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A controlled experiment was conducted to investigate temperature-induced alterations in serum biochemical indices and gut microbiota of endemic fish (Procypris rabaudi) in Jinsha River, a cascade hydropower development river in southwest China. Three temperature treatments (16℃, 20℃, 24℃) were established with exposure durations of 24h and 10d. Results showed that compared to the ambient temperature group, the serum antioxidant enzyme activity was promoted in the low temperature group after 24h but inhibited after 10d. The decrease in lysozyme (LZM) activity in the low temperature group of 10d (67.12%) was more significant than that of 24h group (52.04%) relative to the ambient temperature group. And the concentrations of glucose (GLU) and CORTISOL were increased significantly in the low temperature groups both 24h and 10d compared with those in the ambient temperature groups. The Chao1index of the low temperature group at 24h were 23.22% and 26.36% lower than those of the ambient temperature and high temperature groups, respectively. The alpha diversity was found to stabilize in the low temperature group after 10d. Compared to 24h, the difference in gut microbiota community structure under different temperature conditions after 10d was smaller, and the low temperatures significantly affected the community composition of the gut microbiota. Co-occurrence network analysis revealed that microbial interactions were simplified and weakened in the low temperature group (24h), whereas prolonged thermal adaptation (10d) was associated with network stabilization. Significant correlations were established between Proteobacteria, Bacteroidetes, and Firmicutes with antioxidant enzymes and LZM activity.

, correspAuthors=Juan CHEN, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Dan-li CHEN, Xiao-ling YANG, Shuai-shuai LIU, Bo ZHANG, Ge CUI, Han GAO, Juan CHEN), CN=ArticleExt(id=1241408719502766382, articleId=1241408716931657936, tenantId=1146029695717560320, journalId=1234093305789726721, language=CN, title=水温变化对金沙江特有鱼类岩原鲤血清生化指标及肠道微生物的影响, columnId=1234106394006311784, journalTitle=中国环境科学, columnName=环境毒理与健康, runingTitle=null, highlight=null, articleAbstract=

以西南梯级水电开发河流金沙江中特有鱼类岩原鲤(Procypris rabaudi)为研究对象,开展了控制实验,分别设置24h和10d的3个处理组(16,20,24℃),探究不同水温条件下岩原鲤血清生化指标和肠道微生物的变化.结果表明,相较于常温组,24h和10d低温组对岩原鲤血清抗氧化酶活力分别起到了促进和抑制作用;10d低温组的溶菌酶(LZM)活力相对常温组的下降率(67.12%)较24h(52.04%)更为显著; 24h和10d低温组的葡萄糖(GLU)和皮质醇(CORTISOL)浓度相较于常温组均显著增加. 24h低温组的Chao1指数比常温组和高温组的分别低23.22%和26.36%,10d低温组的alpha多样性趋于稳定;相比于24h,10d后不同水温条件下肠道微生物群落结构差异更小;低温显著影响肠道微生物的群落组成.共现网络显示24h低温组的微生物共生关系简化,微生物间相互作用减弱;而长期水温变化则使微生物的互作关系趋向稳定.相关性分析显示变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)等与抗氧化酶和LZM活力存在显著相关性.

, correspAuthors=陈娟, authorNote=null, correspAuthorsNote=
* 责任作者,教授,
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陈丹丽(1999-),女,广东汕头人,河海大学硕士研究生,主要研究方向为水电开发河流的生物响应机制..

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陈丹丽(1999-),女,广东汕头人,河海大学硕士研究生,主要研究方向为水电开发河流的生物响应机制..

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陈丹丽(1999-),女,广东汕头人,河海大学硕士研究生,主要研究方向为水电开发河流的生物响应机制..

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不同字母表示同一时间下不同处理组间差异显著(P<0.05)

, figureFileSmall=RyGzDKdYFS5cUYmMpiXucw==, figureFileBig=Ao6MCFULduOudO1d4VhOWA==, tableContent=null), ArticleFig(id=1241408726838604750, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408716931657936, language=EN, label=Fig.2, caption=The alpha diversity indices and NMDS analysis of gut microbial community of Procypris rabaudi under different water temperature conditions, figureFileSmall=2gNCOraS9W1Ls2BpVh6cww==, figureFileBig=zmoZD/63DAgytxLbuetpUg==, tableContent=null), ArticleFig(id=1241408726989599713, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408716931657936, language=CN, label=图2, caption=不同水温条件下的岩原鲤肠道微生物群落的alpha多样性指数和NMDS分析

(a)Chao1指数,(b)Shannon指数,(c)Simpson指数,(d)24h NMDS分析,(e)10d NMDS分析;不同字母表示同一时间下不同处理组间显著差异(P<0.05),ns表示没有显著差异; *表示P<0.05,**表示P<0.01,***表示P<0.001

, figureFileSmall=2gNCOraS9W1Ls2BpVh6cww==, figureFileBig=zmoZD/63DAgytxLbuetpUg==, tableContent=null), ArticleFig(id=1241408727350309886, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408716931657936, language=EN, label=Fig.3, caption=Gut microbial community composition of Procypris rabaudi under different water temperature conditions, figureFileSmall=QmfVVfsDQY0JVO2fhAwdHQ==, figureFileBig=kFLIV1pPuKU5WSTKGOtmCg==, tableContent=null), ArticleFig(id=1241408727513886736, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408716931657936, language=CN, label=图3, caption=不同水温条件下的岩原鲤肠道微生物的群落组成

(a)-(c)岩原鲤肠道微生物优势门(top 10)相对丰度及其组间差异,(d)-(f)岩原鲤肠道微生物优势科(top 10)相对丰度及其组间差异; ND表示未检出;不同字母表示同一时间下不同处理组间显著差异(P<0.05),ns表示没有显著差异

, figureFileSmall=QmfVVfsDQY0JVO2fhAwdHQ==, figureFileBig=kFLIV1pPuKU5WSTKGOtmCg==, tableContent=null), ArticleFig(id=1241408727643910179, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408716931657936, language=EN, label=Fig.4, caption=Co-occurrence network and subnetwork topological features of gut microbial community of Procypris rabaudi, figureFileSmall=t8JXcRuophRRRQdJHIXlMw==, figureFileBig=duG6pwdCqQP7RDHzm0pSPQ==, tableContent=null), ArticleFig(id=1241408727874596923, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408716931657936, language=CN, label=图4, caption=岩原鲤肠道微生物群落共现网络和子网络拓扑指标

(a)岩原鲤肠道微生物共现网络,(b)节点数,(c)连接数,(d)平均度,(e)平均路径长度;不同字母表示同一时间下不同处理组间显著差异(P < 0.05),ns表示没有显著差异

, figureFileSmall=t8JXcRuophRRRQdJHIXlMw==, figureFileBig=duG6pwdCqQP7RDHzm0pSPQ==, tableContent=null), ArticleFig(id=1241408728013008972, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408716931657936, language=EN, label=Fig.5, caption=Correlation analysis of gut microbial community of Procypris rabaudi with serum biochemical indices, figureFileSmall=1qtXVudgYz1C6ms/UNF6cA==, figureFileBig=rRD8kqmP7Ozkg//nMLD0Fw==, tableContent=null), ArticleFig(id=1241408728327581803, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408716931657936, language=CN, label=图5, caption=岩原鲤肠道微生物群落与血清生化指标相关性分析

*表示P < 0.05,**表示P < 0.01,***表示P < 0.001

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水温变化对金沙江特有鱼类岩原鲤血清生化指标及肠道微生物的影响
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陈丹丽 1, 2 , 杨晓灵 3 , 刘帅帅 1, 2 , 张波 1, 2 , 崔戈 1, 2 , 高寒 1, 2 , 陈娟 1, 2, *
中国环境科学 | 环境毒理与健康 2025,45(4): 2288-2297
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中国环境科学 | 环境毒理与健康 2025, 45(4): 2288-2297
水温变化对金沙江特有鱼类岩原鲤血清生化指标及肠道微生物的影响
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陈丹丽1, 2 , 杨晓灵3, 刘帅帅1, 2, 张波1, 2, 崔戈1, 2, 高寒1, 2, 陈娟1, 2, *
作者信息
  • 1.河海大学环境学院,江苏 南京 210098
  • 2.河海大学浅水湖泊综合治理与资源开发教育部重点实验室,江苏 南京 210098
  • 3.珠江水资源保护科学研究所,广东 广州 510611
  • 陈丹丽(1999-),女,广东汕头人,河海大学硕士研究生,主要研究方向为水电开发河流的生物响应机制..

通讯作者:

* 责任作者,教授,
Effects of water temperature changes on serum biochemical indices and gut microbiota of Endemic Fish (Procypris rabaudi)in Jinsha River
Dan-li CHEN1, 2 , Xiao-ling YANG3, Shuai-shuai LIU1, 2, Bo ZHANG1, 2, Ge CUI1, 2, Han GAO1, 2, Juan CHEN1, 2, *
Affiliations
  • 1.College of Environment, Hohai University, Nanjing 210098, China
  • 2.Key Laboratory of Integrated Regulation and Resource Department on Shallow Lakes, Ministry of Education, Hohai University, Nanjing 210098, China
  • 3.Scientific Institute of Pearl River WaterResources Protection, Guangzhou 510611, China
出版时间: 2025-04-20
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以西南梯级水电开发河流金沙江中特有鱼类岩原鲤(Procypris rabaudi)为研究对象,开展了控制实验,分别设置24h和10d的3个处理组(16,20,24℃),探究不同水温条件下岩原鲤血清生化指标和肠道微生物的变化.结果表明,相较于常温组,24h和10d低温组对岩原鲤血清抗氧化酶活力分别起到了促进和抑制作用;10d低温组的溶菌酶(LZM)活力相对常温组的下降率(67.12%)较24h(52.04%)更为显著; 24h和10d低温组的葡萄糖(GLU)和皮质醇(CORTISOL)浓度相较于常温组均显著增加. 24h低温组的Chao1指数比常温组和高温组的分别低23.22%和26.36%,10d低温组的alpha多样性趋于稳定;相比于24h,10d后不同水温条件下肠道微生物群落结构差异更小;低温显著影响肠道微生物的群落组成.共现网络显示24h低温组的微生物共生关系简化,微生物间相互作用减弱;而长期水温变化则使微生物的互作关系趋向稳定.相关性分析显示变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)等与抗氧化酶和LZM活力存在显著相关性.

梯级水电开发  /  水温  /  岩原鲤  /  血清生化指标  /  肠道微生物

A controlled experiment was conducted to investigate temperature-induced alterations in serum biochemical indices and gut microbiota of endemic fish (Procypris rabaudi) in Jinsha River, a cascade hydropower development river in southwest China. Three temperature treatments (16℃, 20℃, 24℃) were established with exposure durations of 24h and 10d. Results showed that compared to the ambient temperature group, the serum antioxidant enzyme activity was promoted in the low temperature group after 24h but inhibited after 10d. The decrease in lysozyme (LZM) activity in the low temperature group of 10d (67.12%) was more significant than that of 24h group (52.04%) relative to the ambient temperature group. And the concentrations of glucose (GLU) and CORTISOL were increased significantly in the low temperature groups both 24h and 10d compared with those in the ambient temperature groups. The Chao1index of the low temperature group at 24h were 23.22% and 26.36% lower than those of the ambient temperature and high temperature groups, respectively. The alpha diversity was found to stabilize in the low temperature group after 10d. Compared to 24h, the difference in gut microbiota community structure under different temperature conditions after 10d was smaller, and the low temperatures significantly affected the community composition of the gut microbiota. Co-occurrence network analysis revealed that microbial interactions were simplified and weakened in the low temperature group (24h), whereas prolonged thermal adaptation (10d) was associated with network stabilization. Significant correlations were established between Proteobacteria, Bacteroidetes, and Firmicutes with antioxidant enzymes and LZM activity.

the cascade hydropower development  /  water temperature  /  Procypris rabaudi  /  serum biochemical indices  /  gut microbiota
陈丹丽, 杨晓灵, 刘帅帅, 张波, 崔戈, 高寒, 陈娟. 水温变化对金沙江特有鱼类岩原鲤血清生化指标及肠道微生物的影响. 中国环境科学, 2025 , 45 (4) : 2288 -2297 .
Dan-li CHEN, Xiao-ling YANG, Shuai-shuai LIU, Bo ZHANG, Ge CUI, Han GAO, Juan CHEN. Effects of water temperature changes on serum biochemical indices and gut microbiota of Endemic Fish (Procypris rabaudi)in Jinsha River[J]. China Environmental Science, 2025 , 45 (4) : 2288 -2297 .
特有鱼类是指生活在某些特定水域或地理区域范围内的鱼类[1],具有地理局限性、数量稀少性和独特的生态适应性,在保护和维持生态平衡方面具有重要价值[2].特有鱼类对外界环境变化极其敏感,特别是水温的变化.水温波动可能扰乱特有鱼类的产卵时间,影响其繁殖成功率[3-5],进而导致特有鱼类种类组成和群落结构特征发生改变[6].血清生化指标常用于评估鱼类对水温变化的适应能力及其生理健康状况[7].水温波动会改变鱼类血清中的超氧化物歧化酶(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,进而影响鱼类的抗氧化性能[8].此外,水温变化引起的溶菌酶(LZM)活力、葡萄糖(GLU)和皮质醇(CORTISOL)浓度变化可能会影响鱼类的免疫功能及能量代谢[9].肠道微生物在鱼类的营养物质消化吸收和免疫调节中发挥着重要作用[10],对维持宿主生理健康至关重要.低温环境可能引起氧化应激反应,影响抗氧化性能[11]和免疫系统[12],破坏肠道屏障,导致肠道微生物群落结构失调和多样性降低[13],从而影响鱼类的健康和营养状态.水温适应条件显著影响藏区特有鱼类Oxygymnocypris stewarti幼鱼的肠道热应激反应和肠道菌群结构[14].同样,青藏高原特有鱼类Gymnocypris eckloni在不同温度下也展现出特有的肠道适应机制[15].肠道微生物群落的失调不仅影响鱼类的免疫调节,还可能通过改变能量代谢、影响抗氧化反应等方式,进一步改变血清生化指标[16-17].然而,目前对于特有鱼类肠道微生物的研究仍较为匮乏.
近年来,大型梯级水库建设引发水库底层低温水的释放导致了河流水温持续下降,显著影响着下游以鱼类为主要对象的水生生物的生长繁殖和栖息模式[18].现有研究多集中于探讨水库建设引起的水温波动对河流生态生境和生物种群结构的影响,关于梯级水电开发下水温演变规律对特有鱼类肠道微生物的影响仍知之甚少.金沙江位于中国长江的上游,是我国最大的水电基地,其水电资源丰富,具备巨大的开发潜力[19].然而,其梯级水库群改变了河流水温,破坏了鱼类栖息地,导致鱼类多样性减少及其摄食、繁殖等生理行为受到严重影响[20].岩原鲤(Procypris rabaudi)作为金沙江特有鱼类[21-22],其生存状况在水电开发影响下不容乐观.因此,本研究选取岩原鲤为研究对象,通过控制实验探究不同水温条件下岩原鲤血清中相关生化指标的变化,分析其肠道微生物的多样性和群落组成差异,并解析微生物间的共生关系,揭示水温和时间耦合变化下岩原鲤血清生化指标及肠道微生物群落的潜在关联.研究结果为认知水温变化对特有鱼类生理状态及肠道微生物的影响提供科学依据,为西南梯级水电开发河流特有鱼类生态响应及管理保护提供数据支持.
金沙江特有鱼类岩原鲤(50尾)均取自宜宾市向家坝鱼类增殖放流站周边某养殖场,体重(551±16) g,体长(26.85±0.33) cm.实验前,在20℃条件下,于玻璃纤维钢养殖池(2.5m×1m×0.8m,水深0.5m)中进行驯化处理,驯化周期为7d,驯化及后续实验所用水源取自金沙江部分河段.期间,每日(9:00)饱食投喂1次膨化配合饲料,喂食量为岩原鲤体重的1.5%,每次投喂后及时捞取残余饲料.每天定时(18:00)进行1次完全换水,以保持水质的稳定和清洁,确保实验环境的稳定性.
选取18条体型均匀,健康状况良好的岩原鲤,确保体重和体长差异最小,以避免个体差异的干扰.分别饲养于3个体积均为2m3的玻璃纤维钢养殖池中,水体流速保持在0.1m/s,确保环境一致,最大限度减少环境变量对实验结果的影响.近年来,金沙江梯级水库区年均最低水温为15℃,年均最高水温为25℃[23-24].因此,实验设置3组不同的水温条件:低温组(16℃)、常温组(20℃)和高温组(24℃).使用电子加热棒(LH-2000,韩国亚美光公司)和控温器(811-H,韩国亚美光公司)在30min内将水温从20℃升高至24℃,并保持恒温.降温处理则采用冷水机(10AD,中国捷之迈公司)将水温从20℃降至16℃并维持稳定.
在实验24h和10d后,从每个养殖池中随机抽取3尾岩原鲤进行样品采集.使用浓度为100mg/L的MS-222麻醉剂进行快速麻醉,随后将鱼体置于冰盘上.采用无菌2mL注射器从尾静脉处采集血样,将其置于4℃冰箱中静置2h后进行离心处理:4℃下,以4000r/min的转速离心20min,分离血清.上层血清立即转移至−20℃冰箱中储存,用于血清生化指标的测定.同时,采集肠道组织,使用生理盐水冲洗后液氮速冻,再转移至−80℃超低温冰箱保存,用于后续肠道微生物组分析.
岩原鲤血清中的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)、与溶菌酶(LZM)的活力以及葡萄糖(GLU)与皮质醇(CORTISOL)的浓度采用南京建成生物工程研究提供的试剂盒进行测定:T-SOD试剂盒(A001-1,羟胺法)、CAT试剂盒(A007-1,可见光法)、GSH-PX试剂盒(A005-1,比色法)、LZM试剂盒(A022-1,比浊法)、GLU试剂盒(A110-1,葡萄糖氧化酶法)、CORTISOL试剂盒(A105-1,酶联免疫吸附法).
使用E.Z.N.A. Soil DNA Kit(Omega Bio-tek,Inc.,USA)的DNA提取试剂盒,按照生产商的说明文件提取18个肠道样本中的DNA.随后,采用Illumina MiSeq PE250平台(Illumina,San Diego,CA,USA)进行细菌16S rRNA基因的高通量扩增子测序.参照Chen等[25]的方法,选取带barcode的特异性引物341F (5’-CATAYGGGRBGCASCAG-3’)和806R (5’-GGAATACHVGGGTWTATAAT-3’),对细菌16S rRNA基因的V3-V4高变区进行聚合酶链式反应(PCR)扩增.参照Liu等[26]的方法构建基因文库,各样品的纯化PCR产物归一化至等摩尔数,并在Illumina MiSeq PE250平台上机测序.使用Quantitative Insights Into Microbial Ecology (QIIME 2)对原始测序数据进行质量过滤,将过滤后的高质量序列按照100%的相似性聚类成扩增子序列变异(ASV),选择每个ASV中出现频率最高的序列,利用RDP分类器在Silva数据库(http://www.arb-silva.de)中对这些ASV代表序列进行比对注释,最终以序列最少的样本为标准将ASV进行均一化处理.
研究数据以样品的3个平行重复的平均值±标准偏差进行统计分析.使用SPSS软件(version 26.0,IBM Corporation,NY,USA)的单因素方差分析(One-way ANOVA)分析同一饲养时间下,血清生化指标、肠道微生物alpha多样性指数、微生物共现网络拓扑指标在不同水温下的差异显著性,并通过Waller-Duncan检验进行多重比较,显著性差异的准则设定为P<0.05.利用R(version 4.0.5)软件“vegan”包进行以下分析:①计算肠道微生物的alpha多样性指数(Chao1指数、Shannon指数和Simpson指数);②依据Bray-Curtis距离(ASV数据log转换:log10x+1))来计算肠道微生物的beta多样性,使用非度量多维尺度分析(NMDS)对肠道微生物群落结构的差异进行可视化,并利用相似性分析(ANOSIM)方法评估3个水温组间微生物群落结构差异的显著性.使用“WGCNA”包构建肠道微生物的共现网络(Spearman |r|>0.7,P<0.01),并通过Gephi(version 0.9.2)平台进行网络可视化.同时,利用“igraph”包计算了共现网络中子网络的节点数、连接数、平均度和平均路径长度等拓扑指标.利用R中的“pheatmap”包绘制alpha多样性、相对丰度前5细菌门、前5细菌科和网络拓扑指标与血清生化指标的Spearman相关性热图,进一步探讨岩原鲤肠道微生物与血清生化指标之间的关系.
图1a~1c所示,在抗氧化性能方面,低温组和高温组24h的SOD、CAT及GSH-PX活力分别为460.19,6.64,684.42U/mL和488.93,13.54,731.60U/mL,显著高于常温组(SOD: 379.78,CAT: 1.70,GSH-PX: 646.75U/mL).而10d时,低温组和高温组的SOD、CAT及GSH-PX活力显著低于常温组(SOD:常温(433.14)>低温(318.37)>高温(251.11);CAT:常温(3.88)>低温(1.89)>高温(1.07); GSH-PX:常温(607.79)>低温(387.01)>高温(246.75) U/mL),并且与24h相比,低温组和高温组的3种抗氧化酶活力均呈现下降趋势.免疫能力的分析如图1d所示,在24h和10d时,低温组的LZM活力均显著低于常温组.其中,10d时低温组的LZM活力相对常温组的下降率(67.12%)较24h(52.04%)更为显著.此外,对于能量指标,24h和10d时,低温组和高温组的GLU和CORTISOL浓度均显著高于常温组(图1e,1f).可见,水温变化对岩原鲤的抗氧化能力、免疫功能和能量水平产生了显著影响.
岩原鲤肠道样品测序共获得898260条高质量序列,每个样品鉴定出的ASV数量介于44~94之间.使用alpha多样性指数(Chao1指数、Shannon指数和Simpson指数)评估肠道微生物的丰富度和多样性.alpha多样性(图2a~2c)结果表明,24h时,alpha多样性随水温的降低而显著减少.其中,低温组的Chao1指数比常温组和高温组的分别低23.22%和26.36%. 10d时,alpha多样性趋于平稳,且在低温组、常温组和高温组间无显著差异.基于Bray-Curtis距离的NMDS分析(图2d,2e)探讨了岩原鲤肠道微生物群落结构的空间分布特征.ANOSIM结果显示,低温组、常温组和高温组肠道微生物群落结构在24h和10d间均呈现出极显著差异(P<0.01),10d的组间差异性(r=0.484)小于24h(r=0.789).此外,长期组与短期组相比,肠道微生物的alpha多样性整体降低且不同水温组之间趋于相似,不同水温组微生物群落的NMDS点位也显示出更高的集中度,意味着长期水温胁迫下肠道微生物群落逐渐趋于稳定.
图3a所示,低温组、常温组和高温组岩原鲤肠道微生物优势门(top10)主要是变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes),它们的总体相对丰度占比之和在24h和10d均高于0.95. 24h时,低温组变形菌门的相对丰度显著高于常温组和高温组,而拟杆菌门的相对丰度显著低于其他两个实验组;相反,10d的高温组变形菌门的相对丰度显著高于低温组和常温组,而拟杆菌门的相对丰度显著低于其他两组(图3b,3c).肠道微生物的优势科(top10)主要包括噬几丁质菌科(Chitinophagaceae)、慢性根瘤菌科(Bradyrhizobiaceae)、萨特氏菌科(Sutterellaceae)、伯克氏菌科(Burkholderiaceae)和叶菌科(Phyllobacteriaceae)等(图3d),这些菌群总体相对丰度占比和在24h和10d均高于0.65.绝大多数优势科在两个时间段内,不同水温下的相对丰度均呈现显著差异.例如:噬几丁质菌科、慢性根瘤菌科、喜气单胞菌科(Comamonadaceae)、博氏菌科(Boseaceae)、弧菌科(Vibrionaceae)等(图3e,3f).其中,噬几丁质菌科的相对丰度最高. 24h和10d的噬几丁质菌科在高温组的相对丰度分别为0.25和0.31,显著高于低温组(24h: 0.21;10d: 0.27)和常温组(24h: 0.23; 10d: 0.28).根瘤菌科相对丰度从高温组到低温组也呈现出显著下降的趋势.可见,随着饲养时间的延长,不同水温条件下肠道微生物的优势门和优势科相对丰度存在明显差异.
对岩原鲤肠道微生物的ASVs进行共现网络分析(图4a),结果显示共现网络共有260个节点和2728条边.正相关和负相关的连接数占比分别为86.10%和13.90%.共现网络节点主要的微生物门类包括变形菌门、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)、拟杆菌门和酸杆菌门(Acidobacteria).如图4b~4e所示,低温组在24h的共现网络节点数、连接数、平均度均显著低于常温组和高温组,而平均路径长度较高;而10d时,低温组、常温组和高温组的各项网络拓扑指标无显著差异.这意味着,在短期内,低温组的岩原鲤肠道微生物群落比常温组和高温组具有更为简单且不稳定的共现关系,而在长期低温胁迫下,微生物的互作关系逐渐趋向协调与稳定.
Spearman相关性分析显示,SOD、GSH-PX和CAT活力与拟杆菌门和厚壁菌门显著正相关,与变形菌门显著负相关.LZM活力与Chao1指数、Shannon指数、拟杆菌门、厚壁菌门、节点数、连接数和平均度显著正相关,与变形菌门和平均路径长度显著负相关.GLU浓度与厚壁菌门和噬几丁质科显著正相关.CORTISOL浓度与Chao1指数、厚壁菌门和叶杆菌科显著正相关(图5).
本研究发现短期低温刺激显著提高了岩原鲤的抗氧化能力,而长期低温则表现出相反趋势(图1a~1c),与Lv等[27]的研究结果相似:急性低温刺激下发现大黄鱼(Larimichthys crocea)也出现了增强抗氧化能力的情况.推测岩原鲤在短期低温下能够通过上调抗氧化酶基因的表达[28],提高抗氧化酶的活性来对抗急性低温刺激引发的氧化损伤;而长期低温刺激下其细胞的氧化功能受损,降低酶活性和整体机能[29].本研究还发现,低温刺激抑制了岩原鲤的免疫能力,并且这种抑制作用随着暴露时间的延长而显著加剧(图1d).这可能是由于长期低温刺激造成生理应激的积累,导致免疫系统受损[30].此外,有研究表明,低温能够激发鱼类的应激反应,促使其能量代谢机制的启动——激活糖原分解,增加葡萄糖水平以抵御寒冷[31].皮质醇作为应激激素,有助于调节代谢,提升葡萄糖的可用性,支持鱼类在低温下的生理活动[32].这可能是导致本研究低温组岩原鲤血清GLU和CORTISOL浓度相比于常温组显著增加的原因(图1e,1f).这些结果表明,河流筑坝导致的下游水温降低可能会对岩原鲤的生理功能产生重要影响,尤其是在长期低温环境下,可能加剧其生理应激和免疫抑制.
有研究发现,低温可能导致岩原鲤体内消化酶活性下降,肠道资源利用失衡,增加微生物群落的竞争压力[33],这可能是本研究中低温组中肠道微生物alpha多样性低于常温组与高温组的主要原因(图2a~2c).已有对鲑鱼(Oncorhynchus keta[34]、大鳞大马哈鱼(Oncorhynchus tshawytscha[35]、杜氏黄鳍鱼幼鱼(Seriola dumerili[36]等鱼类的研究报道,鱼类肠道微生物群落结构在不同水温之间差异显著.本研究中岩原鲤肠道微生物群落结构在不同水温条件下也存在显著差异(图2d,2e),可能是由于水温的变化影响肠道微生物的生长速率和代谢活动,影响了肠道微生物群落结构[37].据此推测,大坝泄水造成的低温环境可能对岩原鲤的肠道微生物群落造成负面影响,低温水可能改变特有鱼类肠道微生物群落结构进而可能导致微生物多样性和功能的紊乱.
有研究发现,具备耐热性的变形菌门能够广泛分布于鱼类肠道中的高温环境[38],这可能解释了本研究为何在长期低温刺激下变形菌门的相对丰度较低(图3c).而本研究在长期低温刺激下拟杆菌门相对丰度较高,与Idenyi等[39]的结果相似,发现低温环境下虹鳟(Oncorhynchus mykiss)肠道微生物群中拟杆菌门的丰度显著增加,可能是因为拟杆菌门对低温适应能力强,能够维持较高的活性.已有研究报道噬几丁质菌科[40]和慢性根瘤菌科[41]在鱼类肠道微生物群中存在.本研究进一步发现,这些细菌科在岩原鲤肠道中也是主要的优势微生物群.高温环境下,噬几丁质菌科通过分解几丁质在岩原鲤肠道中发挥了关键的消化作用,并反过来促进了其在肠道内的生长与繁殖[42],从而增加了其相对丰度.这可能解释了为何在高温环境下噬几丁质菌科的相对丰度相对于其他处理组明显增高(图3e,3f).慢性根瘤菌科的丰度也随着水温降低而减少,其倾向于高温环境下表现出较强的适应性[43].
共现网络分析是目前探究微生物间复杂相互作用的有效手段,并能揭示环境变化对微生物群落结构的影响[44].在岩原鲤的肠道微生物共现网络中,正相关连接的比例显著高于负相关连接,说明该微生物群落主要以互利共生关系为主导(图4a).微生物群落共现网络中的子网络拓扑特征可以揭示微生物间相互作用的复杂性和紧密性[45],共现网络的节点数、连接数和平均度越低,平均路径长度越长,表明微生物间的相互作用越弱,网络结构变得更加简单且不稳定.与Mu等[46]的研究结果相似,发现鼠兔(Ochotona curzoniae)在低温胁迫下肠道细菌互作网络的稳定性降低.低温压力导致肠道微生物之间相互作用减弱,抵抗环境干扰的能力下降[47],肠道群落转变为简单的共存关系.这可能也是本研究中短期低温环境下岩原鲤肠道微生物的共存关系显著简化,与常温和高温条件下相比更为简单的原因(图4b~4e).低温水下泄可能减弱微生物之间的相互作用,甚至可能导致部分有益菌群的减少,从而影响鱼类的健康状态.此外,长期水温胁迫下肠道微生物的alpha多样性、群落结构和共现网络拓扑特征均表现出不同水温组间趋于同质化的趋势,表明长期温度压力可能对鱼类肠道微生物产生不可逆的不利影响[48-49].
肠道微生物群的改变可以通过氧化应激、营养代谢和肠道损伤等各种途径影响宿主的生理过程[50].已有研究表明,变形菌门中的病原体(如大肠杆菌和沙门氏菌)能够导致幼年杂交石斑鱼[51]和大口黑鲈(Micropterus salmoides[52]肠道屏障功能受损.Guo等[53]研究发现,肠道内高丰度的变形菌门抑制抗氧化能力,并引发免疫紊乱[54].本研究中变形菌门和SOD、GSH-PX、CAT、LZM活力等呈现显著负相关(图5),推测低温刺激下岩原鲤肠道微生物中变形菌门明显增加可能通过破坏肠道屏障功能,导致肠道通透性增加,进而引发炎症和氧化应激反应.这些变化可能通过影响宿主的抗氧化能力和免疫功能,进一步改变血清生化指标[16-17].本研究中厚壁菌门和拟杆菌门与SOD、GSH-PX、CAT、LZM活力产生显著正相关,可能是由于厚壁菌门和拟杆菌门能产生短链脂肪酸(如醋酸、丁酸)等代谢物,这些代谢物具有抗炎、抗氧化作用并增强肠道屏障功能[16],进而有利于促进肠道健康[55].本研究为水温和饲养时间耦合变化下血清生化指标与岩原鲤肠道微生物之间的潜在关联提供了新的见解.目前低温水下泄对肠道微生物群落的潜在作用的研究较为有限,本研究发现的这些水温驱动的变化对理解水库建设引起的低温水对特有鱼类肠道微生物群落的影响,进而对鱼类健康状况(如消化功能)的影响,具有重要参考价值.但其潜在机制仍值得进一步的深入研究.
4.1 短期低温刺激提高了岩原鲤的抗氧化能力,而长期低温则表现出抑制作用.此外,低温刺激在抑制岩原鲤免疫能力的同时,也促进了能量代谢过程.
4.2 低温组肠道微生物的alpha多样性显著低于常温组和高温组,不同水温条件下岩原鲤肠道微生物群落结构组成存在显著差异.
4.3 低温组肠道微生物网络复杂性和稳定性显著低于常温组和高温组,表明低温环境下共生关系更为简单,群落网络更不稳定.
4.4 相关性分析显示,变形菌门、拟杆菌门和厚壁菌门等与抗氧化酶活力和LZM活力存在相关性.
  • 国家重点研发计划项目(2022YFC3203902)
  • 国家自然科学基金项目(52425902)
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  • 接收时间:2024-09-26
  • 首发时间:2026-03-19
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出版历史
  • 收稿日期:2024-09-26
基金
国家重点研发计划项目(2022YFC3203902)
国家自然科学基金项目(52425902)
作者信息
    1.河海大学环境学院,江苏 南京 210098
    2.河海大学浅水湖泊综合治理与资源开发教育部重点实验室,江苏 南京 210098
    3.珠江水资源保护科学研究所,广东 广州 510611

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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