Dataset: EN584 LV polysaccharide hydrolysis rates
Deployment: EN584

For mesocosm (large volume) incubation experiments (referred to as “LV” incubations), a 30L Niskin bottle rosette was used to collect the water. Separate casts were used to collect surface water, bottom water, and water from the depth at which oxygen showed a minimum, according to the CTD. From each depth, 20L seawater from single Niskin bottles was dispensed using cleaned silicon tubing into a single carboy. Prior to filling, carboys were rinsed 3x with water from the same Niskin bottle used to fill the carboy. Four carboys were filled at each depth. Triplicate 20L carboys were amended with ca. 500 mg (exact mass was recorded for each addition) of HMW Thalassiosira; unamended single carboys were used for controls. All mesocosms were incubated in the dark at near in-situ temperatures. Mesocosms were sub-sampled at the start of incubation (0 days), and then after 2 d, 7d, and 16d for the following assays: bacterial production using 3H-Leucine, dissolved organic carbon (DOC), nutrients, bacterial cell counts, peptidase and glucosidase activity measurements. At the 16d subsampling timepoint, polysaccharide hydrolase activity measurements were initiated, using fluorescently labeled polysaccharides (Arnosti 2003). These polysaccharide incubations were sampled at time points of 0, 2, 5, 10, 17, and 30 days (with the zero-time sample being at the 16-day timepoint of the mesocosm experiment).

The hydrolysis of high molecular weight substrate to lower molecular weight hydrolysis products was measured using gel permeation chromatography with fluorescence detection, after the method of Arnosti [1996, 2003]. In short, the subsample was injected onto a series of columns consisting of a 21 cm column of G50 and a 19 cm column of G75 Sephadex gel. The fluorescence of the column effluent was measured at excitation and emission wavelengths of 490 and 530 nm, respectively. Hydrolysis rates were calculated from the change in molecular weight distribution of the substrate over time, as described in detail in Arnosti [2003].