These data include oceanographic, chemical, and biogeochemical rate measurements relating to oxygen, nitrogen, and sulfur species from Saanich Inlet, British Columbia, Canada, spanning lower oxycline and anoxic depths in the basin. Data and samples were collected aboard the R/V John Strickland during two cruises in August, 2021 and June, 2022. Data from 2021 capture the dynamics of an oxygen renewal event of the deep anoxic basin, which occurred between the second and third sampling days. Data from 2022 were collected during stable, stratified conditions with complete anoxia in the deep basin throughout the sampling period.

Table of Contents

• Coverage
• Dataset Description
  • Methods & Sampling
  • Data Processing Description
• Related Publications
• Parameters
• Instruments
• Deployments
• Project Information
• Funding

Nutrients - Sample collection: Duplicate 15-mL polypropylene tubes were rinsed three times with 0.2-µm filtered seawater before collecting 10 mLs and freezing at -20 degrees C.

Nitrite - Sample collection:  Duplicate 125-mL HDPE bottles were rinsed three times with 0.2-µm filtered seawater before collecting 100 mLs and storing at room temperature in the dark.

Ammonium - Sample collection: Duplicate 50-mL glass tubes were rinsed three times with sample before collecting 40 mLs and adding 10 mLs orthophthaldialdehyde (OPA). Samples were stored in the dark prior to analysis.  

Oxygen - Sample collection:  Duplicate oxygen flasks were sampled from a dedicated Niskin bottle. Flasks were rinsed with sample while inverted then allowed to overfill three times their volume. Flasks immediately received 1-mL each manganous chloride reagent and alkaline sodium iodide flasks through a submerged spigot and were sealed with ground glass stoppers, using caution to avoid introduction of bubbles. After 30 seconds of vigorous shaking, flasks were stored in the dark with a deionized water seal around the stoppers. 

Nitrous Oxide - Sample collection:  Water was sampled through gas-tight Viton tubing into 20-mL glass vials filling from the bottom with three times overflow. Vials were closed bubble-free with deoxygenated butyl rubber stoppers, crimp-sealed and stored inverted in the dark at 4 °C until analysis.

Sulfide - Sample collection:  Water was sampled through gas-tight viton tubing into 12-mL glass exetainer vials, filling from the bottom with three times overflow and no headspace. Samples were immediately amended with N,N-dimethyl-p-phenylenediamine reagent and stored in the dark until analysis (within 6 hours) (following Cline, 1969).

DNA - Sample collection:  2.5 L of seawater were filtered onto 0.2-µm Sterivex filters. After addition of 1.2  mL of lysis buffer, filters were frozen on dry ice until the end of the day, when they were transferred to a -80 C freezer. 

RNA - Sample collection:  Between 2.25-2.5 L of seawater were filtered onto 0.2-µm Sterivex filters within 20 minutes of CTD recovery. Filters were flooded with RNA preservative and frozen on dry ice until the end of the day, when they were transferred to a   -80 C freezer. 

Nutrients - Analysis methods: Nitrite and nitrate plus nitrite were measured on an Astoria Nutrient Autoanalyzer following the methodology of Barwell-Clarke and Whitney 1996. Samples from sulfidic depths were sparged with dinitrogen gas for 20 minutes prior to analysis.

Nitrite - Analysis methods: Using a Greiss reaction method, samples were first treated with 1.0 mL of sulfanilamide solution and allowed to react for 2-8 minutes. After addition of 1.0 mL of naphthylethylenediamine solution and a 1-hour reaction time, samples were measured spectrophotometrically over a 10-cm path length following the methodology of Strickland & Parsons 1972.

Ammonium - Analysis methods: Samples were allowed to react with OPA for 4-8 hours before fluorometric measurement following the methodology of Holmes et al 1999.

Oxygen - Analysis methods: Dissolved oxygen was measured by titration with sodium thiosulfate following the methodology of Codispoti 1988.

Nitrous Oxide - Analysis methods: Nitrous oxide concentrations were measured by cavity ring-down spectrometry following the methodology of Ji & Grundle 2019.

Sulfide - Analysis methods: Samples were analysed spectrophotometrically following the methodology of Cline 1969.

DNA - Analysis methods: DNA was extracted and sent for sequencing of 16S rRNA gene amplicons and metagenomes on Illumina instruments (MiSeq/NextSeq).

RNA - Analysis methods: RNA was extracted, reverse transcribed to cDNA, and sent for metatranscriptomic sequencing on Illumina instruments (MiSeq/ NextSeq).

Coverage: Saanich Inlet — a seasonally anoxic fjord on Vancouver Island, British Columbia, Canada

NSF Award Abstract:

Nitrous oxide (N2O) is a gas produced by microbes in both aquatic and terrestrial environments, and, like other greenhouse gases, it contributes to global warming. Furthermore, N2O can destroy ozone, a gas responsible for protecting the earth from dangerous ultraviolet radiation. In the ocean, N2O production is largely controlled by the amount of available dissolved oxygen, with more N2O being produced under low oxygen concentrations; however, when no oxygen is available, a scenario referred to as anoxia, microbes in the ocean switch from producing N2O to consuming N2O. In recent years, it has become evident that zones of low oxygen are expanding in some areas of the oceans, and this has raised concern that more N2O will be produced. If this occurs, more N2O will be emitted to the atmosphere, and will lead to further global warming and ozone destruction. Because of this, research has largely focused on understanding how much N2O is produced in the ocean under low oxygen conditions. If, however, anoxic zones also increase in size, this could act to balance out, at least to some degree, the predicted increase in N2O production caused by the expansion of zones where oxygen is present but in low concentrations. This study aims to simultaneously measure N2O production and consumption, in both low oxygen and anoxic zones and identify the microbes responsible for N2O production and consumption. Our results will: 1) lead to a much better understanding of how N2O consumption in anoxic zones could help to balance out an increase in N2O production if low oxygen zones in the ocean continue to expand, 2) help to inform models aimed at predicting oceanic N2O production and emissions to the atmosphere under future ocean conditions, and 3) allow us to better understand the microbes involved in N2O production and consumption. Our study will support a postdoc and undergraduate students who will work at the interface of marine chemistry and community genomics. The PIs plan to specifically consider applications from underrepresented minorities and students at institutions with limited opportunities. The PIs also plan a number of other educational/outreach programs ranging from teacher-training workshops, teacher internships, and academic and public lecture series.

The oceanic production of the potent greenhouse and ozone destroying gas nitrous oxide (N2O) increases as dissolved oxygen (DO) concentrations transition from oxic to hypoxic. Marine DO concentrations have decreased globally with climate change and oceanic hypoxic zones have expanded and predicted to continue expanding. This increase is cause for concern that N2O production in the ocean will increase in the future which would lead to higher emissions to the atmosphere. As a result, much research has focused on quantifying the oxygen thresholds that correspond to large increases in N2O production. In contrast, relatively few studies have aimed to quantify the capacity for net N2O consumption, resulting from microbial N2O reduction to N2 under anoxic conditions, to buffer against predicted N2O production increases if anoxic zones expand in conjunction with hypoxic zones. To this end, this study aims to simultaneously quantify N2O production and consumption from oxic-hypoxic-anoxic water column zones, in order to determine the potential for N2O consumption to counteract predicted increases in N2O production. Our field work be conducted in Saanich Inlet, a British Columbian fjord which is an ideal natural laboratory for our study, as it is characterized by a well-established oxycline and anoxic zone. Specifically, we aim to 1) measure bulk N2O concentrations, and, using 15N tracer techniques, quantify N2O production and consumption rates as DO concentrations decrease from oxic to anoxic conditions, 2) quantify the magnitude by which N2O consumption in the anoxic zone balances increased N2O production in the overlying hypoxic region, and 3) definitively link observed N2O production and consumption rates to the microorganisms mediating this process, focusing specifically on distinguishing N2O consumption via denitrifier (NO3- to N2) versus non-denitrifier (N2O to N2 only) taxa. Ultimately, our results will provide quantitative information on N2O consumption rates over fluctuating ocean conditions, thereby helping constrain models of oxygen effects on net N2O production and ocean-to-atmosphere greenhouse gas fluxes. Furthermore, this work will identify the taxonomic breadth of microbes capable of N2O reduction and their linkage to actual N2O reduction rates, thereby providing a quantitative understanding of whether or not the detection of specific bio-signatures is predictive of marine N2O dynamics.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.