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  • 月刊，1984年创刊
    主 管：内蒙古自治区教育厅
    主 办：包头医学院
    主 编：赵云山
    特邀主编：高长青
    编辑出版：包头医学院学报编辑部
    国际刊号：ISSN 1006－740X
    国内刊号：CN 15－1182/R
    邮发代号：16-292
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2021 Vol. 40, No. 4
Published: 2021-07-25
Research and Application
Overview and Specific Topi
 
       Research and Application
475 Combined Effects of High Temperature and Ammonia Stress on Osmoregulation of Pacific White Shrimp Litopenaeus vannamei
XIONG Dalin, DUAN Yafei, CHEN Chengxun, SUN Yongxu, DONG Hongbiao, ZHAN Aijun, ZHANG Jiasong
DOI: 10.16378/j.cnki.1003-1111.19281
The changes in Na+/K+-ATPase and Ca2+-ATPase activities in gill and the contents of free amino acid (FAA) in muscle and gill were monitored in Pacific white shrimp Litopenaeus vannamei with body weight of (6.77±0.51) g reared in 200 L fiberglass barrels at initial stocking density of 50 per barrel under two temperatures (normally 30 ℃ and acute increase from 30 ℃ to 33 ℃ within 1 h) × 4 ammonia nitrogen concentrations (0, 5, 15, and 25 mg/L) adjusted with ammonium chloride solution for 72 h to evaluate the effects of ammonia-nitrogen stress on the physiological osmoregulation of Pacific white shrimp under acute warming temperature. The results showed that the activities of Na+/K+-ATPase and Ca2+-ATPase were found to be significantly increased in 25 mg/L and 5 mg/L ammonia groups at 30 ℃(P<0.05), significant increase in activities of Na+/K+-ATPaes and Ca2+-ATPase in all ammonia groups at 33 ℃(P<0.05). In the same ammonia concentration, there were significant differences in the activities of Na+/K+-ATPase and Ca2+-ATPase between 30 ℃ and 33 ℃ groups, significantly lower enzyme activity in the 33 ℃ group than that in the 30 ℃ group at 25 mg/L ammonia concentration (P<0.05). In gill and muscle, the total contents of FAA were significantly lower in the 33 ℃ group than that in 30 ℃ group at ammonia concentration of 15 mg/L and 25 mg/L (P<0.05). Under ammonia stress, alanine level in muscle was increased significantly in 30 ℃ and 33 ℃ groups with increasing ammonia concentration (P<0.05), and significantly higher at 33 ℃ than at 30 ℃ group (P<0.05).The findings indicated that ammonia stress sharply affected Na+/K+-ATPase and Ca2+-ATPase activities as well as the contents of FAA under acute warming temperature,leading to dysfunction of the physiological osmoregulation of Pacific white shrimp.
2021 Vol. 40 (4): 475-482 [Abstract] ( 343 ) HTML (1 KB)  PDF (1120 KB)  ( 305 )
483 Nitrogen and Phosphorus Budgets of Polyculture System of Mud Crab Scylla paramamosain, Tiger Shrimp Penaeus monodon and Razor Clam Sinonovacula constricta
ZHAO Yanfei, ZHONG Shengping, WANG Xianfeng, HUANG Lingguang, LIU Xujia, XIONG Xiangying
DOI: 10.16378/j.cnki.1003-1111.20076
The ternary polyculture models of mud crab Scylla paramamosain, tiger shrimp Penaeus monodon and razor clam Sinonovacula constricta were investigated in land-based perigrid experiments in seawater ponds at stocking density of 35 individuals of mud crab and 300 individuals of tiger shrimp (as a control, CS) with 5 different proportions of razor clam including 250 (CSR1), 500 (CSR2), 1000 (CSR3), 2000 (CSR4), and 4000 (CRS5) individuals. In the feeding trial, budgets and utilization rates of nitrogen and phosphorus were analyzed by one-way ANOVA and multiple comparisons. The results showed that the feed was the main input of nitrogen (accounting for 84.02%—86.31%) and phosphorus (accounting for 98.08%—98.83%) in the enclosures, the main output of nitrogen (accounting for 44.21%—56.40%) and phosphorus (accounting for 50.38%—60.96%) were in the sediments. Under different models, there were differences in sizes, survival rate, specific growth rate, yield and output value of the cultured organisms, in which the maximal benefits were observed in CSR3 and CSR4 groups. The nitrogen utilization rates of each group ranged from 12.35% to 32.66%, and CSR5>CSR3, CSR4>CSR1, CSR2>CS, showed significant differences (P<0.05). The phosphorus utilization rates were ranged from 9.38% to 23.10%, CSR3, CSR4, CSR5>CS, CSR1, CSR2, with significant differences (P<0.05), indicating that the addition of an appropriate amount of razor clam in the polyculture system of shrimp and crab improved the breeding efficiency and the utilization rates of nitrogen and phosphorus significantly. In conclusion, CSR3 treatment group was the most reasonable, followed by CSR4. Under the conditions of this experiment, the optimal collocation was crab at 35 individuals/25 m2, shrimp at 300 individuals/25 m2 and razor clam at 1000 individuals/25 m2.
2021 Vol. 40 (4): 483-491 [Abstract] ( 194 ) HTML (1 KB)  PDF (1099 KB)  ( 197 )
492 Enrichment and Release of Inorganic Arsenic in Edible Parts of Swimming Crab Portunus trituberculatus
ZHAO Yanfang, NING Jinsong, KANG Xuming, DING Haiyan, ZHAI Yuxiu, SHANG Derong, SHENG Xiaofeng
DOI: 10.16378/j.cnki.1003-1111.19175
Mature female swimming crab Portunus trituberculatus with carapace width of (140.02±15.14) mm after temporary held were reared in a 500 L polyethylene aquarium, and exposed to 0.01 mg/L trivalent inorganic arsenic and 0.10 mg/L pentavalent inorganic arsenic prepared by mixed sodium arsenate and arsenic trioxide stock solution. The mature female swimming crab in the control group was exposed to clean and filtered fresh seawater with background inorganic arsenic content of (1.18±0.16) μg/L, and without inorganic arsenic addition. In the semi-static exposure 7-day enrichment test, the water was changed every 24 hours, and 3 samples of swimming crab were randomly taken on the 1, 3, 5, and 7 days, and the hepatopancreas, gonad and muscle tissues were immediately dissected and stored at -80 ℃ ultra-low temperature refrigerator for determination of the contents of total arsenic in 3 edible parts. In the 28-day release test, the swimming crab was exposed to natural sea water completely changed every 24 hours and 3 crab individuals were randomly sampled on the 7, 14, 21, and 28 days for determination of the contents of total arsenic in the 3 edible parts above in order to explore the accumulation and releasing of different inorganic arsenic in the edible tissues. Results showed that the bioaccumulation of inorganic arsenic was low in the three edible tissues, first increase in the content of total arsenic and then decrease in the hepatopancreas during the accumulation, without obvious change in the content of total arsenic in the muscle and gonad. During the releasing period, there was decrease in total arsenic content in the gonad and hepatopancreas, without obvious change in total arsenic content in the muscle. There were different main forms of arsenic in the three edible tissues of swimming crab in the blank control group, with arsenobetaine AsB and dimethylarsinate DMA forms in the muscle and gonad, and AsB in the hepatopancreas in the control group. The findings indicated that swimming crab had higher bioavailability of As(Ⅲ)than As(Ⅴ), and that the inorganic arsenic was mainly accumulated in the hepatopancreas, and not in the muscle and gonad.
2021 Vol. 40 (4): 492-500 [Abstract] ( 188 ) HTML (1 KB)  PDF (1359 KB)  ( 150 )
501 Landmark-Based Morphometric Identification of Mitten Crab Eriocheir from Suifen River
JIANG Xiaodong, CHENG Yongxu, YANG Yuhong, WU Xugan
DOI: 10.16378/j.cnki.1003-1111.20016
This study was conducted to compare the morphological characters of wild mitten crab Eriocheir collected from natural waters in Mudanjiang City, Heilongjiang Province, Zhenjiang City, Jiangsu Province, Beihai City, Guangxi Province, and Hyogo Prefecture in Japan, 15 male and 15 female samples at each sampling site, including Suifen River (SF) population from September to November 2017 and October 2018 with the wild Chinese mitten crab Eriocheir sinensis in Changjiang River (YZ) as well as the wild Japanese mitten crab E. japonicus in the Hepu Basin (HP) and the Hyogo of Japan (JP) in order to explore the current status of the wild mitten crab Eriocheir population in the Suifen River Basin. (1) The principal component analysis showed that SF and YZ individuals were slightly overlapped and divided into an independent group by the first two principal components, and that HP and JP individuals were divided into another independent group. (2) The thin-plate splines showed that morphological differences between populations mainly focused on frontal and lateral spines. (3) The stepwise discriminant analysis revealed that discriminative accuracies of SF and YZ individuals were 87%—93% and that the discriminative accuracy of HP and JP individuals was both 100%. (4) In the cluster analysis, SF and YZ individuals were clustered firstly, whereas HP and JP individuals formed another distinct cluster. In conclusion, the wild mitten crab in the Suifen River had closer morphological characters to the Chinese mitten crab based on landmark-based morphometrics analysis, but it still needs to be confirmed by molecular biological technique.
2021 Vol. 40 (4): 501-507 [Abstract] ( 170 ) HTML (1 KB)  PDF (3288 KB)  ( 180 )
508 Effect of Acute High Temperature Stress on Tissue Damage and HSPs Gene Expression of Largemouth Bass Micropterus salmoides “Youlu No.3”
LU Jian, ZHANG Jiajia, ZHOU Guoqin, WANG Peipei, QING Hui
DOI: 10.16378/j.cnki.1003-1111.20095
Juvenile largemouth bass Micropterus salmoides “Youlu No.3” with body weight of (9.87±1.2) g held at 25 ℃ (control group) was directly exposed to the water environment at 28, 31, 34, and 37 ℃ and sampled at 0 h, 6 h, and 48 h to investigate the effects of temperature on structural damage of liver and gill tissues and expression of HSPs gene in liver and gill of largemouth bass “Youlu No.3”. The results showed that no differences in structure of the liver and gill tissues were observed in the 28 ℃ and 31 ℃ groups between the treated and control largemouth bass “Youlu No.3”. However, there was slight cell edema in liver and gill tissues at 34 ℃, and the liver cells showed increase in vesicles structure, the messy and blurred, and the nucleus dissolved boundaries between cells at 37 ℃. Meanwhile, the gill lamellae became overall edema and severely bending, some swollen gill lamellae due to too many red blood cells. The expression levels of HSPs gene were found to be varied with changes in different water temperatures. At 6 h, the mRNA expression levels of HSP70 and HSC70 in liver and gill tissues were increased with the increase in stress temperature, with the maximal expression levels of HSP70 and HSC70 at 37 ℃. At 48 h, the mRNA expression levels of HSP70 and HSC70 in liver and gill tissues were decreased to some extent compared to that at 6 h (except for HSP70 at 31 ℃ gill tissue). The mRNA expression levels of HSP70 and HSC70 in liver and gill tissues showed a trend of first increasing and then decreasing, with the minimal level in the 37 ℃ group. The findings indicated that high temperature stress led to some stress damage to largemouth bass “Youlu No.3”. In aquaculture practices, attention should be paid to changes in temperature to reduce the impact of temperature stress on the immune function of fish.
2021 Vol. 40 (4): 508-515 [Abstract] ( 200 ) HTML (1 KB)  PDF (3580 KB)  ( 300 )
516 Biochemical and Enzymatic Characteristics of Phenoloxidase in Sea Cucumber Apostichopus japonicus
XIAO Yao, JIANG Jingwei, LI Shilei, DONG Ying, ZHOU Zunchun
DOI: 10.16378/j.cnki.1003-1111.20021
In order to understand the biochemical and enzymatic characteristics of phenoloxidase (PO) in the coelomocytes of sea cucumber Apostichopus japonicus, the coelomocytes of females and males were used as samples, and then using the method of linear-gradient native-PAGE combined with catechol staining, the four kinds of POs were purified and characterized, and the biochemical and enzymatic properties of different POs were determined by spectrophotometry. The optimum pH for the activity was 9.0 in AjPO4, 8.0 in AjPO5, 9.0 in AjPO6 and 8.0 in AjPO7. The activities of AjPO4, AjPO5 and AjPO6 in boiling water were stable, indicating that the heat resistance of AjPO4, AjPO5 and AjPO6 was better than that of AjPO7. The four POs catalyzed catechol, L-dopa, dopamine and hydroquinone, among which the optimal substrate was catechol, but failed to catalyze tyrosine, indicating that the four POs belonged to laccase-type phenoloxidase, and had the highest affinity to catechol.The four AjPOs were activated by Mn2+ and Fe2+, indicating that the function of PO system in sea cucumber was regulated by the divalent metal ions in vivo. In addition, the activity of the four POs was inhibited not only by citric acid, sodium sulfite and ascorbic acid, but also by sodium diethyl dithiocarbamate (DETC), a copper chelating agent, suggesting that all the four POs be copper-containing enzymes. The enzymology characteristics of AjPOs provide references for further understanding of the characteristics and functional mechanism of AjPO system.
2021 Vol. 40 (4): 516-522 [Abstract] ( 208 ) HTML (1 KB)  PDF (5968 KB)  ( 337 )
523 Distribution of Sex Steroids in Tissues of Adult Yesso Scallop Patinopecten yessoensis
GUO Zhenyi, XIE Xinran, LI Yajuan, LIU Liangjie, ZHANG Lijing, WU Shaoxuan, ZHANG Lingling, WANG Shi
DOI: 10.16378/j.cnki.1003-1111.19259
Sex steroid hormones play an important role in sex differentiation, reproduction and development of animals. However, it is still unclear whether there are the synthesis, transformation, and transportation of sex steroid hormones in mollusks. In this study, we used enzyme-linked immunosorbent assay (ELISA) to analyze the distribution of progesterone, testosterone and estradiol in tissues of adult female and male Yesso scallop Patinopecten yessoensis. Results showed that progesterone accounted for about 80% of total hormone content in hepatopancreas, significantly higher than that in the other tissues, suggesting that progesterone may be synthesized or absorbed in hepatopancreas; the maximal level of steroid hormones was observed in the hemolymph, suggesting that steroid hormones may be accumulated in hemolymph; the levels of the three hormones were significantly positively correlated, indicating potential conversion of the three hormones in the scallop. No much difference in the hormone levels was found between male and female scallops in most tissues, but estradiol/testosterone was significantly higher in females than that in the gonad and hepatopancreas in males, indicating that the ability to produce estradiol was higher in the female gonads and hepatopancreas. The finding provides an important reference for further understanding of the synthesis and transformation of sex steroid hormones in mollusks.
2021 Vol. 40 (4): 523-529 [Abstract] ( 180 ) HTML (1 KB)  PDF (1431 KB)  ( 185 )
530 Morphometric Differences between Mandarinfish Siniperca chuatsi and Bigeye Mandarinfish S. kneri and their Intermediate Form in the Middle Reaches of the Changjiang River
ZHU Shuqin, ZHAO Jinliang, ZHOU Yunhong, WU Hongyan
DOI: 10.16378/j.cnki.1003-1111.19298
In order to identify and distinguish the “intermediate form” samples collected from Poyang Lake and Dongting Lake, which have the characteristics of both mandarinfish Siniperca chuatsi and bigeye mandarinfish S. kneri, the countable characteristics(numbers of fins and gill rakes) and morphological framework characteristics of “intermediate form”, mandarinfish and bigeye mandarinfish were compared and determined, and analysis of variance, principal component analysis, discriminant analysis, and STRUCTURE cluster analysis were performed. The results showed that: (1) the intermediate form was between mandarinfish and bigeye mandarinfish, in terms of eye size, relative position of the posterior edge of the maxilla to the posterior edge of the eye, and the bulge degree of the head and back. (2) There were no significant differences in the countable traits (number of fin ray, and number of gill raker) between the intermediate form and mandarinfish and bigeye mandarinfish. (3) Analysis of variance showed that in quantitative traits, the difference between the intermediate form and mandarinfish and bigeye mandarinfish mainly lied on: the relative position of the maxillary bone to the eye, the eye diameter, the protruding degree of the head and back, and the length of the caudal peduncle; principal component analysis showed some frame traits of the back of the head and the anterior part of the back of the head and body height were the main components of morphometric difference between the intermediate form and mandarinfish and bigeye mandarinfish. The discriminant accuracy of mandarinfish, bigeye mandarinfish and intermediate form was 90%, 81.8% and 85.4%, respectively, with the average discriminatory accuracy of 85.6%. STRUCTURE cluster analysis showed that the morphometric characteristics of the intermediate form were between that of mandarinfish and bigeye mandarinfish, and inclined towards mandarinfish. The findings can provide some basic data for understanding of intraspecific variation or interspecific hybridization of mandarin fishes in the Changjiang River.
2021 Vol. 40 (4): 530-536 [Abstract] ( 268 ) HTML (1 KB)  PDF (1976 KB)  ( 207 )
537 Investigation and Analysis on Death of Marine Life in Local Area of Dalian Shallow Sea in Spring 2020
WU Jinhao, SONG Guangjun, SONG Lun, WANG Bai, ZHANG Yimin, YIN Minghao, WANG Zhaohui, ZHOU Zunchun
DOI: 10.16378/j.cnki.1003-1111.20041
The field investigation and sample collection were carried out after the death of marine life in Dalian shallow sea area. Through the statistics and analysis of species identification, the causes of death of marine life were discussed from several aspects. The results showed that there were 52 species of marine organisms in 10 categories to be collected from 4 locations, showing mollusks (16 species)> arthropods (15 species)> annelids (7 species)> echinoderms (5 species)> chordates (4 species)> other phyla (1 species each). There were 38 dead species, among which the death amount of Protankyra bidentata, Upogebia major and Solen stritus was particularly obvious. According to the analysis, water pollution, disease and low oxygen level were unlikely to lead to the massive death of marine organisms. The extreme sea conditions under strong wind conditions had changed the habitat environment of marine organisms, making them unable to escape. The joint action of large-scale cooling and snowfall in a short time led to the occurrence of the disaster event. The findings can provide a technical reference for the government to carry out the prevention and treatment of similar sudden ecological environment disasters.
2021 Vol. 40 (4): 537-543 [Abstract] ( 153 ) HTML (1 KB)  PDF (1750 KB)  ( 312 )
544 Inhibition of Black Tea and Green Tea Treated by Different Methods to Alga Alexandrium tamarense
CHEN Xuehao, WU Yuqiang, YANG Zhi, ZHOU Lihong
DOI: 10.16378/j.cnki.1003-1111.20042
Harmful algae bloom will threaten the balance of marine ecosystem, and it is of great significance to find algal inhibiting materials to control red tide. The short-term and long-term inhibitory effects of black tea and green tea on toxic algae Alexandrium tamarense were studied by reasonable application methods including comparing the algal inhibition effect of dry tea powder, water extract and tea residue. The results showed that the 5 h inhibition rates (IRs) of 1 mg/mL tea extract were very high [(99.45±0.39)% (green tea) and (100.00±0.00)% (black tea)], and 1h-IC50 of tea extracts were 0.85 mg/mL green tea and 0.87 mg/mL black tea, respectively. Green tea residues had stronger algal inhibition than black tea residues did, and the lower the concentration was, the greater the difference of the algal inhibition was; the maximal IRs of 0.80 mg/mL green tea and black tea residue were (94.56±1.03)% and (22.17±1.66)%, respectively; 1h-IC50 of green tea and black tea residues were 3.39 mg/mL and 8.96 mg/mL, respectively. The high algal inhibition rate (green tea powder≥(94.23±0.64)%, black tea powder ≥(88.31±2.35)%) of 1 mg/mL tea powder was kept from 24 to 120 hours, and the IR of green tea powder was higher than that of black tea powder before 24 hours, without significant difference between them after 48 hours. In terms of the 72-hour average IR for each concentration, the average IR of green tea extract was 1.95—3.72 times as that of black tea extracts at a concentrations of 0.63 mg/mL or less; the average IR of green tea residue was 1.19—4.24 times as that of black tea residue at concentration between 0.80 mg/mL and 8.00 mg/mL. It was found that green tea had stronger inhibitory effect on A. tamarense than black tea did, and green tea residue was a promising algal inhibiting material.
2021 Vol. 40 (4): 544-552 [Abstract] ( 137 ) HTML (1 KB)  PDF (2384 KB)  ( 151 )
553 Eye Development-Related Genes Involved in Embryonic Development Period of Japanese Spineless Cuttlefish Sepiella japonica
LI Shaogang, LI Yongqin, XU Lele, CHEN Daohai, WANG Liyun
DOI: 10.16378/j.cnki.1003-1111.20052
Genes of SIX Homeonox 3 (six3), Eyes Absent (eya), and Paired Box 6 (pax6) involved in the eye development were selected from the transcriptome database established from Japanese spineless cuttlefish Sepiella japonica embryonic stage by Illumina HiSeqTM 4000 high-throughput sequencing. The relative transcriptive expression level of these three genes in three embryo periods (eye primordium formation period, functional organ differentiation period, and larval period), and eight stages of Japanese spineless cuttlefish were detected by the RT real-time PCR. The results showed that the expression level of eye development-related genes of Japanese spineless cuttlefish was different during the three embryonic development periods. The relative transcriptive level of six3 and eya genes was increased significantly from the eye primordium formation period to the larval period. However, the relative transcriptive level of pax6 gene was increased from the eye primordium formation period to the functional organ differentiation period, and subsequently decreased gradually till to the larval period. It is proposed that six3, eya, and pax6 genes play different regulatory functions in the retinal determination gene network network, and have important significance for regulation of the eye development in embryo of Japanese spineless cuttlefish.
2021 Vol. 40 (4): 553-560 [Abstract] ( 148 ) HTML (1 KB)  PDF (1918 KB)  ( 236 )
561 Antimicrobial Susceptibility in Escherichia coli Isolated from Fishery Products in Zhanjiang
DING Xiuqiong, HUANG Wu, LIU Jianfang, LEI Xiaoling, MAI Jiahui, HUANG Jianyong, NIE Fanghong
DOI: 10.16378/j.cnki.1003-1111.20013
Antimicrobial susceptibility of 52 susceptible Escherichia coli strains isolated from fresh samples such as oyster Crassostrea ariakensis, green mussel Perna viridis, Manila clam Ruditapes philippinarum, hard clam Meretrix meretrix, and scallop Mimachlamys nobilis, and frozen samples of fish fillets and cooked shrimp to 19 antibacterial drugs including penicillins, quinolones, monocyclic lactams and sulfonamides by using K-B paper diffusion method, and then extended-spectrum β-lactamase (ESBLs) and plasmid-mediated quinolone resistance (PMQR) genes were amplified by PCR to detect the resistance of E. coli in fishery products in Zhanjiang. Results showed that all of the 52 E. coli strains were resistant to more than one antibiotic, with higher resistance rate of erythromycin(90.4%), sulfamethoxazolethe(55.8%), trimethoprim (53.8%), and cefotaxime (51.9%) than that in other antibiotics. About 76.9% of E. coli showed multiple resistant of up to 9 categories of antibiotics, and 10 strains were confirmed to be ESBLs bacteria,with detection rate of 19.2% (10/52), 8 strains carrying gene blaTEM, 7 strains carrying gene blaSHV, without blaCTX and blaOXA genes. It was confirmed that 15 strains were PMQR bacteria, with detection rate of 28.8% (15/52), the all carrying qnrA, aac (6′)-Ib-c and qepA genes, 9 strains carrying qnrS gene, 8 strains carrying qnrD gene, and only 1 strain carrying oqxB gene, without qnrB, qnrC and oqxA genes. Both ESBLs gene and PMQR gene were carried by 2 E. coli isolates. The findings revealed that E. coli from fishery products in Zhanjian area was resistant to a wide range of antibiotics and that combination of ESBLs gene with PMQR gene enhanced the drug resistance of bacteria.
2021 Vol. 40 (4): 561-570 [Abstract] ( 148 ) HTML (1 KB)  PDF (1067 KB)  ( 268 )
571 Safety Assessment and Acute Toxicity of Four Kinds of Heavy Metal Ions to Manila Clam Ruditapes philippinarum
CHEN Zhi, WEI Chengchen, HU Fengxiao, HUANG Jian, LI Weipeng, LIAN Chenyang
DOI: 10.16378/j.cnki.1003-1111.20075
The acute toxic effects of four kinds of heavy metal ions on Manila clam Ruditapes philippinarum were investigated by toxicology experiment, in order to evaluate the tolerance of Manila clam to Cd2+, Mn2+, Zn2+ and Cu2+. The results showed that the mortality of Manila clam was positively proportional to the concentration of heavy metal ions, with significant dose-response relationship. The medium lethal concentration (LC50) values of Cd2+ treatment was 38.62 mg/L for 24 h, 29.83 mg/L for 48 h, 8.11 mg/L for 72 h, and 2.67 mg/L for 96 h with safe concentration of 0.03 mg/L, whereas the LC50 of Mn2+ was 712.58 mg/L for 24 h, 421.57 mg/L for 48 h,188.74 mg/L for 72 h, and 61.27 mg/L for 96 h with safe concentration of 0.61 mg/L. The LC50 of Zn2+ was 141.27 mg/L for 24 h,72.20 mg/L for 48 h, 34.91 mg/L for 72 h and 21.57 mg/L for 96 h with safe concentration of 0.22 mg/L. The LC50 of Cu2+ was 29.88 mg/L for 24 h, 12.06 mg/L for 48 h,3.62 mg/L for 72 h and 0.79 mg/L for 96 h with safe concentration of 0.01 mg/L. The results showed that the toxicity of four heavy metal ions to Manila clam was in the order of Cu2+>Cd2+>Zn2+>Mn2+, the lower safe concentration of Cu2+ than the standard of fishery water quality, so special attention should be paid to nursery and breeding processes.
2021 Vol. 40 (4): 571-576 [Abstract] ( 165 ) HTML (1 KB)  PDF (1487 KB)  ( 180 )
577 Effects of Different Intensive Grass Carp Culture Modes on Aeromonas Community
XU Xiandong, FU Huiyun, OUYANG Min, WAN Guoyuan, RAO Yi, HUANG Jiangfeng, ZHOU Zhiyong, LIU Lihui
DOI: 10.16378/j.cnki.1003-1111.20061
The abundance and diversity of Aeromonas bacteria were investigated in water samples collected at 15 cm of the surface at both ends and the middle of the pond, and sediment samples in three culture modes of grass carp Ctenopharyngodon idellus ponds including intensive formulated feed pond, ryegrass and other forages mountain pond, and grass carp, fish and vegetable symbiosis (aquaponic) pond from May to July 2017 (the period of vigorous breeding and feeding) by RS plate separation method and by 16S rDNA sequencing. The results showed that a total of 54 strains of bacteria were isolated, and divided into 10 genera and 22 species, with 23 Aeromonas strains, 8 Pseudomonas strains, 6 Citrobacter strains, 7 Enterobacter strains, 3 Pantoea strains and 3 Serratia strains. There was 1 strain of bacteria in genera such as Pectinobacterium, Salmonella, Kluyverella and Leclercia, with the maximal number of Aeromonas veronii (12 strains), followed by A. hydrophila (10 strains). There were 6 strains of E. tabaci, 5 strains of C. freundii, and 1 or 2 of other strains. The analysis of bacterial species and quantity in different modes revealed that there was the most bacterial species and quantity (12 species and 21 strains), and the least pathogenic bacteria in the aquaponics system. The least bacterial species (5 species and 13 strains) and higher proportion of pathogenic bacteria was observed in the intensive grass carp culture pond with commercial feed. The distribution characteristics of bacteria showed that A. veronii was mostly distributed in water, and A. hydrophila was mostly found in sediment.
2021 Vol. 40 (4): 577-582 [Abstract] ( 158 ) HTML (1 KB)  PDF (3083 KB)  ( 157 )
583 Analysis of Gastric Evacuation Characteristics and Associated Mathematical Models in Juvenile Megalobrama pellegrini
QU Huantao, LIAO Jianxin, DAI Wei, LIU Yong, LU Xuebao, YANG Yuanjin
DOI: 10.16378/j.cnki.1003-1111.19160
Gastric evacuation rate was determined for juvenile Megalobrama pellegrini, an endangered endemic fish species mainly found in the upper Yangtze River basin in China, and 300 experimental fish with boby weight of (38.17±2.45) g were removed from various intervals (0, 2, 4, 6, 8, 12, 16, 20, and 24 h) after satiation feeding at (15.0±0.3) ℃. The wet mass of the chyme was determined, and three mathematical models were used to get the best fitting data of gastric evacuation in juvenile M. pellegrini. By further comparing the residual squares (RSS) and the residual standard deviation (SDR), the square root model was shown to be the best model for expression gastric evacuation rate in this fish. Its regression equations for the wet mass of the chyme was expressed as y0.5=0.951-0.036t (r2=0.899, P<0.001). According to the square root model, the gastric evacuation rate of juvenile M. pellegrini was 0.036 g/h, and 80% of the theoretical gastric emptying time was 14.83 h. Based on the above data, the optimal feeding interval could be 15 h for juvenile M. pellegrini at (15.0±0.3) ℃.
2021 Vol. 40 (4): 583-588 [Abstract] ( 161 ) HTML (1 KB)  PDF (1094 KB)  ( 181 )
589 Molecular Detection and Early Warning of Streptococcus agalactiae and S. iniae
CUI Miao, WU Min, LIU Ru, LI Jingjing, ZHANG Huijie, XU Delin, ZHANG Qizhong
DOI: 10.16378/j.cnki.1003-1111.19191
Two pairs of specific primers designed according to the 16S rRNA partial gene sequence of Streptococcus agalactiae and the Sip gene sequence of S. iniae, respectively, were applied to establish a rapid and sensitive PCR detection system, aiming to detect trivial amounts of S. agalactiae and S. iniae simultaneously. Two distinct DNA fragments of 870 bp and 614 bp covering the 16S rRNA partial gene sequence and the Sip gene sequence were amplified from the genomic DNA of S. agalactiae and S. iniae, respectively. No cross-reactivity was observed between the two Streptococcus species or with other fish/nonfish pathogens, including Edwards ellatarda, Aeromonas veronii, Vibrio vulnificus, A. sobria, V. alginolyticus, and Escherichia coli. The two primer pairs were capable of detecting genomic DNA of S. agalactiae and S. iniae in concentrations as low as 9.84×10-5 ng/μL and 9.30×10-5 ng/μL, respectively, and of detecting the bacterial cells of S. agalactiae and S. iniae in amount as low as 2.76×103 cfu/mL and 2.51×103 cfu/mL. The duplex PCR designed in this study had strong specificity, high sensitivity, high detection rate, and was used for rapid identification of lower amount of S. agalactiae and S. iniae. The finding provides a valuable tool for early warning detection of the pathogens leading to streptococosis.
2021 Vol. 40 (4): 589-595 [Abstract] ( 185 ) HTML (1 KB)  PDF (1852 KB)  ( 270 )
596 Early Development of Vertebral Column and Appendicular Skeleton in Butterfish Scatophagus argus
DENG Pingping, SHI Yonghai, XU Jiabo, JIANG Fei, YUAN Xincheng
DOI: 10.16378/j.cnki.1003-1111.20015
The early development of vertebral column and appendicular skeleton was observed in butterfish Scatophagus argus from newly hatching larvae through 45 days post-hatching (dph) by double staining technique under a stereo microscope. It was observed that the notochord was a cone without segmentation from 1 to 8 dph, and segmented untill 8 dph. Segmented hard bone rings appeared at 28 dph, and 22—24 vertebrae were formed in the spine at 38 dph. The neural arches and haemal arches were appearance at 11 dph marked by the beginning of the vertebral column development. Some bone rings appeared clearly at 28 dph. Vertebral column was ossified from anterior to posterior, and then completely ossified at 38 dph. There were 10—12 cervical vertebra segments in trunk vertebra, and 11—12 cervical vertebra segments in the caudal vertebra. The appendicular skeletons were developed from the pectoral fins, ventral fins, caudal fins, anal fins to dorsal fins. The hypuralat was observed in the caudal fin at 11 dph, and appendicular pectoral fins were formed at 5 dph, pectoral girdle and cleithrum were ossified at 38 dph. Cartilages of appendicular of the ventral fins were formed at 5 dph, and the ventral fins were completely ossified at 33 dph. The caudal fin was completely ossified at 41 dph. and cartilages of appendicular anal fins were formed at 13 dph. The frontal fin ray of dorsal fins ws transformed 4 spines at 22 dph, 10—11 dorsal spines at 21 dph, and the dorsal fins began to ossify at 38 dph. The development of the dorsal fins began with appearance of 9 hyaloid in pterygiophores at 13 dph, and began to ossify at 33 dph. The butterfish had upturned notochord at 15 dph, with capacity of active feeding, becoming juveniles at water temperature of 25.5—29.1 ℃ and a salinity from 25 to 1.7. The butterfish had ctenoid scales at 20 dph, and moved from the gradually desalted sea water to fresh water, when the first inverted dorsal spine, number of gluteal spines and the presence of pectineal scales were significant markers for the classification of Scatophagidae in the Perciformes. The research on early development of the vertebral column and the appendicular skeleton in butterfish S. argus is important for functional adaptation during early development environment, production operation and classification.
2021 Vol. 40 (4): 596-602 [Abstract] ( 162 ) HTML (1 KB)  PDF (3839 KB)  ( 293 )
603 Identification of a Pathogenic Aeromonas media Isolated from Schizopygopsis younghusbandi
ZENG Benhe, YANG Chengnian, QIU Yulin, WANG Jinlin, WANG Wanliang, WANG Jian, ZHU Chengke
DOI: 10.16378/j.cnki.1003-1111.19157
To identify a pathogenic bacterium found in Schizopygopsis younghusbandi, a dominant pathogenic strain named LS-01 was isolated from the liver of the dying fish, and were identified according to gram staining, physiological, biochemical and molecular biological methods, and drug resistance was carried out meanwhile. The results showed that strain LS-01 was a short rod-shaped gram-negative bacterium and the physiological analysis based on 16S rRNA and gyrB gene revealed that the strain LS-01 was similarity of up to 99% with Aeromonas media. Artificial infection experiments showed that the strain LS-01 had median lethal concentration of 3.98×106 cfu/mL. In addition, drug sensitivity test showed that A. media LS-01 was highly sensitive to 12 drugs, such as ceftazidine, cefoperazone, levofloxacin and florfenicol.
2021 Vol. 40 (4): 603-609 [Abstract] ( 150 ) HTML (1 KB)  PDF (1500 KB)  ( 166 )
610 Screening and Identification of a Marine Actinomycete with Antibacterial and Ammonia-Nitrogen Degradation Functions
TANG Manli, LI Shuxin, PENG Yun, CHEN Yingying, WANG Yanan, MA Guizhen, BAO Zenghai
DOI: 10.16378/j.cnki.1003-1111.20044
Using Vibrio parahemolyticus and V. vulculus as indicator bacteria, the bacteriostatic activity of 37 strains of marine actinomyces in sterile fermentation broth was determined by Oxford cup method, and the bacteriostatic activity of the strains was screened out and their bacteriostatic spectra were determined. The strains with strong bacteriostatic activity were inoculated into the culture medium with a mass concentration of ammonia nitrogen of 50 mg/L and fermented for 7 d. The concentration of ammonia nitrogen in the fermentation broth of different strains was determined by indophenol blue spectrophotometry, and the excellent strains of actinomycetes with functions of bacteriostatic and degrading ammonia nitrogen were screened out. The selected strains were identified through morphological observation, physiological and biochemical test and 16S rDNA sequence analysis. The results showed that 15 strains of actinomycetes had inhibitory effects on both V. parahaemolyticus and V. vulnificus, in which strain F28 had the strongest inhibitory effect on V. parahaemolyticus and V. vulnificus, with diameter of the inhibitory ring of (31.57±0.63) mm and (18.93±0.92) mm, respectively. In addition, the strain had a strong inhibitory effect on all 10 pathogenic bacteria tested, with a wide inhibitory spectrum. Strain F28 had strong ammonia nitrogen degradation ability with degradation rate of 46.86%. Combined with the results of bacteriostatic activity, it was concluded that strain F28 as Streptomyces mediolani had both bacteriostatic and ammonia-nitrogen degradation functions, and potential development and application prospects.
2021 Vol. 40 (4): 610-617 [Abstract] ( 152 ) HTML (1 KB)  PDF (3106 KB)  ( 347 )
618 Molecular Cloning, Partial Sequence and Tissue Expression of Dmrt1 Gene in Siamese Fighting Fish Betta splendens
HUANG Yang, LI Jinhai, WANG Wenji, GUO Yuwen, LI Guangli, CHEN Huapu
DOI: 10.16378/j.cnki.1003-1111.19238
Doublesex and Mab-3 related transcription factor 1 (Dmrt1), a member of DMRT family, plays critical roles in sex determination, sex differentiation and gonadal development. The molecular cloning of a partial sequence of Dmrt1 was performed in the brain tissue of Siamese fighting fish Betta splendens by RACE, and Dmrt1 gene expression pattern was detected in all tissues and embryonic development of Siamese fighting fish by semi-quantitative RT-PCR and real-time quantitative RT-PCR. Siamese fighting fish had Dmrt1 consisting of 1037 nucleotides, with a 798 bp length of open reading frame (ORF), and coding 265 amino-acids. Phylogenetic analysis showed that Dmrt1 in Siamese fighting fish was more closely related to climbing perch Anabas testudineus. In addition, tissue-specific and sexual dimorphism expression profile of Dmrt1 was detected in Siamese fighting fish. In males, Dmrt1 was expressed in the testis and spleen exclusively. In females, however, Dmrt1 was highly expressed in the heart, weakly expressed in the brain, pituitary, spleen, muscle, kidney and intestine, without expression in ovary and liver. Moreover, during the period of embryonic development, Dmrt1 was predominately expressed in the fertilized eggs, and weakly from the gastrula stage to heart beating stage. The findings indicate that Dmrt1 may play an important role in male germ cell development, and reproductive regulation in spermatocyte maturation in Siamese fighting fish.
2021 Vol. 40 (4): 618-623 [Abstract] ( 173 ) HTML (1 KB)  PDF (2574 KB)  ( 473 )
       Overview and Specific Topi
624 Advances on Immunological Mechanisms of Important Immune Organs against Pathogenic Microorganisms in Fish: a Review
LUO Zhiwen, DONG Zhixiang, LIN Lianbing, ZHANG Qilin
DOI: 10.16378/j.cnki.1003-1111.19236
2021 Vol. 40 (4): 624-634 [Abstract] ( 294 ) HTML (1 KB)  PDF (1904 KB)  ( 424 )
635 Research Perspectives of Oral Vaccine Delivery System in Fisheries: a Review
YUAN Gaoliang, ZHU Lei, KONG Xianghui
DOI: 10.16378/j.cnki.1003-1111.19288
2021 Vol. 40 (4): 635-642 [Abstract] ( 226 ) HTML (1 KB)  PDF (1051 KB)  ( 191 )
643 A Review of Research Advancement on Fisheries Biology of Japanese Spanish Mackerel Scomberomorus niphonius
QU Junyue, YANG Guangmingmei, FANG Zhou, CHEN Xinjun
DOI: 10.16378/j.cnki.1003-1111.19184
2021 Vol. 40 (4): 643-650 [Abstract] ( 235 ) HTML (1 KB)  PDF (2254 KB)  ( 373 )
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