant acceleration swimming capacity had repeatability among four acceleration rates. Ucat was positively correlated to the anaerobic metabolism contribution (UcatUgt) by the acceleration swimming test at three acceleration rates with the exception of 0.333 cm/s2. However, the aerobic metabolic capacity (Ugt) was negatively related to the anaerobic metabolic capacity (UcatUgt) in the juvenile crucian carp by the Ucat test. The study suggested that no methodological difference could be found in aerobic capacity by the test of Ucrit and EPOC, and that correlation among traits of energy metabolism and constant individual difference in acceleration swimming performance could be found in the juvenile crucian carp. Performance trade-off was detectable between aerobic metabolic capacity and anaerobic metabolic capacity in this species.
THE EFFECT OF ACCELERATING FLOW ON SWIMMING BEHAVIOR OF JUVENILE ARISTICHTHYS NOBILIS AND HYPOPHTHALMICHTHYS MOLITRIX DURING DOWNSTREAM MIGRATION
Min-Ne LI, Hai-Feng ZHU, Zhi-Jun JIN, Jia LUO, Jia-Wei XU, Zhi-Min LI, Wei-Zhong MA, Xiao-Tao SHI
2018, 42(3): 571-577   doi: 10.7541/2018.071
[Abstract](19) [FullText HTML](14) [PDF 591KB](3)
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Accelerating flow generally exists during fish downstream migration passage, which will affect swimming behavior of individual fish. In this study, the swimming behavior of juvenile silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis) during downstream migratory were tested under two types of accelerating flows (Ⅰ: 22—55 cm/s, Ⅱ: 25—60 cm/s). The objective of this experiment was to quantify the swimming behavior, including downstream pass rate, the way of downstream movement, and the time spent in each area. The preferred flow velocity was measured by the percentage of residence time (Ft). The result showed that the fish performed two kinds of downstream behavior in accelerating flow, which were " direct” and " non-direct”. The direct downstream rate of silver carp and bighead carp were 35.71% and 30% in Ⅰ, and 45.83% and 59.09% in Ⅱrespectively. During the non-direct downstream movement, silver carp spent significantly longer time in the flow less than 30 cm/s under both I and II flow conditions (P<0.05), which indicated that the preferred water velocity of silver carp in the tests was less than 30cm/s. The time spent by bighead carp was significant longer in the flow velocity less than 30 cm/s and 50—55 cm/s under flow conditionⅠ than in flow with other velocity (P<0.05), and under flow condition II, the time was longer in the flow velocity less than 30 cm/s (P<0.05), which implicated that the preferred water velocity of bighead carp in the test was less than 30 cm/s. Based on the results from two types of accelerating flow conditions, it showed that there was significant difference (P<0.05) in the residence time between silver carp and bighead carp in the flow with velocities less than 30 cm/s and 50—55 cm/s, but there was no significant difference (P>0.1) in medium and high velocity conditions. The study demonstrated that the accelerating flow can affect the swimming behavior of individual fish during downstream migration, and there is some interspecific difference in the ways of downstream migration and preferred velocity between silver carp and bighead carp.
EFFECTS OF TEMPERATURE, pH AND SALINITY ON THE SURVIVAL OF JUVENILE PERCOCYPRIS PINGI RETRODORSLIS
Fang-Peng JIN, Guang-Hua LI, Lin LI, Xin QIN, Kun LI, Jun-Jie WU, Hai-Tao GAO, Yi-Dan XIA, Feng LIU
2018, 42(3): 578-583   doi: 10.7541/2018.072
[Abstract](29) [FullText HTML](18) [PDF 520KB](9)
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To study the tolerance of Percocypris pingi retrodorslis to temperature, pH, and salinity, the juvenile Percocypris pingi retrodorslis were used as experimental objects. A single factor static acute toxicity test was used to study their tolerance to temperature, pH and salinity. The experimental results showed that their tolerance on temperature ranged from 0 to 32℃, and the lethal high and low temperature was 32℃ and 1℃, respectively. The optimum growth temperature was between 8 and 27℃. In addition, the optimum growth pH was 5.0—9.0. However, the juvenile fish began to die when pH was higher than 9.5 or lower than 4.7 (within 96h after treatment). The effects of different alkalinity on the survival of juvenile fish have significant difference (P<0.05), after the treatment for 24h, 48h, 72h and 96h, but there was no significant difference effects on the survival at different acidity level (P>0.05). TheLC50 of acidity for the juvenile fish was 3.90, 3.96, 4.15 and 4.40 after treated for 24h, 48h, 72h and 96h, respectively. The LC50 of alkalinity was 11.20, 11.10, 10.98 and 10.89 after the treatment of 24h, 48h, 72h and 96h, respectively. The juvenile fish began to die when salinity was higher than 7.5, and the effects of different salinity on the survival of juvenile fish were significantly different (P<0.05). TheLC50 of salinity for juvenile fish was 10.30, 9.25, 9, 8.85 and 8.82 g/L at 12h, 24h, 48h, 72h and 96h after treatment, respectively, and two safe concentration (SC) levels were 0.89 g/L and 2.385 g/L. The Percocypris pingi retrodorslis has been listed as endangered species, and our data will provide a theoretical foundation for the artificial breeding and trans regional breeding of Percocypris pingi retrodorslis.
DIAGNOSIS AND TREATMENT OF THE BACTERIAL DISEASES IN THE YANGTZE FINLESS PORPOISE
Zhi-Gang LIU, Ai-Fang ZHENG, Min-Min CHEN, Yu-Xi LIAN, Hu-Yan JIANG, Dao-Ping YU
2018, 42(3): 584-592   doi: 10.7541/2018.073
[Abstract](46) [FullText HTML](26) [PDF 1082KB](2)
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Pathogens are often found with Yangtze finless porpoises (Neophocaena asiaeorientalis asiaeorientalis; YFPs) that live under captive and semi-natural conditions. To investigate the diagnosis and treatment of bacterial diseases of Yangtze finless porpoise, the pathogenic bacterium Staphylococcus aureus and Morganella morganii were isolated from naturally infected Yangtze finless porpoise that live under semi-natural conditions in Anqing Xijiang River. The drug-sensitivity tests of the pathogenic bacterium of the Yangtze finless porpoise demonstrated that pathogens are Staphylococcus aureu and Morganella morganii. The outcomes of treatments based on drug test results were effective. This study provides a good reference for the diagnosis and prevention of bacterial diseases of the Yangtze finless porpoise and dolphin.
THRESHOLDS FOR THE HEARING OF MARBLED ROCKFISH SEBASTICUS MARMORATUS
Xu-Guang ZHANG, Hong-Yi GUO, Jia-Kun SONG
2018, 42(3): 593-598   doi: 10.7541/2018.074
[Abstract](19) [FullText HTML](14) [PDF 1162KB](2)
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The thresholds for the hearing of marbled rockfish (Sebasticus marmoratus) were studied using the auditory evoked potential (AEP) technique. The AEP responses to seven different sonic stimulations, whose tone burst frequencies are between 100—1000 Hz, from eight fishes were recorded by two subcutaneous electrodes (Tucker Davis Technologies). Our results showed that marbled rockfish were sensitive to the low frequency sounds ranging from 100 to 300 Hz, and the most sensitive sound frequency is 150 Hz, corresponding to the lowest hearing threshold of 70 dB re 1 μPa. The dominant frequency of the fish sound matches with their sensitive sound frequency range, which indicates that marbled rockfish are able to detect their own sounds, and the importance of acoustic communication among them. Meanwhile, low-frequency anthropogenic noise may affect their acoustic communication.
STUDIES ON CRYOPRESERVATION AND TRANSPLANTATION OF THE MALE GERM CELL IN AMERICAN SHAD (ALOSA SAPIDISSIMA)
Xu-Ling WU, Xiao-You HONG, Kai-Bin LI, Xin-Ping ZHU, Hong-Yan XU
2018, 42(3): 599-605   doi: 10.7541/2018.075
[Abstract](17) [FullText HTML](15) [PDF 911KB](1)
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Germ cells, different from somatic cells, carry the hereditary information and can transmit it to next generation. The germ cells cryopreservation and culture are the key techniques for cell engineer breeding and genetic resource preservation. This study investigated the effects of cryopreservation of the testicular blocks or dissociated cells on the viability of male germ cells of American shad after 250 days using Hochest33342/Propidium Iodide (PI) staining. The results showed that the overall viability of germ cells from the testis blocks cryopreservation was higher than that of dissociated cells cryopreservation, suggesting that the tissue frozen could efficiently cryopreserve the germ cells at different stages in American shad. Moreover, the thawed germ cells of American shad were stained with PKH26 before transplantation into newly-hatched zebrafish fries (1 days post hatching). At the 5th day post transplantation, the donor cells were still detected in some recipients and some donor cells were co-localized with the endogenous PGCs of recipient, implying that the germ cells of American shad after a long-term cryopreservation still possess the properties of germ cells and could integrate to the genital ridge of zebrafish host. The findings of this study provide technological and theoretical basis for further investigations on germ cell development, culture and genetic resource preservation in American shad or other anadromous fish in wild field.
COMMUNITY STRUCTURE OF SNAILS AND ITS RELATIONSHIP WITH ENVIRONMENTAL FACTORS ALONG ZHEJIANG COAST IN SPRING
Zhan-Hui LU, Wen-Bin ZHU, Kai-Da XU, Yong-Dong ZHOU, Qian DAI, Kan-Er LU
2018, 42(3): 606-615   doi: 10.7541/2018.076
[Abstract](26) [FullText HTML](15) [PDF 2455KB](5)
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Based on the survey data collected in April 2015 via bottom trawl along Zhejiang coast, this study identified the species composition and dominant species, as well as analysed the resource biomass and density, and community diversity. In addition, the relationship between the community structure of snails and environmental factors was also explored in this study. The results showed that 27 snail species were identified, and they belong to 2 orders, 15 families and 21 genera. The dominant species were Turritella bacillum and Siphonalia spadicea; The average biomass and density were 246.64 kg/km2 and 27.75×103 ind./km2, respectively, whose distributions appeared to be inhomogeneous. However, the density in the northern area was lower than that in the middle-southern area. The average values for species abundance index (D), species diversity index (H′) and species evenness index (J′) were 0.36, 0.56 and 0.50, respectively, indicating the snail community diversity was on a low level. Species diversity index (H′) display spatial distribution pattern, but the other two indexes remained almost constant in the study area. The ABC curve showed that snail community was unstable in spring. According to the CCA analysis, the distribution of snail community was affected mainly by depth, bottom temperature and bottom dissolved oxygen in the surveyed area.
EFFECT OF CADMIUM ON THE FEEDING CAPACITY AND PHYSIOLOGICAL STATUS OF DAPHNIA MAGNA
Qian WANG, Hu-Fei GUO, Lan WANG
2018, 42(3): 616-621   doi: 10.7541/2018.077
[Abstract](20) [FullText HTML](16) [PDF 568KB](2)
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To investigate the effects of cadmium (Cd2+) on the feeding capacity and its mechanism of water flea, Daphnia magna, three experimental groups with different Cd2+ concentrations (0.01 mg/L, 0.05 mg/L and 0.09 mg/L) and one control group were set up in this study, and these groups were exposed for 24h and 48h, respectively. The effects of Cd2+ levels on the feeding and filtration rate, tissues accumulation of Cd2+, total antioxidant capacity (T-AOC), contents of malonyldialdehyde (MDA), and the activities of acetylcholine esterase (AChE) in D. magna were analyzed. The results showed that (1) the feeding and filtration rates of D. magna decreased with increasing Cd2+ concentrations. Especially, when the D. magna was exposed to 0.09 mg/L Cd2+ for 48h, the feeding and filtration rates dropped dramatically (P<0.05,P<0.01), with the feeding rate decreasing by 43.89% compared to the control group. (2) With the increase of Cd2+ concentration, the accumulation of Cd in tissues was significantly higher in the experimental groups than that in the control group. (3) The total antioxidant capacity (T-AOC) and content of MDA in the water flea increased gradually with the concentration level of Cd2+. In the group with Cd2+ concentration of 0.09 mg/L, the levels of T-AOC and MDA increased significantly (P<0.05) compared to the control group. T-AOC was negatively correlated with the feeding rate whose correlation coefficientR2 were 0.9521 and 0.9389, respectively. (4) The activity of AChE decreased with increasing Cd2+ concentration, especially when D. magna were exposed to higher concentrations of 0.05 and 0.09 mg/L (P<0.05). The results indicated that Cd2+ stress not only caused oxidative damage to water flea, but also induced the abnormal conduction function to the nerve system, which would result in inhibition of feeding behavior in D. magna.
RELATIONSHIP BETWEEN PHYTOPLANKTON MORPHOLOGY-BASED FUNCTIONAL GROUPS AND ENVIRONMENTAL FACTORS OF DIFFERENT HABITAT IN THE LAKE XIANNÜ AND INFLOW RIVERS
Chen YU, Meng ZHANG, Hong-Wen CHEN, Min ZHOU, Tao WANG, Chao LI, Min ZHANG
2018, 42(3): 622-634   doi: 10.7541/2018.078
[Abstract](43) [FullText HTML](14) [PDF 1112KB](4)
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Phytoplankton is an essential component for the functioning of material cycle and energy flow which accounts for half of earth’s primary production. Meanwhile, phytoplankton community structure and distribution are important indicators of water environment and its quality. Identifying the main factors driving phytoplankton community structure is essential for better understanding and therefore managing freshwater ecosystem. Hence, we investigated phytoplankton community structure, morphology-based functional groups and temporal and spatial distribution of these groups among Lake Xiannü and inflow rivers during 2014—2015. There were 82 species of 7 phyla recorded, including 15 species of Cyanophyta, 33 species of Chlorophyta, 23 species of Bacillariophyta, 3 species of Pyrrophyta, 5 species of Euglenophyta, 2 species of Cryptophyta and 1 species of Chrysophyta. The average phytoplankton density and biomass were in ranges of 7.95×106—2.19×109 ind./L and 10.52—792.91 mg/L, respectively. Non-flagellated, with siliceous exoskeletons phytoplankton (Group Ⅵ, diatom) was the dominant species in river habitat in spring and autumn. Unicellular flagellates of medium to large phytoplankton (Group Ⅴ, Cryptophyta and Pyrrophta) was the dominant species in lake zones. Although biomass of Group Ⅵ phytoplankton was relatively high in different habitats, large filaments with aerotopes phytoplankton (Group Ⅲ) and large mucilaginous colonies (Group Ⅶ), such as Cyanophyta, increased significantly between summer and autumn. Analysis of the relationship between phytoplankton morphology-based functional groups and environmental factors found that the biomass of Group Ⅴ and Ⅵ was affected by the TN and TP concentration in winter and spring. In summer and autumn, phytoplankton of Group Ⅲ and Ⅶ were affected by multiple factors including water temperature, turbidity and TN and TP concentrations.
CREATION OF HIGH ARGININE PRODUCING STRAIN OF SYNECHOCYSTIS SP. PCC 6803 BY UV MUTAGENESIS
Shui-Ling JI, Hong GAO
2018, 42(3): 635-640   doi: 10.7541/2018.079
[Abstract](16) [FullText HTML](34) [PDF 496KB](3)
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Arginine has been widely used in medicine and food industry. Synechocystis sp. PCC 6803, a unicellular cyanobacterium, synthesize amino acids and other compounds using nitrate or nitrite as the nitrogen source. In flue gases released by industrial production, NOx (nitrogen oxides) can react with water to form nitrate and nitrite, which can be utilized by cyanobacteria. Hence, creating high arginine producing strain of Synechocystis sp. PCC 6803 has a strong application prospects, in denitrification of industrial flue gases. This study reported the successful creation of a high arginine producing strain #13807-111-55 of Synechocystis sp. PCC 6803 by 0.8 g/L D-arginine and 0.2 g/L 6-azaruacil resistant mutants with UV mutagenesis. Compared with the parent strain #13807, extracellular arginine production per OD730 of #13807-111-55 increased 62.3-fold, reaching (0.76±0.1) mg/(L·OD730). Total arginine production per OD730 of #13807-111-55 increased about 6.0-fold, reaching (0.82±0.08) mg/(L·OD730). Extracellular arginine production per OD730 of the mutant was significantly higher than that of intracellular, showing that the strain was the arginine-secreting mutant with strong production potential.
THE RECENT DIATOM FLORA IN LAKE KANAS, XINJIANG: UNUSUAL SPECIES AND NEW RECORDS IN CHINA
Xue-Ru LIN, Rioual Patrick, Zhi-Juan BAI, Wei PENG, Ming-Jie Sun, Xiao-Zhong HUANG
2018, 42(3): 641-654   doi: 10.7541/2018.080
[Abstract](18) [FullText HTML](14) [PDF 5560KB](4)
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To assess the diatom diversity of Lake Kanas, the largest freshwater lake in the Altay region, we analyzed a total of 36 diatom samples: 19 samples from the surface sediment in the lake, and 17 samples were the water of the lake, rivers and wetlands. In all, 123 taxa belonging to 46 genera were observed. Some of the 123 taxa listed are illustrated with LM and SEM photos. Among them, 13 are newly recorded species in China. They are Amphora indistincta Levkov, A. neglecta f. densestriata Foged, Asterionella edlundii Stoermer & Pappas, Discostella stelligeroides (Hustedt) Houk & Klee, Fragilaria capucina fo. sublanceolata-baikali Flower & Williams, Gomphonema makarovae Lange-Bertalot, Hippodonta geocollegarum Lange-Bertalot, Metzeltin & Witkowski, H. pseudorostrata Metzeltin Kulikovskiy & Lange-Bertalot, Nitzschia metzeltinii Lange-Bertalot, Placoneis opportuna (Hustedt) Chudaev & Gololobova, Sellaphora pseudobacillum (Grunow) Lange-Bertalot & Metzeltin, S. madida (Kociolek) Wetzel and Staurosira aff. sviridae Chudaev & Gololobova.
RESEARCH PROGRESS ON GENETIC TRANSFORMATION TECHNOLOGY OF EUGLENA
Qing SHAO, Yong-Guang JIANG, An-Ping LEI, Zhang-Li HU, Jiang-Xin WANG
2018, 42(3): 655-662   doi: 10.7541/2018.081
[Abstract](18) [FullText HTML](20) [PDF 502KB](3)
Abstract:
Euglena have important scientific value for studying biological evolution and chloroplast endosymbiosis and provide high value-added dietary supplements as well as high-quality feedstock for biofuel production. Studies of Euglena have been mainly explored by physiological and biochemical methods. With almost completed Euglena genome, we employed diverse OMIC techniques such as single cell RNA-seq, transcriptomic, proteomic, methylomic, and metabolomics to study the chloroplast development and the communication between chloroplast and nucleus. We hooked up some interesting genes and microRNA targets. Little progress has been achieved in genetic engineering of Euglena due to lack of efficient techniques for genetic operation. In here, we reviewed the methods in genetic transformation of Euglena, such as antibiotic biomarkers, biolistic, electroporation, glass beads and Agrobacterium transformation techniques, RNAi, potential CRISPR/cas9, and cpf1 genome editing, to provide a reference for functional genomics and biotechnology of Euglena.

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Journal Introduction

  • Establishment Time:1955  Bimonthly
  • Competent unit:Chinese Academy of Sciences
  • Host unit:Institute of Hydrobiology, Chinese Academy of Sciences ; Chinese Society for Oceanology and Limnology
  • Editor-in-Chief:GUI Jian-Fang
  • ISSN 1000-3207
  • CN 42-1230/Q

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