Professional Documents
Culture Documents
Food Control
journal homepage: www.elsevier.com/locate/foodcont
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
173
Table 1
A matrix of the bacteriology and parasitology of sachet water.
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
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
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
Table 4
Parasitological organisms identied.
Parasite
Live Rotifers
Sarcocystis
Microsporidia
Cyclospora
Cryptosporidium oocysts
Small round worm
Unidentied protozoa
CharcoteLeyden crystals
Debris and other particles
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)
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