Sampling Device-Dependence of Prokaryotic Community Structure on Marine Particles: Higher Diversity Recovered by in situ Pumps Than by Oceanographic Bottles

10 pages, 3 figures, 2 tables, supplementary material https://www.frontiersin.org/articles/10.3389/fmicb.2020.01645/full#supplementary-material.-- Data Availability Statement. The raw sequence data have been deposited in the Figshare data repository, together with the non-rarefied OTU table, the taxonomy table and the environmental data used in this study, 10.6084/m9.figshare.12333107Microbes associated with sinking marine particles play key roles in carbon sequestration in the ocean. The sampling of particle-attached microorganisms is often done with sediment traps or by filtration of water collected with oceanographic bottles, both involving a certain time lapse between collection and processing of samples that may result in changes in particle-attached microbial communities. Conversely, in situ water filtration through submersible pumps allows a faster storage of sampled particles, but it has rarely been used to study the associated microbial communities and has never been compared to other particle-sampling methods in terms of the recovery of particle microbial diversity. Here we compared the prokaryotic communities attached to small (1–53 μm) and large (>53 μm) particles collected from the mesopelagic zone (100–300 m) of two Antarctic polynyas using in situ pumps (ISP) and oceanographic bottles (BTL). Each sampling method retrieved largely different particle-attached communities, suggesting that they capture different kinds of particles. These device-driven differences were greater for large particles than for small particles. Overall, the ISP recovered 1.5- to 3-fold more particle-attached bacterial taxa than the BTL, and different taxonomic groups were preferentially recovered by each method. In particular, typical particle-attached groups such as Planctomycetes and Deltaproteobacteria recovered with ISP were nearly absent from BTL samples. Our results suggest that the method used to sample marine particles has a strong influence in our view of their associated microbial communitiesVP received funding from Edith Cowan University (G1003456) and from the School of Science at Edith Cowan University (G1003362) to support this work. CR-G and JG were supported by the grants CTM2015-70340-R and RTI2018-101025-B-I00 of the Spanish Ministry of Science, Innovation and Universities and by the Generalitat de Catalunya Consolidated Research Group 2017SGR/1568. PM acknowledges the support of the Generalitat de Catalunya (MERS 2017 SGR – 1588)With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)Peer reviewe

Impact of particle flux on the vertical distribution and diversity of size-fractionated prokaryotic communities in two East Antarctic polynyas

Antarctic polynyas are highly productive open water areas surrounded by ice where extensive phytoplankton blooms occur, but little is known about how these surface blooms influence carbon fluxes and prokaryotic communities from deeper waters. By sequencing the 16S rRNA gene, we explored the vertical connectivity of the prokaryotic assemblages associated with particles of three different sizes in two polynyas with different surface productivity, and we linked it to the magnitude of the particle export fluxes measured using thorium-234 (234Th) as particle tracer. Between the sunlit and the mesopelagic layers (700 m depth), we observed compositional changes in the prokaryotic communities associated with the three size-fractions, which were mostly dominated by Flavobacteriia, Alphaproteobacteria, and Gammaproteobacteria. Interestingly, the vertical differences between bacterial communities attached to the largest particles decreased with increasing 234Th export fluxes, indicating a more intense downward transport of surface prokaryotes in the most productive polynya. This was accompanied by a higher proportion of surface prokaryotic taxa detected in deep particle-attached microbial communities in the station with the highest 234Th export flux. Our results support recent studies evidencing links between surface productivity and deep prokaryotic communities and provide the first evidence of sinking particles acting as vectors of microbial diversity to depth in Antarctic polynyas, highlighting the direct influence of particle export in shaping the prokaryotic communities of mesopelagic waters

Review of the analysis of 234Th in small volume (2–4 L) seawater samples: Improvements and recommendations

The short-lived radionuclide 234Th is widely used to study particle scavenging and transport from the upper ocean to deeper waters. This manuscript optimizes, reviews and validates the collection, processing and analyses of total 234Th in seawater and suggests areas of further improvements. The standard 234Th protocol method consists of scavenging 234Th from seawater via a MnO2 precipitate, beta counting, and using chemical recoveries determined by adding 230Th. The revised protocol decreases sample volumes to 2 L, shortens wait times between steps, and simplifies the chemical recovery process, expanding the ability to more rapidly and safely apply the 234Th method

Review of the analysis of Th-234 in small volume (2-4 L) seawater samples: improvements and recommendations

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Clevenger, S. J., Benitez-Nelson, C. R., Drysdale, J., Pike, S., Puigcorbe, V., & Buesseler, K. O. Review of the analysis of Th-234 in small volume (2-4 L) seawater samples: improvements and recommendations. Journal of Radioanalytical and Nuclear Chemistry, 329(1), (2021): 1–13, https://doi.org/10.1007/s10967-021-07772-2.The short-lived radionuclide 234Th is widely used to study particle scavenging and transport from the upper ocean to deeper waters. This manuscript optimizes, reviews and validates the collection, processing and analyses of total 234Th in seawater and suggests areas of further improvements. The standard 234Th protocol method consists of scavenging 234Th from seawater via a MnO2 precipitate, beta counting, and using chemical recoveries determined by adding 230Th. The revised protocol decreases sample volumes to 2 L, shortens wait times between steps, and simplifies the chemical recovery process, expanding the ability to more rapidly and safely apply the 234Th method.The authors would like to acknowledge support from the National Aeronautics and Space Administration (NASA) as part of the EXport Processes in the Ocean from RemoTe Sensing (EXPORTS) program awards 80NSSC17K0555; and the Woods Hole Oceanographic Institution’s Ocean Twilight Zone study for KOB and SJC

Small phytoplankton drive high summertime carbon and nutrient export in the Gulf of California and Eastern Tropical North Pacific

Summertime carbon, nitrogen, and biogenic silica export was examined using ²³⁴Th:²³⁸U disequilibria combined with free floating sediment traps and fine scale water column sampling with in situ pumps (ISP) within the Eastern Tropical North Pacific and the Gulf of California. Fine scale ISP sampling provides evidence that in this system, particulate carbon (PC) and particulate nitrogen (PN) concentrations were more rapidly attenuated relative to ²³⁴Th activities in small particles compared to large particles, converging to 1–5 µmol dpm⁻¹ by 100 m. Comparison of elemental particle composition, coupled with particle size distribution analysis, suggests that small particles are major contributors to particle flux. While absolute PC and PN export rates were dependent on the method used to obtain the element/²³⁴Th ratio, regional trends were consistent across measurement techniques. The highest C fixation rates were associated with diatom-dominated surface waters. Yet, the highest export efficiencies occurred in picoplankton-dominated surface waters, where relative concentrations of diazotrophs were also elevated. Our results add to the increasing body of literature that picoplankton- and diazotroph-dominated food webs in subtropical regions can be characterized by enhanced export efficiencies relative to food webs dominated by larger phytoplankton, e.g., diatoms, in low productivity pico/nanoplankton-dominated regions, where small particles are major contributors to particle export. Findings from this region are compared globally and provide insights into the efficiency of downward particle transport of carbon and associated nutrients in a warmer ocean where picoplankton and diazotrophs may dominate. Therefore, we argue the necessity of collecting multiple particle sizes used to convert ²³⁴Th fluxes into carbon or other elemental fluxes, including <50 µm, since they can play an important role in vertical fluxes, especially in oligotrophic environments. Our results further underscore the necessity of using multiple techniques to quantify particle flux given the uncertainties associated with each collection method.This is the publisher’s final pdf. The article is copyrighted by American Geophysical Union and published by John Wiley & Sons, Inc. It can be found at: http://agupubs.onlinelibrary.wiley.com/agu/journal/10.1002/%28ISSN%291944-9224

Quantifying 210Po/210Pb disequilibrium in seawater: A comparison of two precipitation methods with differing results

The disequilibrium between lead-210 (210Pb) and polonium-210 (210Po) is increasingly used in oceanography to quantify particulate organic carbon (POC) export from the upper ocean. This proxy is based on the deficits of 210Po typically observed in the upper water column due to the preferential removal of 210Po relative to 210Pb by sinking particles. Yet, a number of studies have reported unexpected large 210Po deficits in the deep ocean indicating scavenging of 210Po despite its radioactive mean life of ∼ 200 days. Two precipitation methods, Fe(OH)3 and Co-APDC, are typically used to concentrate Pb and Po from seawater samples, and deep 210Po deficits raise the question whether this feature is biogeochemically consistent or there is a methodological issue. Here, we present a compilation of 210Pb and 210Po studies that suggests that 210Po deficits at depths \u3e 300 m are more often observed in studies where Fe(OH)3 is used to precipitate Pb and Po from seawater, than in those using Co-APDC (in 68 versus 33% of the profiles analyzed for each method, respectively). In order to test whether 210Po/210Pb disequilibrium can be partly related to a methodological artifact, we directly compared the total activities of 210Pb and 210Po in four duplicate ocean depth-profiles determined by using Fe(OH)3 and Co-APDC on unfiltered seawater samples. While both methods produced the same 210Pb activities, results from the Co-APDC method showed equilibrium between 210Pb and 210Po below 100 m, whereas the Fe(OH)3 method resulted in activities of 210Po significantly lower than 210Pb throughout the entire water column. These results show that 210Po deficits in deep waters, but also in the upper ocean, may be greater when calculated using a commonly used Fe(OH)3 protocol. This finding has potential implications for the use of the 210Po/210Pb pair as a tracer of particle export in the oceans because 210Po (and thus POC) fluxes calculated using Fe(OH)3 on unfiltered seawater samples may be overestimated. Recommendations for future research are provided based on the possible reasons for the discrepancy in 210Po activities between both analytical methods

High particulate organic carbon export during the decline of a vast diatom bloom in the Atlantic sector of the Southern Ocean

Carbon fixation by phytoplankton plays a key role in the uptake of atmospheric CO2 in the Southern Ocean. Yet, it still remains unclear how efficiently the particulate organic carbon (POC) is exported and transferred from ocean surface waters to depth during phytoplankton blooms. In addition, little is known about the processes that control the flux attenuation within the upper twilight zone. Here, we present results of downward POC and particulate organic nitrogen fluxes during the decline of a vast diatom bloom in the Atlantic sector of the Southern Ocean in summer 2012. We used thorium-234 (234Th) as a particle tracer in combination with drifting sediment traps (ST). Their simultaneous use evidenced a sustained high export rate of 234Th at 100 m depth in the weeks prior to and during the sampling period. The entire study area, of approximately 8000 km2, showed similar vertical export fluxes in spite of the heterogeneity in phytoplankton standing stocks and productivity, indicating a decoupling between production and export. The POC fluxes at 100 m were high, averaging 26±15 mmol C m−2 d−1, although the strength of the biological pump was generally low. Only <20% of the daily primary production reached 100 m, presumably due to an active recycling of carbon and nutrients. Pigment analyses indicated that direct sinking of diatoms likely caused the high POC transfer efficiencies (~60%) observed between 100 and 300 m, although faecal pellets and transport of POC linked to zooplankton vertical migration might have also contributed to downward fluxes

Distribution and export of particulate organic carbon in East Antarctic coastal polynyas

Polynyas represent regions of enhanced primary production because of the low, or absent, sea-ice cover coupled with the proximity of nutrient sources. However, studies throughout the Southern Ocean suggest elevated primary production does not necessarily result in increased carbon export. Three coastal polynyas in East Antarctica and an off-shelf region were visited during the austral summer from December 2016 to January 2017 to examine the vertical distribution and concentration of particulate organic carbon (POC). Carbon export was also examined using thorium-234 (234Th) as a proxy at two of the polynyas. Our results show that concentrations and integrated POC stocks were higher within the polynyas compared to the off-shelf sites. Within the polynyas, vertical POC concentrations were higher in the Mertz and Ninnis polynyas compared to the Dalton polynya. Similarly, higher carbon export was measured in the diatom-dominated Mertz polynya, where large particles ( \u3e 53 μm) represented a significant fraction of the particulate 234Th and POC (average 50 % and 39 %, respectively), compared to the small flagellate-dominated Dalton polynya, where almost all the particulate 234Th and POC were found in the smaller size fraction (1 – 53 μm). The POC to Chlorophyll-a ratios suggest that organic matter below the mixed layer in the polynyas consisted largely of fresh phytoplankton at this time of the year. In combination with a parallel study on phytoplankton production at these sites, we find that increased primary production at these polynyas does lead to greater concentrations and export of POC and a higher POC export efficiency

Small Phytoplankton Drive High Summertime Carbonand Nutrient Export in the Gulf of California and Eastern Tropical North Pacific

Summertime carbon, nitrogen, and biogenic silica export was examined using 234Th:238U disequilibria combined with free floating sediment traps and fine scale water column sampling with in situ pumps (ISP) within the Eastern Tropical North Pacific and the Gulf of California. Fine scale ISP sampling provides evidence that in this system, particulate carbon (PC) and particulate nitrogen (PN) concentrations were more rapidly attenuated relative to 234Th activities in small particles compared to large particles, converging to 1–5 µmol dpm−1 by 100 m. Comparison of elemental particle composition, coupled with particle size distribution analysis, suggests that small particles are major contributors to particle flux. While absolute PC and PN export rates were dependent on the method used to obtain the element/234Th ratio, regional trends were consistent across measurement techniques. The highest C fixation rates were associated with diatom‐dominated surface waters. Yet, the highest export efficiencies occurred in picoplankton‐dominated surface waters, where relative concentrations of diazotrophs were also elevated. Our results add to the increasing body of literature that picoplankton‐ and diazotroph‐dominated food webs in subtropical regions can be characterized by enhanced export efficiencies relative to food webs dominated by larger phytoplankton, e.g., diatoms, in low productivity pico/nanoplankton‐dominated regions, where small particles are major contributors to particle export. Findings from this region are compared globally and provide insights into the efficiency of downward particle transport of carbon and associated nutrients in a warmer ocean where picoplankton and diazotrophs may dominate. Therefore, we argue the necessity of collecting multiple particle sizes used to convert 234Th fluxes into carbon or other elemental fluxes, including \u3c50 \u3eµm, since they can play an important role in vertical fluxes, especially in oligotrophic environments. Our results further underscore the necessity of using multiple techniques to quantify particle flux given the uncertainties associated with each collection method

Intercalibration studies of \u3csup\u3e210\u3c/sup\u3ePo and \u3csup\u3e210\u3c/sup\u3ePb in dissolved and particulate seawater samples

Documented is an intercalibration (IC) exercise for both 210Po and 210Pb in seawater aliquots distributed between up to eight international laboratories that followed individual protocols. Dissolved and particulate samples were provided by GEOTRACES during two IC cruises at baseline stations in the North Atlantic and North Pacific oceans. Included were surface and/or deep dissolved and particulate samples at each site, plus complete profiles analyzed by the laboratory of the lead author. An unspecified solid phase standard was also distributed with 210Po and 210Pb in secular equilibrium to confirm spike calibrations. The 210Po activities reported n = 8) for the standard were very similar with a relative standard deviation (RSD) of 3.6% and mean value indistinguishable from the certified value, confirming accurate calibration of Po spikes. For seawater samples, the agreement was strongly dependent for both nuclides on the activity of the samples. The agreement was relatively good for dissolved seawater samples (RSD = 9% to 29%, n = 4), moderate for the particulate samples (RSD = 12% to 80%, n = 8), and poor for particulate dip blanks (RSD = 50% to 200%, n = 8). Noted is the higher apparent affinity of 210Po versus 210Pb for polysulphone filter material. Some lack of reproducibility between labs may have been caused by unspecified differences in individual lab protocols and calculations. A minimum sample activity of 0.1 dpm for both nuclides is recommended for an adequate reproducible sample activity. It is suggested that a consistent set of procedures and calculations be used to optimize future 210Po and 210Pb analyses in seawater samples