Volume 31,2024 Issue 12

研究论文

• Cloning and expression characterization analysis of nanos2 gene in Ctenopharyngodon idella and Squaliobarbus curriculus

  HUANG Ling, YE Huan, YUE Huamei, HOU Yanling, QU Ziling, RUAN Rui, LI Chuangju

  2024,31(12):1411-1421, DOI: 10.12264/JFSC2024-0254

  Abstract: (124) HTML (205) PDF (8.66 M)(129)

  Abstract:

  The grass carp (Ctenopharyngodon idella) represents an economically important fish species in freshwater aquaculture within China. However, the urgency to develop superior varieties has become paramount due to significant germplasm degeneration in grass carp. Germline stem cell transplantation emerges as a promising and potent technique for reducing the sexual maturity cycle in fish. Consequently, our objective was to shorten the generation interval for obtain functional gametes of grass carp by employing barbel chub (Squaliobarbus curriculus) as a surrogate host. The accurate identification of germ cells between donor and recipient species is a critical step in the successful establishment of germline stem cell transplantation. However, the method of identification of germ cells in grass carp and barbel chub is not clear. In the present study, the full-length cDNA of reproduction-related gene nanos2 were cloned from grass carp and barbel chub, which are 649 and 636 base pairs, encoding 145 and 144 amino acids, respectively. Sequence analysis revealed that the grass carp Nanos 2 (CiNanos2) amino acid sequence exhibits a high degree of sequence identity to that of barbel chub (91.67%), and to that of zebrafish (Danio rerio) (65.49%). Phylogenetic tree analysis showed that grass carp is clustered together with barbel chub, indicating the closest genetic relationship between them. The expression of nanos2 transcripts in grass carp and barbel chub was predominantly observed in the gonads, with significantly higher levels detected in the testis compared to the ovary, suggesting that nanos2 might plays a crucial role in the development of gonad. Species-specific and common primers were designed based on the alignment of nanos2 sequences between grass carp and barbel chub for PCR analysis. The results demonstrated that the grass carp-specific primers had exclusively target product (179 bp) in the gonad of grass carp, while the barbel chub-specific primers had only expected product (265 bp) in the gonad of barbel chub gonads. Additionally, the common primers were able to amplify indiscriminately in the gonads of both grass carp and barbel chub, producing a target product of 251 bp, implying that the germ cells of grass carp and barbel chub could be efficiently distinguished via species-specific primers by PCR. Our study laid the foundation for further investigating the mechanism of the nanos2 gene in gonad development of grass carp and barbel chub. Meanwhile, it provided an effective method for monitoring the chimerism and development of grass carp germ cells in the gonads of barbel chub.

• Transcriptome analysis of heat tolerance-related biological processes and pathways during heat-hardening in Mytilus galloprovincialis

  ZHANG Wanliang, DONG Zhenyu, WANG Hao, WEI Bingqi, LIN Huajian, WANG Weifeng, GUO Baoying

  2024,31(12):1422-1431, DOI: 10.12264/JFSC2024-0113

  Abstract: (69) HTML (138) PDF (1.77 M)(99)

  Abstract:

  Heat hardening in Mytilus galloprovincialis is a survival strategy for enhancing thermal tolerance in response to extreme low tides during consecutive low-tide periods in spring. Transcriptome sequencing was used to analyze the gill tissues of mussels in the heat-hardening (H) and control (N) groups to uncover the mechanisms of thermal tolerance enhancement under heat hardening. The results showed that, compared to the control group, the heat-hardening group had 651 differentially expressed genes, including 159 significantly upregulated and 492 significantly downregulated genes. Gene ontology (GO) analysis revealed significant enrichment of differentially expressed genes in biological processes such as metabolism, substance transport, and signal transduction. Gene set enrichment analysis (GSEA) of significantly enriched items in the GO analysis revealed the upregulation of biological processes related to macromolecule metabolism, nucleic acid metabolism, and metabolism of compounds containing nuclear bases, which are associated with macromolecule metabolism, substance metabolism, and energy metabolism. Kyoto encyclopedia of genes and genomes (KEGG) analysis revealed that differentially expressed genes were significantly enriched in signaling pathways related to metabolic regulation, such as Rap1, apoptosis-multiple species, and MAPK. These findings indicate that thermal hardening in Mytilus galloprovincialis may enhance tissue functional capacity by activating various macromolecular metabolic processes while also strengthening the ability of gill tissues to actively remove damaged cells, thus enabling early physiological regulation and adaptive responses to subsequent high-temperature stress, leading to better functional performance. This study explored the biological processes and metabolic pathways associated with thermal hardening in Mytilus galloprovincialis, providing a foundation for investigating the molecular mechanisms underlying heat-hardening in this species.

• Cloning and expression analysis of sequence encoding Rheb in Eriocheir sinensis

  TIAN Zhihuan, JIAO Chuanzhen

  2024,31(12):1432-1440, DOI: 10.12264/JFSC2024-0212

  Abstract: (75) HTML (168) PDF (1.90 M)(107)

  Abstract:

  Ras homologue enriched in the brain (Rheb) is an important positive regulator on the upstream of the mammalian mechanistic target of rapamycin (mTOR) signaling pathway. The cDNA sequence encoding Rheb in Eriocheir sinensis (EsRheb) was cloned using RT-PCR in order to investigate the role of this gene in the regulation of growth, development, and nutrient metabolism of Eriocheir sinensis via the mTOR signaling pathway. The sequence and structural characteristics of EsRheb were analyzed using bioinformatics. The tissue distribution of EsRheb mRNA and the effects of animal starvation and eyestalk ablation on EsRheb transcription were detected using qRT-PCR. The results showed that EsRheb cDNA encoded 182 amino acids; its amino acid sequence, conserved domain, and functional sites were highly similar to the Rheb of a wide range of species; and its three-dimensional structure was highly consistent with that of human Rheb. EsRheb mRNA expression was high in the hepatopancreas, stomach, myocardium, skeletal muscles, and Y organ, but was much lower in the eye stalk. Starvation treatment of animals significantly decreased the transcriptional expression of EsRheb in the skeletal muscle but had no significant effect on its expression in the hepatopancreas. The removal of the eye stalk significantly increased the transcriptional expression of EsRheb in the Y organs and skeletal muscles. This study provides a basis for elucidating the function of EsRheb in regulating the mTOR-mediated growth and metabolism of E. sinensis.

• Effects of hypoxia stress on hematological physiology and biochemistry, gill histiology and hepatic antioxidant capacity of black scraper Thamnaconus septentrionalis

  LIU Zhifei, WANG Feng, CHEN Shuaiyu, CHEN Xiatian, JIA Yudong

  2024,31(12):1441-1452, DOI: 10.12264/JFSC2024-0220

  Abstract: (88) HTML (156) PDF (6.39 M)(107)

  Abstract:

  In the present study, the hypoxia tolerance and alternation of hematological, physiological, and biochemical parameters, gill histology, and hepatic antioxidant capacity subjected to hypoxia stress were investigated in black scraper Thamnaconus modestus. The closed static water method was used to analyze the dissolved oxygen (DO) levels at critical oxygen tension (P_crit) and loss of equilibrium (LOE) in black scraper. Subsequently, changes in hematological, physiological, and biochemical parameters, gill histology, and hepatic antioxidant enzyme activity were detected at P_crit, LOE, reoxygenation 3 hours (R3) and 6 hours (R6). The results showed that the DO concentrations of the black scraper [(101.23±14.28) g] at P_crit and LOE were (4.0±0.2) mg/L and (2.2±0.2) mg/L, respectively. Hypoxia significantly increased glucose and cortisol levels. White and red blood cell numbers also increased with the highest value observed at LOE (P＜0.05). The hemoglobin concentration and hematocrit showed an upward trend at LOE, but the difference was not significant (P＞0.05). Meanwhile, hypoxia significantly increased the length, width, basal epithelial thickness, and perimeter of the secondary lamellae, and decreased interlamellar distance and the proportion of available gas exchange (P＜0.05). Hypoxia stress also induced branchial clubbing, fusion, and matrix hyperplasia of the secondary lamellae. Hepatic superoxide dismutase, catalase, glutathione peroxidase activity, and malondialdehyde levels significantly increased under hypoxia (P＜0.05), peaking at LOE. Reoxygenation treatment for 6 hours alleviated the stress mentioned above effects. These results indicated that hypoxia stress lead to significantly alterations in the black scraper's hematological, physiological and biochemical profiles, gill morphology, and hepatic antioxidant capacity, which are substantially alleviated after 6 hours re-oxygenation. These findings provide valuable insights into the physiological mechanisms underlying hypoxia tolerance and support the development of efficient and healthy breeding practices for black scraper.

• Transcriptomic responses to chronic heat stress in black rockfish (Sebastes schlegelii)

  TAN Jie, ZHANG Zheng, LIU Changlin, WANG Liang, XIAO Wenjie, JIANG Da

  2024,31(12):1453-1463, DOI: 10.12264/JFSC2024-0226

  Abstract: (71) HTML (124) PDF (1.51 M)(94)

  Abstract:

  Black rockfish (Sebastes schlegelii) inhabit the coastal regions of the northwestern Pacific Ocean, including the East China Sea, Yellow Sea, and the coastal areas of Korea and Japan. It is highly favored by consumers because of its delicate flavor and high nutritional content. Black rockfish is among the most economically cultivated marine species in northern China. The juveniles were raised in offshore net cages. The optimum temperature for black rockfish growth ranges from 18 ℃ to 24 ℃. In summer, the seawater temperature sometimes exceeds 28 ℃ in black rockfish farms. As global warming intensifies, long-term exposure to high temperatures in summer will affect the growth and survival of black rockfish cultured in net cages, indicating that the effects of chronic heat stress on black rockfish merit further research. However, the molecular mechanisms underlying the responses of black rockfish to chronic heat stress remain largely unknown. Understanding these mechanisms will improve fish welfare and farm production. Marker genes to monitor heat stress are required to identify heat-resistant fish. In this study, we conducted RNA-seq analysis to characterize the genes and pathways involved in chronic thermal stress responses in the gills, liver, and intestines of black rockfish. Healthy black rockfishes were cultured at a normal temperature (18 ℃) and a high temperature (27 ℃). For the chronic heat stress treatment, water temperature was increased from 18 ℃ to 27 ℃ at a constant rate of 1 ℃ per day, and maintained for 10 days. The gill, liver, and intestinal tissues were used as experimental materials in both the heat stress and normal groups. Total RNA was extracted, and 18 mRNA libraries were constructed and sequenced using the Illumina HiSeq-4000 technology platform. Differentially expressed genes were analyzed using edgeR. Bioinformatic analysis was performed on the GO and KEGG functions of the differentially expressed genes, and the key differentially expressed genes were further validated using RT-qPCR. In total, 306 annotated differentially expressed genes (DEGs) were identified in the gill, of which 96 and 210 were up-and downregulated, respectively. In total, 806 annotated differentially expressed genes (DEGs) were identified in the liver, of which 382 were upregulated and 424 were downregulated. A total of 343 annotated differentially expressed genes (DEGs) were identified in the intestine, among which 162 and 181 were up-and downregulated, respectively. A Venn diagram showed that 12 DEGs were shared among three tissues. And 40, 53, and 49 DEGs were shared between liver and gill, liver and intestine, gill and intestine, respectively. Furthermore, GO functional enrichment analysis revealed that the DEGs were mainly enriched in proteolysis, extracellular region, structural molecule activity, receptor regulator activity, and receptor-ligand activity in the gill; in lipid metabolic process, cellular amino acid metabolic process, cytoplasm, and oxidoreductase activity in the liver; and in oxidation-reduction process, extracellular region, and cofactor binding in the intestine. Among the up-regulated genes under heat stress included the heat shock proteins 90α, period circadian protein homolog 2, serpin H1, and the down-regulated genes included period circadian protein homolog 1. Ten DEGs were subjected to reverse transcription quantitative PCR (RT-qPCR) for relative quantification to assess the differences in gene expression between the normal and high temperature groups. The expression trends observed in the RT-qPCR analysis were consistent with those identified in the RNAseq data, which confirmed the reliability of the transcriptomic sequencing results. These results provide abundant data for further studies on the molecular mechanisms of the chronic heat stress response in Sebastes schlegelii.

• Histological observation of the visual organ of Brachymystax tsinlingensis Li 1966 at the early developmental stage

  ZHANG Lin, GUO Xingchen, SONG Rongqun, WANG Zhenlu, YE Huan, XIONG Dongmei, LI Zhigang, SHAO Jian

  2024,31(12):1464-1473, DOI: 10.12264/JFSC2024-0244

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  Abstract:

  Brachymystax tsinlingensis Li (1966) is sensitive to light and susceptible to death by strong light during long-term artificial breeding, especially during early developmental stages. The development of visual organs and the characteristics of retinal development were systematically observed using histological methods to analyze the physiological causes of this phenomenon and improve the efficiency of large-scale artificial breeding. The results showed that the embryo entered the organ differentiation stage, and the optic cup was formed 177 hours post-fertilization (HPF). At 213 HPF, a primitive retina and crystal sac were formed, and the cuboidal epithelium constituted the cornea. The crystal sac had developed into the lens, the cells of the neural layer of the retina had increased notably, and the staining of the pigment cell layer deepened at 357 HPF. At 452 HPF, the sclera appeared, and the retina differentiated into four layers (pigment epithelium, outer nuclear layer, inner nuclear layer, and optic ganglion cell layer). After 508 h, the fertilized egg broke the membrane and hatched larvae. The visual organs of hatchlings were relatively well-developed, and the lens is fully differentiated and has the same structure as that of the adult; the retina has been differentiated into 8 layers, and the pigment granules in the pigment layer are clearly visible; the sclera consists of elastic fibers and cartilage; the iris is in the form of a single-layer ring; and the choroid is present. At 5 days after hatching (DAH), a small number of cone cells appeared, and the visual cell layer was pure cone cell structure. At 10 DAH, the retina differentiated into a complete 10-layer structure, ON/SC was 1.27＞1, a few optic rods began to appear in the visual cell layer, and the inner nuclear layer was differentiated into three clear layers with the initial visual imaging system. At 12 DAH, ON/G was 1.07 ＞ 1, the retinal network was less convergent, and the number of nuclei in the outer nuclear layer was comparable to the number of ganglion cells, which was sufficient to receive signals from the visual cells. At 16 DAH, the mean value of the retinal bright-adapted pigmentation index (PI) was 0.75, whereas the mean value of the dark-adapted PI was significantly reduced to 0.55 (P ＜ 0.05). At 21 DAH, the number of optic rod cells increased, and the cornea differentiated into a complete 5-layer structure; at 41 DAH, the iris differentiated, the choroid was perfected, and all parts of the visual organs were fully developed. Therefore, the retinal structure and visual characteristics of B. tsinlingensis show strong light sensitivity, and vision plays a major role in its behavior and feeding activities. Reducing environmental light appropriately during early breeding of B. tsinlingensis could reduce the stress response and improve the survival rate of seedlings. The results of this study provide a reference for the proliferation, conservation, and utilization of B. tsinlingensis resources.

• Swimming ability and behavior of Brachymystax tsinlingensis Li, 1966 larvae

  YANG Hongcan, LIU Peng, WANG Yizhou, LI Junyi, ZHOU Qiong, WANG Zhenlu, SHAO Jian

  2024,31(12):1474-1483, DOI: 10.12264/JFSC2024-0207

  Abstract: (69) HTML (124) PDF (1.58 M)(101)

  Abstract:

  This study used artificially bred Brachymystax tsinlingensis Li (1966) larvae [body weight: (0.06± 0.02) g, body length: (1.85±0.11) cm] as experimental subjects to explore the swimming ability and behavioral characteristics of B. tsinlingensis larvae and scientifically solve the problems of water velocity environment selection, during processes such as seedling cultivation and artificial release. The induced velocity, critical swimming speed, and burst swimming speed were measured using the increasing velocity method at (11.4±0.17) ℃, and the sustained swimming ability and durable swimming ability were measured using the fixed velocity method. The results showed that the induced velocity, critical swimming speed, and burst swimming speed were (0.03±0.01) m/s, (0.14±0.03) m/s, and (0.22±0.03) m/s, respectively. Relative induced velocity, relative critical swimming speed and relative burst swimming speed were (1.62±0.29) BL/s, (7.55±1.61) BL/s, and (11.48±1.79) BL/s, respectively. The maximum sustained swimming speed and the maximum durable swimming speed were 0.13 m/s and 0.25 m/s, respectively. This was similar to the mean critical swimming speed and mean burst swimming speed. Continuous and endurance experiments showed that the swimming duration of B. tsinlingensis larvae was negatively correlated with flow rate (lgT=−24.48X+7.09, R² =0.79, P＜0.01), tail beat frequency was positively and linearly correlated with flow rate (TFB=0.70U+5.54, R² =0.89, P＜0.01), and stride length was linearly and positively correlated with flow rate (SL=0.09U+0.11, R² =0.99, P＜0.01). Therefore, artificial breeding and release were performed at a flow rate of 0.04–0.14 m/s. The results of this study contribute to improving the survival rate of artificial cultivation and release effectiveness of B. tsinlingensis and provide a reference for the selection of resource conservation and utilization environments.

• Differences in water quality, bacterial structure and function of particle-associated bacteria and surrounding bacteria in small greenhouse farms of Litopenaeus vannamei under different yield levels

  HUANG Yong, WANG Baojie, LIU Mei, REN Ziwen, JIANG Keyong, ZHONG Chen, XU Kefeng, GAO Yan, WANG Lei

  2024,31(12):1484-1496, DOI: 10.12264/JFSC2024-0190

  Abstract: (59) HTML (125) PDF (2.35 M)(94)

  Abstract:

  In recent years, the number of small greenhouse farms in China has expanded rapidly. Within the aquaculture process of these small greenhouses, microorganisms play a key role in decomposing residual bait and feces in the water column. However, the lack of nutrient-degrading microbial taxa in the water column, combined with excessive baiting by farmers, often leads to the accumulation of inorganic nitrogen. This accumulation negatively affects the yield and economic benefits of cultured species. The present study was conducted to investigate the differences in water quality, as well as the structure and function of particle-associated bacteria (PAB) and non-mobile free-living bacteria (NFB) in small greenhouse farms. This investigation focuses on the turbidity of the water column caused by the increasing concentration of suspended particles during the aquaculture process, in order to reveal the factors influencing the culture yield. Differences in the structure and potential function of the two bacteria were analyzed by high-throughput sequencing of 16S rRNA. Additionally, real-time fluorescence quantitative PCR was used to detect the abundances of ammonia-oxidizing bacteria amoA and nitrite-oxidizing bacteria nxrB, both of which are associated with nitrogen transformation in suspended particles. The results of this study showed that the concentration of suspended particles was significantly higher in high-yield small greenhouses compared to low-yield small greenhouses (P＜0.001). Additionally, the concentration of suspended particles in the water column was closely related to shrimp production. The Chao1 and Shannon indices of PAB were found to be higher than those of NFB in the aquaculture water column of small greenhouses, indicating greater diversity and abundance of PAB. Notably, Flavobacteriaceae were significantly enriched in PAB, indicating that they were the dominant functional microorganisms in PAB. The abundance of biosynthesis, material, and energy metabolism functional pathways in PAB was significantly higher than that in NFB (P＜0.05). Among these ways, nitrification-denitrification was notably upregulated in PAB, indicating that PAB is more capable of material synthesis and nutrient metabolism. Real-time fluorescence quantitative PCR results revealed that the abundances of major functional genes of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria were higher in the PAB of high-yield small greenhouses than in those of low-yield small greenhouses, indicating that nitrifying bacteria were more mature in the PAB of high-yield small greenhouses. This study demonstrated that PAB play an essential role in the removal of nitrogenous pollutants from aquaculture water in small greenhouse farms, which further affects shrimp production by regulating water quality. The results of this study provide an important theoretical basis for water quality control and efficient shrimp culture in small greenhouse farms.

• Effects of purification units on phytoplankton in water column in recirculating pond aquaculture systems

  ZHENG Yingqiang, DAI Lili, ZHANG Hui, PENG Liang, TAO Ling, LI Gu, CHAI Yi, LIAO Yongling

  2024,31(12):1497-1510, DOI: 10.12264/JFSC2024-0234

  Abstract: (56) HTML (144) PDF (1.94 M)(94)

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  We quantitatively analyzed the changing patterns of phytoplankton after being treated by purification units in this study with three groups of recirculating pond aquaculture systems—a composite pond recirculating aquaculture system (A), a paddy field-fish pond recirculating aquaculture system (B), and a combined wetland-pond recirculating aquaculture system (C)—to explore the influence law of aquaculture water purification on phytoplankton community structure and diversity. The removal efficiency of different pollutants varied among the three systems, with system A having the highest removal efficiency for potassium permanganate index, total nitrogen, and suspended solids, whereas system C had the lowest efficiency for these contaminants. Compared to the inflow water, the Chl a concentrations in the effluent water were decreased by 52.08%, 67.61%, and 27.06% in systems A, B, and C, respectively. During the study period, 138 species from seven phyla of phytoplankton were identified, which were dominated by species from Chlorophyta, Cyanophyta, and Bacillariophyta. The number of phytoplankton species varied between the different systems; however, the dominant species and their dominance were not significantly different. Compared to the inflow water, the density and biomass of different phytoplankton species in the purification units were substantially decreased in systems A and B, and the algal removal rates exhibited an increasing trend along the cross-sections of the purification unit. Phytoplankton density fluctuated greatly in the purification unit of system C. Overall, phytoplankton species from different phyla were similarly affected by the purification units. The diversity of phytoplankton in the purification units was increased compared to the inflow water in the three systems, with the H′ and J index values increasing significantly in systems A and B (P＜0.05). Pearson correlation and redundancy analyses indicated that the densities of phytoplankton were significantly positively correlated with factors such as total phosphorus, total nitrogen, and potassium permanganate index in the water (P＜0.05), but there were differences in the factors influencing phytoplankton among different systems. This study revealed that aquacultural water purification significantly affects the phytoplankton community. Optimizing the purification unit environment may be useful for improving the purification efficiency of pollutants and phytoplankton.

• Stock discrimination of Larimichthys crocea from Naozhou and Minyuedong groups based on fish morphology, otolith morphology and chemistry

  YANG Min, QIN Jiao, GU Long, LI Ruojing, ZHAO Na, ZHANG Bo

  2024,31(12):1511-1523, DOI: 10.12264/JFSC2024-0193

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  Larimichthys crocea, an economically important fish species along the coast of China, has faced severe threats to its resources in recent years due to overfishing and loss of genetic diversity. To characterize the phenotypic traits of the less-studied Naozhou group of L. crocea, a combination of fish morphology, otolith morphology, and otolith chemistry was employed to discriminate the two stocks of L. crocea, the Naozhou and Min-yuedong groups sampled in Zhanjiang and Zhangzhou, respectively. The results of the principal component analysis indicated that the two populations could not be separated by the first two principal components of fish morphology, but could be distinctly separated by otolith morphology. Analysis of variance (ANOVA) indicated significant differences in fish and otolith morphologies between the two stocks (P＜0.05). The deformation diagram of fish morphology showed that Naozhou individuals were generally more elongated, with relatively shorter pectoral fin bases than Min-Yuedong individuals (P＜0.05). A deformation diagram of otolith morphology revealed that the otoliths of the Naozhou stock were more compressed overall, exhibiting a thicker neck in the sulcus and a relatively deeper first posterior dorsal depression than those of the Min-Yuedong stock (P＜0.001). Otolith chemical analysis revealed that the Ba/Ca and Sr/Ca ratios in the otoliths of the Naozhou stock were significantly higher than those in the Min-Yuedong stock, whereas the Mn/Ca ratio was significantly lower (P＜0.0001). There were no significant differences in the Mg/Ca ratios (P＞0.05). The Sr/Ca ratios exhibited the most pronounced differences between the two stocks from the core to the edge of the otolith when compared to the ratios of Ba, Mg, and Mn with Ca, with virtually no overlap at the measurement points. Discriminant analysis results indicated that the discrimination accuracy based on fish morphology (85.2%) was lower than that based on otolith morphology (100%) and chemistry (100%). The morphological differences between the two groups may be related to the differences in water flow and temperature between their habitats, with the Naozhou group inhabiting lower-flow and higher-temperature environments. Variations in otolith chemistry can be partly explained by salinity and temperature. Fish morphology, otolith morphology, and microchemical analyses demonstrated sufficient efficacy in the stock identification of the Naozhou and Min-yuedong groups of L. crocea. These findings provide a scientific basis for stock discrimination and fishery management of L. crocea.

• The trophic structure of main fishery organisms in the southwestern continental shelf of the Nansha Islands in spring

  QIU Xingyu, LIU Qingxia, CHEN Zuozhi, CAI Yancong, DAI Shouhui, HUANG Honghui

  2024,31(12):1524-1538, DOI: 10.12264/JFSC2024-0202

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  Based on the fishery biological survey data of the southwestern continental shelf of the Nansha Islands area in spring 2023, the trophic positions of major fishery organisms and the overlap of trophic niche among different functional groups were investigated by applying the carbon and nitrogen stable isotope techniques, and trophic continuum profiles were constructed. The results showed that the δ¹³C values of the fishery organisms ranged from −20.31‰ to −17.56‰, with a mean value of (−19.07±0.72)‰, δ¹⁵N values ranged from 7.79‰ to 10.80‰, with a mean value of (9.28±0.77)‰. Using zooplankton as the baseline organism, the trophic level of the fishery organisms ranged from 2.95 to 4.29. Compared with other marine areas in the South China Sea, the food chain length of the main fishery organisms in the southwestern continental shelf of the Nansha Islands area was relatively short, the initial food source was relatively single, the width of the niche was small, the degree of redundancy of consumers in the food web was high, and competition for food among species was intense. The results of cluster analysis of δ¹³C and δ¹⁵N values of fishery organisms were classified into four trophic groups, namely, zooplankton feeding group, mixed feeding group, large predator feeding group, and benthic feeding group, and the differences among the trophic groups were significant (P＜0.01). The trophic niches of fishery organisms were analyzed according to the δ−space (95% confidence interval) standard ellipse and structural parameter indexes, and it was found that the SEAc=0.66‰² . NR (δ¹⁵N range) and CR (δ¹³C range) of the mixed feeding group were 1.93 and 1.64, respectively, which indicated that the food resources were widely utilized and the food chain was long in this community. The niche overlap rates of the mixed-feeding group with plankton, large carnivores, and benthic feeding groups were 7.32%, 5.42%, and 0.60%, respectively. Compared to the pelagic zone in the central and western parts of the South China Sea, the overlap rate of trophic groups on the southwest shelf of Nansha was relatively low, indicating that the trophic niches of fishery organisms in different layers of the South China Sea have distinct spatial differentiation characteristics.

• Study On The Sources Of Occurrence Floating S.horneri In The Coastal Waters Of Gouqi Island

  zhuweidong, yangqifan, wanghuijie, miaohang, liufengyu, qinpengjie, biyuanxin

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  为探究枸杞岛近岸海域漂浮铜藻(Sargassum horneri)发生量及其来源，本研究以枸杞岛筏架定生和底栖铜藻为研究对象，根据定生铜藻生长、气囊形成时间及生物量变化规律，从源头上解析漂浮铜藻发生时间、生物量及漂浮路径。结果表明：(1)来源于筏架定生的漂浮铜藻形成时间比来源于底栖的漂浮铜藻早5—6个月，枸杞岛近岸海域约99.4 %的漂浮铜藻生物量来源于筏架定生铜藻，而底栖铜藻形成的漂浮铜藻生物量仅占0.6 %。(2)筏架定生铜藻脱离附着基后形成的漂浮铜藻株数约22×106 株，生物量约141938 t，经估算仅枸杞岛筏架定生来源的漂浮铜藻生物量约占东海区漂浮铜藻总量的23.4 %，来源于底栖铜藻占0.13 %。(3)结合我国不同区域定生铜藻生物量、漂浮铜藻出现时间、潮流及长江环锋影响铜藻漂浮路径等因素分析，我国漂浮铜藻可能有两条路径来源，北面来源为辽东半岛和山东附近海域，南面为长江河口以南区域。以上研究结果为解析枸杞岛近岸海域漂浮铜藻发生量来源及发生可能性上提供了参考，同时也为“金潮”溯源提供了一定依据。

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• Optimization of anesthetic for safe transportation of juvenile Stichopus monotuberculatus and evaluation of anesthetic effect

  Fenghaoran, Huanglianghua, Zhengbojun, Huangmaoning, Wujunhao, Xuqiang

  Abstract: (2) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  花刺参（Stichopus monotuberculatus）因其对环境变化及人为刺激极为敏感，人工繁育的苗种在运输过程中有应激性强、死亡率高的问题，开发一种适宜的麻醉方法可减轻幼参在运输过程中的应激反应和死亡率。本研究比较了五种常见的渔用麻醉剂：丁香油、2-苯氧乙醇、硫酸镁、MS-222和氯化镁对花刺参幼参的麻醉效果。通过建立基于行为学指标的麻醉及恢复阶段评价方法，将整个麻醉过程分为轻度麻醉、中度麻醉、深度麻醉、轻度苏醒、中度苏醒和完全苏醒六个阶段。研究结果表明，丁香油、硫酸镁和氯化镁均能将花刺参幼参麻醉。其中，浓度0.2ml/L的丁香油对花刺参幼参的麻醉效果最好，0.02ml/L和0.04ml/L的浓度可提升花刺参幼参长时间运输的存活率。浓度0.5ml/L的丁香油对幼参的麻醉时间最短，为6.04±1.24min。应用浓度0.1mol/L的氯化镁麻醉剂，幼参进入深度麻醉时间最慢，为33.63±7.92min，而应用浓度0.25mol/L硫酸镁麻醉剂，幼参麻醉后达到完全苏醒时间最慢，为60.39±5.28min。高浓度的氯化镁和硫酸镁麻醉前后花刺参幼参体长无明显变化；MS-222和2-苯氧乙醇对花刺参幼参不具有麻醉作用。本文筛选出适宜花刺参幼参的麻醉剂为丁香油，浓度为0.02ml/L，可保证幼参60h长途运输后的有效恢复和高存活率，为花刺参苗种的长距离安全运输提供重要的技术保障。

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• Comparison of marine environmental factors and pearl production between spring and autumn nucleus insertion models

  Liao Yongshan, Wu Xuhui, Tao Chuangui, Wang Yabing, Deng Yuewen, Wang Qingheng

  Abstract: (9) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为了探究马氏珠母贝(Pinctada fucata martensii)春季与秋季植核育珠模式各自的优势，同时查明育珠效果与主要环境因子之间的关系，本研究调查了2020年4月至2022年7月连续四季育珠生产的产能指标和海区环境因子。实验设春季植核组(S)和秋季植核组(F)，分别在2020年4月(S20)、2021年的4月(S21)和2020年10月(F20)、2021年10月(F21)植核，逐月调查育珠贝存活率，测定养殖海区水温、溶解氧、盐度等主要环境因子；育珠9个月后采收珍珠，统计留核率、优珠率、珍珠层厚度等产能指标。结果显示，实验期间养殖海区水温为17.23~33.23 ℃，具有明显的季节波动，S组和F组水温大于或接近30 ℃的月份分别为5个月和2个月；海区溶解氧含量为4.44~8.08 mg/L，存在着明显的季节变化；除10月盐度偏低外，其余月份盐度稳定在30‰~35‰间；pH总体稳定，在8.05~8.28之间波动；海区透明度为80~200 cm，总体呈现为夏秋季节低，冬春季节高。育珠期结束后，各组育珠贝总体存活率由高到低依次为F21>F20>S21>S20，F组显著高于S组(P<0.05)；S组中，水温与阶段性存活率呈显著负相关(P<0.05)，其他环境因子与阶段存活率相关性不显著(P>0.05)，F组的环境因子与阶段性存活率之间的相关性均不显著(P>0.05)。各组留核率、商品珠率和优珠率差异不显著(P>0.05)；S组的珍珠层厚度显著大于F组(P<0.05)，提高约20%。结果表明, 秋季植核育珠生产模式可显著提高育珠贝的成活率，增加珍珠产量, 春季植核育珠生产模式珍珠质量更好；夏季高温是造成S组死亡率较高的主要原因。本研究可为海水珍珠健康养殖提供数据支撑和理论基础。

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• Characteristics of the cyp19a Gene and Functional Analysis in the Sexual Gland Development of Grass Carp (Ctenopharyngodon idellus)

  xiaoxiaofang, lishengjie, liuqingshan, dujinxing, leicaixia, zhutao, tianjing, songhongmei

  Abstract: (10) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为探究cyp19a基因在草鱼(Ctenopharyngodon idellus)性别分化和性腺发育过程中的表达模式和作用机制，本研究通过RACE技术克隆cyp19a基因全长cDNA序列，采用qRT-PCR技术检测cyp19a基因在草鱼发育不同时期性腺组织和成鱼各组织中的表达模式，合成cyp19a-dsRNA并对雌性草鱼进行RNA干扰实验，对照组注射等量生理盐水，检测性腺组织中性别相关基因的表达量和血清性类固醇激素含量。结果显示，草鱼cyp19a的cDNA序列全长为2268bp，其中开放阅读框(Open reading frame，ORF)长1554bp，5’非编码区(Untranslated region，UTR)48bp，3’UTR 617bp，编码517个氨基酸。草鱼cyp19a基因在性腺中表达量最高，其次是肌肉、脑和心脏组织。不同发育时期的性腺中，cyp19a在卵巢中的表达水平均显著高于精巢(P < 0.05)，且在4龄时达到峰值，表明该基因在草鱼卵巢的发育和成熟过程中起关键作用。按5μg/g注射 cyp19a-dsRNA沉默干扰后5天，dsRNA注射组cyp19a表达量与对照组雌鱼相比显著降低(P < 0.05)，靶基因沉默效率在1d时为59.72%，之后逐步上升。与对照组雌鱼相比，dsRNA注射组在1～3d时foxl2a和foxl2b表达量显著上调(P < 0.05)，dmrt1表达量下调(P > 0.05)，amh表达量显著下调(P < 0.05)，并在干扰后4～5d时foxl2a、foxl2b、dmrt1和amh的表达量逐步恢复至接近对照组水平(P > 0.05)；此外，注射组血清中雌二醇(estradilol，E2)含量相比对照组雌鱼显著下降(P < 0.05)，睾酮(testosterone，T)含量显著上升(P < 0.05)。综上，草鱼cyp19a可能通过负反馈调控foxl2a和foxl2b表达以此调节草鱼E2水平，正向调控dmrt1和amh表达量而调节其性类固醇激素含量，促进卵巢和卵细胞的分化、发育和成熟，研究结果为深入探索草鱼生殖调控机制奠定基础。

  • 1
• Characterization of AGL gene expression in glycogenolysis of the Jinjiang oyster Crassostrea ariakensis

  ZHAO Liyan, LI Zhuanzhuan, MA Peizhen, LIU Zhihong, SUN Xiujun, ZHOU Liqing, WU Biao

  Abstract: (13) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  糖原是影响牡蛎品质的关键指标，而糖原脱支酶（AGL）在糖原分解过程中发挥重要作用。为明确AGL基因对近江牡蛎（Crassostrea ariakensis）糖原代谢的影响，本研究在分析AGL基因序列特征和进化特点基础上，体外构建含有该基因的质粒并转化到HT115（DE3）大肠杆菌（Escherichia coli）中，使其表达AGL基因的双链RNA（dsRNA），通过饲喂该菌株实现RNA干扰，并对干扰前后近江牡蛎AGL基因表达量和糖原含量的相关性进行了研究。结果表明，AGL基因编码区序列长度为4719 bp，编码1572个氨基酸，含4个结构域，预测蛋白分子量为178.23 kDa，理论等电点为6.21，基因进化关系与物种传统分类关系一致；RNA干扰15 d和30 d，与对照组（饲喂含EGFP dsRNA的大肠杆菌）相比，实验组（饲喂含AGL dsRNA的大肠杆菌）牡蛎的AGL基因表达量均显著降低（P < 0.05），糖原含量显著升高（P < 0.05），干扰AGL基因能够显著影响到糖原水平的变化；干扰30 d，实验组和对照组牡蛎AGL表达量比干扰15 d时均显著增加（P < 0.05），糖原含量显著降低（P < 0.05），这可能是由于此时期性腺发育需要消耗糖原所致；同时，进一步的相关性分析结果表明AGL基因表达量与糖原含量呈显著负相关关系（P < 0.05）。本研究明确了近江牡蛎AGL基因序列结构特征以及其表达量与糖原含量的相关性，丰富了AGL基因功能研究的科学数据，为深入研究牡蛎糖原代谢的分子机制提供了参考。

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• Study on the annual ovary development and large-scale induced spawning of hooksnout carp Opsariichthys bidens

  DENG Xiang, ZOU Cong, SHEN Fan, QIAO Hao-Feng, CHEN Yu-Pei, HU Chen-Hao, BIE Han-Qing, FAN Qi-Xue, Shen, Zhigang

  Abstract: (50) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为建立马口鱼(Opsariichthys bidens)规模化人工繁育技术体系，对马口鱼卵巢发育的周年变化进行了研究观察，对其规模化人工繁殖技术进行了系统研究。结果显示，湖北地区雌性马口鱼性腺指数(GSI)周年变化范围为1.16%-11.65%。繁殖期为4月中旬至7月中旬，雌鱼产卵前平均绝对繁殖力为6165.4粒。秋季水温逐渐降低时(9-10月)，马口鱼卵巢GSI逐渐降低，卵巢转至Ⅲ期并以该时期延续。第二年2月，马口鱼GSI达到极低水平(1.16%~1.65%)，卵巢中Ⅲ时相卵母细胞数量比例在2月水温进一步降至8.0 ℃以下后开始增加。规模化人工繁殖试验中发现，LRH-A2 (5 μg/kg)与HCG (1000 IU/kg)组合使用催产效果最佳，催产率达87.0%。结果表明，0.3% NaCl溶液为最佳授精介质，人工授精最佳精卵比为5.0×104-1.0×10?：1，平均受精率为66.6%，平均孵化率为80.1%。5月中旬之前催产效应时间为20.0-26.0 h，5月中旬后效应时间为16.0-17.0 h，积温为400.00-430.00 ℃?h。水温20.0-25.0 ℃时，孵化用时为69.0-90.0 h；水温大于26.0 ℃时，孵化时长为42.5-48.0 h，孵化积温为1200.00-1300.00 ℃?h。研究还成功地进行了当年繁殖马口鱼亲本的第二次人工繁殖，发现再次催产效果与第一次无明显差异。研究揭示了池塘培育条件下马口鱼卵巢发育与成熟规律，明确了其一年多次产卵的生物学特性，成功开展了马口鱼大规模人工催产与孵化，为马口鱼遗传改良、相关研究工作及产业发展奠定了重要基础。

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• Habitat suitability and release area selection of Hexagrammos otakii in coastal Dalian

  Feng Chao, DONG Jing, LIU Xiuze, Dezhong Qu, JI Guang, WANG Aiyong, WANG Xiaolin, WANG Bin, XU Yanzhao, Zhang Depu

  Abstract: (38) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  根据2015—2023年5—10月的渔业调查资料, 利用同步获得的底层环境因子作为自变量, 包括水深、温度、盐度、溶解氧和叶绿素a, 结合增强回归树模型(boosted regression tree, BRT)建立各月的栖息地适宜性指数 (habitat suitability index, HSI), 分析大连海域大泷六线鱼的栖息地适宜性, 识别其资源与环境因子的关系, 探究其栖息地的时空变化, 为该物种的栖息地保护和放流区域选址提供依据。研究显示, 水深是影响大泷六线鱼栖息地适宜性最重要的环境因子, 其他因子如水温、溶解氧和叶绿素a的影响因月份而异。适宜栖息地的时空分布表明, 大泷六线鱼的生殖洄游习性以及作为岩礁性、冷温性底栖鱼类的生态习性是适宜栖息地时空分布变动的重要原因。总体上大连黄海海域相比于大连渤海海域具有更大面积的适宜栖息地, 本文结果结合以往研究表明, 如以5—6月作为大连海域大泷六线鱼的放流时间, 其5月适宜的放流区域宜为大连黄海海域的大连湾三山岛至金普新区沿岸及长海县岛礁周边水域和大连渤海海域的长兴岛以北近岸水域；6月适宜的放流区域在大连黄海与5月保持不变, 大连渤海海域宜为旅顺老铁山岛礁海域及金州湾以西海域。本文结果初步表明, 包括HSI模型在内的物种分布模型可对渔业生物的适宜栖息地进行可视化, 展现了其在确定适宜放流区域等放流策略制定中可视化和量化的优势。

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• Hydrodynamic characteristics of a submersible cage in the waves and currents

  LI Xiaoran, CUI Yong, GUAN Changtao, WANG Gang, WU Jintao, GUO Yinuo

  Abstract: (44) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  网箱养殖正迈向深远海域发展，新型潜降式网箱能够在台风来临前自动实现潜降避浪，既解决了传统重力式网箱在巨浪强流下易破坏的问题，而相较于大型养殖平台又降低了制造成本。本文所研究的新型潜降式网箱由四点锚泊系统、潜降系统等部分组成，其在极端海况下不同状态时的水动力特性尚未深入研究。因此，针对波流联合条件下网箱处于漂浮及潜降状态时的水动力特性开展研究，不仅有助于揭示潜降系统对网箱动力响应的影响机制，更有助于优化网箱结构，为降低成本和减少能源损耗提供科学依据。本研究基于势流理论和有限元参数化建模技术开展数值模拟试验，结果显示：最大锚绳张力随波流参数的增大而增大，工程中应选用高强度材料制作锚碇锚绳，垂荡、纵荡、纵摇同波高均存在显著正相关关系，纵摇在漂浮时与周期呈负相关，潜降时与周期呈正相关，说明波陡对于新型潜降网箱漂浮稳定性有显著影响，应对网箱所处海域的波陡幅值进行实时监测。比较网箱在潜降过程中锚绳的力学特性和运动响应变化，计算出锚绳张力平均减幅达到59%，垂荡、纵荡和纵摇平均减幅分别达到70%、57%和49%；网箱各运动响应峰值均较小，垂荡峰值仅为0.549m，纵荡峰值仅为1.055m，纵摇峰值仅为7.116°，表明网箱具有较好的抗风浪性能。

  • 1
• Analysis of Gut Microbiota and Environmental Safety of Gene-Edited Half-Smooth Tongue Sole(Cynoglossus semilaevis)

  wuyinqi, chenzhangfan, lipengfei, cuizhongkai, dinglanqing, chengfangzhou, yanxv, zhouzebin, chensonglin

  Abstract: (33) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  半滑舌鳎(Cynoglossus semilaevis)是我国特有的一种海水养殖鱼类,但其雄鱼生长缓慢、个体小的问题严重制约了半滑舌鳎养殖业的推进和发展。本实验室利用TALEN基因组编辑技术,成功获得半滑舌鳎dmrt1纯合突变雄鱼,生长速度较普通雄鱼快2倍以上,为半滑舌鳎基因编辑育种技术奠定了基础。为评估基因编辑后半滑舌鳎的健康状况与环境安全性,本研究运用高通量测序技术对相同养殖条件下的半滑舌鳎dmrt1纯合突变雄鱼和普通雌雄鱼的肠道微生物菌群进行了对比分析。另外,针对环境安全风险,本研究检测了与dmrt1纯合突变雄鱼共同饲养的普通鱼基因型以及饲养环境的水体eDNA。结果表明,半滑舌鳎dmrt1纯合突变雄鱼的肠道微生物菌群Alpha多样性及组成结构与普通雌雄鱼均不存在显著性差异,肠道微生物功能预测与普通雌雄鱼相比亦无差异。与dmrt1纯合突变雄鱼共同饲养的普通雌雄个体中均无法检测到编辑后的基因。另外,对半滑舌鳎dmrt1纯合突变雄鱼的养殖水环境及其排水路径中的5个采样点的水样进行了扩增与测序,结果并未发现编辑后的目的基因。研究表明,dmrt1基因编辑并不会导致半滑舌鳎dmrt1纯合突变雄鱼的肠道微生物菌群发生显著性变化,亦不会对同一环境下养殖的其他个体或养殖的水体环境产生影响,不存在环境风险。

  • 1
• The research on the response of hemolymphocytes to nucleus-inserting stimulation and biomineralization function of Hyriopsis cumingii

  SUN Junlin, CHEN Yiwen, CHEN Xiaofeng, LI Jiale, BAI Zhiyi, LI Wenjuan

  Abstract: (32) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为深入研究淡水珍珠贝三角帆蚌(Hyriopsis cumingii)插核刺激对血淋巴细胞的影响及其在珍珠矿化中的功能，本研究通过光学和电子显微镜，探讨了三角帆蚌血淋巴细胞的形态和分类，根据细胞复杂性、细胞大小、核质比将其分为4种类型。通过吉姆萨染色和流式细胞术分析发现，插核刺激后血淋巴细胞的颗粒复杂度增加，细胞直径变大，颗粒细胞比例显著升高(P＜0.05)，透明细胞比例显著降低(P＜0.05)，同时通过组织切片观察，插核部位附近有大量血淋巴细胞聚集且颗粒细胞在插核部位被招募。血淋巴细胞和血清中的Ca2+含量在插核后2 d、5 d和10 d显著上升(P＜0.05)，20 d和50 d显著下降(P＜0.05)，揭示了血淋巴细胞在数量组成、分布和Ca2+水平上均响应插核育珠过程。通过体外碳酸钙结晶实验表明血淋巴细胞和血清能够影响方解石的形貌，但未改变晶体晶型。综上所述，血淋巴细胞对三角帆蚌插核刺激有迅速响应且对生物矿化过程有调控作用，本研究为进一步探究血淋巴细胞在贝类生物矿化中的调控作用奠定了理论基础。

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• Effects of Ammonia Nitrogen on Serum and Intestinal Immune Responses in Rainbow Trout(Oncorhynchus mykiss) Infected with IHNV

  LIU Xuejiao, LI Jie, SHAN Caixia, HAI Qiang, LIU Hao, LIU Zhe, ZHU Lirui, WANG Jianfu

  Abstract: (36) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为探究氨氮胁迫对虹鳟IHNV的易感性影响及其机制。选用体重为13.5±0.5 g的虹鳟幼鱼,分别设置对照组(C 组)和三个氨氮胁迫组(L组,2 mg/L、M 组,4 mg/L、H 组,6 mg/L),进行为期10 d的氨氮胁迫,之后腹腔注射TCID50为10-7.875^/ml的IHNV 0.25ml,分别在攻毒后的0、3、5、7和9 d检测其血清中的碱性磷酸酶(AKP)、谷丙转氨酶(ALT)和丙二醛(MDA)的活力,肠道中免疫相关基因Mx、IgM、IRF7和IFN-α的表达量,以及攻毒后第7 d其肠道组织学变化和菌群差异。结果显示：IHNV攻毒后虹鳟鱼种的死亡率与氨氮胁迫浓度呈正相关；随着氨氮浓度升高,虹鳟肠上皮细胞空泡化程度加重,绒毛面积减小；攻毒后其血清中AKP活力显著降低(P < 0.05),而ALT和MDA活力表现出先升高后降低的趋势,肠组织中Mx、IRF7、IFN-α和IgM表达量均表现为先升高后降低的趋势；攻毒后7 d,在门水平上,随着氨氮浓度升高其肠道内假单胞菌门(Pseudomonadota)和支原体门(Mycoplasmatota)丰度显著升高(P < 0.05),而蓝藻门(Cyanobacteriota)和芽孢杆菌门(Bacillota)丰度显著降低(P < 0.05)；在属水平上,相较于C组,氨氮胁迫组中有害菌群丰度升高,而有益菌群丰度则表现出下降趋势。以上结果表明,氨氮胁迫会增加虹鳟对IHNV的易感性,降低其抗氧化能力和免疫力,并破坏其肠道组织及菌群稳态。本研究为虹鳟养殖中生态防控IHN提供理论依据。

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• Study on artificial propagation and early development of Garra mirofronits

  wangyuesong, yangyang, ganxing, yinhuazheng, pengli, zhangzhiwei, chengrui, chenfeng, shishaolong, mashu, zhangzhiming

  Abstract: (38) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为研究奇额墨头鱼的早期发育时序特征，丰富其繁殖生物学、发育生物学研究资料，为人工繁育和苗种规模化生产提供技术支撑。利用野外采集的奇额墨头鱼(Garra mirofronits)性成熟个体，通过人工催产及干法授精获得其受精卵，详细观察并记录其早期发育过程。共开展3次人工繁殖实验，平均催产率为100 %、受精率(89.93±0.72) %、孵化率(87.83±1.00) %。受精卵呈浅灰色，卵膜薄而透明，无黏性，卵径(1.12±0.04) mm，吸水膨胀后卵膜径达(3.37±0.09) mm，膨胀系数300.89%。水温18~21 ℃下，经24~26 h孵化出膜，孵化积温为496.04 ℃·h，历经受精卵、胚盘期、卵裂期、囊胚期、原肠胚期、神经胚期、器官形成期、心脏搏动期和出膜期等9个时期。初孵仔鱼全长为(4.50±0.07) mm，肌节32对，卵黄囊仔鱼期历时5 d，全长特定生长率为9.22 %·d?1; 仔鱼期共历时16 d，全长特定生长率为7.02 %·d?1。奇额墨头鱼早期发育时序特征遵循硬骨鱼类发育的基本规律，符合漂流性卵胚胎发育的一般特点，为典型产漂流性卵鱼类。

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• Study on temporal characteristics and environmental factors of Coilia coilia migration into and out of the Poyang Lake during the early period of fishing ban

  yang yanping, wang sheng, xuan zhongya, ma fengjiao, peng legen, wen sixin, shen yingdong, liu kai

  Abstract: (44) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为掌握禁捕初期鄱阳湖刀鲚(Coilia nasus)繁殖群体及幼鱼出入湖时序特征，支撑刀鲚资源养护和鄱阳湖生物完整性评价，为鄱阳湖刀鲚栖息地识别积累数据，2023年4—10月刀鲚出入湖汛期内，在鄱阳湖江湖通道湖口水域设置固定站位，跟踪监测刀鲚繁殖群体入湖和幼鱼出湖资源动态，分析刀鲚出入湖时序特征及环境影响因子。就入湖群体而言，4—7月单位捕捞数量变幅为19~971 ind./(1000m2·h)，均值为294±201 ind./(1000m2·h)；单位捕捞重量变幅为0.83~84.25 kg/(1000m2·h)，均值为25.92±18.10 kg/(1000m2·h)。体长变幅为150.6~382.1 mm，均值为298.2±29.1 mm；体质量变幅为10.6~238.1 g，均值为89.9±29.8 g；雌雄比例为2.2：1，性腺发育至Ⅰ—Ⅴ期的个体比例依次为0.09%、46.01%、38.0%、13.08%和2.06%。时间特征显示，5月为刀鲚繁殖群体入湖高峰期，单位捕捞数量和重量分别为514±168 ind./(1000m2·h)和47.04±16.08 kg/(1000m2·h)，极显著高于其他月份(P<0.01)，体长、体质量随洄游汛期后移呈极显著减小(P<0.01)，其中5月体长均值最大(301.5±26.4 mm)，7月最小(292.2±30.6 mm)；4月体质量均值最大(98.49±34.02 g)，7月最小(82.0±25.2 g)；同期雌性个体比例持续上升，雄性比例持续下降，性腺发育至Ⅱ期的个体比例由4月的81.14%下降至7月的22.22%，Ⅲ期及以上个体比例则由4月的18.77%上升至7月的77.78%。就出湖群体而言，7—10月单位捕捞数量变幅为11~8571 ind./(1000m2·h)，均值为1316±1908 ind./(1000m2·h)。时间特征显示出湖高峰期为7月和9月，其中7月单位捕捞数量均值最大, 为2845±2940 ind./(1000m2·h); 其次为9月, 单位捕捞数量均值为1769±1862 ind./(1000m2·h); 10月最小, 单位捕捞数量均值为285±327 ind./(1000m2·h)，环境因子分析显示鄱阳湖刀鲚幼鱼出湖群体单位捕捞数量的时序变化与水位差之间的变化趋势一致。研究结果表明鄱阳湖刀鲚汛期提前，持续时间延长，种群小型化趋势改善，资源恢复趋势显著。

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• Analysis of genetic diversity and selection characteristics based on WGS technology in cultured Topmouth culter (Culter alburnus) population

  Huali Zhao, Di-an Fang, Tianyou Li, Jia Yuan, Baoxing Liu, Dongpo Xu

  Abstract: (55) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为评估养殖群体翘嘴鲌（Culter alburnus）的遗传多样性水平，探究其遗传特征及群体间的遗传差异，本研究利用全基因组重测序（Whole genome resequencing, WGS）技术，对来自浙江太湖（TH）、广东清远（QY）和江苏扬州（YZ）的三个养殖群体进行了深入分析。总共鉴定出23,156,699个SNPs和5,838,147个Indels位点，SNP变异位点主要位于基因间区域（47.21%）和内含子区域（39.37%）。三个群体的SNP标记均显示为低度多态性（PIC值<0.25）。太湖、清远、扬州三个群体平均近交系数分别为0.1923、0.0631和 -0.0280。群体结构分析表明，太湖群体形成了独特聚类，而清远和扬州群体在遗传上则受到了多个祖先群体的影响。连锁不平衡衰减图（LD decay）显示太湖群体的衰减距离最大，衰减速度最慢。遗传分化指数（Fst）和基因流（Nm）均表明太湖群体与其他群体间存在明显分化。通过基于遗传分化系数 （Fst）和核苷酸多样性（π）的选择性扫描分析，以清远和扬州群体为参考群体，在太湖群体中分别鉴定到了572个和602个选择性基因，涉及能量代谢、氧化应激和炎症反应等生物过程，最多选择的区域位于chr 3和chr 7上。综上所述，三个翘嘴鲌养殖群体的遗传多样性水平均较低，其中太湖群体的遗传多样性最低，受到较强的人工选择和近亲繁殖影响，而清远群体与扬州群体则显示出频繁的基因交流，可能存在着种质混杂现象。本研究可为翘嘴鲌养殖群体的育种选育及种质资源的保护和管理提供基础数据支撑。

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• Mechanisms of α-Ketoglutarate's Effect on Antioxidant Enzyme Activity, Digestive Enzyme Activity, and Gut Microbiota in Carassius auratus Exposed to Carbonate-Alkaline Stress

  HAN Lin, LIU Wenzhi, YUAN Fangying, ZHAO Yan, SUN Yanchun

  Abstract: (38) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为探究碳酸盐碱暴露下α-酮戊二酸(α-ketoglutarate, AKG)对鲫抗氧化酶、消化酶活性和肠道菌群的调控机制, 本研究结合生化分析与16S rRNA测序技术, 设计了5个实验组: 淡水组(C组)、两种盐碱暴露组(20 mmol/L NaHCO3, T组; 40 mmol/L NaHCO3, F组), 以及两种AKG调控组(20 mmol/L NaHCO3, TA组; 40 mmol/L NaHCO3, FA组)。研究结果表明, 随着碳酸盐碱浓度的增加, 超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的活性显著降低, 丙二醛(MDA)含量显著升高。此外, 鲫肠道的消化酶(淀粉酶AMS、胰蛋白酶CHY、脂肪酶LPS)活性亦出现不同程度的下降。外源补充AKG后, 鲫肠道的抗氧化酶和消化酶活性得到了显著恢复, 丙二醛(MDA)含量显著降低。与C组相比, T组和F组的变形菌门(Proteobacteria)丰度显著增加, 而厚壁菌门(Firmicutes)的丰度显著下降; T组和F组的优势菌属分别为鞘氨醇单胞菌属(Sphingomonas)和希瓦氏菌属(Shewanellaceae)。然而, 在AKG调控组(TA组和FA组)中, 变形菌门(Proteobacteria)丰度大幅下降, 厚壁菌门(Firmicutes)的丰度上升, TA组的优势菌属转为芽孢杆菌属(Bacillus), 而FA组的优势菌属仍然为希瓦氏菌属(Shewanellaceae)。综上所述, 本研究发现碳酸盐碱暴露导致了鲫肠道氧化应激反应, 并削弱了肠道菌群中有益菌的丰度。补充AKG能够通过增强抗氧化酶和消化酶活性、调节肠道有益菌群的组成, 有效缓解了碳酸盐型盐碱生境对鲫的毒性作用。本研究为精准调控营养功能饲料的创制和提高盐碱水资源的利用效率提供了新的理论依据和实践思路。

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• Path analysis of the effects of morphological traits on body weight of different genders of Diptychus maculatus in Muzhati River

  haohuimin, wangbo, zhanglirong, wangzhengwei, liyanhui, niezhulan

  Abstract: (48) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为探究木扎提河不同性别斑重唇鱼形态性状对体重的影响，以123尾雌鱼和124尾雄鱼为研究对象，分别测定体重、12个传统形态性状及21个框架形态性状，通过相关性分析、回归分析及通径分析研究形态性状对体重的影响。结果表明：（1）斑重唇鱼雌雄各形态性状与体重之间均呈极显著正相关（P＜0.01），其中雌性个体的全长（X1）和体长（X2）与体重相关系数最大，均为0.930，雄性个体的吻端到腹鳍基部起点的距离（X24）与体重的相关系数最大，为0.939；（2）斑重唇鱼雌雄形态性状与体重的回归方程：Y雌= -34.488+0.128X1+0.562X4+0.145X2+0.241X27-0.231X34+0.579X3 （R2=0.929），Y雄= -18.753+0.139X24+0.337X3+0.143X23+0.218X31+0.212X28+0.321X8+0.096X29 （R2=0.945），式中X1~X4分别为全长、体长、体宽、体高，X8为眼间距，X23为吻端到背鳍起点的距离，X27~X29分别为背鳍起点到胸鳍起点、腹鳍起点和臀鳍起点的距离，X31为背鳍基部末端到腹鳍起点的距离、X34为尾鳍背部起点到臀鳍起点的距离；（3）通径分析表明，对雌鱼体重直接作用最大的是全长，其次是体长，通径系数分别是0.313和0.299；对雄鱼体重直接作用最大的是X24，其次是X23，通径系数分别是0.236和0.226。该研究充实了斑重唇鱼形态学相关数据，为其后期选育工作提供基础理论依据。

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• Development and experiment of real-time monitoring software of farmed fish omnidirectional sonar based on YOLOv8

  Sun Pengqi, Hu Jiangzhen, Huang Xiaohua, Sun Jialong, Li Gen, Tao Qiyou, Yuan Taiping, Pang Guoliang, Hu Yu

  Abstract: (44) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  针对水产养殖鱼群数量监测效率低、精确度不足的问题, 本研究以罗非鱼(Oreochromis sp.)为研究对象, 提出了一种基于全向扫描声呐与 YOLO (You Only Look Once) 模型的实时鱼群监测方法。本方法利用全向扫描声呐采集水下鱼群影像数据, 通过 YOLOv8 算法与实时监测技术实现目标识别与分析, 并结合基于欧氏距离和空间分析算法, 合并与排除异常数据点, 最终获得鱼群数量与空间分布。实验针对不同鱼群数量（50条、100条、150条、200条）进行了评估, 监测精确度分别为 93.5%、94.5%、89.6%和85.8%, 整体平均精确度达 90.9%。结果表明, 该方法显著提高了养殖鱼群数量监测的实时性和精确度, 为水产养殖中鱼群数量监测提供了一种高效的解决方案, 对优化水产养殖管理、提高生产效率及促进生态养殖可持续发展具有重要意义。

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• Effects of dietary protein and lipid levels on growth and hepatic health in Sander lucioperca juvenile

  ZHAO Jie, ZHENG Xianhu, SUN Zhipeng, WANG Liansheng, FAN Ze, LIU Yang, GAO Jiamin, PAN Yadan, XU Yihua, LU Cuiyun

  Abstract: (68) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为探究不同饲料蛋白质和脂肪水平及其互作对梭鲈（Sander lucioperca）幼鱼生长、肝脏组织形态、抗氧化酶活性及免疫相关基因表达的影响，实验设计2个蛋白质水平（51%、54%）和3个脂肪水平（12%、15%、18%）共6种饲料，对梭鲈幼鱼（1.16±0.01 g）进行8周养殖实验。结果表明：饲料脂肪水平升高显著提高梭鲈幼鱼终末体质量（FBW）和特定生长率（SGR），但饲料蛋白质和脂肪水平对FBW和SGR无显著交互作用。饲料蛋白质水平为51%时，梭鲈幼鱼FBW和SGR在脂肪水平为18%时最大；饲料蛋白质水平为54%时，梭鲈幼鱼FBW和SGR在脂肪水平为15%时最大。随着饲料脂肪水平的增加，全鱼粗蛋白质含量逐渐降低（P<0.05），粗脂肪含量呈先增加后降低。饲料蛋白质水平为54%，脂肪水平为18%时观察到明显的肝细胞空泡变性，其他饲料组无异常。饲料蛋白质水平为51%时，随着饲料脂肪水平的升高，过氧化氢酶、谷胱甘肽和超氧化物歧化酶活性显著升高；饲料蛋白质水平为54%时，随着饲料脂肪水平的升高，过氧化氢酶活性显著升高。随着脂肪水平的升高肝脏补体基因（c3、c4和c5）和抑炎细胞因子相关基因（tgf-β）的表达水平显著升高，促炎细胞因子相关基因（il-1β、il8和tnf-β）的表达水平显著降低。综上，适宜饲料脂肪水平会提高梭鲈幼鱼生长性能，节约蛋白质用量，但过高脂肪水平则会增加肝脏病变的风险，建议梭鲈幼鱼饲料蛋白质和脂肪水平为51%和18%。

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• Comparative analysis of the differences in gut microbial diversity between water mold infection and healthy states in the giant whiskered schizothoracine

  WANG Yuya, Liu Yixuan, WANG Wanliang, CAO Pengxi

  Abstract: (34) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  巨须裂腹鱼(Schizothorax macropogon)是国家二级保护鱼类,近年来其资源量急速锐减,具有较高生态价值和经济价值,目前研究多集中于资源调查、个体生物学和群体遗传学研究,有关肠道微生物研究较少。本研究旨在探讨巨须裂腹鱼患水霉病组和健康组肠道菌群群落结构组成和多样性特征,并分析其潜在功能。研究采用高通量测序技术,提取获得患病巨须裂腹鱼与健康巨须裂腹鱼两组鱼类的肠道内容物,分析了肠道微生物的OUT数量、菌群组成、丰度及Alpha多样性,并预测期功能。经α和β多样性分析,健康组巨须裂腹鱼的肠道微生物群落丰富度和多样性高于患病组,且肠道微生物群落之间具有显著差异(P <0.05); 健康组与患病组巨须裂腹鱼肠道微生物在群落组成上具有相似性,但相对丰度具有明显差异。细菌共获得了7个优势菌门,包括蓝菌门(Cyanobacteriia)、变形菌门(Proteobacteria)和梭杆菌门(Fusobacteriota)等,与健康组相比患病组特有属为拟杆菌属(Bacterodies)和变形杆菌属(Proteocatella)等；真菌共获得5个优势菌门,包括子囊菌门(Ascomycota)、担子菌门(Basidionmycata)和罗兹菌门(Rozellomycota)等,健康组的特有属为角担菌属(Ceratorhiza)；患病组的特有属为Boeremia和曲霉属(Tausonia)。LDA判别图显示了健康组和患病组巨须裂腹鱼肠道之间存在38个显著差异的细菌菌群,45个显著差异的真菌菌群；巨须裂腹鱼肠道微生物的共现性网络具有较高的连接性,且协同作用大于拮抗作用。根据功能预测分析,健康组巨须裂腹鱼肠道细菌功能菌群与患病组之间具有显著差异(P <0.05),且健康组的相对丰度高于患病组。本研究揭示了巨须裂腹鱼肠道微生物的物种组成和多样性特征,深入了解了巨须裂腹鱼与其肠道微生物之间的关系,为西藏地区特有鱼类肠道菌群的研究提供了基础的理论数据。

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• Characteristics and influencing factors of CH₄ and NO₂ emissions from pond culture and rice-crab co-culture models of Chinese mitten crab (Eriocheir sinensis)

  xuke, xiaolin, sunyunfei, chengyongxu

  Abstract: (55) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为研究中华绒螯蟹(Eriocheir sinensis)池塘养殖与稻蟹共作两种模式下水-气界面温室气体的排放特征及其影响因素,本实验使用静态箱-气相色谱仪法分别测定两种养殖模式下的温室气体排放通量,同时采集两种养殖模式下相关环境因子,并进行对比分析。结果表明,稻蟹共作模式N2O排放通量是池塘养殖的2.8倍,波动范围为34.94~126.90 μg.m-2^.h-1^,平均值为85.41±5.78 μg.m-2^.h-1^,CH4排放通量是池塘养殖的1.19倍,波动范围为1.32~39.50 mg.m-2^.h-1^,平均值为13.69±1.42 mg.m-2^.h-1^{。池塘养殖和稻蟹共作的全球增温潜势分别为}10.73±1.16 t.hm-2^和13.07±1.62 t.hm-2^,两种模式的排放高峰期均出现在8月高温期。多元逐步回归表明,池塘养殖模式CH4的排放量主要受水温和总氮 TN 的影响,N2O排放量则主要受到硝态氮 NO3-^-N 的影响；稻蟹共作模式CH4排放量主要受到总磷 TP 含量的影响,而N2O排放量则主要受到溶解氧 DO 和水温的影响。本研究为中华绒螯蟹不同养殖模式下温室气体的排放提供一定数据支持,同时为稻蟹共作养殖模式的推广示范提供了一定的理论依据。

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• The analysis and comparison of circadian behavioral rhythm between Hippocampus kuda and Oryzias melastigma

  lijiawei, Lin Penghong, Xu Yongjian

  Abstract: (45) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为了探究不同鱼类昼夜行为节律差异及其变化规律，提升人工养殖技术。本文针对管海马和海水青鳉在人工养殖条件下的昼夜行为节律进行分析与比较，结果表明：在光强2000lx、温度25±1℃、盐度25±0.5及光周期14L:10D的条件下，管海马和海水青鳉的行为节律均表现出昼强夜弱。管海马在08:00-10:00间活动节律最强，行为组成最丰富。其中，宏观行为组成包括游动占比47.40%±1.1%、爬行3.0%±0.6%、移动4.80%±0.7%以及摆动32.50%±2.1%和静止12.30%±0.8%等；细微行为组成包括：鳃盖运动38.77±1.14次/min、吻盖运动38.77±1.14次/min、眼球运动108.50±5.52次/min、背鳍运动116.87±0.64次/min和胸鳍运动67.81±2.27次/min。次强节律时间段为12:00-14:00，主要为摆动行为，占比为59.20%±5.1%。海水青鳉也在08:00-10:00时间段有最强烈的活动节律，其中，游动占比88.0%±1.1%、鳃盖运动407.88±3.8次/min、颌运动407.88±3.8次/min、眼球运动7.38±0.9次/min、背鳍运动373.17±2.5次/min、胸鳍运动400.83±6.8次/min、尾鳍运动416.58±7.7次/min。因此，不管是管海马还是海水青鳉，作为视觉性鱼类，均存在明显的昼夜行为节律表现。其中，8:00-10:00时间段是两种鱼类行为的活跃期，活动节律强、行为组成丰富，因此可以选择该时间段作为相关行为学研究的适宜观察时段。

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• Evaluation of the Effectiveness of Molecular Marker-Based Release for Chinese Shrimp in the Yellow Sea off the Southern Coast of the Shandong Peninsula

  qiujingyaun, lvding, huyulong, wangweiji, lvguanzheng, shanxiujuan

  Abstract: (39) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  中国对虾(Fenneropenaeus chinensis)是我国黄渤海重要的增殖放流物种,其放流效果的评估对黄渤海中国对虾资源的养护具有重要意义。为开展此项研究,本研究先后于2023年春季在山东半岛南部海阳黄海水产有限公司,收集生产完成的中国对虾亲虾404尾,作为放流苗种；当年9月份中国对虾秋汛期间在黄岛、海阳和崂山海域,回捕中国对虾个体共计461尾；于2024年春季,在崂山和日照海域,分别采集中国对虾生殖洄游个体110尾和399尾。实验采用微卫星分子标记和线粒体COI基因序列信息相结合的亲子溯源方法,对山东半岛南部黄海海域中国对虾放流效果进行了评估。结果显示,2023年秋汛回捕样本以及2024年生殖洄游的回捕样本,共计监测到放流个体112尾,其中,秋汛回捕样本和生殖洄游样本中各监测到56尾来自放流的个体。上述结果表明,山东半岛南部黄海海域增殖放流的中国对虾,无论对当年黄海秋汛生物量和翌年生殖洄游的亲虾资源量均有显著贡献。

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• pIgR gene in Aristichthys nobilis: Molecular characterization and immune response to Aeromonas hydrophila challenge

  Xia Hu, Ding Cheng, Liu Liang-guo, YANG Xin-Yue, BU Jian-Chao, Chen Fu-yan, and YANG Pin-Hong

  Abstract: (72) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  多聚免疫球蛋白受体(polymeric?immurnoglobulin?receptor，pIgR)是一种重要的鱼类黏膜免疫因子，在介导多聚免疫球蛋白向黏膜免疫组织的黏液中分泌和转运中起着关键的作用。本研究利用PCR与RACE技术克隆获得鳙（Aristichthys nobilis）pIgR基因cDNA全长序列，鳙pIgR基因cDNA序列全长1885 bp，其开放阅读框长1008bp，编码336个氨基酸，鳙pIgR具有1个信号肽序列，之后为胞外区、跨膜区和胞内区，胞外区包括2个免疫球蛋白样功能域（Ig-like domains, ILD），其三级结构呈扭曲的“L”型。系统进化树分析发现鳙pIgR基因与同属鲤形目鱼类的pIgR聚为一支。实时荧光定量PCR检测结果显示鳙pIgR基因的表达量在肝脏中最高，其次为脾、肠、头肾、鳃及皮肤，在中肾、心及脑中具有较低的表达量，其表达水平最低的组织是肌肉。鳙被嗜水气单胞菌感染5 d，通过组织切片与HE染色观察pIgR基因表达水平高的免疫相关组织病理变化，发现肠绒毛宽度增宽，黏液细胞增多，肠壁增厚，表现出明显的肠道炎症症状；头肾与脾脏组织出现大量的炎症细胞，脾脏中观察到大量的空泡细胞；肝脏肝窦増宽；鳃丝变形缩短，鳃丝细胞凋落。鳙被嗜水气单胞菌感染，头肾、脾脏、鳃、肠和肝中pIgR基因的表达量在感染28 d内呈先上升后降低的趋势，鳃和肠中pIgR基因的相对表达量在感染5 d达到峰值，头肾、脾脏与肝中分别在感染7 d和14 d达到最大值，且鳙pIgR基因在肠（18.63倍）和鳃（16.53倍）中达到峰值时的相对表达量高于其在头肾（9.38倍）、脾脏（15.98倍）和肝（12.28倍）中在峰值时的相对表达量。实验结果表明嗜水气单胞菌感染鳙不仅导致其免疫相关组织发生病变，诱导产生炎症反应，且对呼吸系统和消化系统也具有影响，在鳙抵御细菌感染的免疫反应中pIgR可能在黏膜免疫系统中起着关键的作用。

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• Analysis for ovarian metabolome between hybrid sturgeon population (Huso dauricus♀×Acipenser schrenckii♂) with differential fertility

  LI Yutao, WANG Ruoyu, MA Bo, CAO Dingchen, SUN Zhipeng, WANG Nianmin, XU Shijian, ZHANG Ying

  Abstract: (84) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  本研究旨在通过鉴定高/低繁殖力雌性杂交鲟成年个体性腺组织间差异代谢物并对其进行信号通路富集分析以确定影响鲟鱼繁殖力的关键代谢物种类及生理代谢途径。本研究测量90尾处于性成熟期的雌性杂交鲟个体（达氏鳇 Huso dauricus♀×施氏鲟 Acipenser schrenckii♂）体重及卵重后计算相对卵重系数（卵重/体重），定义相对卵重系数小于0.15为低繁殖力个体，大于0.20则为高繁殖力个体。本试验共计选取高/低繁殖力杂交鲟个体各8尾，采集卵巢组织进行非靶向代谢组学分析。结果表明，与低繁殖力组相比，高繁殖力组杂交鲟个体相对卵重系数显著提高76.92%（P<0.05），说明本试验设定的分组标准合理 。另外发现，在高/低繁殖力组杂交鲟卵巢组织中共计鉴定到14大类小分子代谢物，其中主要代谢物类别为脂质/类脂质分子、有机酸及衍生物以及有机杂环化合物。基于差异代谢物筛选阈值，本试验在高/低繁殖力组之间共计鉴定到11种差异代谢物，分别隶属于五大类别。与低繁殖力组相比，高繁殖力组个体卵巢中（-）维生素B2、维生素B2、亚精胺、腺苷以及溶血磷脂酰肌醇22：6等8种差异代谢物显著下调（P<0.05）。随后对差异表达代谢物进行通路富集分析，发现氨基酸代谢相关通路（“精氨酸和脯氨酸代谢”、“β-丙氨酸代谢”）和维生素代谢相关通路（“维生素B2代谢”、“维生素消化与吸收”）被显著富集（上述4条通路P值分别为0.0019、0.0247、0.0001、0.0004）。另外发现，差异代谢物腺苷可参与14条代谢途径，推测其可能对于杂交鲟繁殖力具有核心调控作用。本研究初步阐明了影响雌性杂交鲟产卵量性状的关键分子信号通路并筛选了高繁殖力表型的候选生物标志物，这将为早期选育高繁殖力杂交鲟群体提供理论基础。

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• Molecular cloning and mRNA expression analysis of Msck8 gene in largemouth bass (Micropterus salmoides)

  yuerongyan, qufengyu, wuhaiqing, linshimei, chenyongjun, luoli, zhouxinghua, heyuanfa

  Abstract: (158) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  细胞角蛋白8(cytokeratin 8, ck8)是一种细胞骨架蛋白,属于角蛋白家族成员,在调控上皮细胞粘附与运动、介导免疫反应等过程中发挥重要作用。为了探究ck8在大口黑鲈(Micropterus salmoides)免疫反应中的作用,本研究利用cDNA末端快速扩增(RACE)技术克隆获得大口黑鲈Msck8基因的全长序列2 424 bp,其中5"非编码区有570 bp,3"非编码区有195 bp,开放阅读框有1 659 bp,可编码552个氨基酸,预测其分子量为61.30 kD,理论等电点为5.17。系统进化树中大口黑鲈与小口黑鲈Msck8蛋白聚为一支,显示了较高的同源性(99.10%)。荧光定量PCR分析显示,Msck8在健康大口黑鲈9个组织中均有表达(P<0.05),其表达量由高到低依次是心脏、肌肉、鳃、肾脏、肝脏、后肠、前肠、中肠和脾脏。经脂多糖免疫刺激后,Msck8在大口黑鲈心脏、肌肉和后肠组织中的表达量均呈时序性变化(P<0.05),心脏和肌肉中均在6 h出现表达高峰,后肠中于48 h出现表达高峰。此外,大口黑鲈经高水平豆粕饲料响应后发现,豆粕替代50%鱼粉组的Msck8基因表达水平显著低于鱼粉组(P<0.05)。综上所述,Msck8可能在鱼体抵御细菌入侵和营养调控的炎症反应中发挥重要作用。

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• Effects of phytosterols on growth, digestion, intestinal, liver morphology, antioxidant and immune levels in Procambarus clarkia

  YANG Zhigang, WANG Xiao, Ren shengjie, LIU Yongjian, GU Xiaolong, WANG Aiming

  Abstract: (310) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  本实验旨在探究饲料中添加植物甾醇对克氏原螯虾（Procambarus clarkii）生长消化、抗氧化、免疫及肠肝形态的影响。在基础饲料中分别添加0%、0.10%、0.19%、0.38%、0.76%的植物甾醇，制备5组等氮等脂饲料，分别命名为CON、P1、P2、P3和P4饲料组，对体均重为（9.37 ? 0.02 g）的克氏原螯虾进行6周的养殖实验。结果显示：P1和P2组的增重率、特定生长率显著高于CON组（P < 0.05），且P2组实验虾的生长性能最佳。P3组实验虾肠道蛋白酶活性显著高于CON组，脂肪酶活性显著低于CON组（P < 0.05）。肝胰腺中P1组酸性磷酸酶和碱性磷酸酶活性最高，血淋巴中P2组酸性磷酸酶和碱性磷酸酶活性最高，但与CON差异均不显著（P > 0.05）。随着植物甾醇水平提高，肝胰腺和血淋巴总超氧化物歧化酶活性与过氧化氢酶活性呈上升趋势，丙二醛含量呈下降趋势。相较CON组，P1组肠道结构更加健康完整，植物甾醇水平到0.19%及以上时，克氏原螯虾的肝胰腺与肠道组织形态出现不良影响。随着植物甾醇水平提高，肝胰腺的NF-κB相对表达水平升高。P1组Hsp70相对表达水平显著高于其他组（P < 0.05）。研究表明，本实验条件下添加0.10%?0.19%植物甾醇可以促进克氏原螯虾的生长消化、改善肠肝组织形态，提高克氏原螯虾抗氧化和免疫能力。

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• Effects of two Bacillus probiotics on growth, digestion, immunity and intestinal microbial communities of largemouth bass (Micropterus salmoides)

  WANG Shigui, YAN Fajun, ZHANG Minglei, MAO Shuquan, ZHU Yongan, FU Chaosong, SONG Liping, LIU Feng

  Abstract: (1674) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  本文选取热带芽孢杆菌(Bacillus tropicus)和传染病研究所副芽孢杆菌(Metabacillus idriensis)为潜在益生菌，探讨它们对大口黑鲈(Micropterus salmoides)生长、消化、免疫及肠道菌群的影响。将初始体重为(92.95±2.36)g的大口黑鲈随机分为5组，每组设3个重复，分别饲喂基础饲料(对照组)和4种试验饲料[于基础饲料中分别添加104和106 CFU/g的B. tropicus(标记为BT4、BT6)或M. idriensis(MI4、MI6)]，饲喂期为60 d。生长试验结果表明，各菌处理组大口黑鲈的终末体重、增重率及特定生长率均高于对照组，但差异不显著(P>0.05)，而饲料系数均显著低于对照且随菌株添加浓度增加呈显著降低变化(P<0.05)。消化方面结果显示，各菌处理组的肠道淀粉酶和蛋白酶活性均高于对照组，其中BT6和MI6对肠道消化性能具有显著提高效果(P<0.05)。免疫方面结果表明，各菌处理组的血清酸性磷酸酶、碱性磷酸酶及溶菌酶以及肠道溶菌酶、超氧化物歧化酶活性均得到显著的提升(P<0.05)，说明菌株对血清和肠道免疫功能均有显著提高作用，但比较发现，总体以BT6与MI6的免疫增强效果最佳。肠道菌群高通量测序结果显示，支原菌属(Mycoplasma)、邻单胞菌属(Plesiomonas)和鲸杆菌属(Cetobacterium)为大口黑鲈肠道优势菌群；各组比较发现，BT6、MI6与对照之间在OTU数量、多样性指数及菌群结构上差异最明显，BT6可降低主要优势菌支原菌属(P<0.05)和多数劣势菌群的丰度来提高菌群均一性进而提高菌群多样性，而MI6可促进多数劣势类群的生长来提高菌群丰富度和均一性进而提高菌群多样性，这反映了不同菌株对肠道菌群作用方式的差异。本研究表明，热带芽孢杆菌和传染病研究所副芽孢杆菌皆可作为益生菌株应用于大口黑鲈养殖，在本实验条件下最佳使用量均为106 CFU/g，它们具有促进大口黑鲈生长、增强消化和免疫力、促进肠道菌群平衡及提高肠道菌群多样性等作用。本研究为大口黑鲈的益生菌开发与应用提供理论支撑。

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• 1.3 RT-qPCR

  zhanghui, zenglin, xiongyifei, songwei

  Abstract: (716) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  毒物兴奋效应是指有机体经历低剂量胁迫后,可以提高其对相同或不同高剂量胁迫因子的耐受性。由此探讨盐度驯化改善大黄鱼(Larimichthys crocea)盐度胁迫耐受性的作用机制,实验设置六个盐度梯度(25、25+12、25+40、20、20+12、20+40,分别记为C组、CL组、CH组、A组、AL组、AH组),大黄鱼盐度暴露24 h后取样。结果显示,CL vs C中大黄鱼肝脏的丙二醛(malondialdehyde,MDA)、脂质过氧化物(Lipid oxidation,LPO)、超氧化物歧化酶(superoxide dismutase,SOD)、溶菌酶(lysozyme,LZM)活力显著上升,过氧化氢酶(catalase,CAT)活力显著降低(P<0.05),碱性磷酸酶(alkaline phosphatase,AKP)的活力变化不显著(P>0.05)。CH vs C中MDA和LPO含量显著上升,SOD、 LZM的活力显著上升,CAT和AKP显著降低(P<0.05)。A vs C中MDA和LPO含量显著上升,SOD、LZM和AKP的活力未发生显著变化(P>0.05),CAT活力显著降低(P<0.05)。 AL vs CL与AH vs CH中MDA和LPO含量显著降低,SOD、 CAT 、LZM和AKP的活力显著上升(P<0.05)。铜锌超氧化物歧化酶(Cu/Zn-SOD)、锰超氧化物歧化酶(Mn-SOD)、CAT、非特异性免疫酶［c型溶菌酶(c-type LZM)、g型溶菌酶(g-type LZM)和AKP］基因表达水平与其酶活成正相关；核转录因子NF-E2相关因子2(NF-E2-related factor 2,Nrf2)和核转录因子-κB(nuclear transcription factor-κB,NF-κB)基因表达水平均分别与其目的基因(抗氧化酶基因和非特异性免疫酶基因)表达水平呈正相关(P<0.05),表明Nrf2和NF-κB可能分别参与了抗氧化和非特异性基因表达调控。综上所述,基于毒物兴奋效应原理,盐度驯化可以通过提高大黄鱼抗氧化和非特异性免疫能力来缓解盐度胁迫所带来的氧化损伤,从而改善大黄鱼的盐度耐受性。

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• Development of vertebral column and appendicular skeleton in Collichthys lucidus larvae and juveniles

  WU Chenyan, TANG Pengyan, ZHONG Junsheng, WANG Xiaodong, HUANG Jieli

  Abstract: (914) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  本研究基于2014年6-7月在杭州湾北部水域用大型仔稚鱼网（口径1.3m、网目0.5mm）进行表层拖网所采集的棘头梅童鱼（Collichthys luncidus）仔稚鱼样本，采用软骨-硬骨双染色技术，探讨了棘头梅童鱼仔稚鱼（体长范围为3.3-12.7mm）脊柱及附肢骨骼的早期发育和骨化特征。研究结果表明：脊柱骨化之前不分节，而是由前向后逐渐长出的一个个骨环把脊柱分节，硬骨化出现的顺序大体上为由前向后。髓弓在脊柱骨化之前完全形成，相对于一些低等鱼类髓弓等的生成方向是从中间向头尾和从后向前进行，棘头梅童鱼的髓弓由前向后生长，属于相对高等鱼类的生长类型。胸鳍支鳍骨是最先发育的附肢骨骼，为能够自由地游泳摄食和从内源性营养转变为外源性营养提供了保障。后颞骨的形状十分特殊，具有四枚向不同方向突起的棘；背鳍支鳍骨由体中部开始向前后生长，尾鳍支鳍骨的骨化晚于其他附肢骨骼，侧尾下骨与其相邻的两枚尾下骨的基部融合，为其游泳提供了强劲的动力。该研究结果不仅可以为早期形态学分类提供骨骼发育学方面的科学依据，对棘头梅童鱼人工繁殖过程中结合骨骼发育进行技术上的契合也具有重要的参考价值。

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• Detection of Nile tilapia specific antibody secreting cells induced by human IgG by ELISPOT assay

  Huang Yu, Pang Bijian, Li Qi, Wu Zaohe, Jian Jichang

  Abstract: (1159) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  通过酶联免疫斑点法(ELISPOT)检测了经人IgG免疫后尼罗罗非鱼(Oreochromis niloticus)头肾、外周血和脾脏中的特异性抗体分泌细胞(antibody secreting cell,ASC)数量。结果表明：首次免疫1天后可在头肾中检测出ASC,而外周血和脾脏第3天才能检测出ASC；头肾、外周血以及脾脏中的ASC均在第12天达到峰值,随后头肾,外周血中的ASC数量显著减少,而在脾脏中ASC数量减少不显著。二次免疫1天后在头肾、外周血和脾脏中均可检测到ASC,ASC数量均在第9天达到峰值,时间早于首次免疫,且在首次免疫和二次免疫中,头肾组织的ASC数量均是三个组织中最高的。酶联免疫吸附试验(ELISA)分别对首次免疫和二次免疫后血清中的抗体水平检测发现,其变化趋势与ASC数量变化规律相同。研究结果表明,尼罗罗非鱼在初次免疫后产生了免疫记忆,在二次免疫过程中产生了更多的ASC和抗体,头肾是ASC的主要来源组织。该结果为鱼类的免疫记忆研究提供重要的科学依据。

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• Effects of Cu acclimation on oxidative damage in the large yellow croaker under cold stress based on transcriptome analysis

  Zeng Lin, Wang Yong Hong, songwei

  Abstract: (711) HTML (0) PDF (0.00 Byte) (0)

  Abstract:

  为探讨铜驯化对低温胁迫下大黄鱼氧化损伤和基因表达水平的影响,本实验将体质量为(48.92±3.62) g的大黄鱼暴露在铜浓度为0和10 μg L^?1的水体中14 d,再暴露在温度为8^°C的水体中24 h。结果显示,低温胁迫显著增加了活性氧(ROS)和脂质过氧化物(LPO)含量。尽管铜驯化对ROS和LPO含量不产生影响,但铜驯化显著增加了低温胁迫下大黄鱼ROS和LPO含量,表明铜驯化加剧了低温胁迫对大黄鱼的氧化损伤。从铜驯化vs.对照组、低温胁迫vs.对照组和铜驯化+低温胁迫vs.低温胁迫中分别筛选到2 288个、1 425个和1 382个差异基因。GO和KEGG分析发现差异基因主要富集在与脂肪酸代谢、糖类有氧代谢、谷胱甘肽代谢、内质网应激、自噬和凋亡等相关的通路中。聚类分析表明,低温胁迫上调了不饱和脂肪酸合成、内质网应激、自噬和凋亡等相关通路中的大部分基因表达,而铜驯化则对低温胁迫下大黄鱼的这些基因表达调控产生了拮抗效应,表明铜驯化通过抑制不饱和脂肪酸合成、内质网应激、自噬和凋亡来降低大黄鱼的低温胁迫耐受性。研究结果为深入研究铜污染物对大黄鱼低温胁迫耐受性的影响及其分子机制提供科学依据。

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