Microbiology – Water Analysis

Chapter 4 – Part 3: Water Analysis

INTRODUCTION ...................................................................................................................................................................

TOTAL COLIFORMS ..............................................................................................................................................................

E. COLI .................................................................................................................................................................................

HPC/TBC/TPC.......................................................................................................................................................................

SAMPLE SUBMISSION ..........................................................................................................................................................

 Microbiology – Water Analysis

Water Analyses
Introduction

There are three main types of microorganisms that can be found in drinking water: bacteria, viruses and
protozoa. These can exist naturally or can occur as a result of contamination from human or animal
waste. Some of these microorganisms are capable of causing illness in humans or livestock. Surface
waters such as rivers, lakes/dams and reservoirs are more likely to contain microorganisms than
groundwater sources unless the groundwater sources are under the direct influence of surface water or
have become otherwise contaminated.

The aim of drinking water treatment is to remove or kill these organisms to reduce the risk of illness in
the consumer. This is achieved by using a multi-barrier approach including the protection of the water
source (where possible), appropriate and effective treatment methods, well maintained distribution
systems and routine verification of the water quality.

All drinking water supplies should be filtered and or disinfected unless proven be safe all of the time.
Even though the water supply source may be clean contamination of the supply may occur at any point
between source and end-point eg breaks in pipelines, uncovered or unsealed reservoirs or problems with
header tanks etc. Sampling points therefore need to take this into consideration. A standard procedure
therefore would be to collect samples from source/s, reservoirs or holding tanks, header tank and end-
point. The results from this would then show if the entire system was at an acceptable level or indicate if
a problem was evident at a particular point.

The processes used to produce potable water are not necessarily used to produce bacteria- free water but
are rather concerned with removing microorganisms that are a potential health threat and making water
aesthetically pleasing.

There are two practical and fundamental measurements used to assess the bacterial quality and safety of
drinking water for livestock (or human) health namely the detection of total coliforms and E. coli.

Total coliforms:
The total coliform group is comprised of various bacterial genera with similar growth,
morphological and biochemical characteristics. They range from being specifically faecal
associated such as E. coli to being widely distributed in water, soil and vegetation. Many total
coliforms are not specific to any one source and are present in both faecal and non-faecal
environments.

The presence of total coliforms in a water distribution system as opposed to the natural
environment results from inadequately treated source water and indicates water treatment failure.

E. coli:
E. coli is a member of the total coliform group of bacteria and is the only member that is found
exclusively in the faeces of humans and animals. Its presence in water indicates not only recent
faecal contamination but also the possible presence of other intestinal disease-causing microbes.
 Microbiology – Water Analysis

A further measurement of water quality involves the Total Plate Count (TPC) or Heterotrophic Plate
Count (HPC) or Total Bacterial Count (TBC) in a water sample. This test is principally used to indicate
the effectiveness of water treatment processes such as filtration and disinfection (ie an indirect indication
of pathogen removal).

HPC (or TPC or TBC):

The HPC value alone does not directly relate to any health risk but does indicate if there are
significant changes in a system over time or a deterioration in water quality before coliform
bacteria are detected. A rising plate count may give the earliest signs of pollution in eg boreholes.

The sample for HPC may be incubated at 22° C or 37° C or both and for 24 or 48 hours. The
results at both temperatures may indicate the risk of the presence of opportunistic pathogens
whereas the warmer incubation temperature affords more information as to potentially dangerous
pollution as the organisms developing at this temperature are chiefly those of soil, sewage or
intestinal origin.

Within water distribution systems we may have the occurrence of Biofilms which represent a
specific form of bacterial colonization. The persistence of potential pathogens is considerably
increased if they form a new biofilm or colonise an existing one. The potential for biofilm
formation and growth is particularly high in narrow-gauge plumbing stystems and the colony count
is directly correlated with the water volume flowing through the system. Nutrients supplied via
drinking water systems for livestock may aid in the rapid development of biofilms.

From a practical point of view it is thus not the absolute numbers of colony count bacteria
enumerated from a supply that are of importance but whether there are significant changes in those
numbers over time. One is thus looking for no abnormal change over time in the count at either of
the incubation temperatures.

Hence this test is most useful for indicating the improvements brought about by changes in the
filtration and or disinfection systems used or in the efficacy of reservoir, tank or pipe-line flushing.
Tests on water samples collected before and after any changes would reflect the changes in water
quality achieved.

In summary water may be tested for the following:

a) Standard Microbiological Safety Requirements: (total coliforms and faecal E.coli).

Both values should be zero cfu per 100 ml sample.

b) Heterotrophic Plate Count (HPC) or Total Plate Count (TPC): principally used to indicate the
effectiveness of water treatment processes such as filtration and disinfection.
Values are acceptable up to about 100 cfu/100 ml.

b) Physical, organoleptic and chemical requirements (aesthetic, macro and micro chemical and mineral
levels, heavy metals.

c) Protozoan parasites (eg Giardia, Cryptosporidium).

 Microbiology – Water Analysis

Sample submission

Figure 11 Figure 12

 Specific sterile, plastic bottles designed for water analysis should be pre-ordered from the laboratory
prior to the collection of samples.
 If sampling from a tap let the water run for a few seconds before collecting the sample and replace
the cap immediately.
 Bottles must be filled to the top.
 Each container must be clearly labeled as to where the sample was collected and on what date. This
information should be written on the body of the bottle with a waterproof permanent ink marker.
Avoid only labeling the lid as the lid and bottle could become separated during analysis, placing the
test results under question.
 Samples should be transported to the laboratory at 4°C in leak proof sealed containers. Ensure that
the submission form is sealed in the plastic sample envelope to avoid leakage or the smudging of any
details completed on the submission form.

Further Reading

1. Plumblee K H. 2004. Clinical Veterinary Toxicology. Mosby, St Louis.

2. SANS 241

3. Guidelines for Canadian Drinking Water Quality. 2006.

4. WHO: 2003. Heterotrophic Plate Counts and Drinking Water Safety.