Abstract:

This dataset was from 2 flask experiments (short term and long term) using natural seawaters from the Galveston Bay, Texas for triplicate treatments for control and oil (4ppm). Included are concentrations characterizing the colloidal EPS (< 0.2 um and > 3kDa) for proteins, neutral sugars, uronic acids, colloidal organic carbon, and colloidal total nitrogen. Permeate from the 3kDa membrane was tested for dissolved organic carbon and dissolved total nitrogen. The long-term experiment was sampled daily for 3 days from January 22-25, 2019. The short-term experiment was sampled for selected hourly intervals (0, 1, 2, 3, 4, 8) on July 18, 2019.

Suggested Citation:

Kamalanathan, Manoj, Kathy Schwehr, Christian Taylor, Charles Bergen, Nicole Patterson, Noah Claflin, Peter Sanstchi, and Antonietta Quigg. 2020. Microcosm: Flask experiment of phytoplankton-bacteria interaction in a natural community using control and oil treatments; non-radioactive experiment; extracellular polymeric substances (EPS) assays for protein, neutral sugar, uronic acid, organic carbon, and total nitrogen. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/n7-n5p6-p566

Publications:

Kamalanathan, M., Schwehr, K. A., Labonté, J. M., Taylor, C., Bergen, C., Patterson, N., … Quigg, A. (2021). The Interplay of Phototrophic and Heterotrophic Microbes Under Oil Exposure: A Microcosm Study. Frontiers in Microbiology, 12. doi:10.3389/fmicb.2021.675328

Purpose:

To track the characterization changes of extracellular polymeric substances (EPS), this experiment ran in parallel without radio-labelled tracers and compliments the measurements made in a parallel experiment conducted using radio-tracers.

Data Parameters and Units:

There are 2 files: short-term and long-term colloidal extracellular polymeric substances (EPS). The headers are- Time (hours or Days), Sample, Colloidal Neutral Sugars (ug/mL), Colloidal Proteins (ug/mL), Colloidal Uronic acids (ug/mL), Colloidal Organic Carbon (ug/mL), Colloidal Total Nitrogen (ug/mL), Dissolved Organic Carbon (ug/mL), Dissolved Total Nitrogen (ug/mL)

Methods:

Time interval samples of 10 mLs were filtered using a 0.2 um filter, and the resulting filtrate was then ultra-filtered via Amicon Ultra-4 Centrifugal Filter Unit with a 3 kDa cut-off membrane (Millipore, USA). The retentate (3 kDa- 0.2 µm) was concentrated to 2 mL for determination of protein, uronic acid and neutral sugars, which were assumed to represent the major components of extracellular polymeric substances (EPS). Protein content in EPS was measured based on a modified bicinchoninic acid method (Smith et al. 1985) using the Pierce protein assay kit, with bovine serum albumin as the standard. The neutral sugar concentration was determined using the anthrone method (Morris 1948), with glucose as the standard. Uronic acid was estimated by adding sodium borate (75 mM) in concentrated sulfuric acid and m-hydroxydiphenyl, with glucuronic acid as the standard (Hung and Santschi 2001). Colloidal organic carbon and colloidal total nitrogen were measured against potassium hydrogen phthalate (KHP) and potassium Nitrate (KNO3) standards on a Shimadzu TOC analyzer.

Instruments:

Shimadzu TOC analyzer

Provenance and Historical References:

Morris Daniel L. 1948. Quantitative determination of carbohydrates with Dreywood’s anthrone reagent. Science 107:254-5. DOI: 10.1126/science.107.2775.254 Hung Chin-Chang, Peter H. Santschi. 2001. Spectrophotometric determination of total uronic acids in seawater using cation-exchange separation and pre-concentration by lyophilization. Analytica Chimica Acta 427(1):111–117. DOI:10.1016/S0003-2670(00)01196-X Smith PK, R.I. Krohn, G.T. Hermanson, A.K. Mallia, F.H. Gartner, M.D. Provenzano, E.K. Fujimoto, N.M. Goeke, B.J. Olsen, and D.C. Klenk. Measurement of protein using bicinchoninic acid. [published correction appears in Analytical Biochemistry 1987 May 15;163(1):279]. Analytical Biochemistry 1985;150(1):76-85. doi:10.1016/0003-2697(85)90442-7

Individual Files: