氨氮胁迫下不同食性鱼类仔鱼抗氧化和非特异性免疫的差异响应

ANTIOXIDANT AND NON-SPECIFIC IMMUNE RESPONSES OF FOUR SPECIES OF FISH LARVAE UNDER AMMONIA STRESS

  • 摘要: 为探究氨氮胁迫对不同食性鱼类的影响, 研究选取不同食性的4种鱼类(鲢Hypophthalmichthys molitrix、草鱼Ctenopharyngodon idella、团头鲂Megalobrama amblycephala和黄颡鱼Pelteobagrus fulvidraco)仔鱼为研究对象, 探讨不同浓度氨氮(0、1、2和3 mg/L)短期胁迫(96h)对其生长、抗氧化和非特异性免疫响应的影响及其分子机制。结果显示: (1)氨氮胁迫导致4种仔鱼的体长生长速度呈现剂量依赖性减缓; (2)不同浓度氨氮导致4种仔鱼体内T-AOC、CAT和GPx含量显著降低(P<0.05), 2和3 mg/L氨氮暴露显著降低了草鱼、团头鲂和黄颡鱼仔鱼体内SOD活力(P<0.05), 仅检测到黄颡鱼gpx及鲢sod转录水平出现显著性下调(P<0.05); (3)在不同浓度氨氮胁迫下, 4种仔鱼相关免疫基因转录水平均呈现一定的上调, 仅鲢il1β转录水平显著下降(P<0.05), 相对地草鱼仔鱼LYZ含量显著性下降, 黄颡鱼仔鱼LYZ含量在2 mg/L氨氮组显著性上升(P<0.05)。双因素方差分析显示, 氨氮对所有抗氧化酶、免疫指标及免疫相关基因有显著影响, 不同种仔鱼之间T-AOC、CAT、GPx和C3及基因cuznsodgpxil1βc3之间差异显著(P<0.05), 但鱼种和氨氮互作效应仅对C3和基因gpxtnfαil1βc3影响显著(P<0.05)。研究表明, 高浓度氨氮急性胁迫引起仔鱼生长迟缓和氧化应激, 降低了其抗氧化性能, 削弱了其非特异免疫防御机能。相比较而言, 肉食性的黄颡鱼仔鱼对氨氮的耐受性较其他几种仔鱼弱, 草食性的团头鲂和草鱼仔鱼居中, 而鲢仔鱼对氨氮的耐受性较强。

     

    Abstract: Ammonia, a ubiquitous pollutant in the aquatic system, has been proved to be high toxic to fish. The early life state of fish is acturally more sensitive to the influence of external environmental factors than adult fish since its antioxidant and immune systems are in lower level. However, there are limited information on difference responses of different species of fish larvae to ammonia stress. Aim to explore the effects of ammonia on the antioxidant and non-specific immune response of different species of fish larvae, four different feed-habits species of fish larvae (omnivory silver carp Hypophthalmichthys molitrix, herbivorous grass carp Ctenopharyngodon idella, Wuchang bream Megalobrama amblycephala and predacity yellow catfish Pelteobagrus fulvidraco) were selected and exposed to different concentrations of total ammonia nitrogen (0, 1 mg/L, 2 mg/L and 3 mg/L) for 96h. The results showed that acute ammonia exposue caused significant decrease of body length on four species of fish larvae in a dependent-concentration manner (P<0.05). Meantime, ammonia exposure significant decreased levels of total antioxidant capacity (T-AOC), catalase (CAT) and glutathione peroxidase (GPx) in four species of fish larvae (P<0.05). Also, 2 and 3 mg/L ammonia significantly reduced the activities of superoxde dismutase (SOD) in the larves of grass carp, Wuchang bream and yellow catfish (P<0.05). As for antioxidative-related genes, significant decrease of transcriptional levels were only detected in yellow catfish gpx and silver carp sod after ammonia exposure (P<0.05). In terms of immune parmameters, ammonia exposure significantly up-regulated transcriptional levels of immune-related genes in four species of fish larvae except silver carp interleukin1β (il1β) (P<0.05). By contrast, significant decrease of lysozyme contents were observed in grass carp larvae after ammonia exposure, and significant increase of lysozyme contents were detected in yellow catfish larvae exposed to 2 mg/L ammonia (P<0.05). The results of two-way ANOVA also confirmed that ammonia could caused to varying degree changes for all parameters including antioxidant enzymes, immune indexes and immune-related gene expression in four species of fish larvae (P<0.05). There were significant differences on levels of T-AOC, CAT, GPx, C3 and the gene expression levels of cuznsod, gpx, il1β, c3 among four species of fish larvae. However, interactive effects of ammonia and species were only observed on C3 contents and the expression of genes gpx, tnfα, il1β and c3 (P<0.05). In summary, ammonia exposure caused oxidative stress, decreased the antioxidant capacities and disturbed innate immune responses in four species of fish larvae, which led to growth retardation. Among four species of fish larvae, the predacity yellow catfish was the weakest to ammonia tolerance, the herbivorous Wuchang bream and grass carp is the next, and omnivory silver carp were the strongest.

     

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