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Calculate the growth rate constant:

During the exponential (or logarathmic) growth phase, a bacterial culture mimics a first-order chemical reaction, i.e. the rate of increase of cells is proportional to the number of bacteria present at that time. The constant of proportionality, , is an index of the growth rate and is called the growth rate constant: Rate of increase of cells = x number of cells The value of can be determined from the following equation: ln Nt - ln N0 = (t - t0) in other words: the natural log of the number of cells at time t minus the natural log of the number of cells at time zero (t0) equals the growth rate constant multiplied by the time interval. For most purposes, it is easier to use log10 values rather than natural logs, so the above equation can be converted as follows: log10 N - log10 N0 = (/2.303) (t - t0) or alternatively: = ( (log10 N - log10 N0) 2.303) / (t - t0) By measuring the increase in the number of cells during a certain time period, the growth rate constant () can be calculated. In the experiment you have just done:

t0 = 1.5h: 1 N = 8.4x10 , log10 N = 1.92 t = 8.5h: 8 N = 3.39x10 , log10 N = 8.53

Therefore in this case:

= ( (log10 N - log10 N0) 2.303) / (t - t0) = ( (8.53 - 1.92) 2.303) / (8.5 - 1.5)

= (6.61 x 2.303) / 7 = 15.22 / 7 = 2.18 hour

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Note that and g are related to each other: = ln2/g = 0.693/g

Calculate the mean generation time: The mean generation time or "doubling time" (g) is the average time required for all the components of the culture to double. This is calculated from the following equation: log10 Nt = log10 N0 + g log102 or alternatively: g = (log10 Nt - log10 N0) / log102 Again, for the experiment you have just done:

t0 = 1.5h: 1 N = 8.4x10 , log10 N = 1.92 t = 8.5h: 8 N = 3.39x10 , log10 N = 8.53

g = (log10 Nt - log10 N0) / log102 = (8.53 - 1.92) / 0.301 = 21.96 generations in 7 hours = 7 / 21.96 = 0.32 hours (x 60 = 19.2 minutes) Note that and g are related to each other: = ln2/g = 0.693/g

Calculation of Generation Time When growing exponentially by binary fission, the increase in a bacterial population is by geometric progression. If we start with one cell, when it divides, there are 2 cells in the first generation, 4 cells in the second generation, 8 cells in the third generation, and so on. The generation time is the time interval required for the cells (or population) to divide. G (generation time) = (time, in minutes or hours)/n(number of generations) G = t/n t = time interval in hours or minutes B = number of bacteria at the beginning of a time interval b = number of bacteria at the end of the time interval n = number of generations (number of times the cell population doubles during the time interval) b = B x 2n (This equation is an expression of growth by binary fission) Solve for n: logb = logB + nlog2 n = logb - logB log2 n = logb - logB .301 n = 3.3 logb/B G = t/n Solve for G G=t 3.3 log b/B

Example: What is the generation time of a bacterial population that increases from 10,000 cells to 10,000,000 cells in four hours of growth?

G = t_____ 3.3 log b/B G = 240 minutes 3.3 log 107/104 G = 240 minutes 3.3 x 3 G = 24 minutes