Discussions

The aim of this experiment was to isolate and identify colonies using three pure culture
techniques: pour-, spread- and streak-plate and also to calculate quantitatively the number
of viable cells via continuous cell dilution and colony counter. After incubation, all the pure
cultures showed changes to its composition, turbidity and colour. Most turned hazy and
spots of colonies were observed. It should be noted that the incubation was done for more
than 24 hours, and this might have an effect on the results.
From results, microbial growth can be seen in all the cultures. It was observed cloudshaped, highly turbid object in the Petri dish. Pour-plate methods cause the culture to have
colonies on the surface as well as subsurface. In this experiment we were able to detect only
one colony on the surface. As for Dilution, All of the dilutions pour on the Petri dish were no
microbial growth except for the culture with dilution factor 1 : 10000 contain the most
microbial for this experiment. Approximating the number of colony around 1200, it was safely
above the 300 count, and thus the CFU calculation could be carried out and determined to
be 1.2X108 CFU/mL. This result is logical since the number of colony should be increasing
with decreasing dilution factor of pour-plate because generally we have no idea of the
number of bacteria in a sample, a dilution series is to ensure that we will obtain a dilution
containing a reasonable number of bacteria to count which is the high concentration of
dilution inhabit the microbial growth. The correct concentration or in this experiment the
dilution with 1: 10000 factor will cause the growth of bacteria as its reach its optimum
amount for suitable growth. However, this theory is not valid and remain unsure because
there is no supporting information on this. however, this experiment prove that on a certain
amount dilution factor for pour-plate method, the bacteria able to growth based on the
optimum concentration of bacteria.

As for the streak-plate method, colonies were very much present on the Petri dish.
Comparing to the clear, visible agar before the incubation, colonies are seen on the trails of
the streaks done during culturing. However, the number of colonies appeared to be below
than 30 colonies, thus it was written down as TNTC.
Apart from that, the results from the spread-plate method seem more promising
compared to the rest as all the colonies were visible not included in experiment b. However,
the colonies were all attached and connected to each other, making it hard to identify and
individually count them. The result show 53 number of colony was on the plate and it was
safely below the 30 count

Even though most of the results were designed as TNTC and should be discarded,
isolation of a single microorganism was successfully achieved as no other microorganism
was detected in the culture. This is confirmed by the non-existence of other colour or shapes
in the culture. Most probably, too much inoculum was placed initially during the preparation
of the culture. Other than that the colony were left to incubate for more than 24 hours, which
might be the cause of its numerous cells. However, after being left for one more day in the
incubator, the cells seem to have maintained its composition as not much change occurred
to it when observed. More solid knowledge is needed to explain this.
Another issue that should be addressed is the method of counting the cells on the
colony counter. We were unaware of the proper method that should be used to calculate the
number of cells directly on the Petri dishes. The proper method should be to count with the
colonies using a general imaginary grid lines so as not to miss out any colony. The difference
between two or more colonies that have grown into contact with each other should also be
recognized. In the case of TNTC, only a fraction of the Petri dish should be taken into
account and this should be synchronized with all the other cultures so the CFU can be
properly aligned with each other in terms of the determination of the number of cells. This
experiment mainly may involve in parallax and human error.