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Microbiological quality of groundwater sources used by rural communities in

Limpopo Province, South Africa

Article in Water Science & Technology · February 2006

DOI: 10.2166/wst.2006.890 · Source: PubMed

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 THE MICROBIOLOGICAL QUALITY OF PRIVATE AND
COMMUNAL BOREHOLES IN THE TSHITALE-HLANGANANI
REGION OF THE LIMPOPO PROVINCE, SOUTH AFRICA
Potgieter N*, Mudau LS and Maluleke FRS
*
Dept of Microbiology, University of Venda for Science and Technology, Private Bag
X5050, Thohoyandou, 0950, Limpopo Province, Republic of South Africa.
nataspo@yebo.co.za

ABSTRACT

A cross-sectional study was carried out to assess the microbiological quality of 97 private
and 97 communal boreholes in the rural Thitale-Hlanganani area of the Limpopo Province,
South Africa. Both bacterial and viral indicator microorganisms were used and included
total coliform bacteria, faecal coliform bacteria, faecal streptococci bacteria, heterotrophic
bacteria, Clostridium perfringens (vegetative cells and spores) and somatic
bacteriophages. The South African criteria guidelines of good (negligible risk of microbial
infection; fit for human consumption), marginal (slight risk of microbial infection; must be
treated before consumption), and poor (risk of infectious disease transmission; not fit for
human consumption) for water used for human consumption, with the relevant counts for
each indicator organism, was used to group the private and communal boreholes
according to the relevant indicator organism. Results indicated that although the majority
of boreholes were placed into the good category, some boreholes did however fall into the
marginal and poor categories for each indictor organism. This indicated the potential
health risk present to the consumers using these boreholes for domestic water sources.
Observations at the boreholes identified various potential sources of pollutions that could
affect the microbiological quality of the borehole water. In conclusion, this study indicated
the need for more intense monitoring of privately owned boreholes and the education of the
rural communities on the installation and the maintenance of both private and communal
boreholes.

INTRODUCTION

The World Health Organization (WHO) estimate that 1,5 billion people in the world do not
have access to safe water and that an estimated 1,8 million people in developing
countries, die every year from diseases associated with unsafe water and inadequate
sanitation (1; 2; 3). South Africa (SA) is a developing country and was rated number 26
worldwide during 1997 in terms of water availability per person out of 149 countries (4).

The backlog of water and sanitation in rural villages in SA is addressed through

Reconstruction and Development (RDP) programs on water and sanitation, using
intersectoral approaches within communities (5; 6; 7). The sole and most cost effective
source of potable water supply in SA is groundwater (8). Ground water has the potential
of serving communities in areas where water infra-structure does not exist and where
water delivery is difficult due to arid conditions (9). It is estimated that almost two thirds of
the rural population in South Africa depends on groundwater sources for domestic
purposes (9). The majority of rural communities use groundwater without any treatment
(10). In addition, the number of on-site sanitation systems in the villages makes the
underground water aquifers vulnerable to contamination and the microbiological quality of
borehole water is not always guaranteed to be microbiologically safe for human
consumption (11). Consequently, the presence of viruses, bacteria, protozoa and
helminthes can cause diarrhoeal diseases. In addition, the presence of certain chemicals
such as iron, copper, zinc, cobalt, magnesium, selenium and chromium in ground water
can also be detrimental to human health (10). Therefore, the quality of borehole water for
domestic use needs to be monitored and the health risk associated with contaminated
drinking water should be considered, hence the need for this study.

MATERIALS AND METHODS

Study site and water sampling procedures

The study was carried out in the Tshitale–Hlanganani area, in the Vhembe District
Municipality of the Limpopo Province of South Africa. This area is predominantly rural with
a low social economic income (12). Groundwater is used as a major source of water
supply (13). The majority of borehole pumps used in the area are either engine or
electricity driven and a few pumps are operated manually. Only a few of the electric and
engine driven pump the underground water directly to communal stand pipes. With the
majority of the boreholes, the electric and engine driven pumps distributes the
groundwater via storage tanks and reservoirs to the stand pipes (13).

A simple random selection method was used to select 30 of 40 villages in the study area.
In these 30 villages, a total of 97 communal boreholes and 97 privately owned boreholes
were identified to be assessed in this study. Water samples were collected monthly from
both private and communal boreholes between August 2002 and August 2003 to include
dry and rainy seasons. Each borehole water sample (2 l) was taken using the standard
collection technique as specified by SABS (14), and transported on ice to the laboratory.
In locations where hand water pumps were used, the water samples were taken directly
from the hand pump. In locations where the borehole was connected to a tap or
distributed via storage tanks or reservoir, water samples were collected from the nearest
tap from the borehole (14; 15).

Detection of indicator organisms

The membrane filtration technique (16) was used and all test were performed in duplicate.
A volume of 100 ml of each water sample was passed through 0.45µm pore size, 47 mm
diameter sterile filter membranes (Millipore, SA) and the membranes placed on the
relevant substrate medium before incubation (16). The average number of colony forming
units per 100 milliliter (cfu.100ml-1) was calculated and recorded.

For the detection of total coliform bacteria in the water samples, m-Endo (DIFCO) agar
plates were prepared in 90 mm Petri dishes according to the manufacturer’s specifications
(Merck, SA). After placing the membranes the bacteria onto the agar media, the plates
were incubated aerobically for 24 hours at 370C. All colonies with a golden metallic sheen
were counted as total coliforms. For the detection of faecal coliform bacteria in the water
samples, m-FC (DIFCO) agar plates were prepared in 90mm Petri dishes according to the
manufacturer’s specifications (Merck, SA). After placing the membranes with bacteria
onto the agar media, the plates were incubated aerobically for 24 hours at 44.50C. All dark
blue colonies were counted as faecal coliforms. For the detection of faecal enterococci in
the water samples, m-Enterococcus (DIFCO) agar plates were prepared in 90mm Petri
dishes according to the manufacturer’s specifications (Merck, SA). After placing the
membranes with the bacteria onto the agar media, the plates were incubated aerobically
for 48 hours at 370C. All red-pink colonies were counted as faecal enterococci.
Clostridium perfringens counts (vegetative cells and spores) were determined using
specific perfringens selective OPSP medium (Oxoid, SA) with supplements. The OPSP
agar plates were prepared in 90 mm Petri dishes according to the specifications of the
manufacturer’s (Oxoid, SA). After placing the membranes with bacteria onto the agar
plates, the plates were incubated in anaerobic conditions at 37˚C for 48 h using
anaerogens sachets in order to produce anaerobic environment. Colonies appearing as
dark brown to black were counted.

Somatic phages were assayed according to the methods described by Grabow et al., (17)
with Escherichia coli strain WG5 (18) as host. Presence-absence test was carried out
according to the method described by Uys (19). Briefly: A volume of 500 ml of each water
sample was poured into a sterile plastic 1l water collection bottle to which 5g Trypticase
peptone, 4g Sodium Chloride and 5ml of a Calcium-Glucose solution were added. The
presence-absence sample one milliliter of the specific host culture was added to each of
the water sample and incubated at 37˚C for 24 h. The presence of somatic coliphages
were determined by spot plating 5 µl of the presence-absence sample to a prepared phage
plates containing a lawn of the WG5 host. The spot plates were incubated overnight at
37˚C and the bacteriophages allowed to produce zones of lysis or plaques where the
suspensions have been spotted.

Classification criteria for borehole water quality

Both private and communal borehole water results were grouped into categories according
to the total number of a specific indicator organism detected. The classification system
described by DWAF (15) and SABS (16) was used to categorize each borehole:
1. good (negligible risk of microbial infection; fit for human consumption)
2. marginal (slight risk of microbial infection; must be treated before consumption)
3. poor (risk of infectious disease transmission; not fit for human consumption)

Table 1 Categories used for water quality assessment (15; 16)

Indicator Water quality assessment criteria

Organism
Good Marginal Poor
Total coliform 10 cfu.100 ml-1 11-100 cfu.100 ml-1 > 100 cfu.100 ml-1
Faecal coliform 0 cfu.100 ml 1-10 cfu.100 ml-1 > 10 cfu.100 ml-1
Faecal enterococci 0 cfu.100 ml 1 cfu.100 ml-1 > 1 cfu.100 ml-1
Clostridium perfringens 0 cfu.100 ml 1 cfu.100 ml-1 > 1 cfu.100 ml-1
Somatic coliphages 0 cfu.10 ml-1 1 cfu.10 ml-1 > 1 cfu.10 ml-1
Cfu = colony forming units
good = fit for human consumption
poor = poses a health risk

Statistical analysis

Data was presented and entered into Microsoft Excel 2000. STATA version 7 Shapiro
Facia was used to determine the normality assumption for continuous variables. For
comparison of categorical data, Fisher exact test or chi-square test was used (if no
expected cell values were less than five). The Mann-Whitney test (sample size, mean and
standard deviation) was also used.

RESULTS

Total coliform

The total coliforms results for water samples tested in private and communal boreholes
during dry and rainy seasons were showed in Figure 1. During the dry seasons, 70% of
private and 78% of communal borehole water samples were classified as good (10 cfu.
100 ml-1); 27% of private and 20% of communal borehole water samples were classified
as marginal (11-100 cfu. 100 ml-1); whereas 3% of private and 1% of communal borehole
water samples were classified as poor (>100 cfu. 100 ml-1). During the dry seasons no
significant differences (P=0.117) were seen between private and communal borehole
water samples. During the rainy seasons 50% of private and 65% of communal borehole
water samples were classified as good (10 cfu.101); 35% of private and 25% communal
borehole water samples were classified as marginal (11-100 cfu.100 ml-1); whereas 15%
private and 20% of communal borehole water samples were classified as poor (>100
cfu.100 ml-1). However, during the rainy seasons significant statistical differences
(P=0.006) could be detected between the private and communal borehole water samples,
implying that contamination was more prevalent in private boreholes than in communal
borehole water.

100
90 Private
Communal
80
70
60
% 50
40
30
20
10
0
Good Marginal Poor Good Marginal Poor

Dry Rainy

Figure 1 Total coliform counts during dry and rainy seasons

Faecal coliforms

The faecal coliforms results for water samples tested in private and communal boreholes
during dry and rainy seasons were showed in Figure 2. During the dry seasons 60%
private and 75% of communal borehole water samples tested were classified as good (0
cfu.100 ml-1); 30% of private and 15% communal borehole water samples were classified
as marginal (1-10 cfu.100 ml-1); whereas 10% of both private and communal borehole
water samples were classified as poor (>10 cfu.100 ml-1). During rainy season 55%
private and 70% communal borehole water samples tested were classified as good (0
cfu.100 ml-1); 25% private and 15% communal borehole water samples were classified as
marginal (1-10 cfu.100 ml-1); whereas 25% private and 15% communal borehole water
samples were classified as poor (>10 cfu.100 ml-1). During both the dry (P=0.006) and
rainy seasons (P<0.0001) the difference in faecal coliform contamination was significant,
although during the rainy seasons the contamination was more prevalent in private
borehole water samples compared to communal borehole water samples.

100
Private
90
Communal
80
70
60
% 50
40
30
20
10
0
Good Marginal Poor Good Marginal Poor

Dry Season Rainy Season

Figure 2 Faecal coliform counts during dry and rainy seasons

Faecal enterococci

The results of faecal enterococci present in water samples tested in private and communal
boreholes during dry and rainy seasons were showed in Figure 3. During dry season
about 70% private and 78% communal borehole water samples were classified as good (0
cfu.100 ml-1); 8% of private and 10% communal borehole water samples were classified as
marginal (1 cfu.100 ml-1); whereas 22% private and 12% communal borehole water
samples were classified as poor (>1 cfu.100 ml-1). During the dry seasons, the
contamination prevalent in the private borehole water samples were slightly higher
compared to contamination seen in communal borehole water samples (P=0.037). During
the rainy seasons, 70% of private and 72% communal borehole water samples tested
were classified as good (0 cfu.100 ml-1); 8% of water samples in both private and
communal boreholes were classified as marginal (1 cfu.100 ml-1); whereas 22% private
and 20% communal borehole water samples were classified as poor (>1 cfu.100 ml-1).
The prevalence of faecal enterococci in both private and communal borehole water
samples did not show any significant difference (P=0.778) during the rainy seasons.
 100
Private
90
Communal
80
70
60
% 50
40
30
20
10
0
Good Marginal Poor Good Marginal Poor

Dry Season Rainy Season

Figure 3 Faecal enterococci counts during dry and rainy seasons

Clostridium perfringens

The results for Clostridium perfringens in borehole water samples tested from private and
communal borehole in dry and rainy seasons were shown in Figure 4. During the dry
seasons 78% private and 75% communal borehole water samples tested were classified
as good (0 cfu.100 ml-1); 1% of private and 8% communal borehole water samples were
classified as marginal (1 cfu.100 ml1); whereas 21% private and 17% communal borehole
water as poor (>1 cfu.100 ml-1). During dry season (P<0.0001) there was a significant
difference in private and communal borehole water implying that contamination was more
in communal borehole water than in private boreholes. In rainy season about 90% of
private and 95% communal borehole water samples were classified as good (0 cfu.100 ml-
1
); 1% private and 2% communal borehole water as marginal (1 cfu.100 ml-1) whereas 9%
private and 3% communal borehole water as poor (>1 cfu.100 ml-1). In rainy season there
was a significant difference (P=0.047) between private and communal borehole water
although the difference is small implying that during the rainy season the contamination is
almost equal for both the private and the communal borehole water.
 100
Private
90
Communal
80
70
60
% 50
40
30
20
10
0
Good Marginal Poor Good Marginal Poor

Dry Season Rainy Season

Figure 4 Clostridium perfringens counts during dry and rainy seasons

Somatic coliphages

The results for the presence-absence tests carried out for somatic coliphages during dry
and rainy season is shown in Figure 5. In dry season about 97% of private and 95%
communal borehole water tested had no somatic coliphages and classified as good (0
cfu.100 ml-1); 3% of private and 5% communal borehole water tested positive for the
presence of somatic coliphages and classified as poor (>1 cfu.100 ml-1). During the rainy
seasons there was no significant difference (P=0.814) in the presence of somatic
coliphages in both private and communal borehole water samples. During the dry
seasons there was significant difference (P=0.014) in the presence of somatic coliphages
between the private and communal borehole water samples.

Private
100
90 Communal
80
70
60
% 50
40
30
20
10
0
Absent Present Absent Present

Dry Season Rainy Season

Figure 5 Somatic bacteriophage counts during dry and rainy seasons

DISCUSSION AND CONCLUSION

During rainy season pollutants can enter into groundwater through percolation and
seepage. The rain can wash particles which have been deposited on nearby surfaces into
the water source through leaching processes (20). This also depends on the type of soil
as well as the physical forms of pollution (20). In dry seasons the water table becomes
very low and the rate of evaporation increases and this affects the oxygen content which in
turn decreases the multiplication of bacteria (21). Low temperatures could also reduce the
amount of oxygen available and hinder the bacterial process (22). Aerobic bacteria can
survive in optimal temperatures which is not too low or too high. During times of
unfavorable growth conditions, anaerobic bacteria may form spores (22). Therefore the
contamination of borehole water will depend on the seasonal variation and the resistance
of particular bacteria to environmental conditions.

In both private and communal boreholes observations, it was found that some of the
boreholes were drilled next to the sanitary facilities i.e. pit latrines and septic tanks. This
posed a health risk because of increased contamination by human faeces as the
wastewater gain access into the groundwater during leaching process. The distance
between the sanitary facilities and borehole water were in most cases very close to each
other. This indicated that there was still a lack of knowledge by the community members
on the principles of groundwater protection. The observations also indicated that
Environmental Impact Assessment (E.I.A) was not done when a site for drilling borehole
was selected. This could be seen by the number of boreholes drilled next to the potential
sources of pollutants.

When communal borehole water is drilled E.I.A is either done or not done, and especially
for private boreholes it is rare for E.I.A to be conducted. This results in the risk of borehole
water contamination. Placing borehole water next to sanitary facilities indicates that there
is a lack of knowledge by most of the community on the importance of groundwater
protection. This was shown by the number of boreholes drilled next to pit latrines.

Lack of sanitary facilities and availability of none V.I.P pit latrines in most of the
households in rural villages where research was conducted, contributed to borehole water
contamination. Consequently, this posed a threat to communal boreholes drilled inside the
stream on other places surrounded by households without pit latrines and some with none
V.I.P toilets. Some of the community members used the bushes nearby the stream and
during rainy days everything was swept into the stream causing ground water
contamination.

In this study, some of the communal and private boreholes were found to be contaminated
with indicator bacteria which indicated that some of these boreholes could have been a
health hazard to their users. The findings in the study indicated that communal and private
borehole water has been contaminated as a result of faecal pollution. Comparison of the
private and communal boreholes showed that there was a slight difference in the
microbiological quality of the two groups with organisms such as Clostridium perfringens
more prevalent in dry season in communal boreholes and rainy seasons in private
boreholes and total coliforms more prevalent during the rainy seasons in private boreholes
as well as in communal boreholes. Both private and communal boreholes have also
shown the faecal contamination by somatic coliphages, faecal coliforms and faecal
enterococci during dry and rainy seasons.
This study have shown the microbiological status of water in the private boreholes found in
the Limpopo Province and have shown the need for education of the owners of these
boreholes together with the rest of the communities on the installation and maintenance of
these boreholes as well as education on basic hygiene and sanitation practices when
using boreholes and collecting water from them. The need for E.I.A before a borehole is
drilled needs to be emphasized.

ACKNOWLEDGEMENTS

The assistance of undergraduate students, Mr. P Ramudingana and Me. AL Nemarude,

are appreciated.

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