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Impact of Bisphenol A on the Physicochemical and Bacteriological Characteristics of

Water in Storage Tanks from various Locations in Salem University Lokoja, Kogi
State, Nigeria
1, 2*UDOCHUKWU, U; 2OLANNYE, PG
1
Department of Biosciences, Salem University, P.M.B. 1060, Lokoja, Kogi State, Nigeria
2
Department of Microbiology, University of Benin, Benin City, Edo State, Nigeria
*Corresponding Author Email: rev.dr.ud@gmail.com

ABSTRACT: This study examined the impact of Bisphenol A (BPA) on the physicochemical and bacteriological
characteristics of water in storage tanks in Salem University Lokoja. Borehole water samples were collected from three
(3) locations within the University environment and stored in a jerry can for analysis. Total Heterotrophic Bacteria Count
(THBC) in water samples ranged from 1.00± 0.30 x104 at week 0 to 8.95±1.00 x104 cfu/ml at week 3, while the total
coliform count (TCC) also ranged from 1.30±0.15 x104 to 7.11±0.82 x104 cfu/ml. TCC and THBC were found to be
higher than the NSDWQ Standard. The identified isolates from the borehole samples were Escherichia coli, Pseudomonas
aeruginosa, Streptococcus faecalis, Bacillus cereus, Staphylococcus epidermidis and Serretia spp. Bisphenol A (BPA)
was not detected in week 0, after week 3, components of Bisphenol A detected were methyl chloride, Benzene and
Dichlorobenzene and their highest values were 0.054±0.033, 0.021±0.020 and 0.055±0.062 mg/l respectively. The pH,
Turbidity, Total suspended solids, BOD and conductivity reduced as storage increased. Magnesium and calcium for
sample B were found to have the highest value of 0.31 and 1.73 mg/l respectively. Storage of water for a long period of
time should be discouraged as it could trigger increased leaching of BPA into the water which will affect its
physicochemical and microbiologically quality.

DOI: https://dx.doi.org/10.4314/jasem.v24i2.5

Copyright: Copyright © 2020 Udochukwu and Olannye. This is an open access article distributed under the
Creative Commons Attribution License (CCL), which permits unrestricted use, distribution, and reproduction in
any medium, provided the original work is properly cited.

Dates: Received: 16 November 2019; Revised: 11 January 2020; Accepted: 22 February 2020

Keywords: Bacteria, Bisphenol A, Storage Water Tanks, Drinking Water Quality.

Good quality water guarantees public health, or weeks increases bacteria load and leaching of
protection of the environment and sustainable Bisphenol A and as such reduces the quality of water
development (Ranjini et al., 2010). Water of good (Jagals et al., 1999: Adeghe and Emejulu, 2016).
quality is of basic importance to human physiology According to (Eniola et al., 2007), storage generally
and man's continued existence depends very much on reduces the numbers of bacteria which is in contrast to
its availability (Lamikara, 1999; FAO, 1997). By (WHO, 2005) which identified water as a major
2025, one-third of the population of the developing components for bacteria growth. Several technologies
world will face severe water shortage (Sota et al., for the treatment of household water in developing
2005). Contaminated water is a global public health countries have been developed to improve the
threat placing people at risk of a host of diarrhoeal and chemical and bacteriological quality of the water and
other illness as well as chemical intoxication. The to reduce waterborne diseases (Mintz et al., 1995).
major risk to human health is faecal contamination of These technologies include physical methods such as
water supplies (Okonko et al., 2009). A significant boiling, heating, sedimentation, filtration, exposure to
proportion of the world's population use potable water ultraviolet radiation from sunlight and chemical
for drinking, cooking, personal and home hygiene disinfection with agents such as sodium hypochlorite,
(WHO, 2005). Before water can be described as chlorine (Mintz et al., 1995; Sobsey, 2002). Bacteria
potable, it has to comply with certain physical, reactivate faster in dechlorinated water than in
chemical and bacteriological standards which are chlorinated water (Sobsey, 2002). BPA is widely used
designed to ensure that the water is potable and safe for mass production of plastic (polycarbonates) and
for drinking (Atuanya et al., 2016). Over 50,000 epoxy resin (Morrissey et al., 1987; Staples et al.,
people die daily due to water borne diseases (Marque 1998). For many years, BPA was treated as a non-toxic
et al., 2003). About 2.3 billion people Worldwide have compound with no negative impact on humans and
mortality and morbidity associated with water related animals. BPA-based products were commonly used to
ailment. In order to improve the bacteriological quality mention lacquers for cans and vessels for storage of
of water consumed by members of rural households, it food, water, and medicines (Staples et al., 1998). Since
is essential to address the quality of stored drinking the second half of the 90-ties, numerous reports have
water and the conditions under which the water arrived stating a negative influence of BPA on human
supplies are stored. Storing of water in tanks for days health (Biles et al., 1997; Del-Olmo et al., 1997). BPA

*Corresponding Author Email: rev.dr.ud@gmail.com

Impact of Bisphenol A on the Physicochemical and Bacteriological…. 224

has been classified as xenobiotic endocrine disruptor, column temperature was initially held at 40oC for 1
disrupting the balance of the hormonal system minute, raised at 120oC at the rate of 25oC/minute, then
(Moriyama et al., 2002). Stored water and Bisphenol to 60oC at the rate of 10oC/minute, and finally to 300oC
A are like two components that go together when at the rate of 5oC/minute, held at the final temperature
subjected to high temperature and over a long period for 15 minute. Detector temperature was kept at 280oC.
of storage (Del-Olmo et al., 1997). Therefore, auditing Helium was used as a carrier gas at a constant flow of
and monitoring of chemical and bacteria quality of 1ml/minute. Mass spectrometry was acquired using
drinking water is an essential aspect of water quality. the electron ionization (EI) and selective ion
Therefore, this study examined the physicochemical monitoring (SIM). Fifty ml (50± 0.01 ml) water was
and bacteriological characteristics of water in storage measured, and 100ml of dichloromethane (DCM) via
tanks from various locations in the Salem University separating funnel and shacked for 30mins for BPA
Lokoja, Kogi State. extraction (Dean and Xiong 2000). This separating
funnel was clamp and a mixture was allowed to
MATERIALS AND METHOD separate out. After separation the DCM portion was
Sample collection and borehole water physiochemical collected. The process was repeated three times for
analysis: Borehole Water sample was collected from complete extraction (FAO, 1997). Blanks were
storage tank in three (3) different locations within the prepared following the same procedure without the
University premises: Hostel (Sample A), the Cafeteria sample. The standard sample used for quality control
(Sample B) and at the College of Natural and Applied was prepared by adding standard solution (BPA) to
Sciences (CNAS) (Sample C). The samples were DCM. All extracts were separated, and activated
spread into three different jerry can which represents copper was added to the combine extract for
our storage tank. The samples were kept outside and desulphurization. After subsequent filter over
sealed to avoid contamination. A glass jar was used at anhydrous sodium sulphate, the solution was
the point of collection for analysis. The samples were concentrated to 1.0ml using a rotary evaporator, an
analysed for their physicochemical, bacteriological internal standard mixture (vinyl chloride) solution was
and Bisphenol A composition. The physicochemical run with the extract for quality control check using
parameters analysed were pH, chloride, sodium, Hewlett Packard 5890 series II gas chromatograph
electricity conductivity, Iron, total dissolved, with mass selective detection (GC-MS) (Dean and
biochemical oxygen demand, total suspended solid, Xiong, 2000).
calcium, manganese, zinc, copper, sulphide and
phosphorus according the method described by RESULT AND DISCUSSION
(Aydin, 2007) The physicochemical analysis result is shown in
(Table 1). The result showed that all physicochemical
Isolation and Identification of bacteria: Nutrient agar parameters for sample A were compliance with the
was used in the isolation and enumeration of bacteria Nigerian Standard of Drinking Water, while the
using the pour plate method. The pure culture was then ammonium (NH3+) and calcium (Ca2+) content for
transferred into nutrient agar slants for biochemical sample B and C exceeded the maximum limit of the
test. Nutrient and MacConkey agar were used to Nigerian Standard of Drinking Water. Also the iron
enumerate the bacteria in the water samples (Fe2+) content for sample B exceeded the required
(Cheesbrough, 2000). Identification of isolates was limit. The pH value for sample B (6.6 ± 0.06 to
based on cultural, morphological and biochemical 6.47±0.97) exceeded the maximum limit of the
characteristics following standard methods (Garrity et Nigerian Standard of Drinking Water. For total
al., 2005; Holt et al., 2000). heterotrophic bacteria count, Sample B had the highest
count which ranged from 6.57 ± 0.93 to
Analysis of Bisphenol A (BPA) in borehole water: 7.4 ± 0.67cfu/ml between week 0 and week 1 while
Hewlett Packard 5890 series II gas chromatograph sample C had the highest count for week 2 and 3 which
equipped with an Agilent 7683B injector (Agilent were 8.95 ± 1.00cfu/ml and 4.26 ± 0.42cfu/ml
Technologies, Santa Clara, CA, USA), a 30m, 0.25mm respectively (Table 2). The coliform bacteria count
i.d. HP-5MS capillary column (Hewlett–Packard, Palo showed that Sample B recorded the highest count for
Alto, CA, USA) coated with 5% phenyl- week 0 which was 4.01 ± 0.57cfu/ml while sample C
methylsiloxane (film thickness 0.25um) and an had the highest count for week 1, 2 and 3 which are
Agilent 5975 mass selective detector (MSD) was used 6.16 ± 0.61, 3.09 ± 0.31 and 7.11 ± 0.82cfu/ml (Table
to separate and qualify the (BPA) compounds. The 3). Possible bacteria pathogens identified from the
samples were injected in the split less mode at an water sample were Escherichia coli, Pseudomonas
injection temperature of 300oC. The transfer line and aeruginosa, Serretia spp. and Staphylococcus
iron source temperature were 280oCand 200oC. The epidermidis (Table 4). The Distribution of bacterial

UDOCHUKWU, U; OLANNYE, PG
Impact of Bisphenol A on the Physicochemical and Bacteriological…. 225

isolate in borehole water samples was investigated, samples were lower than NSDWQ Standard OF
and it was observed that Escherichia coli had the 500mg/L. Total dissolved solids in drinking water has
highest percentage occurrence with 30.5% and been associated with sewage urban runoff, natural
Serretia spp. (11.1%) with least percentage occurrence sources and industrial waste water (NSDWQ, 2007).
(Table 5). The Bisphenol A contingents were This finding is contrary to the observation of (Ballester
identified from the water samples. It was observed that and Sunyer, 2000) whose TDS were in line with
hexane, vinyl chloride, toluene and NSDWQ standard. Total suspended solid (TSS)
tetrachloroethylene were not detected in the water ranged from 0.25-2.68 mg/l which decreased as
samples. Dichlorobenzene and methylene chloride had storage increased. The biological oxygen demand
the highest value of 0.055 ± 0.062 and 0.054 ± 0.033 (BOD) reduced also followed suit as there is no
mg/l respectively while benzene had the lowest value standard limit to BOD. Cadmium and lead were not
of 0.021 ± 0.020 mg/l (Table 6). The physiochemical detected in the water samples at week zero, but at week
parameters were critically analyzed for a period of 3 components of cadmium and lead were detected
3weeks. There was gradual increase in the pH of water (Table 1). The presence of sodium, calcium and
samples from week 0 to week 3, the pH of all samples magnesium salts in water helps in reducing incidence
ranged from 5.33 to 6.6. The pH of Sample B is in line of cardiac disease (Mintz et al., 1995). The iron
with Nigeria Standard of Drinking Water Quality content of the water samples in this study is in line with
(NSDWQ) of 6.5-8.5. This shows that as storage NSDWQ standard of 0.3 mg/l (NSDWQ, 2007). The
increases pH also increases this is in agreement with chlorine content of the stored water ranges from
Aydin. (2007) who recorded increased pH as storage 4.31 ±0.71 to 19.68 ± 0.90 mg/l and 7.02 ± 0.86 to
increases. The pH of Sample A and C for both week 0 22.78 ± 0.86 mg/l for week 0 and week 3 respectively.
and week 3 are below (NSDWQ). The pH of Sample
B is in line with NSDWQ standard (NSDWQ, 2007). Chlorine content in Sample A and B reduced as storage
The pH of water samples at week 0 were suitable for increased which was the same for Sample C which
bacteria proliferation, a neutral pH will support growth increased with time. Conductivity, sulphates, nitrates
of a large number of bacteria (Madigan et al., 2000). and phosphates reduced as storage increased in all the
The turbidity of all water samples ranges from 0.07 to samples. The physiochemical parameters were
0.6 ntu. There was decreased in turbidity as storage compared to Nigeria Standard of Drinking Water
increased, low turbidity is often expected not to exceed Quality. The total bacterial counts for all the water
5 ntu (EPA, 2003). samples were generally high and exceeded the limit of
1.0x102 cfu/ml which is the standard limit of
The total dissolved solids in Sample A and C reduced heterotrophic count for drinking water (NSDWQ,
as storage increased, an increase was observed in 2007).
Sample B. Total dissolved solids (TDS) of all the .

Table 1: Physicochemical parameters of borehole water sample During Storage over a period of 3 weeks.

Note: NTU =Nephalometric Units, NS = No Standard, NSDWQ = Nigerian Standard of Drinking Water
Table 2: Total Heterotrophic Bacterial Count in Borehole Water during Storage in Jerry can (104 cfu/ml).

UDOCHUKWU, U; OLANNYE, PG
Impact of Bisphenol A on the Physicochemical and Bacteriological…. 226

Table 3: Total Coliform bacteria Count in Borehole Water During Storage in Jerry Can (104 cfu/ml).

Table 4: Cultural, morphological and biochemical characterization of Bacteria isolates in borehole water samples

Table 5: Distribution of Bacterial isolate in Borehole Water Samples

Table 6: Bisphenol A Composition in Borehole Water samples over time

UDOCHUKWU, U; OLANNYE, PG
Impact of Bisphenol A on the Physicochemical and Bacteriological…. 227

The count is indicative of the presence of organic and fit for drinking. High temperature and a prolonged
dissolved salts in the water. From the result above, it storage have been implicated in the increased release
was observed that in all water samples the mean total of BPA in water tanks. Hence plastic borehole
heterotrophic bacteria count ranges from 1.00 ± 0.30 overhead tanks should not be directly exposed to the
x 104 to 2.13 ± 0.21 x 104 cfu/ml in Sample A, 3.63 ± atmosphere as direct rays of the sun leads to fast
0.31 x 104 to 8.70 ± 1.01 x 104 cfu/ml in Sample B and leaching of toxic chemicals. This research has shown
2.61 ± 0.82 x 104 to 8.95 ± 1.00 x 104 cfu/ml in that there is need for an improvement in disinfection
Sample C which had the highest count (Table 2). The and cleaning of storage tanks, hence drinking water
total coliform counts for all samples were exceedingly should be stored and used within days. There is also
higher than the NSDWQ of maximum contamination need for public awareness programmes to educate the
level (MCL) for coliform bacteria in drinking water public on the possible health implications of drinking
(NSDWQ, 2007). The high coliform count obtained in water which has been stored for a prolonged time.
the samples may be an indication that the water
sources are faecally contaminated (Obi and Okocha, REFERENCE
2007). None of the water samples complies with Atuanya, E. I., Adeghe, M. O. and Udochukwu, U.
NSDWQ standard for coliform in water. Bacteria (2016). Bioavailability of Plastic Contaminants
isolated from all water samples include bacillus and Their Effects on Plastic Bottled and Sachet
cereus, serretia spp., Escherichia coli, Pseudomonas Drinking Water Supplies. British Microbiology
aeruginosa, Streptococcus faecalis, and Research Journal, 14(4): 1-10.
Staphylococcus epidermidis which are also of public
health significance. Staphylococcus epidermidis is Adeghe, O.M. and Emejulu, M.J. (2016). Evaluation
known to produce enterotoxin (Aydin, 2007). The of Bisphenol A, Physiochemical Properties and
presence of coliform indicates that all water samples Microbial Characterization of Borehole Water
are not fit for drinking and the observation in this study Stored in Plastic Containers. Journal of Applied
suggest that high heterotrophic count in water reflects Science and Environmental Management, 20
high coliform count. Presence of high coliform count (4):1119-1124
in borehole water samples could be attributed to the
proximity of the borehole near to a septic tank at a Aydin, A. (2007). The microbial and physicochemical
distance less than the 30m recommended by NSDWQ. quality of ground water in west Thrace Turkey.
The results above shows that the coliform population Polish Journal of Environmental Studies, 16 (3):
increased with time. There was alternate increase and 377–383.
decrease of Bacteria count in samples C as storage
progresses this is in contrast to Orji et al. (2006) who Ballester, F. and Sunyer, J. (2000). Drinking water and
recorded a direct increase of Bacteria count. Bisphenol gastrointestinal disease, need of better understand
A analysis at week zero shows that no BPA and an improvement in public health surveillance.
contingents were detected in all the water samples, but Journal of Epidemiology of Community Health,
at week 3 there were detectable quantities of BPA 54:3-5.
components. This indicates that temperature and
storage duration is a major factor in Bisphenol A Biles J.E., Mc Neal T.P., Begley T.H. (1997).
degradation in storage tanks (Table 6). The Determination of bisphenol A in reusable
composition of Bisphenol A found in week 3 is enough polycarbonate food-contact plastics and migration
to cause serious health hazards when accumulated in to food-simulating liquids. Journal Agricultural
the body. The BPA detected in this research work is in Food Chemistry, 45: 3541–3544.
line with Vandenberg et al. (2007) who recorded
increase in Bisphenol A contingents as storage time Carwile J.L., Luu H.T., Bassett L.S., Driscoll D.A.,
increases. The presence of potential pathogens such as Yuan C., Chang J.Y., Ye, X., Calafat A.M.,
Escherichia coli, Pseudomonas aeruginosa, Serretia Michels K.B. (2009). Polycarbonate bottle use
spp. and Staphylococcus epidermidis (Table 4) in the and urinary Bisphenol A concentrations.
water samples indicates the absence of sanitation on Environmental and Health Perspectve, 117:
the part of the users. 1368–1372.

Conclusion: This study shows that storage of water Cheesbrough M. (2000). Bacteriological testing of
overtime leads to increased leaching of BPA which water: In District Labouratory Practicein
will encourage the proliferation of bacteria and also Tropical Countries. Part 2:149-154.
affect its physicochemical properties. In comparism to
NSDWQ standard, all water samples studied were not

UDOCHUKWU, U; OLANNYE, PG
Impact of Bisphenol A on the Physicochemical and Bacteriological…. 228

Del-Olmo M., Gonzalez-Casado N.A. and Navas J.L. Marque, S., Jacqmin-Gadda, H., Dartigues, J.F. and
(1997). Determination of bisphenol A (BPA) in Commenges D. (2003). Cardio-vascular mortality
water by gas chromatography-mass spectrometry. and calcium and magnesium in drinking water: an
Anal Chimica Acta 346: 87–92. ecological study in elderly people. European
Journal of Epidemiology, 18: 305–309.
Dean, J.R., and Xiong, G. (2000)."Extraction of
organic pollutants from environmental matrices Mintz, E. D. Reiff, E. M, Tauxe and R.V. (1995). Safe
selection of extraction technique", Trends in water treatment and storage in home: A practical
Analytical Chemistry, 19(9): 553-564. new strategy to prevent water borne diseases.
JAMA, 273: 948-953.
Dolinoy, D.C, Huang D. and Jirtle R.L. (2007).
Maternal nutrient supplementation counteracts Moriyama K., Tagami T., Akamizu T., Usui T., Saijo
bisphenol A-induced DNA hypomethylation in M., Kanamoto N., Hataya Y., Shimatsu A.,
early development. Processes of National Kuzuya H., Nakao K. (2002). Thyroid hormone
Academic Science USA, 104 (32): 56-61. action is disrupted by Bisphenol A as an
antagonist. Journal Clinical Endocrinologic
Eniola K.I.T., Olayemi A.B., Awe S., Adegoke A., Metabolism, 87: 5185–5190.
Osanoto I.B., Abolade G.O. and Kayode-Isola
T.M. (2006). Effect of Storage on Bacteriological Morrissey R.E., George J.D., Price C.J., Tyl R.W.,
Quality of Well Water. African Journal of Marr M.C. and Kimmel C.A. (1987). The
Clinical and Experimental Microbiology, developmental toxicity of bisphenol A in rats and
1:567pp. mice. Fundamental Application on Toxicology, 8:
571–582
Food and Agriculture Organisation (1997). Chemical
analysis manual for food and water, 5th Ed, FAO NSDWQ, (2007). Nigeria Standard for Drinking
Rome. pp 20-26. Water Quality .Nigeria Industrial Standard
Approve by Standard Organization of Nigeria
Garrity, G.M., Brenner, D.J., Krieg, N.R., and Staley Governing Council. ICS13.060.20:15-19.
J.T. (2005). Bergey’s Manual of Systematic
Bacteriology, 2nd ed., Springer, New York Obi, C.N. and Okocha, C.O. (2007). Microbiological
123pp. and physicochemical analyses of selected
borehole waters in World Bank Housing Estate,
Holt, J.G., Krieg, N.R., Sneath, P.H.A., Staley, J.T. Umuahia, Abia state Nigeria. Journal of
and Williams S.T. (2000). Bergey’s Manual of Engineering and applied Sciences, 2 (5): 920-929.
Determinative Bacteriology, 9th ed. Lippincott,
Williams and Wilkins, Philadelphia, PA 234pp. Okonko, I.O, Adejoje, O.D, Ogunnusi, T.A, Fajobi, E.
and Shittu, O.B. (2009). Microbiological and
Jagals, P., Bokaka, T. C. and Grabow W. O. K (1999). physicochemical analysis of different water
Changing consumer water patterns and their effect samples used for domestic purposes in Abeokuta
on microbiological water quality as a result of an and Ojota, Lagos Nigeria .African Journal of
engineering intervention. Water South Africa. 25 Biotechnology, 7 (5): 617-621.
:297-300.
Orji, M. U., Ezenwaje, E. E; Anyaegbunam, B. C.
Kawamura, Y., Koyano, Y., Takeda, Y. and Yamada, (2006). Spatial appraisal of shallow water well
Y. (1999). Journal Food Hygiene Society. Jpn, water pollution in Awka, Nigeria. Nigeria journal
206p. microbiology, 20:1389-1394
Lamikara, A. (1999). Essential microbiological for
students and practitioners of pharmacy, medicine Rajini K. Roland P. John C.and Vincent R (2010)
and microbiology, 2nd Edition, Amkra books, .Microbiological and physicochemical analysis of
406pp. drinking water in George Town. Nature and
Science, 8(8)261-265.
Madigan, M. T., Martinko, J. M. and Parker, J. (2000).
Brock Biology of Microorganisms. Prentice Hall Sobsey, M.D. (2002).Drinking water and health
Press, USA.p.32. research:a look to the future in the United States
and globally. Journal of Water and Health, 4: 17-
21.

UDOCHUKWU, U; OLANNYE, PG
Impact of Bisphenol A on the Physicochemical and Bacteriological…. 229

US Environmental protection Agency (USEPA)

Soto A.M, Justicia H, W ray J. W, Sonnenschein, C. (1995). Guidelines for water supply 7th edition.
(2005) .p-Nonyl-phenol: anestrogenic xenobiotic NewYork: WHO press pp.144-162?
released from "modified" polystyrene. Vandenberg L.N, Schaeberle C.M, Rubin B.S,
Environmental Health Perspectives, 92:167-173. Sonnenschein C, Soto A.M. (2007).The male
mammary gland: a target forth exenoestrogen
Staples C.A., Dorn P.B., Klecka G.M., O’Block S.T., bisphenol A. Reproductive Toxicology. 37:15-23.
Harris L.R. (1998). Quantitative determination of
bisphenol-A in river water by coolon-column WHO. (2005). Financial management of water supply
injection-gas chromatography-mass and sanitation. World Health Organization.
spectrometry. International journals on Geneva, p.56.
Environmental Analysis of Chemical 69:83–98.

UDOCHUKWU, U; OLANNYE, PG