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Biogeosciences, 17, 5209–5221, 2020 https://doi.org/10.5194/bg-17-5209-2020 © Author(s) 2020. This work is distributed under the Creative Commons Attribution 4.0 License. Methane paradox in tropical lakes? Sedimentary fluxes rather than pelagic production in oxic conditions sustain methanotrophy and emissions to the atmosphere Cédric Morana1,2, Steven Bouillon1, Vimac Nolla-Ardèvol1, Fleur A. E. Roland2, William Okello3, Jean-Pierre Descy2, Angela Nankabirwa3, Erina Nabafu3, Dirk Springael1, and Alberto V. Borges2 1KU Leuven, Leuven, Belgium 2Université de Liège, Liège, Belgium 3National Fisheries Resources Research Institute, Jinja, Uganda Correspondence: Cédric Morana ([email protected]) Received: 21 April 2020 – Discussion started: 5 May 2020 Revised: 20 August 2020 – Accepted: 8 September 2020 – Published: 29 October 2020 Abstract. Despite growing evidence that methane (CH4) for- 1 Introduction mation could also occur in well-oxygenated surface fresh wa- ters, its significance at the ecosystem scale is uncertain. Em- Emissions from inland waters are an important component of pirical models based on data gathered at high latitude pre- the global methane (CH4) budget (Bastviken et al., 2011), in dict that the contribution of oxic CH4 increases with lake particular from tropical latitudes (Borges et al., 2015). While size and should represent the majority of CH4 emissions in progress has been made in evaluating the CH4 emission large lakes. However, such predictive models could not di- rates, much less attention has been given to the underlying rectly apply to tropical lakes, which differ from their temper- microbial production (methanogenesis) and loss (methane ate counterparts in some fundamental characteristics, such as oxidation) processes. It is generally assumed that CH4 in year-round elevated water temperature. We conducted stable- lakes originates from the degradation of organic matter in isotope tracer experiments, which revealed that oxic CH4 anoxic sediments. Because most methanogens are consid- production is closely related to phytoplankton metabolism ered to be strict anaerobes, and net vertical diffusion of CH4 and is a common feature in five contrasting African lakes. from anoxic bottom waters is often negligible (Bastviken et Nevertheless, methanotrophic activity in surface waters and al., 2003), physical processes of CH4 transport from shal- CH4 emissions to the atmosphere were predominantly fu- low sediments are usually invoked to explain patterns of local elled by CH4 generated in sediments and physically trans- CH4 concentration maxima in surface waters (Fernández et ported to the surface. Indeed, CH4 bubble dissolution flux al., 2016; Peeters et al., 2019; Martinez-Cruz et al., 2020). In- and diffusive benthic CH4 flux were several orders of mag- deed, CH4-rich pore water is regularly released from littoral nitude higher than CH4 production in surface waters. Micro- sediment into the water column during resuspension events bial CH4 consumption dramatically decreased with increas- associated with surface waves (Hofmann et al., 2010). ing sunlight intensity, suggesting that the freshwater “CH4 The view that CH4 is formed under strictly anaerobic paradox” might be also partly explained by photo-inhibition conditions has been challenged by several recent studies, of CH4 oxidizers in the illuminated zone. Sunlight appeared which have proposed that acetoclastic methanogens directly as an overlooked but important factor determining the CH4 attached to phytoplankton cells are involved in epilimnetic dynamics in surface waters, directly affecting its production CH4 production (Grossart et al., 2011; Bogard et al., 2014) by photoautotrophs and consumption by methanotrophs. and are responsible for distinct near-surface peaks of CH4 concentration in certain thermally stratified, well-oxygenated waterbodies (Tang et al., 2016). It has also been shown that Cyanobacteria (Bizi˘ c´ et al., 2020) and widespread marine Published by Copernicus Publications on behalf of the European Geosciences Union. 5210 C. Morana et al.: Sedimentary fluxes rather than pelagic production phytoplankton (Klintzsch et al., 2019) are able to release a the neighbouring Lake Edward (−0.28971◦ N, 29.73327◦ E), 2 substantial amount of CH4 during a culture study, and this a holomictic, mesotrophic large lake (2325 km ). CH4 production mechanism might be linked to photosyn- Water samples from pelagic stations of Lake Katinda thesis. From a model-based approach, epilimnetic CH4 pro- (0.3 km from shore, 14 m deep), Lake George (2 km from duction was shown to sustain most of the CH4 oxidation in shore, 2.5 m deep) and Lake Edward (15 km from shore, 20 m 14 Canadian lakes (DelSontro et al., 2018) and would even deep) were collected in April 2017 (rainy season) and Jan- represent up to 90 % of the CH4 emitted from a temperate uary 2018 (dry season). Pelagic sites of Lake Kyamwinga lake (Donis et al., 2017). Further, empirical models based (0.5 km from shore, 40 m deep) and Lake Nyamusingere on data gathered in boreal and temperate lakes predict that (0.2 km from shore, 3 m deep) were sampled only once, in the contribution of oxic CH4 increases with lake size (Gün- April 2017 and January 2018, respectively. thel et al., 2019) and should represent the majority of CH4 2 emissions in lakes larger than 1 km . Still, aerobic CH4 pro- 2.2 Environmental setting of the study sites duction has so far only been documented in temperate and boreal lakes so that such predictive models could not di- Conductivity, temperature and dissolved-oxygen concentra- rectly apply to tropical lakes, which differ from their temper- tion measurements were performed with a Yellow Spring In- ate counterparts in some fundamental characteristics, such as strument EXO II multi-parametric probe. Samples for par- year-round elevated water temperature. Primary production, ticulate organic carbon (POC) concentration were collected methanogenic and methanotrophic activities, and cyanobac- on glass fibre filters (0.7 µm nominal pore size) and anal- terial dominance are potentially much higher in tropical lakes ysed with an elemental analyser coupled to an isotope ratio due to favourable temperature (Lewis, 1987; Kosten et al., mass spectrometer (EA–IRMS; Morana et al., 2015a). Pig- 2012). It has also been shown that CH4 emissions are pos- ment concentrations were determined by high-performance itively related to temperature at the ecosystem scale (Yvon- liquid chromatography (Descy et al., 2017) after filtration Durocher et al., 2014) of water samples through glass fibre filters (0.7 µm nominal Here, we tested the hypothesis that phytoplankton pore size). metabolism could fuel CH4 production in well-oxygenated Water samples for determination of dissolved CH4 con- waters in five contrasting tropical lakes in East Africa cover- centration were transferred with tubing from the Niskin bot- ing a wide range of sizes, depths, and productivity (Lake Ed- tle to 60 mL borosilicate serum bottles that were poisoned ward, Lake George, Lake Katinda, Lake Nyamusingere and with 200 µL of a saturated solution of HgCl2, closed with Lake Kyambura). Phytoplankton activity could provide di- a butyl stopper and sealed with an aluminium cap. The verse substrates required for CH4 production mediated by concentrations of dissolved CH4 were measured with the methanogenic Archaea, or alternatively CH4 could be di- headspace equilibration technique (20 mL headspace) using rectly released by phytoplankton cells. Additionally, the sig- a gas chromatograph with flame ionization detection (GC– nificance of epilimnetic CH4 production at the scale of the FID, SRI8610C). 13 aquatic ecosystem was assessed by quantifying CH4 release Samples for δ C–CH4 determination were collected in from sediments, CH4 production and oxidation rates in the 60 mL serum bottles following the same procedure as sam- water column, and CH4 diffusive and ebullitive emissions to ples for CH4 concentration determination. In the labora- 13 the atmosphere. tory, δ C–CH4 was measured as described in Morana et al. (2015b). Briefly, a 20 mL helium headspace was created in the serum bottles, then samples were vigorously shaken 2 Material and methods and left to equilibrate overnight. The sample gas was flushed out through a double-hole needle and purified of non-CH4 2.1 Site description volatile organic compounds in a liquid N2 trap, CO2 and H2O were removed with a soda lime and magnesium perchlo- The sampled lakes cover a wide range of size (<1 to rate traps, and the CH4 was converted to CO2 in an online 2300 km2), maximum depth (3–117 m), mixing regimes, combustion column similar to that in an elemental analyser phytoplankton biomass and primary productivity (Ta- (EA). The resulting CO2 was subsequently pre-concentrated ble S1 in the Supplement). Oligotrophic Lake Kyamwinga in a custom-built cryo-focussing device by immersion of a ◦ ◦ (−0.18054 N, 30.14625 E) and eutrophic Lake Katinda stainless-steel loop in liquid N2, passed through a micro- (−0.21803◦ N, 30.10702◦ E) are stratified, small but deep packed GC column (HayeSep Q 2 m, 0.75 mm ID; Restek), tropical lakes located in western Uganda. Neighbour- and finally measured on a Thermo Scientific Delta V Advan- ◦ ◦ ing Lake Nyamusingere (−0.284364 N, 30.037635 E) tage isotope ratio mass spectrometer (IRMS). CO2 produced is a small but shallow and polymictic eutrophic lake. from certified reference standards for δ13C analysis (IAEA- 2 13 Lake George is a larger (250 km ), hypereutrophic, shallow CO1 and LSVEC) were used to calibrate δ C–CH4 data. Re- lake located at the Equator (−0.02273◦ N, 30.19724◦ E). A producibility of measurements estimated based on duplicate single outlet (Kazinga Channel) flows from Lake George to injection of a selection of samples was typically better than Biogeosciences, 17, 5209–5221, 2020 https://doi.org/10.5194/bg-17-5209-2020 C. Morana et al.: Sedimentary fluxes rather than pelagic production 5211 0.5 ‰ or better than 0.2‰ when estimated based on multiple collected, taking care to avoid disturbance at the sediment– injection of standard gas.