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Spatial distribution of sediment bacterial communities from São Francisco River headwaters is influenced by human land-use activities and seasonal climate shifts

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Abstract

Riverbed sediments are dynamic freshwater environments colonized by a great diversity of microorganisms which play important roles in supporting freshwater ecosystem by performing a vast array of metabolic functions. Recent evidence generated by HTS approaches has revealed that the structure of sediment microbial communities is influenced by natural seasonal variations in water such as temperature or streamflow as well by disturbances caused by local human activities. Here, a spatiotemporal analysis of sediment microbial distribution from São Francisco River headwaters section was conducted using Illumina 16S rRNA-V4 region amplicon sequencing in order to accomplish three major goals: (i) to investigate whether the diversity and composition of bacterial communities accessed in riverbed sediments vary in response to distinct land-use activities; (ii) to estimate whether the diversity patterns vary between the dry and wet seasons; and (iii) to evaluate whether the diversity of bacterial metabolic functions, predicted by PICRUSt2 approach, varies similarly to the estimated taxonomic diversity. Our findings revealed that bacterial communities in the sediment show differences in diversity and taxonomic composition according to the anthropic activities performed in the local environment. However, the patterns in which this taxonomic diversity is spatially structured show differences between the dry and wet seasons. On the other hand, the most changes in predicted bacterial metabolic functions were verified between sediment samples accessed in portions of the river located in protected and unprotected areas. Our findings contributed with new evidence about the impact of typical land-use practices conducted in countryside landscapes from developing countries on riverbed bacterial communities, both in their taxonomic and functional structure.

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References

  1. Gutknecht JL, Goodman RM, Balser TC (2006) Linking soil process and microbial ecology in freshwater wetland ecosystems. Plant Soil 289:17–34. https://doi.org/10.1007/s11104-006-9105-4

    Article  CAS  Google Scholar 

  2. Shi J, Zhang B, Liu J, Fang Y, Wang A (2022) Spatiotemporal dynamics in microbial communities mediating biogeochemical cycling of nutrients across the Xiaowan Reservoir in Lancang River. Sci Total Environ 813:151862. https://doi.org/10.1016/j.scitotenv.2021.151862

    Article  CAS  PubMed  Google Scholar 

  3. Wang J, Chen Y, Cai P, Gao Q, Zhong H, Sun W, Chen Q (2022) Impacts of municipal wastewater treatment plant discharge on microbial community structure and function of the receiving river in Northwest Tibetan Plateau. J Hazard Mater 423:127170. https://doi.org/10.1016/j.jhazmat.2021.127170

    Article  CAS  PubMed  Google Scholar 

  4. Sigee D (2005) Freshwater microbiology: biodiversity and dynamic interactions of microorganisms in the aquatic environment. John Wiley & Sons

    Google Scholar 

  5. Cottrell MT, Waidner LA, Yu L, Kirchman DL (2005) Bacterial diversity of metagenomic and PCR libraries from the Delaware River. Environ Microbiol 7:1883–1895. https://doi.org/10.1111/j.1462-2920.2005.00762.x

    Article  CAS  PubMed  Google Scholar 

  6. Liu J, Chen X, Shu HY, Lin XR, Zhou QX, Bramryd T, Shu W-S, Huang LN (2018) Microbial community structure and function in sediments from e-waste contaminated rivers at Guiyu area of China. Environ Pollut 235:171–179. https://doi.org/10.1016/j.envpol.2017.12.008

    Article  CAS  PubMed  Google Scholar 

  7. Williamson CE, Dodds W, Kratz TK, Palmer MA (2008) Lakes and streams as sentinels of environmental change in terrestrial and atmospheric processes. Front Ecol Environ 6:247–254. https://doi.org/10.1890/070140

    Article  Google Scholar 

  8. Vörösmarty CJ, McIntyre PB, Gessner MO, Dudgeon D, Prusevich A, Green P, Glidden S, Bunn SE, Sullivan CA, Liermann R, Davies PM (2010) Global threats to human water security and river biodiversity. Nature 467:555. https://doi.org/10.1038/nature09440

    Article  CAS  PubMed  Google Scholar 

  9. Oribhabor BJ, Udoidiong OM, Udoh DF, Akpan BE (2013) Evaluation of the ecological impact of human settlement on the water quality of Lower Cross River, Nigeria. J Ecosyst 2013:840295. https://doi.org/10.1155/2013/840295

    Article  Google Scholar 

  10. Weijters MJ, Janse JH, Alkemade R, Verhoeven JT (2009) Quantifying the effect of catchment land use and water nutrient concentrations on freshwater river and stream biodiversity. Aquat Conserv 19:104–112. https://doi.org/10.1002/aqc.989

    Article  Google Scholar 

  11. Villéger S, Blanchet S, Beauchard O, Oberdorff T, Brosse S (2011) Homogenization patterns of the world’s freshwater fish faunas. PNAS 108:18003–18008. https://doi.org/10.1073/pnas.1107614108

    Article  PubMed  PubMed Central  Google Scholar 

  12. Taylor KG, Owens PN (2009) Sediments in urban river basins: a review of sediment–contaminant dynamics in an environmental system conditioned by human activities. Journal Soils Sediment 9:281–303. https://doi.org/10.1007/s11368-009-0103-z

    Article  Google Scholar 

  13. Keesstra SD, Geissen V, Mosse K, Piiranen S, Scudiero E, Leistra M, van Schaik L (2012) Soil as a filter for groundwater quality. Curr Opin Env Sust 4:507–516. https://doi.org/10.1016/j.cosust.2012.10.007

    Article  Google Scholar 

  14. Vignesh S, Dahms HU, Emmanuel KV, Gokul MS, Muthukumar K, Kim BR, James RA (2014) Physicochemical parameters aid microbial community? A case study from marine recreational beaches, Southern India. Environ Monit Assess 186:1875–1887. https://doi.org/10.1007/s10661-013-3501-z

    Article  CAS  PubMed  Google Scholar 

  15. Wang P, Chen B, Yuan R, Li C, Li Y (2016) Characteristics of aquatic bacterial community and the influencing factors in an urban river. Sci Tot Environ 569:382–389. https://doi.org/10.1016/j.scitotenv.2016.06.130

    Article  CAS  Google Scholar 

  16. Tian W, Zhang H, Guo Y, Wang Z, Huang T (2022) Temporal and spatial patterns of sediment microbial communities and driving environment variables in a shallow temperate mountain river. microorganisms 10:816. https://doi.org/10.3390/microorganisms10040816

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Winter C, Hein T, Kavka G, Mach RL, Farnleitner AH (2007) Longitudinal changes in the bacterial community composition of the Danube River: a whole-river approach. Appl Environ Microbiol 73:421–431. https://doi.org/10.1128/AEM.01849-06

    Article  CAS  PubMed  Google Scholar 

  18. Gibbons SM, Jones E, Bearquiver A, Blackwolf F, Roundstone W, Scott N, Hooker J, Madsen R, Coleman ML, Gilbert JA (2014) Human and environmental impacts on river sediment microbial communities. PLoS One 9:e97435. https://doi.org/10.1371/journal.pone.0097435

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Lliro M, Inceoglu O, Garcia-Armisen T, Anzil A, Leporcq B, Pigneur L, Viroux L, Darchambeau F, Descy J, Servais P (2014) Bacterial community composition in three freshwater reservoirs of different alkalinity and trophic status. PLoS One 9:1–27. https://doi.org/10.1371/journal.pone.0116145

    Article  CAS  Google Scholar 

  20. Sorokin DY, Berben T, Melton ED, Overmars L, Vavourakis CD, Muyzer G (2014) Microbial diversity and biogeochemical cycling in soda lakes. Extremophiles 18:791–809. https://doi.org/10.1007/s00792-014-0670-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Wu H, Hao B, Cai Y, Liu G, Xing W (2021) Effects of submerged vegetation on sediment nitrogen-cycling bacterial communities in Honghu Lake (China). Sci Tot Environ 755:142541. https://doi.org/10.1016/j.scitotenv.2020.142541

    Article  CAS  Google Scholar 

  22. Wang P, Zhao J, Xiao H, Yang W, Yu X (2018) Bacterial community composition shaped by water chemistry and geographic distance in an anthropogenically disturbed river. Sci Tot Environ 655:61–69. https://doi.org/10.1016/j.scitotenv.2018.11.234

    Article  CAS  Google Scholar 

  23. Lu Z, Liu Z, Zhang C, Wei Q, Zhang S, Li M (2021) Spatial and seasonal variations of sediment bacterial communities in a river-bay system in South China. Appl Microbiol Biotechnol 105:1979–1989. https://doi.org/10.1007/s00253-021-11142-z

    Article  CAS  PubMed  Google Scholar 

  24. de Paula M, Silva TAC, Araújo AS, Lacorte GA (2021) Assessments of bacterial community shifts in sediments along the headwaters of São Francisco River, Brazil. Conservation 1:91–105. https://doi.org/10.3390/conservation1020008

    Article  Google Scholar 

  25. Hu A, Ju F, Hou L, Li J, Zhi X, Wang H, Mulla SI, Sun Q, Burgmann H, Yu C-P (2017) Strong impact of anthropogenic contamination on the co-occurrence patterns of a riverine microbial community. Environ Microbiol 19:4933–5009. https://doi.org/10.1111/1462-2920

    Article  Google Scholar 

  26. CBHSF. Comitê de Bacia Hidrográfica do São Francisco (2016) Plano de Recursos Hídricos da Bacia Hidrográfica do Rio São Francisco 2016–2025. RF3 – Resumo executivo. https://cbhsaofrancisco.org.br/documentacao/plano-de-recursos-hidricos-2016-2025/ (Accessed May 18 2020). (Document in Portuguese).

  27. ANA. Agência Nacional de Águas (2015) Water resources in Brazil: Brazilian hydrographic regions. Available at: http://www.snirh.gov.br/portal/snirh/centrais-de-conteudos/conjuntura-dos-recursos-hidricos/conjuntura_informe_2015.pdf (Accessed May 18 2020). (Document in Portuguese).

  28. Silva FAM (2008) Clima do bioma Cerrado. In: Agricultura tropical: quatro décadas de inovações tecnológicas, institucionais e políticas, pp 93–148 Book chapter in Portuguese

    Google Scholar 

  29. Dantas MS, Oliveira JC, Pinto CC, Oliveira SC (2020) Impact of fecal contamination on surface water quality in the São Francisco River hydrographic basin in Minas Gerais, Brazil. J Water Health 18:48–59. https://doi.org/10.2166/wh.2019.153

    Article  Google Scholar 

  30. Gilbert JA, Jansson JK, Knight R (2014) The Earth Microbiome Project: successes and aspirations. BMC Biol 12:69. https://doi.org/10.1186/s12915-014-0069-1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP (2016) DADA2: high-resolution sample inference from Illumina amplicon data. Nat Methods 13:581. https://doi.org/10.1038/nmeth.3869

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Zhang L, Zhao T, Wang Q, Li L, Shen T, Gao G (2019) Bacterial community composition in aquatic and sediment samples with spatiotemporal dynamics in large, shallow, eutrophic Lake Chaohu, China. J Freshwater Ecol 34:575–589. https://doi.org/10.1080/02705060.2019.1635536

    Article  CAS  Google Scholar 

  33. Duarte S, Cássio F, Ferreira V, Canhoto C, Pascoal C (2016) Seasonal variability may affect microbial decomposers and leaf decomposition more than warming in streams. Microb Ecol 72:263–276. https://doi.org/10.1007/s00248-016-0780-2

    Article  CAS  PubMed  Google Scholar 

  34. Chiaramonte JB, Roberto MC, Pagioro TA (2013) Seasonal dynamics and community structure of bacterioplankton in upper Paraná River floodplain. Microb Ecol 66:773–783. https://doi.org/10.1007/s00248-013-0292-2

    Article  PubMed  Google Scholar 

  35. Fazi S, Vázquez E, Casamayor EO, Amalfitano S, Butturini A (2013) Stream hydrological fragmentation drives bacterioplankton community composition. PLoS One 8:1–10. https://doi.org/10.1371/journal.pone.0064109

    Article  CAS  Google Scholar 

  36. Zhang M, Yu N, Chen L, Jiang C, Tao Y, Zhang T, Chen J, Xue D (2012) Structure and seasonal dynamics of bacterial communities in three urban rivers in China. Aquat Sci 74:113–120. https://doi.org/10.1007/s00027-011-0201-z

    Article  CAS  Google Scholar 

  37. Boyero L, Pearson RG, Gessner MO, Barmuta LA, Ferreira V, Graça MA, Dudgeon D, Boulton AJ, Callisto M, Chauvet E, Helson JE (2011) A global experiment suggests climate warming will not accelerate litter decomposition in streams but might reduce carbon sequestration. Ecol Lett 14:289–294. https://doi.org/10.1111/j.1461-0248.2010.01578.x

    Article  PubMed  Google Scholar 

  38. Zoppini A, Amalfitano S, Fazi S, Puddu A (2010) Dynamics of a benthic microbial community in a riverine environment subject to hydrological fluctuations (Mulargia River, Italy). Hydrobiologia 657:37–51. https://doi.org/10.1007/s10750-010-0199-6

    Article  CAS  Google Scholar 

  39. Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Peña AG, Goodrich JK, Gordon JI, Huttley GA (2010) QIIME allows analysis of high throughput community sequencing data. Nat Methods 7:335e336. https://doi.org/10.1038/nmeth.f.303

    Article  CAS  Google Scholar 

  40. QIIME 2 Development Team (2017) QIIME:2 https://docs.qiime2.org

  41. Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, Peplies J, Glockner FO (2013) The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res 41:D590–D596. https://doi.org/10.1093/nar/gks1219

    Article  CAS  PubMed  Google Scholar 

  42. Porter TM, Hajibabaei M (2018) Scaling up: a guide to high-throughput genomic approaches for biodiversity analysis. Mol Ecol 27:313–338. https://doi.org/10.1111/mec.14478

    Article  PubMed  Google Scholar 

  43. Douglas GM, Maffei VJ, Zaneveld JR, Yurgel SN, Brown JR, Taylor CM, Huttenhower C, Langille MGI (2020) PICRUSt2 for prediction of metagenome functions. Nat Biotechnol 38:685–688. https://doi.org/10.1038/s41587-020-0548-6

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Kanehisa M, Goto S (2000) KEGG: Kyoto Encyclopedia of Genes and Genomes. Nucleic Acids Res 28:27–30. https://doi.org/10.1093/nar/28.1.27

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Liu Y, Tong T, Li B, Xie S (2019) Dynamics of bacterial communities in a river water treatment wetland. Ann Microbiol 69:637–645. https://doi.org/10.1007/s13213-019-01454-x

    Article  Google Scholar 

  46. Roberto AA, Van Gray JB, Leff LG (2018) Sediment bacteria in an urban stream: spatiotemporal patterns in community composition. Water Res 134:353–369. https://doi.org/10.1016/j.watres.2018.01.045

    Article  CAS  PubMed  Google Scholar 

  47. Costa EP, Pinto CC, Soares ALC, Melo LDV, Oliveira SC (2017) Evaluation of violations in water quality standards in the monitoring network of São Francisco River basin, the third largest in Brazil. Environ Monit Assess 189:2–16. https://doi.org/10.1007/s10661-017-6266-y

    Article  CAS  Google Scholar 

  48. Moss JA, Nocker A, Lepo JE, Snyder RA (2006) Stability and change in estuarine biofilm bacterial community diversity. Appl Environ Microbiol 72:5679–5688. https://doi.org/10.1128/AEM.02773-05

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Marti E, Balcázar JL (2014) Use of pyrosequencing to explore the benthic bacterial community structure in a river impacted by wastewater treatment plant discharges. Res Microbiol 165:468–471. https://doi.org/10.1016/j.resmic.2014.04.002

    Article  CAS  PubMed  Google Scholar 

  50. Lu X-M, Lu P-Z (2014) Characterization of bacterial communities in sediments receiving various wastewater effluents with high-throughput sequencing analysis. Microb Ecol 67:612–623. https://doi.org/10.1007/s00248-014-0370-0

    Article  CAS  PubMed  Google Scholar 

  51. Drury B, Rosi-Marshall E, Kelly JJ (2013) Wastewater treatment effluent reduces the abundance and diversity of benthic bacterial communities in urban and suburban rivers. Appl Environ Microbiol 79:1897–1905. https://doi.org/10.1128/AEM.0352712

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Kalia A, Gosal SK (2011) Effect of pesticide application on soil microorganisms. Arch Agron Soil Sci 57:569–596. https://doi.org/10.1080/03650341003787582

    Article  CAS  Google Scholar 

  53. Wakelin SA, Colloff MJ, Kookana RS (2008) Effect of wastewater treatment plant effluent on microbial function and community structure in the sediment of a freshwater stream with variable seasonal flow. Appl Environ Microbiol 74:2659–2668. https://doi.org/10.1128/AEM.02348-07

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Crump BC, Adams HE, Hobbie JE, Kling GW (2007) Biogeography of bacterioplankton in lakes and streams of an arctic tundra catchment. Ecology 88:1365–1378. https://doi.org/10.1890/06-0387

    Article  PubMed  Google Scholar 

  55. Lozupone CA, Hamady M, Kelley ST, Knight R (2007) Quantitative and qualitative β-diversity measures lead to different insights into factors that structure microbial communities. Appl Environ Microbiol 73:1576–1585. https://doi.org/10.1128/AEM.01996-06

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Read DS, Gweon HS, Bowes MJ, Newbold LK, Field D, Bailey MJ, Griffiths RI (2015) Catchment-scale biogeography of riverine bacterioplankton. ISME J 9:516. https://doi.org/10.1038/ismej.2014.166

    Article  CAS  PubMed  Google Scholar 

  57. Székely AJ, Berga M, Langenheder S (2013) Mechanisms determining the fate of dispersed bacterial communities in new environments. ISME J 7:61–71. https://doi.org/10.1038/ismej.2012.80

    Article  CAS  PubMed  Google Scholar 

  58. Huang W, Chen X, Wang K, Chen J, Zheng B, Jiang X (2019) Comparison among the microbial communities in the lake, lake wetland, and estuary sediments of a plain river network. MicrobiologyOpen 8:e00644. https://doi.org/10.1002/mbo3.644

    Article  PubMed  Google Scholar 

  59. Abia ALK, Alisoltani A, Keshri J, Ubomba-Jaswa E (2018) Metagenomic analysis of the bacterial communities and their functional profiles in water and sediments of the Apies River, South Africa, as a function of land use. Sci Total Environ 616:326–334. https://doi.org/10.1016/j.scitotenv.2017.10.322

    Article  CAS  PubMed  Google Scholar 

  60. Wu H, Li Y, Zhang J, Niu L, Zhang W, Cai W, Zhu X (2017) Sediment bacterial communities in a eutrophic lake influenced by multiple inflow-rivers. Environ Sci Pollut R 24:19795–19806. https://doi.org/10.1007/s11356-017-9602-4

    Article  CAS  Google Scholar 

  61. Wei Z, Xinlong B, Limin Z, Anqi W, Yongzhen P (2014) Population dynamics of nitrifying bacteria for nitritation achieved in Johannesburg (JHB) process treating municipal wastewater. Bioresource Technol 162:30–37. https://doi.org/10.1016/j.biortech.2014.03.102

    Article  CAS  Google Scholar 

  62. Febria CM, Hosen JD, Crump BC, Palmer MA, Williams DD (2015) Microbial responses to changes in flow status in temporary headwater streams: a cross-system comparison. Front Microbiol 6:522. https://doi.org/10.3389/fmicb.2015.00522

    Article  PubMed  PubMed Central  Google Scholar 

  63. Kolb S, Horn MA (2012) Microbial CH4 and N2O consumption in acidic wetlands. Front Microbiol 3:78. https://doi.org/10.3389/fmicb.2012.00078

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Jani K, Ghattargi V, Pawar S, Inamdar M, Shouche Y, Sharma A (2018) Anthropogenic activities induce depletion in microbial communities at urban sites of the river Ganges. Curr Microbiol 75:79–83. https://doi.org/10.1007/s00284-017-1352-5

    Article  CAS  PubMed  Google Scholar 

  65. Wu J, Xu Z, Li H, Li P, Wang M, Xiong L, Zhang J (2019) Long-term effect of water diversion and CSOs on the remediation of heavy metals and microbial community in river sediments. Water Sci Technol 79:2395–2406. https://doi.org/10.2166/wst.2019.242

    Article  CAS  PubMed  Google Scholar 

  66. Huang S, Chen C, Jaffé PR (2018) Seasonal distribution of nitrifiers and denitrifiers in urban river sediments affected by agricultural activities. Sci Tot Environ 642:1282–1291. https://doi.org/10.1016/j.scitotenv.2018.06.116

    Article  CAS  Google Scholar 

  67. Zhang S, Wang J, Hao X (2020) Fertilization accelerates the decomposition of microplastics in mollisols. Sci Tot Environ 722:137950. https://doi.org/10.1016/j.scitotenv.2020.137950

    Article  CAS  Google Scholar 

  68. Kirchman DL (2018) Processes in microbial ecology. Oxford University Press

    Book  Google Scholar 

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Funding

This work was granted by Instituto Federal de Minas Gerais—Edital de Pesquisa Aplicada no. 169/2015 and by Fundação de Amparo à Pesquisa de Minas Gerais—FAPEMIG (MPR-01008-16).

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GAL and AABS contributed to conception and design of the experiments. MPJ and TACS performed the experiments. MPJ and GAL analyzed the data. MPJ wrote the draft manuscript. All authors contributed to the final version of the manuscript.

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Correspondence to Gustavo Augusto Lacorte.

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Figure S1

Rarefaction curves of observed ASVs for sediment samples from São Francisco River headwaters (PNG 249 kb)

High resolution image (TIF 1841 kb)

Figure S2

Diversity of metabolic functions predicted by PICRUSt2 for bacterial communities from each land-use zone organized by season (PNG 225 kb)

High resolution image (TIF 6404 kb)

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de Paula, M., da Costa, T.A., Silva et al. Spatial distribution of sediment bacterial communities from São Francisco River headwaters is influenced by human land-use activities and seasonal climate shifts. Braz J Microbiol 54, 3005–3019 (2023). https://doi.org/10.1007/s42770-023-01150-8

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