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1.7
Module 1.7 Water Qualities and Testing
Safe Water
Water that is safe can be characterised by the following:
• Pathogen Free
• Low in concentrations of toxic chemicals
• Clear
• Not salty
• Tasteless and colourless
• Non corrosive; does not cause scaling of pipes or staining of clothes
Contaminants
The following are substances that result in poor quality water.
• Inorganic Toxic Compounds
arsenic, mercury, selenium, lead, chromium, cyanide, fluoride, nitrate
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• Organic Toxic Compounds
pesticides, herbicides, dioxins, PCBs
• Turbidity
sediment
• Salinity
Dissolved salts such as chlorides
• Colour, taste, odour
iron, manganese, hydrogen sulfide
• Hardness
Calcium, Magnesium
The World Health Organization (WHO) has produced water quality guideline levels for
use as targets and as an aid for countries who wish to produce their own guidelines. In
many regions, however, the WHO guideline levels may not be achievable in the short
term and, therefore, interim national standards should be set which promote improved
water quality and which are realistic. Setting targets that are too high can be
counterproductive; they may be ignored if they are not attainable. National standards
should reflect national conditions, priorities and capacity to improve water supplies,
especially in small communities where the choice of source and treatment are limited,
and finances are constrained.
The primary objective of water treatment should be the removal of pathogens in the water
to levels that are sub-infectious to the local population. These levels can vary from place
to place, and the question of ‘To what standard should the water be treated?’ is a difficult
one.
To apply the stringent standards recommended by the World Health Organization and by
most governmental regulators may result in:
• Condemnation of acceptable water supplies, thus placing unnecessary stress on
the local population
• Unnecessary expense to bring the water to desired quality, as opposed to investing
in other essential services such as education and health care.
• Ignoring of the rules by the locals, thus bringing similar regulations into disrepute
Cairncross and Feachem emphasize that it is necessary to use a great deal of common
sense in the use and interpretation of bacteriological water quality standards. These
standards or goals must be set realistically, and national authorities must decide
themselves what are reasonable levels to aim for, given the particular environmental and
economic circumstances of the country.
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Water quality testing alone is an inadequate response to the challenges of ensuring water
safety. Occasional tests conducted on a water supply may provide a false sense of
security as water quality can vary widely and rapidly. For these reasons water quality
testing should be accompanied by verification of the state of the source or supply, for
instance, by sanitary inspection as described in the WHO Guidelines for Drinking-water
Quality Volume 3. End-product testing is now only one of several key management
tools in the provision of safe drinking water under the forthcoming Guidelines for
Drinking Water Quality. The development, implementation and use of a Water Safety
Plan that considers the quality and overall management of drinking water from source to
consumer is the goal for all water supplies. In such a plan, now including hazard analysis-
critical control points (HACCP), end product testing is not a critical control point (CCP).
This is a recent paper on the WHO web site that describes risk assessment of water
supply. It is one of the documents that will go into the new WHO water quality
guidelines
http://whqlibdoc.who.int/publications/2001/924154533X_chap9.pdf (Apr 05)
Bacteria Testing
Testing for every conceivable pathogen in water would be both time consuming and
expensive. Instead, the presence or absence of certain indicator organisms is used to
determine the biologic safety of the water. Indicator tests are cheaper, easier to perform
and yield faster results, compared to direct pathogen monitoring. These indicator
organisms possess the following characteristics:
Commonly used indicators are total coliforms, E.coli, and H2S bacteria, which are all
fecal indicators. Thermo-tolerant (i.e. faecal) coliform bacteria (particularly Escherichia
coli) are a sub group of the total coliform group which are produced in the gastro
intestinal tract of warm blooded animals and occur almost entirely in faeces. Among the
coliforms in human faeces, 96.4% are faecal coliforms.
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Contamination Indicators
E.coli (preferred indicator choice) or thermo tolerant coliforms (also called faecal
coliform, acceptable alternative to E.coli indicator) are used as indicators of faecal
pollution. If E.coli is present then it is likely that pathogens are also present. The WHO
recommends that all water intended for drinking should have non-detectable faecal
pollution in any 100 ml sample.
Alternative figures in the following table are often quoted which may be more
appropriate for rural communities:
The previous version (2nd Edition) has Volumes 1, 2 and 3. They are each about 250
pages long! Good bed time reading.
http://www.who.int/water_sanitation_health/dwq/guidelines4/en/ (Apr 05)
Canadian water quality standards summary table (Guidelines booklet must be purchased)
http://www.hc-sc.gc.ca/hecs-sesc/water/pdf/summary.pdf (Apr 05)
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Bacteria Testing
There are three main testing techniques for water analysis of bacteria - Membrane
Filtration, Most probablye Number and Presense/absence. The following are typical
prodecures that are used for these techniques.
Procedure:
1. Disposable Whirlpack Bags are used for sample collection, as shown in Figure 1.
Each new bag is pre-sterilized and sealed. During sampling, a bag is unsealed,
filled up with sample water, and closed. The bag should be refrigerated (i.e.
placed in a cooler) until analysis. Analysis must be performed as soon as
possible, ideally within 2 hours.
3.
Cup where
Figure 1. Collect samples the a Whirlpack bag.
using
sample is poured
Top part:
sterilized
between each
sample
The filter paper is under the cup
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5. Steps five to seven outline the procedure to test for one water sample.
6. Remove the lid from a Petri dish. Place an absorbent pad inside the dish using a
steralized forcep. Pour 2 mL of the media broth evenly over the absorbent pad.
Depending on the type of analysis required, the type of media broth differs. For
thermo-tolerant coliform (i.e. faecal), “m-FC” broth is frequently used. For
E.coli, “Coliblue-24” broth can be used. For total coliform, “m-Endo” is
commonly used. Broths can be purchased as pre-packaged ampoule or the broths
can be prepared from dry powder.
7. Setup the membrane filtration unit. One filter paper, grid side up, is placed into
the assembly using sterile forceps. Pour a pre-measured amount (usually 100mL)
of the water sample into the funnel. Apply a vacuum below the filter using the
pump-syringe attached to the assembly in order to force the water through the
filter paper.
8. Transfer the filter paper, grid side up, on the absorbent pad in the previously
prepared Petri dish using sterile forceps. A slight rolling motion was applied
during the transfer to avoid air to be trapped in between the pad and the filter.
Cover the petri dish with its lid.
9. Invert the petri dish (upside down) and place the petri dish in the incubator.
Incubate at the required temperature for the required time.
10. Remove the Petri dishes from the incubator and count the colonies, using the grid
of the filter to avoid double-counting or missing some colonies. A magnifying
glass and a white fluorescent light source can be very helpful.
11. Report the coliform density as number of colony forming units (CFU) per 100 mL
of sample. Preferably, each plate should have between 20 and 80 colonies for
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easy and accurate counting. Samples producing more than 250 colonies can be
reported as “too numerous to count” (TNTC). Some colonies may overlap thus
creating counting errors. Dilutions of water samples can be made to avoid this.
Dilution water must contain zero coliform and other sources of interference/
contamination. Ideally, non-chlorinated water that has been boiled and cooled
down to room temperature can be used as the dilution water.
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No preparation of petri dishes is required. These kits usually come with prepared media
(M-Endo Broth), Petri Dishes (with Absorbent Pads/Filters), Petri Dish Monitors
(funnels), and Petri Dish Bottom Caps.
Filtration
• Prepare filtration apparatus.
• Lock in a pre-sterilized petri dish with monitor in position. Do not touch the
inside of the petri dish to avoid cross-contamination.
• Mix the water sample thoroughly to obtain a uniform composition. Samples
should be 20 ml or larger. Pour samples directly into the funnel (monitor).
• Apply vacuum to the sample. Hold the lower body of the funnel and swirl liquid
while filtering to ensure uniform mixing and distribution. If sample size is 100 ml,
swirling action is not required.
• Rinse the funnel walls with at least 30 ml of boiled, dechlorinated water. (To
dechlorinate water, use tap water and leave exposed to atmosphere for 48 hours.
Boil for 10 minutes.) Draw rinse buffer through filter. Repeat for a total of three
rinses.
• Turn off the vacuum and lift off the petri dish with funnel. Remove funnel from
the dish. Dispense a 2 ml M-Endo Broth into the dish.
• Reapply vacuum to draw off excess liquid.
• Replace the cover tightly, and then cap the bottom.
Incubation
• Place the petri dish in the incubator and incubate at the required temperature for
the required time.
Total Coliform bacteria - m-Endo 35 oC ± 0.5 oC for 24 hours.
Fecal Coliform bacteria - m-FC 44.5 oC ± 0.2 oC for 24 hours.
• The bacteria will grow and individual colonies can be counted after 24 hours.
Colony counting
• After the required incubation period, take out the dish and remove the cover.
Place the dish in the stereoscope microscope with a daylight white fluorescent
light source. Examine and count all colonies with a golden-green sheen
appearance. (Colors may be different depending on media used)
• Colonies shown in every grid square are to be counted. To facilitate this counting
procedure, proceed from top to bottom and left to right until all grid squares are
covered.
Indicator organism levels in water samples are expressed as the number per 100 ml. The
following relation can be used to enumerate bacterial density:
M = A/B x 100
Where:
M = number of micro organisms/100 ml.
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A = number of bacterial colonies counted.
B = volume of sample filtered, ml.
Presence/Absence Test
The hydrogen sulphide test (H2S) paper strip bacteriological test is used to detect the
presence of certain micro organisms in the water.
This experiment is a test for H2S. In order to check for the presence of these "sentinel" or
"indicator" bacteria in water, we put them in contact with a strip of paper that has been
impregnated with a nutritive substance (food for bacteria) plus a colour indicator that
turns black when it comes in contact with hydrogen sulphide. If the test paper turns black,
it means that H2S was produced, which in turn means that bacteria of faecal origin are
likely present in the water sample.
Arsenic Testing
Arsenic is a toxic substance. Long-term ingestion of arsenic contaminated water can
cause melanosis (pigmentation on skin), keratosis (hardening skin bulges on palms and
feet), cancer of lungs, bladder, kidney, skin, and liver, as well as vascular and
neurological diseases.
WHO set a guideline value of 10 ug/L (equivalent to 10 parts per billion). Many
industrialized countries have also adopted 10 ug/L as their national standard. For
developing countries, most have a national standard of 50 ug/L, partly due to economic
considerations and the lack of tools and technique to measure accurately at such low
concentration. Refer to Table 2 for the current national standard for arsenic for various
countries.
Table 2. National Standards for Selected Countries for Arsenic in Drinking Water
(source: Towards a More Effective Operational Response – Arsenic Contamination of Groundwater in
South and East Asian Countries, Volume I, Policy Report. Page 31. The World Bank. March 2005.)
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For most of the test kits, the arsenic measurement is based on the Gutzeit procedure. In
this procedure, inorganic arsenic is reduced to As(III) and then converted to arsenic gas.
Various chemicals may be added to facilitate the reduction of arsenic to As(III), as well as
to remove interferences. Then the arsenic gas reacts with a paper strip impregnated with
mercuric bromide and produces a yellow to brown stain. The amount of arsenic present
in the water is directly related to the intensity of the colour. The colour developed on the
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paper strip can be compared to a standard colour chart to determine the arsenic
concentration in the sample water. Figure 6 shows an arsenic test kit developed by
Industrial Test Systems, Inc. of USA. Figure 7 shows a colour chart taken from an
arsenic test kit developed by Environment and Public Health Organization of Nepal.
Figure 7. A colour chart corresponding the yellow-brown colour intensity to the arsenic
concentration. Taken from an arsenic test kit developed by Environment and Public
Health Organization of Nepal.
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Qualitative identification involves our senses of smell, taste and sight and includes the
following properties:
• Colour, clarity, smell, taste, scaling, staining, sediment, corrosion
The pH of the water in a stream, river, lake or underground flow will vary depending on a
number of conditions: the source of the water; the type of soil, bedrock and vegetation
through which it travels; the types of contaminants the water encounters in its path; and
even the amount of mixing and aeration due to turbulence in its flow. The effects of a
specific type of water pollution on living plants and animals can vary greatly depending
on the pH.
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Colour
Apparent colour is a measure of the optical effects of both dissolved and suspended
matter. It is measured relative to a set of standard samples (the Hazen scale). Humus and
peat materials carried by many rivers can give their waters a characteristic brown colour.
Color and odor have no direct chemical significance but indicate quality or pollution.
Dark brown or yellowish water usually contains tannins and other vegetable chemicals,
sediments usually cause reddish colors. True color is due to dissolved materials and
apparent color is due to turbidity or suspended particles. Filtration is used to
distinguish between the two colors. Pure water does not have odor or taste.
Turbidity
Turbidity is a measure of the light-scattering properties of water and hence an index of
the amount of suspended particles. The current standard for potable (drinking) water is a
maximum turbidity of 5 ‘nephelometric turbidity units’ (NTU), though less than 1 NTU is
recommended if the water is to be efficiently treated.
Turbidity Tube
The turbidity tube is a simple and easy way to estimate water clarity.
1. Using the knife-cut the 8-foot fluorescent light sleeve into three equal lengths (32
inches).
2. Insert the 1 9/16 to 1 5/8-inch white Plexiglas disc into one end and seal with
Plexiglas sealant. If disc has a center hole, plug it with sealant. (Note: this will
likely have to be treated with sealant more than once to fill all spaces. An easy
way to check to see if more sealant is necessary, is to blow into the tube at the
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opposite end of the disc and feel if air escapes near the end with the disc inserted
into it.)
3. Using the black marker or electrical tape (and razor blade to cut edges smooth),
color half of the white Plexiglas disc or color two opposite quadrants black,
similar to a secchi disc.
4. Drop the white and black disc (target) into the tube.
5. Starting from the top of the target draw a line around the tube, leaving a space
(gap) in the circular line for a label.
Place lines at the heights above the target as shown in the following table:
Line Distance above target (inches) Turbidity Units (roughly NTUs)
1 2.875 200
2 4.5 100
3 7.5 50
4 12.25 20
5 17 15
6 20.75 10
Note that turbidity unit labels are not always equally spaced, therefore if using this
method you cannot estimate NTUs between lines on the turbidity tube.
These directions are based on information from Jim Peterson, UWEX Environmental
Resources Center, UW-Madison.
http://clean-water.uwex.edu/wav/monitoring/turbidity/tubedirections.htm (Apr 05)
This web site also describes how to build and use a turbidity tube.
http://www.environmentmonitor.green.net.au/water/turbid.htm (Apr 05)
Heavy metals
Heavy metal pollution is typically associated with mining activities or discharges from
some industries. Persistent toxicants (heavy metals) in water and sediments affected by
heavy metal pollution can have serious affects on the aquatic ecosystem and can make
water unsuitable for human consumption. Some animals can also ‘bioaccumulate’ metals,
making them unsafe to eat.
Other Testing
Levels of certain compounds and their impacts on water quality are indicated by the
biochemical oxygen demand (BOD) of the water. A high BOD is often accompanied
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by a low DO level.
To determine the content and amount of other contaminants in water, more sophisticated
water testing is required. This test usually can not be performed in a field setting.
Microbial Samples
• Refer to the earlier section on Membrane Filtration Technique
When sampling, make sure that your hands do not touch the lip of the bottle. You should
also pay special care and attention to handling the bottle cap to prevent contamination.
Ask the laboratory to explain the proper way to place the water sample into the bottle.
Use an ice chest to keep the water samples below stream temperature during
transportation. Try to get the samples to the lab within 24 hours. In general, the shorter
the time between collection of a sample and its analysis, the more reliable the results.
Failure to follow proper field handling protocol and laboratory techniques can be a source
of error. Make a record of every sample collected and identify every bottle, preferably by
attaching an appropriately inscribed tag or label. Record sufficient information to provide
positive sample identification at a later date. Include the name of the collector, date, hour,
exact location, water temperature, and any other data that may be needed for correlation,
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Resources
The following is a list of companies that supply water testing equipment and materials.
Many of them have good web sites and will provide you with significant information if
requested. Remember that good testing equipment is very expensive and it takes a
considerable amount of skill to do tests successfully.
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The following site is a short article from the mid 1990’s about low cost methods for
detecting contaminants in water supplies. These methods were developed by the
Canadian International Development Research Council
http://web.idrc.ca/es/ev-5527-201-1-DO_TOPIC.html (Apr 05)
The following is a web site that is set up as a school water testing program that can be
done by students. It may be very useful for teaching water quality.
http://www.aquatox.net/index.cfm?op=ShowNews (Apr 05)
The following is a publication of the WHO entitled “Evaluation of the H2S Method for
Detection of Fecal Contamination of Drinking Water”, Geneva, WHO, 2002.
http://www.who.int/water_sanitation_health/dwq/wsh0208/en/ (Apr 05)
This is an excellent guide to learn more on water samples, and analysis. It include a
description of the contaminant, health impacts, and treatment options.
http://wilkes.edu/~eqc/helpguide.htm (Apr 05)
This site shows the Colilert test system. It used a combination of Presence/Absence tests
and Most Probable Number Tests for E. Coli and Coliform.
http://www.idexx.com/water/products/Colilert/ (Apr 05)
This site shows the Coliplate system which is a Most Probable Number test. Each plate
costs about $ 8 CDN.
http://www.bluewaterbiosciences.com/coliplate.asp (Apr 05)
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