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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:
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.
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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:
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.
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2024,31(12):1432-1440, DOI: 10.12264/JFSC2024-0212
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.
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LIU Zhifei, WANG Feng, CHEN Shuaiyu, CHEN Xiatian, JIA Yudong
2024,31(12):1441-1452, DOI: 10.12264/JFSC2024-0220
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 (Pcrit) 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 Pcrit, 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 Pcrit 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.
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TAN Jie, ZHANG Zheng, LIU Changlin, WANG Liang, XIAO Wenjie, JIANG Da
2024,31(12):1453-1463, DOI: 10.12264/JFSC2024-0226
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.
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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
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.
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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:
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.
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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:
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.
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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:
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.
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YANG Min, QIN Jiao, GU Long, LI Ruojing, ZHAO Na, ZHANG Bo
2024,31(12):1511-1523, DOI: 10.12264/JFSC2024-0193
Abstract:
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.
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QIU Xingyu, LIU Qingxia, CHEN Zuozhi, CAI Yancong, DAI Shouhui, HUANG Honghui
2024,31(12):1524-1538, DOI: 10.12264/JFSC2024-0202
Abstract:
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.
Volume 31,2024 Issue 12
研究论文
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Correlation analysis of bacterial structure and environmental factors during the skin ulcer syndrome outbreak period of Apostichopus japonicus cultured in cage aquaculture system in Fujian Province
Wang jinjin, Wang yingeng, Li bin, Rong xiaojun, Liao meijie, Ge jianlong, Chen Zhengqiang, Liu qingbing, Jiang Minqi
Abstract:
本研究以2023年11月份福建东吾洋的小雷江、文岐、围江海域吊笼养殖区暴发的刺参腐皮综合征为研究对象，对其发病特征、菌群结构及菌群与环境因子的相关性进行解析，以期掌握该养殖模式下相关疾病发生时期的养殖系统的菌群结构特征，为该区域刺参病害防控提供科学依据。本研究通过对患病组织及水样中微生物的16S rDNA可变区V3~V4区域进行高通量测序及菌群结构分析，结果表明检测到的细菌归属于33门，72纲，350 科，各区域患病刺参体壁样品中，位于前三的优势菌门均为变形菌门（Proteobacteria）、拟杆菌门（Bacteroidota）和放线菌门（Actinobacteriota），优势菌科有黄杆菌科（Flavobacteriaceae）、红杆菌科（Rhodobacteraceae）、肠杆菌科（Enterobacteriaceae）和微杆菌科（Microbacteriaceae）等；在3个区域的水体样品中，位于前二的优势菌门均为变形菌门和拟杆菌门，优势菌科有黄杆菌科、红杆菌科、放线菌科（Actinomarinaceae）和蓝细菌（unclassified_Cyanobacteriia）等。α多样性分析显示Chao1和ACE指数在不同区域的刺参体壁和水样中均表现出显著的差异性，PCoA分析显示水体样品聚集在一起外周包围着刺参体壁样品，表明水样菌群与海参健康具有一定的相关性。对所研究样品中科水平上Top 20的菌群进行相关性分析发现，18个菌群间有100对组合具有显著相关性，且毛螺菌科（Lachnospiraceae）、颤螺菌科（Oscillospiraceae）与鼠杆菌科（Muribaculaceae）相互之间存在着极强的相关性，相关性系数达到了0.95以上。菌群与环境因子间相关性分析显示，BOD/Hg/Zn/Ni四种环境因子与微生物的α多样性具有显著的相关性（p < 0.05）。综合上述结果表明，患病刺参个体的体壁组织中存在大量的弧菌，患病组织与水体中的菌群具有显著的相关性，且环境因子对菌群结构及微生态的稳定性具有重要影响。相关研究结果将有助于为刺参腐皮综合征的防控提供科学依据，并进一步推动刺参养殖过程中环境保护及养殖工艺的优化，为科学养殖及病害防控提供理论依据。
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Screening of Genes and SNP Markers Related to the Body Wall Proportion of Sea Cucumber (Apostichopus japonicus) Based on Selective Sweeping Analysis
Song zhiwei, Ge Jianlonng, Wang Jinjin, Liao Meijie, Li Bin, Wang Yingeng, Rong Xiaojun, Niu Licheng, He Yating
Abstract:
刺参(Apostichopus japonicus)出皮率即刺参体壁占总体重的比例, 是刺参重要的经济性状和判别刺参品质的重要指标，开展刺参出皮率的遗传调控机制可为良种选育提供科学依据。 本研究以同一养殖环境下的同一批次养殖刺参群体为基础, 对250个随机选择个体进行出皮率测定, 筛选出高出皮率(H组)和低出皮率(L组)刺参各25头, 基于全基因组重测序和选择消除分析, 筛选与出皮率相关的受选择区域, 挖掘出皮率相关基因和关键通路, 进一步确定出皮率相关SNP位点和优势基因型。全基因组重测序共得到7398410个高质量的SNP, 群体遗传结构分析表明两组间遗传结构无明显分化。基于Fst & Pi的选择消除分析分别筛选出高出皮率组和低出皮率组524个和484个受选择区域, 分别注释到214个和193个候选功能基因。通路富集分析得到1199个预测GO类目和44个KEGG代谢通路, 从其中获得与出皮率差异相关的4个显著差异富集通路: 代谢途径(metabolic pathways)、脂肪酸伸长(fatty acid elongation)、脂肪酸代谢(fatty acid metabolism)和黏蛋白O-聚糖生物合成(mucin type O-glycan biosynthesis)。通过功能基因检索分析筛选出ELP2、TPRA1、G2E3、TIE1、DOCK1 5个可能与高出皮率相关的功能基因, 进一步筛选出在这5个相关基因内与出皮率相关的28个SNP位点, 并获得了与高出皮率相关的优势基因型4个。上述研究结果为探究刺参出皮率遗传调控机制提供了基础数据, 也为培育高出皮率刺参优良品种培育和标记辅助育种技术建立奠定基础。
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Research on the hydraulic characteristics of large land-based high-position circular aquaculture tank for pollution collection
GUI Fukun, CHEN Jianeng, FENG Dejun, NAN Hailin, ZHOU Qixian, ZHANG Qingjing, WU Yanfei
Abstract:
为探究推水器作用下大尺寸陆基高位圆池内的水动力特性。通过进行现场试验,设计了3组推水器布设角度(与池壁的相对角度,θ = 0°、20°、45°),每组角度工况下设置7组推水器数量(n = 2、3、4、6、8、10、12)。利用“小威龙”声学多普勒流速仪测量池内表、中、底三个不同水层(h = 30 cm、60 cm、90 cm)的流速分布,从平均流速、养殖池均匀系数U等方面分析养殖池内水动力特性的变化。结果表明：推水器设置方式对流场和水动力特性影响显著,在布设12个推水器的情况下,当布设角度θ为0°时,池中央存在大面积的低流速区,不同水层的养殖池均匀系数集中在0.15～0.25之间,水体混合效果较差。布设角度θ从20°增大到45°时,池内低流速区面积随着角度增大而减小,布设角度为45°时各个工况下的平均流速都超过了0.047 m/s,养殖池均匀系数都在0.3以上,养殖池流场分布最为均匀,池内高流速区面积显著扩大。布设角度θ为45°时,当推水器布设数量n > 2,推水器数量的变化对池内流场分布的影响没有明显差异。相对于布设数量n上的变化,布设角度θ会对池内流场变化带来更明显的影响。本研究结果可为大尺寸圆形养殖池内推水器布设方式的优化提供理论依据。
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Molecular cloning, interaction with CDX2 and expression regulation of CDK2 in grass carp (Ctenopharyngodon idella)
HE Zhimin, LUO Junhan, WANG CHEN Haoyang, TANG Xiang, JIANG Ran, WANG Qixiang, LIU Jing, LI Yanfang, GAO Xing, ZENG Peng, QING Yanling, LUO Yanan, LIU Zhen
Abstract:
为探讨草鱼(Ctenopharyngodon idella) cdk2的分子特征、CDK2蛋白与CDX2的相互作用及其表达营养调控, 本研究通过克隆获得草鱼cdk2基因序列, 其开放阅读框897 bp, 编码298个氨基酸, 与其他物种CDK2氨基酸序列同源性达到80%以上, 且具有保守S_TKc功能域; 基于cdk2的密码子偏好性及聚类分析均表明, 草鱼与斑马鱼(Danio rerio)的亲缘关系最近; 相对基于cdk2 RSCU的聚类分析, 基于CDK2氨基酸序列的系统进化树分析更符合传统物种的分类; 组织表达分析结果表明, 草鱼cdk2 mRNA在血液组织表达水平最高(P<0.05), 肝脏和脾脏次之。通过分子对接和酵母双杂交分析了草鱼CDK2与其下游磷酸化底物蛋白CDX2的相互作用, 结果表明草鱼CDK2与CDX2具有相互作用。本研究进一步通过养殖实验探究了饲料中添加不同水平的游离氨基酸CAA(游离lys和Glu)、二肽KE (Lys-Glu, KE), 三肽GHK (Gly-His-Lys, GHK)对cdk2的表达调控, 结果表明, 相比对照组, 0.5% GHK对草鱼cdk2 mRNA的表达促进作用最大, 氨基酸CAA相比具有同样氨基酸组成的KE二肽, 更能促进cdk2的表达。综上所述, 草鱼cdk2基因表达具有组织特异性, CDK2与CDX2相互作用, cdk2表达水平受到饲料中氨基酸和小肽水平的影响。本研究为其他硬骨鱼类cdk2分子特征的研究, 鱼类肠道功能基因的调控机制探索研究提供新的线索, 并为后续开展cdk2基因调控氨基酸及小肽的转运吸收分子机制研究提供理论基础。
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Establishment of a highly sensitive one-step PCR assay for detecting white spot syndrome virus (WSSV) in Procambarus clarkii based on the vp19 gene
Chen Nan, Luo Xiaohong, Liao Mingcong, Hu Ruixue, Li Dan, Gu Zemao
Abstract:
白斑综合征病毒（WSSV）是危害克氏原螯虾养殖的重要病原。本研究旨在针对该病毒早期检测和诊断速度慢、灵敏度不高、不便捷等问题，建立新的WSSV一步式PCR检测方法。该方法基于WSSV主要囊膜蛋白VP19的基因保守序列设计引物，建立方法并针对分析特异性、分析灵敏度、诊断特异性、诊断灵敏度开展了与WOAH巢式PCR方法的比较评估。结果表明，本方法对WSSV具有高分析特异性，仅在阳性样本中扩增出196 bp条带，与传染性皮下和造血器官坏死病毒等其它常见水产病原无交叉反应。分析灵敏度测试显示，以WSSV标准质粒为模板时，本方法理论检出限低至1.83 copies/μL；以田间样品混合DNA为模板时，最低检出限达0.82 fg/μL，均优于WOAH推荐巢式PCR方法。同时，我们以WOAH Taqman qPCR方法为标准，比较了本方法和巢式PCR对317份田间样品的检测结果，本方法的诊断特异性为97.01%，诊断灵敏度为95.08%，均优于巢式PCR。综上所述，本研究建立的一步式PCR检测方法具有高特异性和高灵敏度的优点，适用于克氏原螯虾WSSV周年监测和快速诊断，能够为有效防控WSSV相关流行病提供重要的技术支持。
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A study on the impact of hydrological connectivity in the Chongming Dongtan tidal creek system of the Yangtze River Estuary on fish community distribution
feishujie, zhangtingting, zhaofeng, zhangtao, yanggang, wangsikai
Abstract:
水文连通性是指示河口湿地生态系统健康和功能的重要特征之一,其对河口鱼类群落的分布具有重要影响。本文选取了水文动态、景观维度上的2类共12个水文连通性指标,探索了河口湿地潮沟系统的水文连通性的差异性及鱼类群落对不同水文连通性栖息地的偏好,旨在为河口湿地的生态修复、生物多样性保护和水资源管理提供科学依据。结果表明：在不同潮汐条件下,河口湿地潮沟系统的水文连通性对鱼类群落的分布具有显著影响,潮差、淹没时长、节点数、河链数和节点连接率β等被识别为关键影响因素。进一步的典型相关分析分析(CCA),可将潮沟系统内部样点区域分为大潮高连通区、大潮低连通区、小潮高连通区、小潮低连通区和长期淹没区,共5类不同水文连通性区域,并且鱼类物种对不同水文连通区域的偏好各异,这些偏好与鱼类的游泳能力、个体大小和摄食行为等相关。例如,大鳞鲴(Xenocypris macrolepis)、光泽黄颡鱼(Pelteobagrus nitidus)等鱼类对水体水流环境适应力强,偏好利用涨落潮进出潮沟觅食,倾向于在大潮高水文连通区域栖息。中国花鲈(Lateolabrax maculatus)和鲻科(Mugilidae)等是具有较强的溯水能力的鱼类,虾虎鱼科(Gobiidae)偏好深入潮沟系统内部更稳定的水体觅食或栖息,它们倾向于在淹水时长较长的小潮高水文连通区域栖息。刀鲚(Coilia ectenes)、大银鱼(Protosalanx chinensis)等幼鱼,对水深有一定的要求,不喜进入潮沟内部,偏好栖息在潮沟口或近岸的区域。研究结果表明了河口潮沟鱼类在潮汐作用下,对不同水文连通性栖息地的具有主动选择和适应性,为河口湿地的生态修复和鱼类生物多样性保护提供了科学依据,有助于在湿地水资源管理中制定调节水文连通性策略,以促进生态系统的可持续发展。
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Transcriptomics investigation of the mechanism of temperature effects on the embryonic development of Eriocheir sinensis
CUI Chaoyue, GENG Zhi, FENG Guangpeng, ZHANG Tao, ZHAO Feng, YANG Gang, ZHUANG Ping, HUANG Xiaorong
Abstract:
中华绒螯蟹（Eriocheir sinensis）是我国重要的经济蟹类，具有典型的洄游习性，每年需经历数月洄游至长江口进行繁殖活动，温度是影响其胚胎发育的关键环境因子之一。本研究通过设置恒温9℃、恒温15℃以及从9℃逐渐升高至15℃三个实验组，查明每个实验组胚胎发育的时间和有效积温，并利用转录组学技术揭示了温度对中华绒螯蟹胚胎发育的分子调控机制。结果表明，温度显著影响胚胎发育进程及有效积温，在恒温9℃时，胚胎停滞在卵裂期不发育，其有效积温为1961.95 h·℃；温度从9℃逐渐升高至15℃时，胚胎能够发育至原溞状幼体期，且与恒温15℃组相比，胚胎发育时间缩短了144h，有效积温缩短了1303.77 h·℃。温度变化显著影响了胚胎发育相关基因的表达模式，当温度从9℃升高至15℃时，与胚胎发育相关的间隔基因（LOC126998238）显著下调，功能通路主要富集在内质网蛋白靶向；在恒温15℃下，胚胎中的母体效应基因（LOC126984051）则显著上调，功能通路主要富集在小分子代谢。研究表明，在一定范围内，胚胎发育随温度升高而加快，与恒温条件相比，变温能显著影响胚胎发育相关基因的表达及功能通路的富集，从而促进中华绒螯蟹胚胎的发育。
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Evaluation of self and hybrid culture edible rate, quality and toxic heavy metals of male Chinese mitten crab reared in cold regions
Sujunjie, Wangshihui, Zhangshuqi, Zhaozhigang, Luoliang, Zhangrui, Guokun
Abstract:
为筛选适宜在寒地养殖的河蟹种质并评价其品质特征，本研究对三种配组河蟹(辽河自交LL、长江自交YY、辽河♀×长江♂杂交LY)扣蟹进行为期5个月的养殖，获得成体雄蟹并比较了可食率、肥满度、色泽、常规营养成分、脂肪酸组成、游离氨基酸组成、矿物质元素组成以及有毒重金属元素。结果表明：LY的出肉率MY和总可食率TEY显著高于其余两组(P< 0.05)；YY和LY头胸甲红度值、肝胰腺亮度值均显著高于LL (P< 0.05)，LY肝胰腺黄度值显著高于其余两组(P< 0.05)；LY性腺系统和肌肉的粗蛋白含量显著高于其余两组(P< 0.05)；LY肝胰腺中∑PUFA、∑EFA、n-3/n-6 PUFA和DHA/EPA高于其余两组； LY性腺系统中脯氨酸Pro含量、肌肉中丙氨酸Ala和脯氨酸Pro含量显著高于其余两组(P< 0.05)；LY性腺系统和肌肉的总苦味氨基酸∑TBV相对较少；LY性腺系统和肌肉的总矿物质元素∑TME显著高于其余两组(P< 0.05)，LY的铁Fe和锌Zn含量显著高于其余两组(P< 0.05)；LL、YY和和LY中有毒重金属元素含量远低于食品中污染物限量国家标准。综上所述，虽然LL肝胰腺指数HSI和性腺指数GSI较高，LL在肝胰腺和肌肉中的总脂含量和∑EFAA高于其余两组，YY肝胰腺和性腺系统中的灰分含量显著高于其余两组(P< 0.05)，但LY在绝大多数大部分指标上更占据优势，表现出更高的营养价值和食用安全性。
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The Pilot Application of Astaxanthin in Preventing Blue Body Syndrome(BBS) in Litopenaeus vannamei
Wei Xuchong, Yi Huadong, Fu Guangxu, Bi Sheng, Zhang Zhilun, Chen Jiahui, Li Guifeng, Wang Haifang
Abstract:
为探讨虾青素在改善凡纳滨对虾(Litopenaeus vannamei)蓝体综合征(Blue Body Syndrome, BBS)引起的体色异常、生长迟缓和体质虚弱中的作用机制，本研究在前期工作的基础上设计了2种虾青素添加水平饲料：0 mg/kg (对照组, CT)和100 mg/kg (虾青素组, AX)，在循环水养殖系统中投喂初始体重为0.77±0.27 g的幼虾60 d，并对两组对虾的体色、不同组织的虾青素含量、组织结构、生长性能、抗氧化能力及免疫能力进行了系统比较与分析。结果显示：(1) CT组活体对虾体表呈现蓝色，表现出典型的蓝体综合征症状。而AX组活体对虾体表呈现浅褐色，处于正常状态。养殖期间，CT组对虾肝胰腺、甲壳、肌肉3个组织的总虾青素含量降低了21.51%。而AX组的总虾青素含量增加了95.17%，且肝胰腺、甲壳的虾青素含量均显著升高(P<0.05)。实验结束时，AX组的对虾蓝体比率为8.67%，小于CT组的76.00%。结果表明，外源供给虾青素显著增加了对虾的虾青素沉积量，并改善对虾体色，减少了蓝体综合征的发生。(2) 在CT组对虾的不同组织切片中，观察到肝胰腺细胞萎缩变形，肌肉肌纤维细胞排列疏松，肠道单层柱状上皮细胞脱落等病理变化，而AX组对虾的各组织结构未见异常。结果表明，蓝体综合征造成对虾多个重要组织的结构损伤，而外源供给虾青素有助于对虾维持不同组织的正常结构。(3) 与CT组相比，AX组对虾的淀粉酶、脂肪酶和胰蛋白酶活性，生长相关基因igf-1, igf-2的相对表达量，生长性能均显著提高(P<0.05)，表明虾青素通过增强对虾的消化酶活性和促进生长相关基因的表达，改善蓝体综合征引起的生长抑制。(4) 与CT组相比，AX组对虾总抗氧化能力、超氧化物歧化酶和过氧化氢酶活性均显著提高(P<0.05)，丙二醛含量显著降低(P<0.05)，表明虾青素通过提高对虾的抗氧化酶活性，改善蓝体综合征引起的抗氧化功能失调。(5) 与CT组相比，AX组对虾免疫相关基因crustin、penaeidin 3a、proPO、relish的相对表达量均显著上调(P<0.05)，表明虾青素通过促进免疫相关基因的表达，改善蓝体综合征引起的免疫抑制。综上，虾青素摄入不足引起的营养代谢紊乱是凡纳滨对虾蓝体综合征发生的重要原因。虾青素在对虾体内被优先用于强化其抗氧化防御系统，保护不同组织的结构与生理代谢活动，随后进行沉积着色，从而改善蓝体综合征引起的体色异常和生理功能紊乱。
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Research progress on key enzyme genes involved in DHA biosynthesis and their transcriptional regulation in fish
HU Xuesong, ZHONG Dawei, JIA Zhiying
Abstract:
二十二碳六烯酸(docosahexaenoic acid, DHA) (22:6n-3)是近年来备受人们关注的一种n-3型长链多不饱和脂肪酸(long chain polyunsaturated fatty acid, LC-PUFA)，在婴幼儿脑和视力发育中不可或缺，并具有调节血脂、抑制肿瘤生长和预防阿尔茨海默症等生理功能。鱼类是人们获取DHA的重要来源。然而，随着野生海洋鱼类资源的下降和养殖鱼类产量的不断攀升，未来从养殖鱼类中获取DHA的比重预计将显著提升。新近研究表明，基于对DHA合成核心因子及其调控机制的深入解析，通过基因操作或选育等手段有望提高养殖鱼类DHA的内源合成能力，从而减少饲料中鱼油的配比，并增加鱼体DHA含量。在LC-PUFAs合成过程中，碳链延长酶(elongases of very long-chain fatty acid, Elovl)和脂肪酸去饱和酶(fatty acid desaturases, Fads)分别通过延长碳链和引入双键，发挥着重要作用。本文综述了鱼类DHA合成途径及在各节点起关键作用的Elovl和Fads基因家族成员；探讨了这些基因的分工、功能的补偿和多样性；在启动子水平分析了转录因子对上述关键酶基因的不同调控模式及其对DHA合成的影响。本文旨为利用基因编辑或分子模块设计等技术开展养殖鱼类DHA合成能力的遗传改良和精准育种提供理论参考。
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Study on the habitat suitability of Miichthys miiuy along the coast of Zhejiang
shen jia rong, jiang ri jin, cui ming yuan, yin rui, zhang hong liang, li qi qun, zhang yu rong
Abstract:
鮸(Miichthys miiuy) 是浙江近海主要的经济鱼类之一，为了解浙江近海鮸栖息地分布与环境因子的关系，本研究根据2022年4月(春季)、7月(夏季)、11月(秋季)和2023年2月(冬季)在浙江近海开展的渔业资源调查数据和同步采集的环境数据，确定了浙江近海鮸的适宜性指数(suitability index, SI)，通过提升树回归模型(boosting regression tree, BRT)赋予各环境因子权重，利用算术平均法(arithmetic mean model, AMM)和几何平均法(geometric mean model, GMM)分别构建栖息地适宜性指数(habitat suitability index, HSI)模型。结果表明：2022~2023年浙江近海鮸栖息地存在季节性差异。春夏两季，鮸最适栖息于底层盐度范围为26.4~32.1的近岸水域；秋季，鮸最适栖息地向离岸侧移动，最适栖息于20~40 m的近海岛礁水域；冬季，鮸最适栖息于底层温度为11.0~14.1 ℃的近海深水区。浙江近海鮸栖息地时空分布的差异主要受环境因子和鮸生活习性的影响。
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The mechanism of carbonate alkaline exposure on growth performance, biochemical indices and liver metabolism of crucian carp (Carassius auratus)
YUAN Fangying, HAN Lin, LIU Wenzhi, WANG Jing, LI Dongping, WANG Peng, SUN Yanchun
Abstract:
为了探讨碳酸盐碱环境暴露对鲫(Carassius auratus)生长性能与生理代谢的影响机制, 实验设置淡水对照组(Con)、20 mmol/L NaHCO3暴露组(T)和40 mmol/L NaHCO3暴露组(F), 在24±1℃下室内暴露56 d后, 采用生长指标和生化检测方法, 结合UPLC-QTOF/MS非靶向代谢组学技术, 从生长性能、抗氧化能力和代谢物的变化等研究角度, 对鲫进行碳酸盐碱暴露下生长、氧化应激和代谢组学变化特征分析, 以期对碳酸盐碱环境暴露下鲫的生长适应性策略进行评估。研究结果显示, 鲫的增重率、生长率以及饲料转化率等生长参数, 与碳酸盐碱浓度呈现负相关趋势。与此同时, 随着碳酸盐碱浓度的增加, 不仅超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)等抗氧化酶的活力显著增加, 血氨、尿素氮(BUN)、总胆固醇(TC)和甘油三酯(TG)等生化参数的含量也明显升高。代谢组学研究结果显示, 碳酸盐碱暴露下差异代谢物(DEMs)参与多种代谢途径, 包括精氨酸生物合成、不饱和脂肪酸代谢、嘌呤代谢、甘油磷脂代谢、鞘脂代谢和谷胱甘肽代谢等。综合上述研究结果, 我们发现, 在碳酸盐碱暴露下鲫肝靶器官中与生长发育、免疫防护、脂肪酸代谢相关的多条关键代谢通路发生紊乱, 造成鲫的氧化防御系统失衡, 引起肝组织损伤; 并且随着碳酸盐碱浓度的升高, 鲫的免疫防护能力进一步遭到破坏, 肝组织损伤呈加剧趋势, 这可能破坏了细胞膜的稳态, 导致细胞增殖活动受到限制, 进而严重阻碍了鲫正常的生长发育。本研究从生长指标和生化分析视角, 结合代谢组学研究, 阐明了碳酸盐碱环境对鲫生长性能及肝脏代谢的不良反应机制, 为盐碱水域中淡水硬骨鱼类的增养殖活动提供了科学依据。
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Study on chlorophyll-a concentration and phytoplankton carbon sequestration capacity in marine ranching based on satellite remote sensing
Abstract:
海洋牧场基于生态学原理，通过投放人工鱼礁等工程设施，在适宜海域构建兼具环境保护、资源养护和渔业产出功能的生态系统，促进海洋生物增殖和栖息，是扩增渔业水域生态系统碳汇的有效途径。本文结合卫星遥感和原位采样，分析海洋牧场的叶绿素a（Chl-a）浓度及浮游植物固碳的时空变化，并运用广义可加模型（Generalized Additive Model, GAM）探讨环境因子对固碳强度的影响。结果显示，（1）遥感数据与原位数据的Chl-a浓度、海表温度（Sea surface temperature, SST）变化趋势基本一致；（2）海洋牧场建成后，Chl-a浓度在季节和空间上发生显著变化，呈现"M"型双峰结构（高峰分别为夏、秋季过渡期和冬、春季过渡期），整体浓度水平显著高于建成前及对照区；（3）固碳强度存在显著的时空分布差异，夏季（6月）出现高峰，整体水平同样显著高于对照区，海洋牧场全年总固碳量为1.85×10^6 kgC；（4）最优GAM模型中，pH、硅酸盐、溶解氧、盐度和纬度对固碳强度有显著影响。
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Study On The Sources Of Occurrence Floating S.horneri In The Coastal Waters Of Gouqi Island
zhuweidong, yangqifan, wanghuijie, miaohang, liufengyu, qinpengjie, biyuanxin
Abstract:
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:
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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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:
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。
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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:
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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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:
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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Study on artificial propagation and early development of Garra mirofronits
wangyuesong, yangyang, ganxing, yinhuazheng, pengli, zhangzhiwei, chengrui, chenfeng, shishaolong, mashu, zhangzhiming
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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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:
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:
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:
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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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:
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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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:
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:
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:
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:
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:
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:
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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