凡纳滨对虾淡化养殖池塘浮游细菌群落时空变化及其对理化因子的响应

祁峰; 热比古丽·沙吾提; 咸玉兰; 韩军军; 米乃瓦尔·木衣提; 陈韩飞; 孙世萍; 马燕武; 陈朋

doi:10.12131/20220192

凡纳滨对虾淡化养殖池塘浮游细菌群落时空变化及其对理化因子的响应

doi: 10.12131/20220192

祁峰^{1, 2,},
热比古丽·沙吾提¹,
咸玉兰^{1, 2},
韩军军¹,
米乃瓦尔·木衣提¹,
陈韩飞^{1, 2},
孙世萍^{1, 2},
马燕武^{1, 2},
陈朋^{1, 2, ,}

1.
新疆维吾尔自治区水产科学研究所，新疆乌鲁木齐 830099
2.
农业农村部西北地区渔业资源环境科学观测实验站，新疆乌鲁木齐 830099

基金项目: 新疆维吾尔自治区自然科学计划 (自然科学基金) 面上项目 (2020D01A102)

详细信息

作者简介:
祁峰：祁　峰 (1981—)，男，高级工程师，硕士，研究方向为水域生态学。E-mail: 6481135@qq.com

通讯作者:
陈　朋 (1982—)，男，高级工程师，硕士，研究方向为渔业生态学。E-mail: 290189260@qq.com

中图分类号: S 917.1
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出版历程
- 收稿日期: 2022-07-11
- 修回日期: 2022-09-28
- 录用日期: 2022-10-24
- 网络出版日期: 2022-10-28

Spatio-temporal changes of bacterioplankton communities in Litopenaeus vannamei desalinated ponds and their responses to physicochemical factors

QI Feng^{1, 2
,},
SHAWUTI Rebiguli¹,
XIAN Yulan^{1, 2},
HAN Junjun¹,
MUYITI Minawaer¹,
CHEN Hanfei^{1, 2},
SUN Shiping^{1, 2},
MA Yanwu^{1, 2},
CHEN Peng^{1, 2
, ,}

1.
Institute of Fishery Research of Xinjiang Uygur Autonenous Region, Urumqi 830099, China
2.
Scientific Observing and Experimental Station of Fishery Resources and Environment in Northwest China, Ministry of Agriculture and Rural Affairs, Urumqi 830099, China

摘要

摘要: 浮游细菌调控是对虾养殖水体环境控制策略的核心内容，探究浮游细菌群落构建的一般规律，可进一步推动对虾养殖水体水质调控技术的研究。运用16S rRNA高通量测序技术，对凡纳滨对虾 (Litopenaeus vannamei) 淡化养殖池塘进行了12次周际调查。结果显示，48个样品共获得2 854个操作分类单元(Operational taxonomic units，OTU, 97%相似性)，序列比对发现古细菌2门1纲1科1属，细菌30门59纲98目199科433属，其中优势菌群25属。优势菌群在组成上有较高的相似性，但各池优势菌群在分布和相对丰度变动上有较大差异。各池系统发育多样性指数总平均值为77.57，变幅为24.39~111.65；香农多样性指数总平均值为3.96，变幅为2.64~5.06；物种丰富度指数总平均值为716，变幅为229~1 054。非度量多维标度分析 (Non-metric multidimensional scaling, NMDS) 表明各池塘浮游细菌群落在养殖初期差异较大，中、后期差异减小，冗余分析 (Redundancy analysis, RDA) 显示活性磷、碱度、溶解氧和硫化物可显著影响浮游细菌的群落结构。
- 凡纳滨对虾池塘 /
- 浮游细菌 /
- 群落多样性 /
- 群落结构 /
- 理化因子
Abstract: The regulation of bacterioplankton is the core content of the environmental control strategy in shrimp aquaculture. Exploring the general rules of the construction of bacterioplankton community can further promote the research on water quality regulation for shrimp ponds. Using 16s rRNA high-throughput sequencing technology, we conducted 12 weekly surveys in Litopenaeus vannamei desalinated ponds.The results indicate that a total of 2 854 OTUs (97% similarity) were obtained from 48 samples (Archaea belonged to 2 phyla, 1 class, 1 family and 1 genus; and bacteria belonged to 30 phyla, 59 classes, 98 orders, 199 families and 433 genera, among which 25 genera were dominant flora). The dominant flora had high similarity in the composition, but with great differences in the distribution and relative abundance in each pond. The total average phylogenetic diversity index was 77.57, ranging from 24.39 to 111.65; the total average Shannon diversity index was 3.96, ranging from 2.64 to 5.06; the total average species richness index was 716, ranging from 229 to 1 054. NMDS analysis shows that the community structure of bacterioplankton in each pond varied greatly at the early stage of aquaculture, but not so greatly at the middle and late stages. The results of redundancy analysis shows that the labile phosphorus, total alkalinity, dissolved oxygen and sulfide were the main environmental factors affecting the distribution characteristics of community structure of bacterioplankton.
- Litopenaeus vannamei pond /
- Bacterioplankton /
- Community diversity /
- Community structure /
- Phycicochemical factor

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图 1 凡纳滨对虾养殖池塘门水平的浮游细菌组成

Figure 1. Composition of bacteriaoplankon at phylum level in L. vannamei ponds

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图 2 各池塘优势菌群组成及变动

Figure 2. Composition and changes of dominant flora in each pond

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图 3 3口池塘的α多样性系数分析

注：箱体上中下线分别为75、50 (中位数) 和25分位数，轴须线最长不超过1.5倍箱体范围，黑色空心圆表示平均数；差异显著性用* (P<0.05)、** (P<0.01)以及*** (P<0.001) 表示；图中的样本量：A：n=22、B：n=12、C：n=12。

Figure 3. α diversity index analysis of bacterioplankton in three ponds

Note: The upper, middle and lower lines of the box are 75, 50 (Median) and 25 quantiles, respectively. The maximum length of whiskers shall not exceed 1.5 times of the box range. The black hollow circles represent the average values. The significant differences were represented by * (P<0.05), ** (P<0.01) and *** (P<0.001). The numbers of replicated samples in this figure are: A: n=22; B: n=12; C: n=12.

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图 4 凡纳滨对虾养殖池塘浮游细菌群落NMDS分析

Figure 4. NMDS analysis of bacterioplankton community in L. vannamei pond

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图 5 各池浮游细菌属水平上共有和特有属的数量

Figure 5. Number of common and endemic genera of bacterioplankton at genus level in each pond

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图 6 各池浮游细菌群落的线性判别分析 (菌群LDA>4)

Figure 6. Linear discriminant analysis of bacterioplankton community in each pond (Bacterial flora with LDA>4)

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图 7 优势菌群分布和群落多样性 (H') 与主要理化因子之间的关系 (II型标尺)

注：Aci. 不动杆菌属；Aer. 气单胞菌属；Fla. 黄杆菌属；Gem. 芽殖杆菌属；GpI. GpIIa；Ilu. 微酸菌属；Lim. 湖栖菌属；Pol. 多核杆菌；Rho. 红杆菌属；Sed. 沉积物杆状菌属；Spa. 发光细菌属；Sph. 鞘脂菌属。

Figure 7. Species associations of dominan flora and diversity (H) with environmental factors (Scaling II)

Note: Aci. Acinetobacter sp.; Aer. Aeromonas sp.; Fla. Flavobacterium sp.; Gem. Gemmobacter sp.; GpI. GpIIa; Ilu. Ilumatobacter sp.; Lim. Limnohabitans sp.; Pol. Polynucleobacter sp.; Rho. Rhodobacter sp.; Sed. Sediminibacterium sp.; Spa. Spartobacteria genera incertae sedis; Sph. Sphingomonas sp..

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表 1 凡纳滨对虾养殖池塘水体主要理化因子

Table 1. Environmental factors in L. vannamei ponds

环境因子 Environmental factor	池塘A Pond A	池塘B Pond B	池塘C Pond C
水温 Temperature/℃	26.19±2.01	26.15±1.57	26.37±2.00
pH	8.10±0.50	8.29±0.37	8.30±0.38
溶解氧质量浓度 DO/(mg·L⁻¹)	8.00±0.68	8.45±1.17	8.11±1.15
铵态氮质量浓度 NH₄-N/(mg·L⁻¹)	0.45±0.73	0.44±0.39	0.33±0.27
亚硝酸氮质量浓度 NO₂-N/(mg·L⁻¹)	0.01±0.01	0.01±0.01	0.01±0.01
硝酸态氮质量浓度 NO₃-N/(mg·L⁻¹)	0.34±0.44	0.41±0.42	0.48±0.53
活性磷质量浓度 PO₄-P/(mg·L⁻¹)	0.12±0.14	0.37±0.24	0.22±0.28
活性硅酸盐质量浓度 SiO₃-Si/(mg·L⁻¹)	11.2±3.55	12.38±7.38	4.03±2.98
高锰酸盐指数 COD_Mn/(mg·L⁻¹)	8.34±7.19	6.12±3.18	5.34±2.91
叶绿素 a 质量浓度 Chl-a/(mg·L⁻¹)	75.78±131.01	56.13±42.81	58.36±61.48
硫化物质量浓度Sul/(mg·L⁻¹)	0.03±0.04	0.02±0.01	0.02±0.01
矿化度质量浓度 Mineralization degree/(mg·L⁻¹)	1209.44±2445.58	813.39±171.69	814.35±41.91
碱度 ALK/(mg·L⁻¹)	138.18±12.06	115.32±33.82	84.46±14.73
总硬度 Total hardness/(mg·L⁻¹)	362.76±395.28	476.5±227.34	349.36±60.18

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表 2 各样本有效序列数据统计

Table 2. Valid sequences of each sample

样品 Sample	条形码 Barcode	有效序列 Valid sequence/条	碱基数 Base number	平均长度 Mean length/bp	最短序列长度 Min. length/bp	最长序列长度 Max. length/bp
S1	GTAACA	82 736	34 604 966	418.26	363	469
S2	CCAGAC	81 905	34 619 470	422.68	361	476
S3	GGTGAA	56 859	23 683 959	416.54	367	475
S4	TGCATC	85 212	35 667 198	418.57	353	469
S5	TCGACC	83 246	34 547 936	415.01	365	470
S6	GTCGCG	73 829	30 547 465	413.76	362	453
S7	CGGATG	83 662	34 668 763	414.39	356	465
S8	GTGAAA	84 384	34 811 366	412.54	350	466
S9	ATCTTG	100 945	41 590 813	412.01	350	452
S10	TATGCA	73 801	30 659 241	415.43	352	459
S11	GTAACA	86 471	35 924 249	415.45	354	471
S12	GCGAGG	90 432	37 722 526	417.14	351	465
S13	CACGAT	54 439	22 735 194	417.63	373	465
S14	GCGGTA	44 227	18 478 585	417.81	352	471
S15	TATCGA	61 431	25 638 627	417.36	359	473
S16	ATCACG	52 384	21 792 829	416.02	376	471
S17	CGGATG	99 211	41 215 038	415.43	360	473
S18	CGCATA	100 001	41 427 908	414.27	372	476
S19	TGCATC	65 712	27 246 907	414.64	359	476
S20	TCAGTA	76 029	31 690 841	416.83	362	471
S21	CGGCAC	75 202	31 405 455	417.61	364	469
S22	ATCACG	71 605	30 328 221	423.55	352	470
S23	CGGATG	63 358	26 635 233	420.39	367	470
S24	GTGAAA	56 429	23 698 925	419.98	350	471
S25	TCAGTA	92 400	38 487 742	416.53	355	435
S26	GAAGTG	87 305	37 043 958	424.31	368	448
S27	TCGACC	94 075	39 472 386	419.58	360	464
S28	CTTGTA	52 273	22 054 548	421.91	373	472
S29	GTTTCG	44 697	18 684 447	418.02	360	462
S30	ATCTTG	59 737	25 072 871	419.72	365	465
S32	GCCATC	78 462	32 993 239	420.5	357	474
S33	TGTGTT	69 613	29 167 734	419.00	351	474
S34	CTTGTA	56 083	23 551 548	419.94	356	471
S35	GTTTCG	45 701	18 993 334	415.60	359	464
S36	TTCGTA	46 337	19 212 573	414.63	356	468
S37	CCAGAC	51 978	21 598 326	415.53	372	436
S38	AGCAGT	77 233	31 835 991	412.21	350	470
S39	GAGGAA	75 226	31 060 291	412.89	370	468
S40	AAGGTA	46 920	19 633 227	418.44	352	450
S41	ATCACG	43 983	18 242 559	414.76	352	469
S42	TAGGAC	66 639	27 702 230	415.71	356	470
S43	TGGACG	49 206	20 369 621	413.97	357	472
S44	AGAACA	50 322	20 765 128	412.65	356	469
S45	GGTGTG	41 169	17 011 770	413.22	350	471
S46	AACTAT	67 691	28 070 176	414.68	357	468
S47	ACTGCG	60 220	25 559 117	424.43	359	474
S48	TGTGTT	94 261	39 467 506	418.7	355	474
S49	TAGGAC	86 904	36 530 365	420.35	353	467

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参考文献(40)

[1]	CABELLO F C. Heavy use of prophylactic antibiotics in aquaculture: a growing problem for human and animal healthy and for the environment[J]. Environ Microbiol, 2006, 8: 1137-1144. doi: 10.1111/j.1462-2920.2006.01054.x
[2]	MA Z, SONG X, WAN R, et al. A modified water quality index for intensive shrimp ponds of Litopenaeus vannamei[J]. Ecol Indic, 2013, 24: 287-293. doi: 10.1016/j.ecolind.2012.06.024
[3]	GONZALEZ J M, SIMO R, MASSANA R, et al. Bacterial community structure associated with a dimethylsulfo niopropionate-producing North Atlantic algal bloom[J]. Appl Environ Microb, 2000, 66: 4237-4246. doi: 10.1128/AEM.66.10.4237-4246.2000
[4]	WEI C L, ZENG Y H, TANG K X, et al. Comparison of bacterioplankton communities in three mariculture ponds farming different commercial animals in subtropical Chinese coast[J]. Hydrobiologia, 2009, 632: 107-126. doi: 10.1007/s10750-009-9831-8
[5]	CARON D A. Inorganic nutrients, bacteria, and the microbial loop[J]. Microb Ecol, 1994, 28: 295-298. doi: 10.1007/BF00166820
[6]	COTNER J B, BIDDANDA B. Small players, large role: microbial influence on biogeochemical processes in pelagic aquatic ecosystems[J]. Ecosystems, 2002, 5: 105-121. doi: 10.1007/s10021-001-0059-3
[7]	LEMONNIER H, COURTIES C, MUGNIER C, et al. Nutrient and microbial dynamics in eutrophying shrimp ponds affected or unaffected by vibriosis[J]. Mar Pollut Bull 2010, 60: 402-411.
[8]	AMANN R I. Phylogenetic identification and in situ detection of individual microbial cells without cultivation[J]. Microbiol Rev, 1995, 59: 143-169. doi: 10.1128/mr.59.1.143-169.1995
[9]	LI C, LIU J, CHEN X, et al. Determinism governs the succession of disturbed bacterioplankton communities in a coastal maricultural ecosystem[J]. Sci Total Environ, 2022, 828: 154457. doi: 10.1016/j.scitotenv.2022.154457
[10]	LIU C H, CHANG F H, YANG J W, et al. Free-living marine bacterioplankton composition and diversity along the Kuroshio region[J]. Deep-Sea Res I, 2022, 183: 103741. doi: 10.1016/j.dsr.2022.103741
[11]	LU Z B, REN Z M, LIN W H, et al. Succession, sources, and assembly of bacterial community in the developing crab larval microbiome[J]. Aquaculture, 2021, 548: 737600.
[12]	LIU Y P, PAN B Z, ZHU X Z, et al. Patterns of microbial communities and their relationships with water quality in a large-scale water transfer system[J]. J Environ Manage, 2022, 319: 115678. doi: 10.1016/j.jenvman.2022.115678
[13]	靳燕, 邱莹, 董志, 等. 北运河浮游细菌集合群落空间变化的环境解释[J]. 中国环境科学, 2021, 41(3): 1378-1386. doi: 10.3969/j.issn.1000-6923.2021.03.041
[14]	杨淑芳. 凡纳滨对虾养殖池塘水质动态变化及细菌多样性研究[D]. 厦门: 集美大学, 2017: 10-19.
[15]	张皓. 养殖环境微生物群落的动态变化及与水环境的互作[D]. 苏州: 苏州大学, 2015: 16-29.
[16]	YANG W, ZHENG C, ZHENG Z M, et al. Nutrient enrichment during shrimp cultivation alters bacterioplankton assemblies and destroys community stability[J]. Ecotox Environ Safe, 2018, 156: 366-374. doi: 10.1016/j.ecoenv.2018.03.043
[17]	YANG W, ZHU J, ZHENG C, et al. Succession of bacterioplankton community in intensive shrimp (Litopenaeus vannamei) aquaculture systems[J]. Aquaculture, 2018, 497: 200-213. doi: 10.1016/j.aquaculture.2018.07.053
[18]	国家环境保护总局水和废水检测分析方法编委会. 水和废水检测分析方法(第四版)[M]. 北京: 中国环境科学出版社, 2002: 30-152.
[19]	EDGAR R C. Search and clustering orders of magnitude faster than BLAST[J]. Bioinformatics, 2010, 26(19): 2460-2461. doi: 10.1093/bioinformatics/btq461
[20]	AKSNES D L, WASSMANN P. Modeling the significance of zooplankton grazing for export production[J]. Limnol Oceanogr, 1993, 38(5): 978-985. doi: 10.4319/lo.1993.38.5.0978
[21]	FAITH D P. Conservation evaluation and phylogenetic diversity[J]. Biol Conserv, 1992, 61: 1-10. doi: 10.1016/0006-3207(92)91201-3
[22]	SHANNON C E, WEAVER W. The mathematical theory of communication[M]. Urbana, IL: University of Illinois Press, 1949: 55-187.
[23]	杨家新. 微生物生态学[M]. 北京: 化学工业出版社, 2004: 45-134.
[24]	HAN J H, BAEK K, LEE M H. Lacibacter nakdongensis sp. nov., isolated from river sediment[J]. Int J Syst Evol Microbiol, 2017, 67: 352-356. doi: 10.1099/ijsem.0.001631
[25]	WITCZAK P, CYDZIK-KWIATKOWSKA A, ZIELIŃSKA M. Treatment of the liquid phase of digestate from a biogas plant for water reuse[J]. Bio Tech, 2019, 276: 226-235. doi: 10.1016/j.biortech.2018.12.077
[26]	XUAN L, SHENG Z, LU J, et al. Bacterioplankton community responses and the potential ecological thresholds along disturbance gradients[J]. Sci Total Environ, 2019, 696: 134015. doi: 10.1016/j.scitotenv.2019.134015
[27]	YANG Y, GAO Y, HUANG X, et al. Adaptive shifts of bacterioplankton communities in response to nitrogen enrichment in a highly polluted river[J]. Environ Pollut, 2019, 245: 290-299. doi: 10.1016/j.envpol.2018.11.002
[28]	COMTE J, del GIORGIO P A. Composition influences the pathway but not the outcome of the metabolic response of bacterioplankton to resource shifts[J]. PLoS One, 2011, 6: e25266. doi: 10.1371/journal.pone.0025266
[29]	ADAMOVICH B V, MEDVINSKY A B, NIKITINA L V, et al. Relations between variations in the lake bacterioplankton abundance and the lake trophic state: evidence from the 20-year monitoring[J]. Ecol Indic, 2019, 7: 120-129.
[30]	吕锡斌, 吴云成, 陈良强, 等. 赤水河流域浮游细菌群落特征及其与水质的关系[J]. 环境科学学报, 2021, 41(11): 4596-4605. doi: 10.13671/j.hjkxxb.2021.0125
[31]	JING X Y, GOU H, GONG Y, et al. Seasonal dynamics of the coastal bacterioplankton at intensive fish-farming areas of the Yellow Sea, China revealed by high-throughput sequencing[J]. Mar Pollut Bull, 2019, 139: 366-375. doi: 10.1016/j.marpolbul.2018.12.052
[32]	SHADE A, HANNES P, STEVEN D A, et al. Fundamentals of microbial community resistance and resilience[J]. Front Microbiol, 2012, 19(3): 417.
[33]	郭远涛. 吸附固定化对藻菌共生体在畜禽养殖废水处理中的强化效果研究[D]. 南昌: 南昌大学, 2021: 12-36.
[34]	CAVENDER-BARES J, KOZAK K H, FINE P V A, et al. The merging of community ecology and phylogenetic biology[J]. Ecol Lett, 2009, 12: 693-715. doi: 10.1111/j.1461-0248.2009.01314.x
[35]	CHESSON P. Mechanisms of maintenance of species diversity[J]. Annu Rev Ecol Syst, 2000, 31: 343-366. doi: 10.1146/annurev.ecolsys.31.1.343
[36]	FAN L M, HU G D, QIU L P, et al. Variations in bacterioplankton communities in aquaculture ponds and the influencing factors during the peak period of culture[J]. Environ Pollut, 2020, 258: 113656. doi: 10.1016/j.envpol.2019.113656
[37]	IVES A R, CARPENTER S R. Stability and diversity of ecosystems[J]. Science, 2007, 317: 58-62. doi: 10.1126/science.1133258
[38]	YACHI S, LOREAU M. Biodiversity and ecosystem productivity in a fluctuating environment: the insurance hypothesis[J]. Proc Natl Acad Sci USA, 1999, 96: 1463-1468. doi: 10.1073/pnas.96.4.1463
[39]	胡晓娟, 李卓佳, 曹煜成, 等. 强天气干扰条件下粤西凡纳滨对虾养殖池塘细菌群落动态特征[J]. 南方水产科学, 2012, 8(5): 52-59. doi: 10.3969/j.issn.2095-0780.2012.05.008
[40]	CONNELL J H. Diversity in tropical rain forests and coral reefs: high diversity of trees and corals is maintained only in a non-equilibrium state[J]. Science, 1978, 199: 1302-1310. doi: 10.1126/science.199.4335.1302