Abstract:

Various extracellular parameters such as enzyme activities, organic carbon and nitrogen, exopolymeric substances, dissolved inorganic nitrogen and phosphate and estimated oil equivalent data were measured over the course of a 16 day period in a mesocosm experiment with Control and water-accommodated fractions (WAF) treatments. Measured data were analyzed to understand the changes in organic carbon and nitrogen utilization during an oil spill.

Suggested Citation:

Manoj Kamalanathanan, Shawn M. Doyle, Chen Xu, Gerardo Gold-Bouchot, Amanda Achberger, Hernando Bacosa, Laura Bretherton, Maya E. Morales-McDevitt, Terry L. Wade, Kathy Schwehr, Peter Santschi, Jason B. Sylvan, Antonietta Quigg. 2019. Mesocosm experiment: Exoenzyme activity, organic carbon and nitrogen, exopolymeric substances, dissolved inorganic nitrogen and phosphate, and estimated oil equivalent data measured using control and WAF treatments. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/2J7H8GGS

Publications:

Kamalanathan, M., Doyle, S. M., Xu, C., Achberger, A. M., Wade, T. L., Schwehr, K., … Quigg, A. (2020). Exoenzymes as a Signature of Microbial Response to Marine Environmental Conditions. mSystems, 5(2). doi:10.1128/msystems.00290-20

Purpose:

To understand the changes in organic carbon and nitrogen utilization during an oil spill.

Data Parameters and Units:

Beta glucosidase: nMols/hr Lipase: nMols/hr Alkaline phosphatase: nMols/hr Leucine amino-peptidase: nMols/hr Particulate organic carbon (POC): mg/L Particulate inorganic nitrogen (PON): mg/L Colloidal organic carbon (COC): mg C/L Colloidal organic nitrogen (CON): mg N/L Neutral sugars: mg Glucose/L Proteins: mg BSA/L Uronic acid: mg Glucuronic acid/L Estimated oil equivalents (EOE): mg/L Dissolved inorganic nitrogen (DIN): umol/L Dissolved inorganic phosphate (DIP): umol/L Please note: In all the files except EOE.csv, column B, C, & D are replicates of Control and column E, F & G are replicates of WAF.

Methods:

The WAF was made using seawater obtained from NOAA and as described by Wade et al. (2017). Oil Analysis: The total oil concentrations were determined by measuring the Estimated Oil Equivalent (EOE) values according to Wade et al. (2017). Briefly, fluorescence of dichloromethane extractions of 5-10 ml of samples were measured at 260/358 nm excitation/emission using a spectrofluorometer (Shimadzu RF-5300). Exoenzyme Assays: The activities of exoenzymes β-glucosidase, leucine amino-peptidase, alkaline phosphatase, and lipase were measured daily for 16 days according to methods described by Kamalanathan et al. (2018). Substrates 4-Methylumbelliferyl β-D-glucopyranoside, 4-Methylumbelliferyl oleate, 4-methylumbelliferyl phosphate, and leucine-AMC hydrochloride were used for enzymes β-Glucosidase, lipase, alkaline phosphatase and leucine amino-peptidase activity, respectively. The samples were incubated with respective substrates at 0.2 mM (final concentration) for three hours and activities were determined as fluorescence at excitation/emission wavelengths of 365/448 nm for MUF tagged substrates and 380/440 nm for AMC tagged substrates. The measurements were done performed using a BioTek CytationTM 5 imaging reader controlled by Gen5 2.09 software (USA). Particulate Organic Carbon (POC) and Particulate Organic Nitrogen (PON) analysis: POC and PON analysis were performed by filtering water samples through a pre-combusted GF/F membrane (0.7 μm, Whatman, United States). After remove removing the carbonates by HCl-fuming, quantification was performed using a Perkin Elmer Series II CHNS 2400 analyzer with Acetanilide (71.09%) was as an analytical standard (Xu et al., 2011). Nutrient analysis were performed by collecting 30 mL of each treatment were collected as triplicates and filtered under vacuum with a 45 μm Milipore filter and kept frozen until its analysis at Geochemical Environmental Research Group (GERG). All nutrient samples were analyzed with an Astoria Pacifica Auto-Analyzer. The nutrient analysis followed the GERG ARM-SOP-0702. Five standards prepared with specific ranges, a PO3-, NO2-, NO3-, NH4+ and a Certified Reference Material (CRM) were run before each sample run. The CRM was also analyzed between each batch of 12 samples with a blank determination. To determine the spike recovery percent, a CRM and a replicate sample were utilized. Peak heights were analyzed and converted into μmol/l concentrations using the Flow Analyzer Software Package II (FASPACII); which controls, collects and processes data from six digital channels and one analog channel simultaneously from the Astoria Pacifica auto-analyzer. Dissolved inorganic nitrogen was calculated by adding NH4, NO2 and NO3 values. Dissolved inorganic phosphate was calculated using PO3- values. Redfield ratios were also made for each mesocosm. Comparisons between treatments and mesocosms were made, as well as correlations to the other measurements taken.

Provenance and Historical References:

Kamalanathan, M., Xu, C., Schwehr, K., Bretherton, L., Beaver, M., Doyle, S., Genzer, J., Hillhouse, J., Santschi, P.H., & Quigg, A. (2018). Extracellular Enzyme Activity Profile in a Chemically Enhanced Water Accommodated Fraction of Surrogate Oil: Toward Understanding Microbial Activities After the Deepwater Horizon Oil Spill. Frontiers in Microbiology, 9. doi:10.3389/fmicb.2018.00798 Wade, T.L., Morales-McDevitt, M., Bera, G., Shi, D., Sweet, S., Wang, B., Gold-Bouchot, G., Quigg, A., & Knap, A.H. (2017). A method for the production of large volumes of WAF and CEWAF for dosing mesocosms to understand marine oil snow formation. Heliyon, 3(10), e00419. doi:10.1016/j.heliyon.2017.e00419 Xu, C., Santschi, P. H., Hung, C. C., Zhang, S., Schwehr, K. A., Roberts, K. A., Guo, L., Gong, G.C., Quigg, A., Long, R.A., & Pinckney, J. L. (2011). Controls of 234Th removal from the oligotrophic ocean by polyuronic acids and modification by microbial activity. Marine Chemistry, 123(1-4), 111–126. doi:10.1016/j.marchem.2010.10.005

Individual Files: