Introduction

Growth is usually characterized as an increase in size. In the case of bacteria, it is growth by numbers.
Bacterial growth refers to the increase of population of cells by division through binary fission. Growth is
routinely measured either through serial dilution and colony count, or though optical density using a
spectrophotometer. Bacterial growth can be visualized graphically in a growth curve, which shows four
phases: the lag phase, exponential phase, stationary phase, and the death phase. In this lab simulation,
bacterial growth of E. coli will be measured through serial dilution and colony count. The growth of E.
coli will also be used to test the antimicrobial properties of an isolated compound taken from fungi
isolated from the Amazon Rainforest.

Results and Discussion

To measure the effect of the fungal compound on bacterial growth, the number of live bacterial cells was
counted at certain time points and compared with the growth medium without the compound. These
values are shown in Table 1 below.

Table 1. Calculated colony forming units

Incubation time (h) cfu/mL of fungal compound cfu/mL of control sample
sample
24 2,200 1.100.000.000
48 160,000 2.300
72 1.100.000 0
96 44.400.000 0

As mentioned previously, the growth of bacteria occurs by binary fission. Since bacterial growth is the
division of cells from 1 to 2, the growth pattern can be described as exponentially doubling. If it were to
be expressed in an equation, the growth formula can be written as Nt = 2n * N0, where N is the number of
cells at a certain time t and n is the number of generations. If the actual number of cells were to be plotted
in a graph, it would be difficult since the numbers could reach hundreds of thousands to a million in a
certain time interval. To easily visualize large numbers, the logarithm of measurements can be taken, as
shown in Figure 1 below. When the logarithmic scale is used, we transform an exponential function into a
linear graph.
 Figure 1. Growth chart of E. coli over 96 hours.

The straight line in the graph above shows that there is a constant growth rate. This growth rate is defined
as k, which refers to the number of generations per time interval. This remains constant. This is shown by
the equation k= n/t or the number of generations per time. The growth rate constant can also be expressed
in logarithmic terms as k=log2(Nt / N0) / t. This equation allows us to calculate the growth rate constant
using any two of the points on the growth chart shown in Figure 1.

Towards the end of the graph, the last point does not line up with the rest which skews the line. This
indicates that the nutrients are starting to become scarce. As bacteria cannot grow perpetually, the growth
curve has to end, along with the depletion of nutrients, space and oxygen. Once all the nutrients are used
up, the number of viable cells will decrease.
 Figure 2. Overlay of E. coli growth curves with and without the antibacterial compound.

The lag in the graph, shown as a flat line in Figure 2, is called the lag phase. This is where the existing
bacterial cells become accustomed to the growth medium to prepare for the upcoming series of cell
divisions. The exponential phase is shown by the ascending straight line. Because it is straight, there
exists a slope, which is referred to as the growth rate, k. Figure 2 also shows the growth curve of E. coli
when the culture media is treated with the antibacterial compound. It can be observed that the
antibacterial compound decreases the growth rate. This is seen as a decline of the steepness of the slope,
which indicates that the fungal compound has reduced k.

Figure 3. Growth curve of E. coli at room temperature and incubator temperature.

As shown in Figure 3, E. coli grows faster in incubator temperature compared to room temperature. This
is shown in the graph by the blue curve ascending before the red curve. Naturally, E. coli inhabit the
intestines of warm-blooded animals. This is why their growth is much faster in incubator temperature,
which simulates body temperature.

Conclusions and Recommendations

Bacterial growth occurs exponentially since each daughter cell doubles in each generation. The growth
can be visualized graphically using a growth curve, which can be expressed exponentially or
logarithmically. By playing around with the values, we can see that it is easier to express bacterial growth
logarithmically because we can quantify growth using the slope, or the growth constant. With the data
that we have, we can see that the isolated fungal compound has decreased E. coli growth, which is
manifested as a decrease in the growth constant. When comparing growth in room temperature and
incubator temperature, the growth of E. coli was much faster in the incubator compared to room
temperature. This is because their optimal temperature is mammalian body temperature since they
naturally reside in the gut.