Groundwater Quality of Boreholes Located Around Cassava

Processing Mills
Keywords:
Awka-North, groundwater, cassava effluent and cyanogenic glycosides.
ABSTRACT:


The bacteriological and physicochemical quality of groundwater sources
around cassava processing mills in Awka-North L.G.A, Anambra State, Nigeria, were
determined over a period of three months (June-August), using standard
microbiological and physicochemical methods. The result revealed that the highest
total heterotrophic and coliform counts of 5.0x10
3
cfu/ml and 2.20x10
3
cfu/ml were
obtained from Urum and Amanuke borehole water systems respectively, while
Ebenebe recorded the least total heterotrophic count (1.10x10
3
cfu/ml). Isu-Aniocha
and Ugbene haboured no coliforms. Identification tests revealed the isolation of
Pseudomonas (83.33%), Bacillus (66.67%), Micrococcus (33.33%), Proteus (16.67%),
E. coli (50%), Salmonella (33.33%), and Streptococcus (50%). Out of the seven
bacterial genera isolated, the borehole samples from Urum and Amanuke each
haboured 71.42%, Mgbakwu recorded 28.57%, Ugbene and Ebenebe had 42.86% and
57.14%, respectively, while the borehole water samples from Isu-Aniocha recorded
the least 14.29%. The pH of the water was acidic in all the samples. Among the
parameters tested, only pH, CN
-
, Fe and Cu were found to be outside the WHO
recommended limits for drinking water in most of the samples.
056-062 | JRPH | 2013 | Vol 2 | No 1

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www.jhealth.info
Journal of Research in
Public Health
An International Scientific
Research Journal
Authors:
Okechi RN
1
,
Onyekere JA
1
,
Azuwike CO
1
,
Nnokwe JC
2
, Ibe IJ
3
and
Ukagwu N
3
.

Institution:
1. Department of
Biotechnology, Federal
University of Technology,
Owerri, Nigeria.

2. Department of Biology,
Federal University of
Technology, Owerri,
Nigeria.

3. Department of
Microbiology, Federal
Polytechnic, Nekede,
Owerri, Nigeria.


Corresponding author:
Okechi RN.







Email:
popebenn@yahoo.com




Web Address:
http://www.jhealth.info
documents/PH0014.pdf.


Dates:
Received: 26 Nov 2012 Accepted: 03 Dec 2012 Published: 04 Feb 2013
Article Citation:
Okechi RN, Onyekere JA

, Azuwike CO

, Nnokwe JC

, Ibe IJ

and Ukagwu N.


Groundwater Quality of Boreholes Located Around Cassava Processing Mills.
Journal of Research in Public Health (2013) 2(1): 056-062
An International Scientific Research Journal
Original Research
Journal of Research in Public Health
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INTRODUCTION
Groundwater is the water that infiltrates the soil
and is stored in the spaces between particles in the
earth (Enger and Smith, 2000). It is one of the major
sources of drinking water. The composition of
groundwater depends on soil types and geological
materials present in the tested site. The amount of
these substances makes it potable or otherwise
(Patrick and Ford, 1990). For most human uses, as well
as some commercial ones, the quality of the water
is as important as its quantity. Possible sources
of groundwater contamination include agricultural
activities, underground storage tanks, landfills,
septic tanks, surface impoundments, mining wastes, and
land application of treated wastewater, open dumps,
cemeteries, construction excavations etc., (Enger and
Smith, 2000). Some of the health effects associated with
such contamination is infectious hepatitis, cholera,
typhoid and paratyphoid fever and dysentery (White,
1986). Heavy metals from various industrial activities
can equally contaminate groundwater and are known to
have serious health effects to human and animals.
According to (Eggleston et al., 1992), cassava in
the fresh form contains cyanide, which is extremely
toxic to human and animals, therefore a need for
processing it to reduce the cyanide content to safe
levels is inevitable. Some of the processed products
include gari, tapioca, industrial starch, acetone,
and alcohol. Cassava processing generates solid and
liquid residues that are hazardous to the environment
(Cumbana et al., 2007). Many toxic effluents and
compounds that have entered the environment as a
result of mans activities are biodegradable or
potentially biodegradable to less toxic compounds by
microorganisms (Ubalua, 2010).
Awka-North Local Government Area is an
agrarian area in Anambra State, Nigeria, with cassava as
one of the major agricultural products. The area
therefore has many cassava processing mills located
even near residential homes, thus generating and
discharging cassava effluent indiscriminately onto the
surrounding environment. Many boreholes are sunk in
the area, serving as the major sources of drinking water
supply to the people. Effluent from these mills could
possibly be contaminating these groundwater sources.
This study therefore examines these groundwater
sources for possible contamination by the effluent.

MATERIALS AND METHODS
Sample sources
A total of 144 water samples were collected
from six different boreholes located not more than
100 m
2
from each cassava processing mill in six selected
communities in Awka-North L. G. A. of Anambra State,
Nigeria. Twenty four samples each from borehole
systems at Mgbakwu, Isu-Aniocha, Urum, Amanuke,
Ugbene and Ebenebe communities were collected as
shown in Table 1.
Sample collection
Ground water samples were collected biweekly
for a period of three months (June-August, 2011)
in duplicates with sterile 1 L plastic can, using the
method described by (Cheesbrough, 2004) and
thereafter taken to the laboratory for analyses.
Bacteriological analysis
Water samples were aseptically subjected
to 10-fold serial dilution, after which 0.1 ml of
10
-3
dilution was used to inoculate duplicate Nutrient
and MacConkey agar plates for the determination of
Total Heterotrophic Bacterial Counts (THBC) and Total
Coliform Bacterial Counts (TCBC), using spread plate
technique as described by (Cheesbrough, 2004).
Inoculated plates were incubated at 37C and 45C for
18-30 h, for heterotrophic bacterial counts and total
Okechi et al.,2013
057 Journal of Research in Public Health (2013) 2(1): 056-062
coliform counts respectively. Identification of isolates
was carried out according to the methods described by
(Cheesbrough, 2004 and Fawole and Oso, 2004).
Physicochemical analysis
Analyses were based on American Public Health
Association guidelines (APHA, 1998). The pH was
determined in situ with Ph meter (Model 7413).
Percentage organic carbon was determined as
described by (Walkley-Black, 1942). Determination of
cyanogenic glycosides was by alkaline titration method,
also, calcium, and magnesium were determined
titrimetrically. Sodium, potassium, copper, manganese,
iron and zinc were all determined using UNICAM 191
Atomic Absorption Spectrophotometer (AAS).

RESULTS
The analysis of the 144 borehole water samples
revealed that the highest total heterotrophic and
coliform counts of 5.0x10
3
cfu/ml and 2.20x10
3
cfu/ml
were obtained from Urum and Amanuke borehole water
systems while Ebenebe recorded the least total
heterotrophic count of 1.10x10
3
cfu/ml. Isu-Aniocha and
Ugbene haboured no coliform. These are as shown in
Figure 1. Identification test revealed the isolation of
twenty distinct colonies from the six boreholes.
The bacteria belong to the following genera
Pseudomonas (83.33%), Bacillus (66.67%), Micrococcus
(33.33%), Proteus (16.67%), E. coli (50%), Salmonella
(33.33%), and Streptococcus (50%). This prevalence of
the isolates is as shown in Figure 2. The percentage of
the bacterial isolates in the borehole samples studied is
as shown in Table-2. From the table, of the seven
bacterial genera isolated, the borehole samples from
Urum and Amanuke each haboured 71.42%, Mgbakwu
recorded 28.57%, Ugbene and Ebenebe had 42.86% and
57.14%, respectively, while the borehole samples from
Isu-Aniocha recorded the least 14.29%. Table-3 shows
the result of the physicochemical analysis of the
borehole samples from these communities. The pH of
the water was acidic in all the samples. Sample from
Urum recorded the highest values in pH, TDS, %OC, CN
-
,
K, Mn, and Cu, while recording the least values for Na,
Ca, Mg, Fe and Zn. Borehole from Mgbakwu recorded
the least for TDS, CN
-
and Mn. Samples from Isu-Aniocha
recorded highest values for Ca, Mg, and Zn
.
Sample from
Amanuke recorded the highest value for Na, while
sample from Ugbene had the highest value for Fe. Other
parameters tested were within WHO recommended
Okechi et al.,2013
Journal of Research in Public Health (2013) 2(1): 056-062 058
Location Sample collection (rainy season) Number of Months Total number of samples
Urum Biweekly 3 (Jun -Aug ) 24
Mgbakwu Biweekly 3 (Jun -Aug ) 24
Isu-Aniocha Biweekly 3 (Jun -Aug ) 24
Amanuke Biweekly 3 (Jun -Aug ) 24
Ugbene Biweekly 3 (Jun -Aug ) 24
Ebenebe Biweekly 3 (Jun -Aug ) 24
Total 144 samples
Table 1 Sources of Borehole Water Samples
Figure 1 Log of Total Heterotrophic and Coliform
Counts from Different Communities
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limits for drinking water.

DISCUSSION
This study revealed the level of contamination of
the various borehole water samples. The presence of
Pseudomonas, Bacillus, Streptococcus, Micrococcus,
Proteus, Escherichia and Salmonella species in some of
the boreholes water samples analysed is indicative of
the poor microbial quality of the boreholes. The World
Health Organization recommended one E. coli colony
per 100 ml of water sample to be normal (WHO, 2006).
However, this indicator organism was found in large
numbers in three borehole samples with prevalence of
50%. The high total coliform counts present in some
of the borehole samples are indicative of possible
feceal contamination. The presence of this indicator
bacterium may suggest possible presence of pathogens
causing cholera, typhoid, and gastroenteritis. The high
prevalence of E. coli could also be attributed to
contamination of the boreholes by cyanogenic
glycosides from the surrounding mills. Some strains
of E. coli have been reported to degrade cyanide
(Boopathy, 2000 and Ubalua, 2010). Approximately 67%
of the samples haboured Bacillus sps. This may not be
Okechi et al.,2013
059 Journal of Research in Public Health (2013) 2(1): 056-062
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surprising as various species of the bacterium have been
reported to be actively involved in cyanogenesis
(Silva-Avalos et al., 1990 and Huertas et al., 2006).
Bacillus sps were not isolated from the borehole sample
from Mgbakwu, where cyanide was detected (Tables 2
and 3). Bacillus species have been associated with the
production of odour and taste in water (Ihejirika et al.,
2011).
The isolation of Micrococcus and Proteus
species, which are strong proteolytic organisms is not
surprising in view of their wide spread distribution in
both soil and water (Roggers et al., 1977). Although they
are known to be non-cyanogenic and non-pathogenic,
they can however bring about changes in water quality
(Wan, 1997). Three of the borehole samples haboured
Streptococcus sps, an organism known to live as
commensal in and on human body, and its presence in
the waters suggest poor human handling. The isolation
of Salmonella sps from Amanuke and Ebenebe borehole
samples indicates that the direct consumption of such
waters without treatment may be very dangerous. The
genus is implicated in salmonellosis. The typhi and
paratyphi sp cause typhoid and paratyphoid fever which
can be spread through contaminated water
(Willey et al., 2008). Salmonella species have also been
involved in cyanide degradation (Boopathy, 2000).
The very high percentage occurrence of
Pseudomonas sps (83.33%) in the water samples could
be attributed to their wide spread distribution
in aquatic and soil ecosystems as reported by
Roggers et al., (1977). The detection of cyanogenic
glycosides in most of the water samples tested could
equally explain the high percentage occurrence of the
organism, as these compounds could serve as carbon
and nitrogen sources to the organism. Some
Pseudomonas sps have been reported to be involved in
Okechi et al.,2013
Journal of Research in Public Health (2013) 2(1): 056-062 060
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cyanogenic glycosides degradation (Gallagher and
Manoil, 2001). Some species of Pseudomonas are
opportunistic pathogens, which do not normally
exist in animal host, but can establish infections
i n i mmu n e - c o mp r o mi s e d i n d i v i d u a l s
(Roggers et al., 1977). This organism is remarkably
resistance to many commonly used antibiotics, a fact
that makes infection by this organism even more
serious. However, this study was carried out in the rainy
season and the effect of seasonal variation on the
microbial biodiversity was not considered. Also, these
isolates were not tested for cyanogenesis activities, thus
further studies are recommended in these directions.
The borehole samples are all recorded with
acidic pH values. These values were outside the WHO
recommended normal range of 6.5-8.5 for drinking
water as reported by Al-Khatib and Arafat, (2009). The
low pH could have resulted from cyanide content of the
cassava mill effluent dumped carelessly on the
surroundings. Cyanogenic glycoside was detected in all
except the water sample from Mgbakwu. The source of
the cyanide could have the effluent discharged
indiscriminately around the processing mills. Ubalua,
2010 reported high loading rate of the effluent as the
paramount factor for movement into water. Low
concentration of cyanide has been reported in
groundwater with acidic soils (Meeussen et al., 1994).
Soil conditions such as low pH, high negative soil
charges, low clay content and anaerobiosis have been
reported to increase the mobility of cyanide in the soil
(Ubalua, 2010). Similar conditions were observed in
these study areas (Okechi et al., 2011). Iron and copper
were equally found to be above the WHO
recommended standard for drinking water in all the
samples. The high content of these metals in the
samples could be attributed to their ability to form
complexes with cyanide, thus increasing cyanide toxicity
(Cheeke, 1995 and McMahon et al., 1995). Other
parameters tested were however within WHO
recommended limits for drinking water.

CONCLUSION
The isolation of some cyanide degrading
bacteria and coliforms, as well as the low pH and higher
cyanogenic glycosides content of the water samples are
all indicative of the detrimental effects of the effluent
on the groundwater quality of the area, thus the need
for proper waste management practices are inevitable.

REFERENCES
AL-Khatib I and Arafat H. 2009. Chemical and
Microbiological Quality of Desalinated Water,
Groundwater and Rain-Fed Cisterns in the Gaza Strip,
Palestine. Elselvier. 1165-1170.

APHA 1998.Standard Methods for the Examination of
Water and Wastewater. 20
th
edn. American Public
Health Association.

Boopathy R. 2000. Factors Limiting Technologies.
Biores. Technol., 74:63-67.

Cheeke PR. 1995. Endogenous Toxins and Mycotoxins
in Forage Grasses and their Effects on Livestock. J. Ani
Sci., 73:909-918.

Cheesbrough M. 2004. Biochemical Test to Identify
Bacteria: District Laboratory Practice in Tropical
Countries. 2
nd
edn. Cambridge University Press, London.
62-70.

Cumbana AE, Mirione E, Julie Cliff JC and Bradbury JH.
2007. Reduction of Cyanide Content of Cassava Flour in
Mozambique by the Wetting Method. Food Chemistry.
101(3):894-897.

Eggleston G, Bokanga M and Jean YW. 1992. Traditional
African Methods for Cassava Processing, Utilization and
Research Needs. In: M.O. Akoroda and O. B Arene (eds.).
Proceedings of 4
th
Tropical Root Crops. East African
Branch, Kinshasa, Zaire, Dec. 5-8, 1989. 36.
Okechi et al.,2013
061 Journal of Research in Public Health (2013) 2(1): 056-062
Enger ED and Smith BF. 2000. Environmental Science - A
Study of Interrelationships. 7
th
edn. McGraw Hill Co Inc.
New York.

Fawole MO and OSO B A. 2004. Characterization of
Bacteria: Laboratory Manual of Microbiology. 4
th
edn.
Spectrum Books Limited. 24-33.

Gal lagher LA and Manoi l C. 2001.
Ps eudomonas aer ugi nos a PAOI Ki l l s
Caenorhabditis elegans by Cyanide Poisioning. J.
Bacterial., 183(21):6207-6214.

Huertas M-J, Luque-Almagro VM, Martinez-Luque M,
Blasco R, Moreno-Vivian C, Castillo F and
Roldan M-D. 2006. Cyanide Metabolism of
Pseudomons pseudoalcaligens CECT5344: Role of
Siderophores. Bioch Soc Transac 34(1): 152-155.

Ihejirika OC, Ihejirika CE, Nwachukwu MI, Ebe TE and
Uzoka CN. 2011. Water Born Diseases Transmission
Potential of Sachet Waters Sold in Owerri, Eastern
Nigeria. Journal of Research in Public Health.
1(1): 001-006.

McMahon JM, White WLB and Sayre RT. 1995.
Cyanogenesis in Cassava (Manihot esculanta Cranz). J.
Exp Bot., 46: 731-741.

Meeussen JCL, Keizer MG, Van Riemsdijk WH, De Haan
FAM. 1994. Solubility of Cyanide in Contaminated Soils.
J Environ Qual., 23: 785-792.

Okechi RN, Chukwura EI, Azuwuike CO and Ihejirika
CE. 2011. The Impact of Cassava Mill Effluent on the
Total Aerobic Bioload and Physicochemical Properties of
the Soil. Journal of Biodiversity and Environmental
Sciences. 1(6), 112-117.

Patrick RE and Ford JQ. 1990. Groundwater
Contamination in the United States. A Publication of
UNEP-Industry and Environment. 29-30.

Rogers YS, Edward AA and John LI. 1977. General
Microbiology. 4
th
edn. MacMillan Publishers Ltd,
London. 595-596.


Silva-Avalos J, Richmond MG, Nagappan O and Kunz
DA. 1990. Degradation of the Metal-Cyano Complex
Tetracyanonickel-ate (11) by Cyanide Utilizing Bacterial
Isolates. Appl Environ Microbiol., 56:3664-3670.

Ubalua AO. 2010. Cyanogenic Glycosides and the Fate
of Cyanide in the Soil. Australian J. Crop Sc.,
4(4):223-237.

Walkey A and Black P. 1942. A Critical Examination of a
Rapid Method of Determining Organic Carbon in Soils.
Soil Sc., 251-265.

Wan P. 1997. Health Impact of Water Supply and
Sanitation Intervention. Waterfront Issue, 10:10-21.

Willey JM, Sheerwood LM and Woolverton CJ. 2008.
Prescott, Harley, and Kleins Microbiology. 7
th
edn.
McGraw Hill Co. Inc., New York.

White GC. 1986. Chlorination of Potable Water. The
Handbook of Chlorination. 2
nd
edn. Von Nostrand
Reinhold. New York. 256-393.

World Health Organization. 2006. Guidelines for
Drinking Water. First Addendum to the 1
st
edn. Vol.1,
Recommendations, Geneva, 595.
Okechi et al.,2013
Journal of Research in Public Health (2013) 2(1): 056-062 062
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