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Growth rate from biomass observation

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Microbial growth rate
Approach: dilution incubation
Context: incubation, lab
Spatial scale: point sample
Temporal scale: hours to days
Units: cells L-1 h-1
Community captured: bulk, single cell
Co-measurements: cell abundance

Method Overview

The dilution method isolates microbial growth rates from grazing loss by diluting a seawater sample with particle-free (0.2 µm filtered) seawater across a range of dilution factors. Dilution reduces predator-prey encounter rates proportionally, while the intrinsic growth rate of the target cells is assumed to remain constant. By measuring net growth rates across this dilution series via flow cytometry (FCM) or epifluorescence microscopy, growth rate (µ) and grazing rate (g) can be separated as the y-intercept and slope of the net growth rate vs. dilution factor regression[1].

The assay can resolve bulk community rates or, when target populations are distinguishable by their optical signatures, population-specific rates.

Scale of measurement

As a bottle-based incubation, this method yields a single point measurement in space. Incubations typically run over hours to days, providing a time-averaged growth rate for the duration of the experiment.

Data generated

The primary outputs are the intrinsic growth rate µ (d-1) and the grazing rate g (d-1). Cell abundance at each dilution level is tracked by flow cytometry or microscopy over the incubation period. Carbon-based rates require cell-specific carbon content estimates.

Units & currency

Units are cells L-1 h-1.

Sample size

Typical samples are < 1 L in volume per dilution treatment.

Repositories & databases

Limitations

The method assumes that grazing rates scale linearly with dilution, and that the physiological state of grazers and prey does not change across dilution levels during the incubation. Bottle confinement can alter trophic interactions and the physical-chemical environment relative to in situ conditions. Nutrient addition to unamended dilution series bottles is sometimes required to prevent nutrient limitation from confounding the growth rate estimate.

Example Applications & Protocols

Classic examples

  • Landry et al. (1993) Time-dependency of microzooplankton grazing and phytoplankton growth in the subarctic Pacific [1]

Recent applications

  • Heneghan et al. (2024) The global distribution and climate resilience of marine heterotrophic prokaryotes [2]

Common calculations/conversions

  • Carbon content per cell is required to convert cell-based rates to carbon units; typical values are 10–20 fg C cell-1 for heterotrophic bacteria.

References

  1. 1.0 1.1 Landry, M. R., Monger, B. C., & Selph, K. E. (1993). Time-dependency of microzooplankton grazing and phytoplankton growth in the subarctic Pacific. Progress in Oceanography, 32(1–4), 205–222. https://doi.org/10.1016/0079-6611(93)90014-5
  2. Heneghan, R. F., Holloway-Brown, J., Gasol, J. M., Herndl, G. J., Morán, X. A. G., & Galbraith, E. D. (2024). The global distribution and climate resilience of marine heterotrophic prokaryotes. Nature Communications, 15, 6943. https://doi.org/10.1038/s41467-024-50635-z