University of Zimbabwe

Department of Civil Engineering

BSc (Honours) Civil Engineering

Laboratory reports Submission Form

SUBJECT/MODULE: WATER AND PUBLIC HEALTH

LEVEL: 3.2

LAB/EXPERIMENT NAME: COLIFORMS

DATE OF LAB: SEPTEMBER 2019

STUDENT NAME: CHIHANYA TEDDY

REG.NO: R165465W

GROUP: B

LECTURER: DR MISI

RESPONSIBLE LAB.TECH: MR KATIVHU

DATE OF SUBMISSION: 19 September 2019

BACKGROUND

Total Coliforms

The total coliform bacteria are a group of microbes which include a wide variety of aerobic and
facultative, Gram-negative, non-spore-forming bacilli that is able to mature in the presence of
comparatively high concentration of bile salts with the fermentation of lactose and production of
acid or aldehyde within 24 hours at 35-37℃ (Singh and Hussain, 2016; WHO, 2011).
Thermotolerant coliforms and faecal coliforms and some naturally occurring bacteria are a subset
of total coliform bacteria. The presence of total coliforms may or may not necessarily indicate
faecal pollution since it occurs both in faecal matter and natural water and in some cases a high
count of total coliforms may be associated with a low count for thermotolerant coliform. WHO
(2011) suggested that this might be caused by water having a recent contact with some soil or
organic matter which are not necessarily of faecal origin or by nutrient-rich environments
promoting the growth of other types of coliform bacteria.

Even though total coliforms are not quite useful as a faecal contamination indicator, various
researchers have used total coliforms together with faecal coliforms to determine faecal
contamination of groundwater (Pujari et al., 2011; Zingoni et al., 2005; Banerjee, 2011).

Faecal Coliforms

Faecal coliforms are coliform organisms which are capable of fermenting lactose at 44-44.5℃
producing an acid and gas (WHO, 2011). In a guide on water quality studies, Bartram and Pedley
(1996) highlighted that the presence of faecal coliforms nearly nearly always indicates faecal
contamination with more than 95% of faecal coliforms secluded from contaminated water are
E.Coli bacteria. The presence of E.Coli bacteria is a conclusive evidence of recent faecal
contamination hence it is often needless to perform tests to confirm the specific presence of
E.Coli in contaminated water. However, Lawrence et al., (2001) and Lewis et al., (1982)
cautioned that the values of faecal coliform counts in tropical waters should be interpreted with
care since a significant percentage of coliforms of non-faecal origin can ferment lactose at
44.5℃.

Escherichia Coli (E.Coli)

Escherichia coli is present in very large numbers in human and animal faeces, sewage and water
that has had recent contact with faecal matter and is rarely found in the absence of faecal
pollution. The E.coli bacteria is a thermotolerant bacteria and can be distinguished from other
thermotolerant coliforms by the ability to generate indole from tryptophan or by the production
of the enzyme b-glucuronidase (WHO, 2011). A positive E.coli test gives a better confirmation of
recent faecal contamination of water.

The World Health Organization recommends that water must be tested for presence of coliforms
and for the water to be safe to drink it must be free of coliforms. There are several methods used
to conduct this test, the common method used is to pass 100ml of water sample through a filter
membrane capable of capturing the bacteria. The filter is then placed in a petri dish with agar to
grow the bacteria overnight. If there is bacteria, they appear as colonies on the filter. The other
method rely on colour change if there is bacteria in the water sample.

TITLE

Coliforms Test

OBJECTIVE
To determine the presence of fecal colonies and total coliforms in the given water samples.

APPARATUS

1. Electric vacuum
2. Filter support
3. Glass or plastic petri dishes
4. Membrane filters
5. Nutrient absorbent pads
6. Forceps
7. Magnifying lens
8. Samples - Borehole, Tap, River

METHOD

1. The pad was placed into a petri dish using the forceps and 2mls of broth were added to
the pad using a pipette and the Petri dish was closed using a lid.
2. 100mls of water sample was poured into a beaker and then into a filter unit for filtration,
the sample was filtered through a membrane to capture bacteria
3. In order to improve efficiency of the filter unit, a vacuum pump was connected thus
increasing the rate of filtration.
4. When the filtration process was over, the membrane was then placed on a pad soaked
with broth, the lid was replaced, and the Petri dish was put in a rack ready to incubate.
5. The procedure was repeated for the same sample for total coliforms testing.
6. The two Petri dishes were then placed in an incubator and were left to incubate for
approximately 16hours.
7. After the 16hours, the colonies with yellow colour and diameter 1mm approximately
were counted and recorded as colony forming units (cfu) per 100ml of sample.
8. Where no yellow colonies were present, the results were reported as 0cfu per 100ml.
9. The results were tabulated as shown

RESULTS

Sample Borehole water Tap water Deep Well Shallow well River
water
Total coliforms 0 0 144 400 TNTC
Fecal coliforms 0 0 60 * TNTC

CONCLUSIONS
World Health Organization recommends that there should be no coliforms in water. . Total
coliforms do not pose risks to human health. They are used to indicate whether other potentially
harmful bacteria may be present. Fecal coliform and E. coli are bacteria, that when present,
indicate that water may be contaminated by human or animal wastes. The presence of total
coliforms in the well sample make the water contaminated and thus unacceptable for drinking.
Still water and mining borehole are safe for drinking since they do not contain both total and
faecal coliforms.

RECOMMENDATIONS

1. Wash hands with alcohol before handling the apparatus

2. Apparatus must be sterilized before use.

REFERENCES

1. SAZS 560: 1997

2. WHO Guideline for drinking-water quality, fourth edition
3. Lawrence, A.R., Macdonald, D.M.J., Howard, A.G., Barret, M.H., Pedley, S., Ahmed,
K.M., Nalubega, M., 2001. Guidelines for Assessing the Risk to Groundwater from On-
Site Sanitation, Nottingham: British Geological Survey