Patricia Louise Pineda Date performed: July 8,2013

AB-1L Date submitted: July 21, 2013

Exercise 3
Population Growth of a cultured organism, Chlorella vulgaris, grown in media with different
pollutant mixtures.

I. Results

Table 3.1 Initial Optical Densities of C. vulgaris incubated in different pollutant mixtures.

Parameter Control Household waste Petroleum waste Control
20% 40% 20% 40%
Replicate
1

1 0.13 0.014 -2.04 -2.06 0 0.088
2 0.22 -0.09 -1.89 -2.35 0 ---
Replicate
2
1 0.146 -0.006 -2.09 -2.13 0 0.098
2 0.28 -0.114 -1.93 -2.10 0 ---
Ave 0.194 -0.196 -1.9875 -2.16 0 0.093


Table 3.2. Final Optical densities of C. vulgaris incubated in different pollutant mixtures.

Parameter
Trial
Control Household Waste Petroleum Waste
blank With
inoculum
20% 40% 20% 40%

Rep 1 1 0 0.354 0.468 -0.89 -3.92 -1.64
2 0 0.329 0.518 -0.61 -3.28 -1.06
Rep2 1 0 0.323 0.366 -0.88 -4.6 0.58
2 0 0.324 0.354 -1.12 -4.64 0.66
Average 0 0.3325 0.4265 -0.875 -4.11 -0.365



































Figure 3.1 C. vulgaris exposed to 20% petroleum Figure 3.2 C. vulgaris exposed to 40%
waste under HPO (400x) petroleum waste under HPO (400x)




II. Discussion

The organism used in this study was Chlorella vulgaris. It is single celled green alga that
belongs to Division Chlorophyta. It contains green pigments such as chlorophyll a and b. It is
capable of multiplying rapidly with the presence of carbon dioxide, water, sunlight, and a fair
amount of minerals.

The population density of C. vulgaris was measured using a spectrophotometer. A
spectrophotometer is an instrument that measures the amount of photons (the intensity of light)
absorbed after it passes through a sample solution. With the spectrophotometer, the amount of a
known chemical substance (concentrations) can also be determined by measuring the intensity of
light detected. The optical density is directly proportional to the concentration of the sample.

As seen in table 3.2 and 3.3, the averages of the replicates per pollutant and in the control were
all computed. In the 20% concentration of the household waste, the optical density increased while in
the 40%, the OD decreased. For the petroleum waste, the final OD for 20% concentration decreased to a
more negative value compared to its initial OD reading. On the other hand, for the 40% concentration,
the OD increased in the final OD reading.

The medium that provided the highest carrying capacity is the 20% household waste which
contains dishwashing liquid from avocado leftovers. On the other hand, the medium that provided the
lowest carrying capacity is the 20% petroleum waste. Carrying capacity is defined as the maximum
population size that an environment (in this case the pollutant mixture) can sustain or support. It is
capable of providing enough nutrients and other essential materials to support as many organisms
possible. In the case of C. vulgaris, the 20% household waste was able to support the highest population
possible based on the OD reading which is 0.4265. This may be due to the avocado leftovers washed by
the dishwashing liquid. The avocado provided nutrients for the growth of the Chlorella.

Increasing pollutant mixture decreases the optical density of the pollutant and algal mixture for
the household waste. In table 3.2, the average of the OD from the two concentrations decreased into a
negative value for the 40% household waste concentration. On the other hand, for the petroleum
waste, although the average optical densities from the two concentrations increased, the values are still
negative which means that the concentration of the C. vulgaris is low compared to that of the household
waste.

C. vulgaris was able to grow/live in both waste products because it has the capability to remove
pollutants in water. C vulgaris is a microalgae that is capable of phytoremediation. This process involves
the reducing of nutrients in a contaminated medium due to the algaes capability to assimilate the
nutrients in the pollutant to their cells. As long as there is the presence of light (in this experiment, the
light came from an artificial source), then the C. vulgaris will be able to phytoremediate the waste
materials.

Although of course, their performance varied depending on the culture environment. Growth of
Chlorella vulgaris was influenced by growth factors such as sufficient light and nutrients.
Phytoremeditation effects in the petroleum waste were not significant because of the presence of heavy
metals and complex hydrocarbons that the C. vulgaris cannot utilize completely. Its remediation
capacity can be greater observed in the household waste wherein the waste materials are not that
complex to utilize.

References:
Phytoremediation of Partially Treated Wastewater by Chlorella vulgaris
<http://psrcentre.org/images/extraimages/712064.pdf>
Spectrophotometry: Basic Principles
<http://www.chm.davidson.edu/vce/spectrophotometry/Spectrophotometry.html>