硫化物對凡納濱對蝦（Litopenaeus vannamei）的毒性效應研究
發(fā)布時間：2018-07-29 08:39
【摘要】：本文報道了水體硫化物對凡納濱對蝦的毒性效應,從生理生化、代謝組學、腸道健康等方面,探討了硫化物對凡納濱對蝦的影響,研究結果為完善水生動物毒理學提供了基礎資料,同時也將有助于緩解硫化物對凡納濱對蝦負面影響方法的研究。主要結果和結論如下:在鹽度25psu和pH7.96±0.19條件下24h、48h、72h和96h時硫物化對凡納濱對蝦的半致死濃度分別為8301.9、6986、5643.2和4254.8μg/L,安全濃度為425.5μg/L。將凡納濱對蝦暴露在 0、425.5μg/L(1/10LC50-96h)和 851μg/L(1/5LC50-96h)的硫化物濃度下,進行了為期21天的慢性暴露,研究了硫化物對該對蝦生長、能量代謝、血清代謝組學、及其腸道健康和菌群的影響。結果發(fā)現(xiàn),隨著硫化物濃度的升高,凡納濱對蝦的成活率顯著降低,增重率和肝體比各組間并沒有顯著差異,肝胰腺和肌肉中的糖原含量降低,且肝胰腺中細胞色素C氧化酶的活性顯著降低,但血清中的血糖、乳酸含量及乳酸脫氫酶的活性顯著上升。代謝組學分析顯示,在425.5μg/L濃度的硫化物暴露條件下,血清中的丙酮酸、琥珀酸、甘氨酸、丙氨酸和脯氨酸等代謝物顯著下調,在851μg/L濃度的硫化物暴露下,血清中的磷酸鹽、琥珀酸、丙氨酸、絲氨酸和組氨酸等代謝物也顯著下調。下降可能因凡納濱對蝦在硫化物慢性暴露下,干擾了蝦體內的蛋白質合成,增強了糖異生的作用,并加速了 ATP在生物合成及三羧酸循環(huán)中作為底物的消耗,促使蝦體提供額外的能量來緩解硫化物的暴露。此外,凡納濱對蝦在不同濃度硫化物暴露下的血清中天冬氨酸、苯丙氨酸、馬來酸及焦谷氨酸等代謝物均顯著上調,說明在慢性硫化物暴露下,不僅對凡納濱對蝦體內的能量代謝造成干擾,并且對蝦體的健康狀況也造成了一定的影響。隨著硫化物濃度的增加,對蝦腸道損傷加重,炎癥和免疫相關細胞因子產(chǎn)生一系列應激反應以及會對腸道微生物菌群結構造成影響。暴露在硫化物條件下,對蝦肌肉生長抑制素(MSTN)的表達顯著下調,熱休克蛋白因子(HSF1)的表達在硫化物濃度為851μg/L時顯著上升,但在對照組和425μg/L沒有顯著差異,腫瘤壞死因子(TNF-α)和C型凝集素3(CTL3)在425μg/L時的表達都顯著上升,但在851μg/L時的表達都下降。硫化物也改變了腸道微生物的菌群結構,大量致病菌如:藍細菌(Cyanobacteria)、弧菌(Vibrio)、發(fā)光桿菌(Photobacterium)的豐富度隨著硫化物濃度增加顯著增長,同時抵抗外來致病菌的益生菌梭菌(Fusobacteria)和適應硫化物暴露的綠菌(Chlorobi)的豐富度也顯著增加。而硫酸鹽還原細菌即硝化螺旋菌(Nitrospira)在硫化物暴露下消失。一些能適應硫化物暴露相關的微生物如:微桿菌(Microbacterium)/副衣原體(Parachlamydia)和希瓦氏菌(Shewanella)也出現(xiàn)顯著下降現(xiàn)象,下降可能因其未適應高濃度硫化物暴露環(huán)境,具體原因待深入研究。其他微生物在硫化物暴露下也有顯著性差異,由于目前對這些微生物在水生生物毒理方面的功能研究有限,文章中未作深入討論,但本研究結果將為進一步的探究提供基礎研究信息,并對在硫化物暴露下甲殼動物腸道健康的研究提供了新方向。
[Abstract]:The toxic effects of sulfides on Penaeus vannamei were reported in this paper. The effects of sulfides on Penaeus vannamei were discussed from physiological, biochemical, metabolic and intestinal health. The results provided a basic information for improving the toxicology of aquatic animals, and would also help to alleviate the negative effects of sulfide on Penaeus vannamei. The main results and conclusions are as follows: in the conditions of salinity 25psu and pH7.96 + 0.19, the semi lethal concentration of sulphide to Penaeus vannamei at 24h, 48h, 72h and 96h respectively is 8301.969865643.2 and 4254.8 mu g/L, and the safe concentration is 425.5 u g/L. to expose Penaeus vannamei to 425.5 mu g/L (1/10LC50-96h) and 851 micron g/L. The effects of sulfide on the growth, energy metabolism, serum metabolism, and intestinal health and bacteria groups were studied for 21 days of chronic exposure. The results showed that the survival rate of Penaeus vannamei decreased significantly with the increase of sulfide concentration, and there was no significant difference between the weight gain rate and the liver body ratio in the hepatopancreas. The content of glycogen in the glands and muscles decreased, and the activity of cytochrome C oxidase in the hepatopancreas decreased significantly, but the blood sugar, lactic acid content and the activity of lactate dehydrogenase in the serum were significantly increased. The metabolomics analysis showed that pyruvic acid, succinic acid, glycine, alanine and proline in the blood were exposed to the concentration of 425.5 u g/L. The metabolites such as ammonia were significantly down, and the metabolites such as phosphate, succinic acid, alanine, serine and histidine in the serum were also significantly reduced under the exposure to 851 g/L concentration of sulfides. The decrease may interfere with the egg white synthesis in shrimp, enhance the effect of sugar isogenesis, and accelerate the AT The consumption of P as a substrate in biosynthesis and the three carboxylic acid cycle encourages shrimp to provide additional energy to alleviate sulphide exposure. In addition, the metabolites of aspartic acid, phenylalanine, maleic acid and pyroglutamic acid in serum of Penaeus vannamei in different concentrations of sulfide exposure are significantly up-regulated, indicating that under chronic sulfide exposure, It not only interferes with the energy metabolism in the Penaeus vannamei, but also affects the health of the shrimp body. With the increase of sulfide concentration, the damage of the intestinal tract is aggravated, the inflammatory and immune related cytokines produce a series of stress reactions, and it will affect the structure of the intestinal microflora. Exposed to vulcanization The expression of MSTN was significantly down, and the expression of heat shock protein factor (HSF1) increased significantly when the concentration of sulfide was 851 u g/L, but there was no significant difference between the control group and 425 g/L. The expression of tumor necrosis factor (TNF- a) and C type agglutinin 3 (CTL3) increased significantly at 425 u g/L, but at 851 um g/L. The sulfide also changed the structure of the intestinal microflora, and a large number of pathogenic bacteria such as Cyanobacteria, Vibrio, and Photobacterium were increased significantly with the increase of sulphide concentration, while the bacteria resistant to the bacteria of the bacteria (Fusobacteria) and the green adapted to the sulphide exposure. The richness of bacteria (Chlorobi) also increased significantly. The sulfate reducing bacteria (Nitrospira) disappeared under sulphide exposure. Some microbes that can adapt to sulfide exposure, such as microbacilli (Microbacterium) / Chlamydia (Parachlamydia) and sivysiella (Shewanella), have also decreased significantly. It does not adapt to the high concentration of sulfide exposure environment, specific reasons for in-depth study. Other microorganisms have significant differences under sulfide exposure, due to the limited research on the function of these microorganisms in aquatic biological toxicology, but the article is not discussed in depth, but the research results will provide a basic research letter for further research. It also provides a new direction for the research of intestinal health of crustacean under sulfide exposure.
【學位授予單位】：華東師范大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S917.4
本文編號：2152115
[Abstract]:The toxic effects of sulfides on Penaeus vannamei were reported in this paper. The effects of sulfides on Penaeus vannamei were discussed from physiological, biochemical, metabolic and intestinal health. The results provided a basic information for improving the toxicology of aquatic animals, and would also help to alleviate the negative effects of sulfide on Penaeus vannamei. The main results and conclusions are as follows: in the conditions of salinity 25psu and pH7.96 + 0.19, the semi lethal concentration of sulphide to Penaeus vannamei at 24h, 48h, 72h and 96h respectively is 8301.969865643.2 and 4254.8 mu g/L, and the safe concentration is 425.5 u g/L. to expose Penaeus vannamei to 425.5 mu g/L (1/10LC50-96h) and 851 micron g/L. The effects of sulfide on the growth, energy metabolism, serum metabolism, and intestinal health and bacteria groups were studied for 21 days of chronic exposure. The results showed that the survival rate of Penaeus vannamei decreased significantly with the increase of sulfide concentration, and there was no significant difference between the weight gain rate and the liver body ratio in the hepatopancreas. The content of glycogen in the glands and muscles decreased, and the activity of cytochrome C oxidase in the hepatopancreas decreased significantly, but the blood sugar, lactic acid content and the activity of lactate dehydrogenase in the serum were significantly increased. The metabolomics analysis showed that pyruvic acid, succinic acid, glycine, alanine and proline in the blood were exposed to the concentration of 425.5 u g/L. The metabolites such as ammonia were significantly down, and the metabolites such as phosphate, succinic acid, alanine, serine and histidine in the serum were also significantly reduced under the exposure to 851 g/L concentration of sulfides. The decrease may interfere with the egg white synthesis in shrimp, enhance the effect of sugar isogenesis, and accelerate the AT The consumption of P as a substrate in biosynthesis and the three carboxylic acid cycle encourages shrimp to provide additional energy to alleviate sulphide exposure. In addition, the metabolites of aspartic acid, phenylalanine, maleic acid and pyroglutamic acid in serum of Penaeus vannamei in different concentrations of sulfide exposure are significantly up-regulated, indicating that under chronic sulfide exposure, It not only interferes with the energy metabolism in the Penaeus vannamei, but also affects the health of the shrimp body. With the increase of sulfide concentration, the damage of the intestinal tract is aggravated, the inflammatory and immune related cytokines produce a series of stress reactions, and it will affect the structure of the intestinal microflora. Exposed to vulcanization The expression of MSTN was significantly down, and the expression of heat shock protein factor (HSF1) increased significantly when the concentration of sulfide was 851 u g/L, but there was no significant difference between the control group and 425 g/L. The expression of tumor necrosis factor (TNF- a) and C type agglutinin 3 (CTL3) increased significantly at 425 u g/L, but at 851 um g/L. The sulfide also changed the structure of the intestinal microflora, and a large number of pathogenic bacteria such as Cyanobacteria, Vibrio, and Photobacterium were increased significantly with the increase of sulphide concentration, while the bacteria resistant to the bacteria of the bacteria (Fusobacteria) and the green adapted to the sulphide exposure. The richness of bacteria (Chlorobi) also increased significantly. The sulfate reducing bacteria (Nitrospira) disappeared under sulphide exposure. Some microbes that can adapt to sulfide exposure, such as microbacilli (Microbacterium) / Chlamydia (Parachlamydia) and sivysiella (Shewanella), have also decreased significantly. It does not adapt to the high concentration of sulfide exposure environment, specific reasons for in-depth study. Other microorganisms have significant differences under sulfide exposure, due to the limited research on the function of these microorganisms in aquatic biological toxicology, but the article is not discussed in depth, but the research results will provide a basic research letter for further research. It also provides a new direction for the research of intestinal health of crustacean under sulfide exposure.
【學位授予單位】：華東師范大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S917.4
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