Food Control 31 (2013) 172e175

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Food Control
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Microbiological quality of packaged water sold in Accra, Ghana

Abena Safoa Osei a, *, Mercy J. Newman b, J.A.A. Mingle b, Patrick F. Ayeh-Kumi b, Mubarak Osei Kwasi c
a

Ghana Standards Authority, Off Madina Road, Near Gulf House, Okponglo, Accra, Ghana
Department of Microbiology, University of Ghana Medical School, Korle-Bu, Ghana
c
Nugouchi Memorial Institute of Medical Research, Legon, Ghana
b

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 10 July 2011
Received in revised form
18 June 2012
Accepted 31 August 2012

Consumers perceive packaged drinking water, usually distributed as bottled or in polyethylene packages
(sachets), as healthier and safer alternatives to tap water. The objective of this study was to determine
the microbiological quality of different types of packaged drinking water available in Accra, Ghana. Sixty
samples of sachet water and ten of (PET) bottled water were randomly purchased from various locations
in Accra. Bacteriological and parasitological analyses of the packaged (sachet and bottled) and tap water
(as control) were done according to standard procedures.
Fifty-two out of 60 sachet water samples (86.7%) had HPC levels well above the internationally recommended limits. For the bottled water, nine out of the ten (90%) were within the recommended limits
for HPC. Two out of the ve (40%) tap water samples (control samples) had HPC above the recommended
limits of 500 cfu/mL. While none of the bottled water samples showed the presence of protozoa, two out
of the ve tap water and 31 out of the 60 sachet water samples had a wide range of protozoa including
rotifers. The ndings clearly indicate that while PET bottled water sold in Accra may be generally safe, the
same cannot be said for the sachet water; since the study found its microbiological quality not signicantly different from tap water.
2012 Elsevier Ltd. All rights reserved.

Keywords:
Packaged water
Bottled
Sachet
Microbiological
HPC
Protozoa

1. Introduction
Water is essential to the sustenance of life and access to safe
drinking water is crucial for human health and well being. Packaged
water is the fastest growing beverage category in the world. For
some consumers it is a healthy alternative to tap water and other
beverages (WHO, 2004). However, several studies have revealed
that the chemical and microbiological qualities of some packaged
water are in violation of acceptable standards, and may not conform
to National Standards as expected by the consumers (Kean, Walker,
Ogutu, Shackleford, & Verghese, 2004). Packaged water has been
implicated as the source of outbreaks of typhoid and cholera in
countries such as Portugal and Spain (Blake et al., 1977; Warburton,
Dodds, Burke, Jonston, & Laffey, 1992). In Ghana, a study on the
microbiological safety of packaged water revealed that heterotrophic bacteria were found in all three types of packaged water e i.e.
bottled water, sealed sachet water and hand-lled hand-tied bagged
water (Obiri-Danso, Okoree Hanson, & Jones, 2003). Heterotrophic
plate count bacteria (HPC) are commonly used to assess the general
microbiological quality of packaged water and water treatment

* Corresponding author. Tel.: 233 277 451675.

E-mail addresses: safoaosei@gmail.com, safoaosei@yahoo.com (A.S. Osei).
0956-7135/$ e see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodcont.2012.08.025

plant operators use it as a quality control tool (WHO, 2004).

According to the WHO Guidelines, nished treated public water for
distribution must not have HPC exceeding 500 cfu/mL. HPC counts
higher than the recommended 500 cfu/mL is an indication that there
is not enough residual disinfectant such as chlorine (WHO
Guidelines for Drinking Water Quality, 2004). International packaged water quality specications recommend HPC limits from 50 to
100 cfu/mL (Robertson & Brooks, 2003, chap. 12). Since packaged
water is often stored at relatively warm temperatures (room
temperatures) for extended periods mostly with no residual disinfectant, studies show that bacteria growth can increase; especially
levels of HPC bacteria including Pseudomonas can increase faster
when the water is stored in plastic/PVC bottles (Ferreira, Morais, & da
Costa, 1994). Indeed, Barrell, Hunter, and Nicholas (2000) noted that
the normal levels of HPC in packaged water according to holding
time and temperature were as follows: 100 cfu/mL when incubated
at 22
C for 72 h and 20 cfu/mL at 37
C for 48 h. Packaged drinking
water can also be contaminated by other microorganisms such as
viruses and protozoa (WHO, 1979). Protozoan parasites which have
been documented to persist or contaminate packaged water include
oocysts of Cryptosporidium parvum and the cysts of Giardia lamblia
(Medema, Schets, Teunis, & Havelaat, 1998). These organisms are
associated with self-limiting diarrhoea but in the immunosuppressed they could cause persistent diarrhoea which leads to severe

A.S. Osei et al. / Food Control 31 (2013) 172e175

173

dehydration and could be fatal (CDC, 1995). Limits for monitoring of

HPC in packaged water depend on the specic country and the
source of the packaged water. The International Bottled Water
Association (IBWA) (2005) recommends that HPC counts should
not exceed the limit of 30 cfu/mL per sample in 100% of the samples
tested at bottling and not more than 200 cfu/mL per sample in 90% of
the samples tested ve days after bottling. Most countries have
established HPC limits for packaged water based on these guidelines. The objective of this study was to determine the microbiological quality of the different types of packaged drinking water
available in Accra, Ghana.

2.4. Parasitological analysis

2. Materials and methods

Table 1
A matrix of the bacteriology and parasitology of sachet water.

2.1. Samples acquisition

Sixty (brands) samples from sixty different companies of sachet
water and ten samples of bottled water were randomly purchased
from various locations in the Accra Metropolis.
In Accra e Ghana, drinking water is packaged in two main
forms: Sachet water and Bottled water. For the sachet water, the
water is packaged in 500 mL plastic sachets. The manufacturers and
wholesalers distribute this packaged in bigger packages. Each Big
pack is a plastic bag containing thirty (30) e of the 500 mL sachets.
For this study a sample of one bag containing thirty sachets
was sampled from each of the sixty manufacturers (brands) and/or
wholesalers visited e this gave sixty samples each sample was a big
bag of thirty 500 mL sachets. To ensure fair sampling, ve
sachets were taken randomly from each big package (bag) of
thirty sachets, and aseptically pooled before inoculation. Three
sachets from the same sample bag were pooled for the parasitological analysis.
For the bottled water, ten samples were taken from ten different
manufacturers. Bottled water in Accra is mainly packaged in boxes
of twenty-four (24) bottles, twenty (20) bottles or packed by dozen.
Three bottles, randomly sampled from each package (a package of
twenty-four, twenty or twelve) were pooled for inoculation.
2.2. Bacteriological analysis of sachet water
Five sachets were randomly selected from each sample bag of
thirty sachets and pooled for analysis. In pooling, each sachet was
vigorously shaken and swabbed using 70% ethanol. Using a pair of
sterile scissors (sterilized by dipping in 96% ethanol and amed
using a Bunsen burner), the sachet was cut open and a fth of the
water (approximately 100 mL) was poured into a sterile sample jar.
The treatment was repeated for the other four sachets to obtain
a nal volume of 500 mL and thoroughly mixed. Using a sterile
pipette, 10 mL of the sample was pipetted into 90 mL of Maximum
Recovery Diluent to give a one-in-ten (10e1) dilution. For each
sample, 1 mL of the diluted sample was plated on Nutrient Agar.
Plates were incubated at 37
C for 24 h. The cultures were done in
duplicates.
2.3. Bacteriological analysis of bottled water
Three bottles of bottled water were randomly taken from each
sample package and pooled for the analysis. Each bottle was
vigorously shaken and disinfected by swabbing, using 70% ethanol.
The bottle was open and a third of the water (approximately
150 mL) was poured into a sterile sample jar. The treatment was
repeated for the other two bottles to obtain a nal volume of
450 mL and thoroughly mixed. Using a sterile pipette, 10 mL of each
sample was pipetted into 90 mL of Maximum Recovery Diluent
(MRD) to give a one-in-ten (10e1) dilution.

Three sachets (of 500 mL each) of water selected at random from

the sample bag were pooled and 50 mL taken into a centrifuge tube.
These samples were centrifuged at 3000 rpm for 20 min. Much of the
supernatant was discarded, leaving about 2 mL of water with the
sediment. Using a micro-pipette the sediment was re-suspended
and transferred to eppendorf tubes and centrifuged again for
another 5 min at 10,000 rpm. The supernatant was discarded leaving
about 0.5 mL of water with the sediment. This was re-suspended and
a drop each placed on two microscope slides e one for wet mount,

Sample
number

HPC on nutrient
agar (cfu/mL)
Plate 1

Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59

6.1 
2.0 
<10
1.9 
3.0 
6.7 
TNTC
1.6 
8.0 
3.8 
TNTC
9.8 
TNTC
2.5 
2.4 
3.7 
2.8 
TNTC
2.5 
TNTC
TNTC
1.8 
TNTC
3.2 
2.5 
6.5 
TNTC
TNTC
1.0 
9.0 
TNTC
TNTC
TNTC
9.0 
2.2 
TNTC
1.7 
2.5 
TNTC
TNTC
TNTC
7.0 
2.1 
TNTC
6.0 
1.4 
1.5 
6.0 
1.7 
TNTC
TNTC
<10
1.8 
TNTC
2.5 
TNTC
TNTC

Plate 2
2

10
103
102
10
102
103
102
102
102
102
103
102
102
103

103
102
103
102

102
10

10
103
103
103

10
102
10
103
103
10
103

103
102

8.1 
2.8 
<10
2.3 
6.0 
8.3 
TNTC
1.7 
1.0 
7.2 
TNTC
1.0 
TNTC
2.4 
1.7 
3.0 
2.5 
TNTC
2.3 
TNTC
TNTC
1.7 
TNTC
2.9 
2.4 
5.3 
TNTC
TNTC
5.0 
5.0 
TNTC
TNTC
TNTC
5.0 
1.8 
TNTC
1.7 
1.4 
TNTC
TNTC
TNTC
2.0 
1.6 
TNTC
6.0 
1.6 
1.5 
4.0 
1.7 
TNTC
TNTC
<10
2.0 
TNTC
4.0 
TNTC
TNTC

102
103
102
10
102
103
103
102
103
102
103
102
102
103

103
102
103
102

10
10

10
103
103
103

10
102
10
103
103
10
103

103
102

Protozoan
organisms
(/)

Debris &
other
particles
(/)

Comments on
water quality









































Not acceptable
Not acceptable
Acceptable
Not acceptable
Acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Acceptable
Not acceptable
Not acceptable
Not acceptable
Acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable
Not acceptable

174

A.S. Osei et al. / Food Control 31 (2013) 172e175

and the other for staining. Cover slips were placed on the slides for
wet mount and observed immediately under the light microscope e
10 and 40 magnications. The slides for staining were rst airdried, and a second drop was added (to make the smear thick) and
again allowed to dry. The dry smear were xed with absolute
methanol and stained using the cold ZiehleNeelsen method.
2.5. Experimental design and data analyses
Based on the relevant sample size calculations, sixty (60)
samples of sachet water, and ten (10) of bottled water samples were
randomly selected from the database of the Ghana Standards
Authority of registered packaged water producers and suppliers.
Tap water from ve different sources was used as control samples;
four from different homes (located at different parts of the city:
Osu, Cantonments, Airport and Legon) and one from the Water
treatment plant at Kpong Water Supplies e treated water just
before distribution. Differences in means of microbial and protozoa
loads among samples of packaged water were determined using
Analysis of Variance (ANOVA).
3. Results
3.1. Bacteriological analyses
The data for bacteriological analyses are presented in Tables 1e
3. Eight out of the sixty samples (i.e. 13.3%) of sachet water had HPC
levels within the recommended limits (i.e. 1  102 cfu/mL). The
remaining fty-two (86.7%) had HPC levels well above the internationally recommended limits (Table 1). For the bottled water
samples, nine out of the ten (i.e. 90%) were within the recommended limit for HPC. Only one (10%) sample had levels above the
recommended limits (Table 2). Two out of the ve tap water
samples analysed as control samples had HPC above the recommended limit of 500 cfu/mL (WHO, 2004). The remaining three
were within the recommended limits (Table 3).
3.2. Results of parasitological analysis
Of the sixty sachet water samples 31 (51.6%) had different kinds
of protozoa present. Eight (13.3%) samples had live Rotifers in them,
Sarcocystis was identied in four (6.6%) samples and Microsporidia
in three (5%). Oocysts of Cryptosporidium were identied in two
(3.3%) samples; Cyclospora were identied in two (3.3%) samples
and another two (3.3%) showed CharcoteLeyden Crystals. One
sample (1.6%) had a small round worm in it (Table 4). Twenty
(33.3%) samples showed different kinds of protozoan organisms
that could not be identied. Observation under the microscope
Table 2
A matrix of the bacteriology and parasitology of bottled water.
Sample
number

Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample
Sample

HPC on nutrient agar (cfu/mL) Protozoan Debris & Comments

organisms other
on water
Plate 2
(/)
particles quality
(/)

Plate 1

23
24
60
61
62
63
64
65
66
70
23

<10
<10
2.0  10
<10
<10
3.6  102
<10
<10
<10
2.0  10
<10

<10
<10
<10
<10
<10
1.1  102
<10
<10
<10
8.0  10
<10

























Acceptable
Acceptable
Acceptable
Acceptable
Acceptable
Not acceptable
Acceptable
Acceptable
Acceptable
Acceptable
Acceptable

Table 3
A matrix of the bacteriology and parasitology of tap water.
Sample
number

Sample
Sample
Sample
Sample
Sample

HPC on nutrient
agar (cfu/mL)

T1
T2
T3
T4
T5

Plate 1

Plate 2

2.3  102
2.0  10
TNTC
TNTC
<10

1.0  102
3.0  10
TNTC
TNTC
<10

Protozoan
organisms
(/)

Debris &
other
particles
(/)

Comments on
water quality










Acceptable
Acceptable
Not acceptable
Not acceptable
Acceptable

Protozoan organisms All organisms identied in the sample.

Debris & other particles Debris, bacteria, yeast cells, and other non-protozoan
structures seen under the microscopic examination.
TNTC Too Numerous To Count.

Table 4
Parasitological organisms identied.
Parasite

Live Rotifers
Sarcocystis
Microsporidia
Cyclospora
Cryptosporidium oocysts
Small round worm
Unidentied protozoa
CharcoteLeyden crystals
Debris and other particles

No. of samples positive

Total/75 (%)

Sachet
water/60

Bottled
water/10

Tap water/5

8
4
3
2
2
1
20
2
39

0
0
0
0
0
0
0
0
0

2
0
0
0
0
0
0
0
0

10
4
3
2
2
1
20
2
39

(16.6)
(6.6)
(5)
(3.3)
(3.3)
(1.6)
(33.3)
(3.3)
(65)

revealed further that 39 samples (65%) had debris in the water.

Thirteen (21.6%) had only debris and no protozoan organisms were
identied in them. Nineteen (31.66%) samples of the sachet water
had no protozoa or debris.
No protozoan organisms or debris were identied in any of the
ten samples of bottled water (Table 2). Two of the ve tap water
samples had Rotifers in them (40%); neither protozoa nor debris
were seen in the other three samples of tap water (Table 3). The
results of the parasitological test are shown in Table 4.
ANOVA of the HPC data showed that there were no signicant
differences between the microbiological quality of sachet water and
tap water.
4. Discussion and conclusions
From the results of the study, out of the sixty samples of sachet
water, 52 (86.7%) had HPC values higher than 100 cfu/mL when
incubated at 37
C for 24 h. Two out of the ve tap water samples
had excessively high HPC levels, and they were the same two
samples that had protozoan organisms in them. Comparing the
results of the tap water samples with those of the sachet water it
seems plausible that the ltration procedures for a good many of
the sachet water production enterprises were inefcient. The
results indicate that either the ltration processes were inefcient
or the tap water may not have been ltered at all. The latter point is
emphasized by the parasitological results where live rotifers were
found in two tap water samples as well as eight (13.3%) sachet
water samples although rotifers are large enough to be removed by
ltration.
Packaged water (specically sachet water) certied by the
National Standard may still have high HPC and protozoan organisms since the National Standard focuses only on the detection of
coliforms. The WHO Guidelines for drinking water allows individual countries to set HPC limits for packaged water. Different
countries therefore assess their packaged drinking water quality

A.S. Osei et al. / Food Control 31 (2013) 172e175

with the HPC limits. A few African countries have reported studies
on packaged water. In a South African study, out of ten bottled
water samples, 8 (80%) showed HPC within the recommended
limits of less than 100 cfu/mL. The study declared South African
bottled water as generally safe (Ehlers, van Zyl, Pavlov, & Muller,
2004). A similar study in Zimbabwe also declared bottled water
processed and bottled in Zimbabwe as safe. The Zimbabwe study
revealed that 4 (6.7%) out of 60 samples of bottled water exceeded
the recommended HPC limits and 7 (11.7%) exceeded the total
coliform limit. The incidence of Staphylococcus aureus was 3.3% that
of Pseudomonas species was 6.7% and Bacillus species, 5% (Okagbue,
Dlamini, Siwela, & Mpofu, 2002). The study concluded that
although locally produced bottled water is generally safe microbiologically, it is necessary to continue with precautionary measures
as any lapse in hygiene may lead to microbial proliferation. In our
study, tap water from four different homes and one sample from
the distribution source were analysed as control samples. Two of
the tap water samples had HPC higher than the recommended
500 cfu/mL. However the sample obtained directly from the Kpong
Water Supply (just before it enters distribution pipelines) had an
HPC level of <10 cfu/mL, which is within the recommended limits.
This suggests that, for the tap water samples that had high HPC
there was either bacterial re-growth or contamination in the
distribution system; this may happen as some bacteria, e.g.
Legionella and Pseudomonas aeruginosa grow in piped water
distribution systems accounting for the increase in HPC levels in tap
water (Exner, Vacata, & Gebel, 2003; WHO, 2004). Elevated HPC
occurs during distribution especially in stagnant parts of piped
distribution systems (WHO, 2003). In addition, microorganisms
such as Legionella, P. aeruginosa, Aeromonas and Mycobacterium
avium will grow in water and on surfaces in contact with water as
biolms (LeChavallier & McFeters, 1985). The persistence of
pathogens is considerably increased if they form a new biolm or
colonize the existing one (WHO, 2003). Microbial re-growth and
the release of microorganisms from the biolms may account for
the high HPC (over 1000 cfu/mL) in two of the tap water samples.
This could mean that two out of ve tap water samples (40%) of tap
water used by sachet water producers for production may be
originally high in HPC with the presence of protozoan organisms. If
such tap water is efciently ltered bacteria and protozoan
organisms should be removed. The high HPC in most of the sachet
water samples produced with tap water as the source water indicates inefcient ltration procedures. Water storage receptacles
were probably another source of the high HPC. Most of the sachet
water producers used overhead PVC water tanks to ensure
a continuous water supply. These tanks could also be the source of
the high HPC and protozoan (planktonic) organisms. From this
study bottled water in Ghana can be said to be generally safe but
there is still the need to monitor HPC levels to ensure the quality
stays acceptable. However, sachet water can not be said to be
generally safe. The high HPC identied in the sachet water indicates
that generally, there is inefcient ltration, as well as unhygienic
production environment and poor plant sanitation. Sachet water,

175

which is an inexpensive form of packaged water in Ghana, and is

often referred to as pure water may not be any more pure than
tap water.
Acknowledgements
Support from the Ghana Standards Authority and the Noguchi
Memorial Institute of Medical Research at Legon, Accra; Assistance
for identication of Parasitological organisms by Mr. Kantanka
Addo-Osafo e Superintendent Technologist, University of Ghana
Medical School.
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