Abstract:

A mesocosm scale experiment was conducted to monitor the temporal changes in microbial cell abundance as well as the concentration and size of microaggregates that form in response to the presence of oil and chemically dispersed oil. Water samples were collected and viewed under the microscope to quantify cell and microaggregate concentrations and to monitor the size of the microaggregates over time.

Suggested Citation:

Amanda Achberger, Charlie Holmes, Jason Sylvan. 2020. Characterization of cell concentrations and microaggregate abundance and size, LTMOSE, a Long Term Marine Oil Snow Mesocosm Experiment, abundance counts. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N71GNKT1

Publications:

Achberger, A. M., Doyle, S. M., Mills, M. I., Holmes, C. P., Quigg, A., & Sylvan, J. B. (2021). Bacteria-Oil Microaggregates Are an Important Mechanism for Hydrocarbon Degradation in the Marine Water Column. MSystems, 6(5). https://doi.org/10.1128/msystems.01105-21

Purpose:

Monitor the temporal changes in microbial cell abundance as well as the concentration and size of microaggregates that form in response to the presence of oil and chemically dispersed oil.

Data Parameters and Units:

Treatment, Time Point, Cell per mL; Aggregates per mL; Area (µm2).

Methods:

Direct cell counts were performed with an epifluorescence microscope (Zeiss Axio Imager.M2) after staining the 2% formalin-fixed samples with DAPI (45uM final concentration) for 5 minutes in the dark and filtering them onto 25mm, 0.2um black polycarbonate filters. Each filter was mounted on a glass microscope slide with a coverslip using two drops of AF1 antifade. A 100um×100um ocular counting grid was used at 1000× magnification for cell counts. Due to their much larger size, aggregates were instead counted for each field of view at 400× magnification. The area of each aggregate was manually calculated using ImageJ software.

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