Generally, the term “coliforms” or “coliform colonies” is an inclusive one which is including

the bacteria such as Escherichia coli (E. coli), Escherichia freundii (E. freundii), Enterobacter
aerogenes (E. aerogenes) and biochemical intermediates between the genera Escherichia and
Enterobacte (“Bacteriological Methods in Water Quality in Water Quality Control Programs”
, 1978). It is includes all of the aerobic, Gram-negative, nonspore-forming rod-shaped bacteria
which is ferment lactose with gas formation within 48 hours at 35°C as defined in “Standard
Methods for the Examination of Water and Wastewater” (14th ed) (“Bacteriological Methods
in Water Quality in Water Quality Control Programs” , 1978). There are few tests for
compliance with bacterial water quality standards such as potability tests on drinking water and
determination of bacterial quality of environmental water (“Bacteriological Methods in Water
Quality in Water Quality Control Programs”, 1978).

The most common and widespread danger associated with drinking water is
contamination, either directly or indirectly, by sewage, other wastes or human and animal
excrement. Ideally, drinking water should not contain any microorganisms known to be
pathogenic. In this experiment, sample from reverse osmosis and filtered water are tested to
determine the presence of coliform bacteria in a water sample. Reverse osmosis is a technology
that is used to remove a large majority of contaminants from water by pushing the water under
pressure through a semi-permeable membrane. Filtered water is the water that have a reduced
number of microorganisms (Ahmed, Kanwal, Tahir & Rauf,2004). Based on the experiment
conducted earlier, there are no production of gas in presumptive test, this indicated that filtered
water and reverse osmosis water is free from contamination. Based on result, filtered water and
reverse osmosis water is claimed to be potable water. This was shown by MPN which showed
the possible number of organisms present in the sample under analysis was none found in the
MPN table. Some examples of industries that use reverse osmosis water include
pharmaceutical, boiler feed water, food and beverage, metal finishing and semiconductor
manufacturing.

Next, some of the coliform bacteria are also found in the polluted water such as ponds.
However, the green pond has the highest presence levels of coliform bacteria compared to the
light pond as it contains the wastage of animal and human wastes or surface runoffs from land
especially during the rainy season. The Most Probable Number (MPN) technique is used to
estimate the total of fecal coliform bacteria which is E. coli. As a result, the green pond are all
positive, where there are the production of acid and gas in the five Durham tubes indicates the
presence of E. coli in the green pond water. Meanwhile, the light pond only showed positive
results for samples in 50mL of LST broth and 3 from 10mL of LST broth. The number of
organisms present are determined by using the MPN table which are 1, 5, 5 (green pond water)
and 1,3,0 (light pond water). This statement states that more than 23 MPN/100mL and 8
MPN/100mL of green pond and light pond sample. The test is then continued with Brilliant
Green Lactose Bile Broth (BGLB) test. BGLB is used to confirm the total coliforms (Bartram
& Balance, 2007). The results for both two samples are positive and more turbid, even though
one is stored at 37.0°C and the others are at 44.5°C. During Indole test, the Tryptone Broth is
dropped with few drops of Kovacs’ Indole reagent, resulting a formation of pink violet layer
on the top of green pond while in the light pond, there is no formation of pink violet layer
(“Indole Test Reagents”, 2017). The streaking technique is applied on MacConkey Agar to
observe the characteristics of the E. coli presence and incubate at 37.0°C overnight (Kumar, D.
et al, 2013). As a result, the positive characteristics result are obtained from the green pond
water sample which is red, circular, convex, smooth surface, dry and have entire edge.
However, the light pond gave a result of circular, pink, moist and sticky and it confirmed that
the light pond is absence E. coli but presence Klebsiella spp.. MacConkey Agar is a selective
and well isolated colonies of lactose-fermenting bacteria that will appear pink to red in color
and surround by a zone of bile salt precipitation (MacConkey, 2017). In conclusion, both pond
water samples are non-potable water as they are unsafe to drink due to the presence of coliform
bacteria which is E. coli, even though the light pond is absence of E. coli.