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Ecological assessment of groundwater ecosystems disturbed by recharge systems using organic matter quality, biofilm characteristics and bacterial diversity
Jérémy Voisin, B. Cournoyer, L. Marjolet, Antonin Vienney, Florian Mermillod-Blondin
To cite this version:
Jérémy Voisin, B. Cournoyer, L. Marjolet, Antonin Vienney, Florian Mermillod-Blondin. Ecological assessment of groundwater ecosystems disturbed by recharge systems using organic matter quality, biofilm characteristics and bacterial diversity. Environmental Science and Pollution Research, Springer Verlag, 2020, 27 (3), pp.3295-3308. �10.1007/s11356-019-06971-5�. �hal-02386310�
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including natural lakes, ponds, and oceans 51
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96 97 98 99 100 101 102 103
located outside the area of influence of the stormwater plume induced by infiltration which 104
was used as a control well, and a well located in the immediate downstream vicinity of the infiltration 105
basin used as a recharge well because it intersected the stormwater plume induced by the infiltration.
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DOC (dissolved organic carbon) was determined using a carbon analyzer (multi N/C® 3100, Analytik 134
Jena, Jena, Germany) based on thermocatalytic oxidation (850°C) of organic carbon and infrared 135
detection of CO2, after removal of dissolved inorganic C with HCl. Biodegradable DOC (BDOC) was 136
determined by the method of Servais et al. (1987, 1989) following Mermillod-Blondin et al. (2015).
137
Briefly, water samples (100 mL) were filtered through a pre-washed 0.2 µm polycarbonate membrane 138
and incubated with a bacterial inoculum at 20°C for 30 days in the dark. DOC concentrations were 139
measured from filtered (0.2 µm) water samples at the start and the end of the incubation period to 140
determine the initial DOC concentration and the remaining DOC concentration after 30 days of 141
incubation (representing the refractory dissolved organic carbon, RDOC), respectively. BDOC 142
concentration was then calculated as the difference between initial DOC concentration and RDOC 143
concentration.
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174 175 176 177 178 179 180 181 182 183 184 185 186 187
qPCR assays were performed on a Bio-Rad CFX96 real-time PCR instrument with Bio-Rad CFX 188
Manager software, version 3.0 (Marnes-la-Coquette, France). Total Bacteroides DNA targets were 189
PCR amplified according to Layton et al. (2006) using the Brilliant II SYBR Green low ROX qPCR 190
master mix for TaqMan qPCR (Agilent, Vénissieux, France). The human-specific HF183 Bacteroides 191
qPCR assay was performed according to Seurinck et al. (2005), and the assay for the 16S rRNA gene 192
segment for total bacteria was performed according to Park and Crowley (2006) using primers 338F 193
and 518R, and using the Brilliant II SYBR green low ROX qPCR master mix for SYBR Green qPCR.
194
Melting T° was 60°C for all assays. Linearized plasmid DNA containing 16S rRNA genes from the 195
targeted DNAs were run as standards using 10-fold dilutions of the plasmids. These plasmids were 196
obtained from Marti et al. (2017). Presence of inhibitors was checked by spiking known amount of 197
plasmid harboring int2 with 10 times dilution of sample DNA extracts (107 copies of plasmid per l 198
of DNA diluted solution). Number of cycles needed to have a significant signal was compared with 199
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wells where only plasmid harboring int2was added to the qPCR mix. When a higher number of cycles 200
was needed to observe a signal, a dilution by 5 or 10 fold was done and another run of tests was 201
performed to confirm the absence of PCR inhibitions. Negative controls without template DNA were 202
run in triplicate. Each assay was triplicated on distinct DNA extracts, and technical triplicates were 203
performed.
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Data deposition 225
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Sequence data are available at the European Nucleotide Archive (https://www.ebi.ac.uk/ena) under 226
the project accession #PRJEB29925. Sample accession #ERS2912777 (SAMEA5128392) = run2.
227
See Table S1 for barcode information.
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356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374
Marschner and Kalbitz 2003; Li et al. 2013; Shen 375
et al. 2015 or/and Fe and Al
376
oxides/hydroxides, McKnight et al. 1992; Kalbitz et al. 2000; Saidy et al. 2013) are 377
likely involved in this VZT-related effect. Consequently, and according to several studies (e.g., 378
Pabich et al. 2001; Shen et al. 2015), the thicker the VZ, the more efficient is its ability to limit 379
groundwater contamination with DOM.
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Based on groundwater samples from Cape Cod (USA), Pabich et al. (2001) determined a statistical 381
relationship between VZT (in m) and DOC concentration (in ) in groundwater: DOC concen- 382
tration = DOC concentration in surface water * e -0.7*VZT. By applying the same relationship using a 383
mean DOC concentration of 5 in surface water (according to Voisin et al. 2018), we calculated 384
DOC concentrations ranging between 0.9 and 3.05 for groundwaters influenced by recharge 385
in sites with vadose zones < 3 meters. Interestingly, the measured DOC concentrations at these sites 386
were in the same range (i.e., 1.6 2.3 ). One would have expected that AR would have pro- 387
duced higher DOC concentrations than those expected from the model of Pabich et al. (2001) because 388
of the stronger hydrological connectivity between the surface and the aquifer in these AR sites (Foul- 389
quier et al. 2011). Two mechanisms could explain the concordance between our measurements and 390
simulations by the model of Pabich and colleagues: (1) soil and VZ of AR systems were very efficient 391
in DOC retention (Zhang et al. 2012; Mermillod-Blondin et al. 2015) and (2) DOC from infiltrating 392
waters was significantly diluted by groundwater (poor in DOC) in recharge zones of the aquifer (Foul- 393
quier et al. 2010). In some of our AR sites, two studies (Mermillod-Blondin et al. 2015; Voisin et al.
394
2018) tried to determine the respective influences of dilution and retention in organic matter dynamics 395
by using chloride as conservative tracer to evaluate the role of dilution. They found that the dilution 396
of surface water with groundwater could not explain the decrease of DOC concentrations from surface 397
waters to the aquifer. Then, it is expected that abiotic and biotic retention of DOC during water infil- 398
tration through the soil and VZ was the main mechanism involved in DOC dynamics in AR sites.
399
Nevertheless, this retention process was not efficient enough to limit significant DOC and BDOC 400
enrichments of groundwater in AR sites with thin VZ. The concentration of organic carbon was thus 401
a good indicator of the connectivity between surface and groundwater ecosystems.
402
groundwater ecosystems experience strong carbon limitation that severely limit growth of 403
Bengtsson 1989; Kazumi and Capone 1994; Baker et al. 2000; Goldscheider 404
2006 405
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Conflicts of interest. None declared.
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*p < 0.05 **p < 0.01 ***p < 0.001
