c14123361t Project Final

Crispen Munashe Mazodze
C14123361T
BSc (Hon) Chemistry
Level 2.1
Analysis of the microbiological contaminants of the groundwater at selected
industrial and residential areas of Chinhoyi
March 2015
Supervisor: Dr Munyuki
Munashe C Mazodze c14123361t

BSc Chemistry 2.1march 2015
Contents
I Acknowledgements
II Dedication
Chapter 1
1. Introduction
Chapter 2
Literature review
2.0 Introduction
2.1 Definition of ground water
2.2 Sources of groundwater contamination
2.3 Total coliform
2.3.1 Health effects of total coliform
2.4 Biological oxygen demand
2.4.1 Health effects of BOD
2.5 Chemical oxygen demand
2.5.1 Health effects of chemical oxygen demand
Chapter 3
Sampling, sample handling and preservation
3.0 Introduction
3.1 Sampling order
3.2 Equipment decontamination
3.3 Sample handling and preservation
3.4 Sampling points
3.4.1 Catholic university
3.4.2 Alaska
3.4.3 Ruvimbo
3.4.4 Delta beverages
3.4.5 White city
Chapter 4
Experimental design
4.0 introduction
4.1 biological oxygen demand
4.1.1 Instrumentation
4.2. Total Coliform
4.3. Chemical Oxygen Demand
Chapter 5
Results
5.0 Total Coliform
5.1 Biological Oxygen Demand
5.2 Chemical Oxygen Demand
Chapter 6
Discussion
6.1 Total Coliform
6.2 Chemical Oxygen Demand
6.3 Biological Oxygen Demand
Chapter 7
Conclusion
References
Appendix A
Appendix B
Acknowledgements
Special thanks to my supervisor Dr Munyuki for his dedication to the

success of this project and his tireless effort to bring the best out of me time
and again until the end. My sincere gratitude to Mr. Mudziwapasi for
allowing me to work in the Bio-tech Laboratory and helping me with all
experimental procedures outlined in this study. I would also want to thank
my self for working tirelessly for the success of this project.
Dedication
This one goes to my dad and mum for their countless support which made
this project feasible.
Chapter 1
1. Introduction
Solid and waste water disposal and other land use related activities
constitute some of the sources of environmental pollution. One of the
consequences of environmental pollution is the deterioration of ground and
surface water. Mankind depend on water for sustenance therefore it is
important for it be of good quality.
Chinhoyi is located in the north-central part of Zimbabwe. It lies west of
the Hunyani River and lies on the Chirundu highway that goes from Harare
to Lusaka. Chinhoyi is the center of a productive agricultural area. Some of
the crops that are grown include tobacco, corn and beans. Cattle are also
raised for beef. There is some mining activities that are also going on in
Chinhoyi for copper and mica. The town is also the gateway to the Chinhoyi
(limestone) Caves, 8 km west.
The town draws its potable water supply from Hunyani River and supplies
it to all its residents throughout the year but the supplied water is in
adequate. The inadequacy has promoted the adoption and implementation
of water conservation measures with consideration being given to
groundwater as main supplementary source. The Municipality of Chinhoyi
drilled several boreholes around different locations found in Chinhoyi for
use by the public. Some affording local residents also drilled their private
boreholes and wells for their own private use.
Unlike public water systems, which are regulated by the Zimbabwe Water
Authority (ZINWA), private wells are the responsibilities of residents.
ZINWA standards do not apply to private wells. Currently water from these
boreholes is used for domestic and agriculture among other uses. For
residents who own boreholes/wells, good quality drinking water depends
on a multi-barrier approach to contamination that includes well monitoring
and maintenance, locating the well away from points of contamination, and
protecting the watershed from excessive pollutant and sediment runoff.
This study focuses on analyzing the microbiological contaminants of
ground water such as bio oxygen demand (BOD), total coliform and
chemical oxygen demand (COD) and the main objectives of this project are
1. Analyzing the amounts of microbiological components present in
sources of ground water sources under study
2. Establishment of the levels of contamination inflicted by
microbiological components
3. Exploring the feasibility of use of the groundwater for domestic,
commercial and industrial purposes

4. In case of contamination recommend a remedy for sustenance of
man crops and animals
5. Device a way basing on the project data to maintain high quality
groundwater for everyday use
Chapter 2
Literature review
2. Introduction
Typically, groundwater is thought of as liquid water flowing through
shallow aquifers, but, in the technical sense, it can also include soil
moisture, permafrost (frozen soil), immobile water in very low
permeability bedrock, and deep geothermal or oil formation water (Iyer
2003). Groundwater is hypothesized to provide lubrication that can
possibly influence the movement of faults. It is likely that much of Earth's
subsurface contains some water, which may be mixed with other fluids in
some instances Greenburg (2005). Groundwater may not be confined only
to Earth. The formation of some of the landforms observed on Mars may
have been influenced by groundwater. There is also evidence that liquid
water may also exist in the subsurface of Jupiter's moon Europa Greenburg
(2005). Groundwater is a highly useful and often abundant resource.
However, over-use, or overdraft, can cause major problems to human users
and to the environment. The most evident problem (as far as human
groundwater use is concerned) is a lowering of the water table beyond the
reach of existing wells. As a consequence, wells must be drilled deeper to
reach the groundwater. Groundwater is also ecologically important. The
importance of groundwater to ecosystems is often overlooked, even by
freshwater biologists and ecologists. Groundwater sustain rivers, wetlands,
and lakes Harilal (2004).
2.1 Definition of Groundwater

Groundwater is the water found underground in the cracks and spaces in
soil, sand and rock. It is stored in and moves slowly through geologic
formations of soil, sand and rocks called aquifers Eaton et al (2015).
2.2 Sources of Groundwater Contamination

Contamination is defined as the introduction of any undesirable physical,
chemical or microbiological material into a water source. There are two

types of contamination sources: point sources and non-point sources
(APHA 1996), (APHA 1998). Point sources include landfills, leaking
gasoline storage tanks, leaking septic tanks and accidental spills; as the
name implies we can point directly to the source of contamination. A point
source is technically defined as any discernable, confined and discrete
conveyance from which pollutants are or may be discharged. Non-point
sources can be less obvious and can include naturally occurring
contaminants, such as iron, arsenic and radiological and runoff from
parking lots, pesticides and fertilizers that infiltrate the soil and make their
way into an aquifer Mayur (2007).
There are numerous regulations that address both point sources and nonpoint sources of contamination. In addition to these regulations, it is
important for citizens to become aware of how their actions can affect the
contamination of groundwater and take action to reduce any potential
contamination. Many factors can affect how groundwater becomes
contaminated. The depth of a well is an obvious factor because a
contaminant has to travel farther in deeper wells. The soil and formations
the contaminant travels through act as a natural filter APHA (1998).
According to Greenburg (2005) the underlying geologic formations
additionally affect the possibility of contamination in two ways. First, the
geological formation can be a source of contamination; there are various
rock formations that contain minerals such as calcium, magnesium, iron,
arsenic and various radiological contaminants. While some of the minerals
may not cause any known health effects, naturally occurring arsenic and
radiological contaminants are known carcinogens. Second, the formation
can slow the contaminant or it may have the opposite effect, for example,
contaminants can travel more quickly through sand than clay. The amount
of rainfall in a given area will contribute to contamination as well; the more
rain that falls, the more water is filtered down to recharge the aquifer.
Rainfall that seeps into the ground can carry these contaminants into the
aquifer.

2.3.0 Total coliform

Coliforms are bacteria that are always present in the digestive tracts of
animals, including humans, and are found in their wastes Harilal (2004),
Wallace (2000). They are also found in plant and soil material. Water
pollution caused by fecal contamination is a serious problem due to the
potential for contracting diseases from pathogens (disease causing
organisms). Frequently, concentrations of pathogens from fecal
contamination are small, and the number of different possible pathogens is
large Wallace (2000).
As a result, it is not practical to test for pathogens in every water sample
collected EPA (2015). Instead, the presence of pathogens is determined
with indirect evidence by testing for an "indicator" organism such as
coliform bacteria. Coliforms come from the same sources as pathogenic
organisms. Coliforms are relatively easy to identify, are usually present in
larger numbers than more dangerous pathogens, and respond to the
environment, wastewater treatment, and water treatment similarly to many
pathogens Hateras (2000). As a result, testing for coliform bacteria can be a
reasonable indication of whether other pathogenic bacteria are present.
The most basic test for bacterial contamination of a water supply is the test
for total coliform bacteria Hateras (2000). Total coliform counts give a
general indication of the sanitary condition of a water supply. Total
coliforms include bacteria that are found in the soil, in water that has been
influenced by surface water, and in human or animal waste. Fecal
coliforms are the group of the total coliforms that are considered to be
present specifically in the gut and feces of warm-blooded animals Hateras
(2000). Because the origins of fecal coliforms are more specific than the
origins of the more general total coliform group of bacteria, fecal coliforms
are considered a more accurate indication of animal or human waste than
the total coliforms.
Escherichia coli (E. coli) is the major species in the fecal coliform group. Of
the five general groups of bacteria that comprise the total coliforms, only E.
coli is generally not found growing and reproducing in the environment.
Consequently, E. coli is considered to be the species of coliform bacteria
that is the best indicator of fecal pollution and the possible presence of
pathogens Hateras (2000), Wallace (2000).
2.3.1 Health Effects of Total Coliform

Coliform or other bacteria in groundwater will not necessarily make you ill.
However, since these organisms are present, other disease-causing
organisms may also be present Hateras (2000). . Health symptoms related
to drinking or swallowing water contaminated with bacteria generally range
from no ill effects to cramps and diarrhea (gastrointestinal distress)
Hateras (2000). Wallace (2000). The presence of coliform bacteria in
drinking water is generally a result of a problem with water treatment or
the pipes which distribute the water, and possible contamination by
disease-causing microorganisms (although the coliform themselves are not
harmful). Disease symptoms may include diarrhea, cramps, nausea, and
possibly jaundice, and any associated headaches and fatigue Hateras
(2000), Wallace (2000), EPA (2015).
Fecal coliforms or E. Coli, are a particular type of coliform bacteria. Their
presence in drinking water is more serious than other coliform bacteria
because they are disease-causing, and also indicate that drinking water has
been contaminated by sewage or animal wastes that contain other disease
causing microorganisms. This type of contamination can cause severe
diarrhea, cramps, and nausea Wallace (2000), Hateras (2000). Because
fecal coliform contamination is more severe than contamination by other
types of coliform bacteria, Treatment Technique Violations: If water is
inadequately treated, microbiological contaminants in the water may cause
diarrhea, cramps and nausea. Two other common waterborne diseases
are giardiasis and cryptosporidiosis; both cause intestinal illness. E.
coli 0157:H7 has also been associated with drinking contaminated water
and can cause intestinal illness. In very rare cases, it can cause hemolytic
uremic syndrome, a serious kidney condition National Science and Tech
(2003).
2.4.0 Biological Oxygen Demand (BOD)

Most natural waters contain small quantities of organic compounds.
Aquatic microorganisms have evolved to use some of these compounds
as food. Microorganisms living in oxygenated waters use dissolved oxygen
to oxidatively degrade the organic compounds, releasing energy which is
used for growth and reproduction Manivaskam (1986), Manivaskam
(2005). Populations of these microorganisms tend to increase in proportion
to the amount of food available. This microbial metabolism creates an

oxygen demand proportional to the amount of organic compounds useful as
food. Under some circumstances, microbial metabolism can consume
dissolved oxygen faster than atmospheric oxygen can dissolve into the
water or the autotrophic community (algae, cyanobacteria and
saprophytes) can produce Manivaskam (2005). Fish and aquatic insects
may die when oxygen is depleted by microbial metabolism. Biochemical
oxygen demand is the amount of oxygen required for microbial metabolism
of organic compounds in water. This demand occurs over some variable
period of time depending on temperature, nutrient concentrations, and
the enzymes available to indigenous microbial populations. In agreement
with Trivedi (1986) and Manivaskam (2005) the amount of oxygen
required to completely oxidize the organic compounds to carbon dioxide
and water through generations of microbial growth, death, decay, and
cannibalism is total biochemical oxygen demand (total BOD). Total BOD is
of more significance to food webs than to water quality. Dissolved oxygen
depletion is most likely to become evident during the initial aquatic
microbial population explosion in response to a large amount of organic
material. If the microbial population deoxygenates the water, however, that
lack of oxygen imposes a limit on population growth of aerobic aquatic
microbial organisms resulting in a longer term food surplus and oxygen
deficit. Drinking water usually has a BOD of less than 1mg/L but when BOD
value reaches 6mg/L, the water is doubtful in purity WHO (2002). The
determination of BOD is used in studies to measure the self-purification
capacity of water bodies WHO (2002). It serves regulatory authorities as
means of checking on the quality of effluents discharged to water bodies
and it is also useful in design of treatment facilities. It is the only
parameter, to give an idea of the biodegradability of any sample and selfpurification capacity of any water body WHO (2002). BOD is among the
most important method in sanitary analysis to determine the polluting
power, industrial wastes and polluted ground water. It serves as a measure
of the amount of clean diluting water required for successful disposal of
sewage waste by dilution WHO (2002), Manivaskam (2005).
2.4.1 Health effects of biological oxygen demand

The presence of high BOD may indicate fecal contamination or increases in
particulate and dissolved organic carbon from non-human and animal
sources that can restrict water use and development, necessitate expensive
treatment and impair ecosystem health WHO (2002), Manivaskam (1986),
Trivedi (1986). Human ill health due to water quality problems can reduce
work capability and affect children's growth and education Trivedi (1986).
Increased concentrations of dissolved organic carbon can create problems
in the production of safe drinking water if chlorination is used, as
disinfection by-products, such as trihalomethanes and other compounds
toxic to humans, may be produced. Increased oxygen consumption poses a
potential threat to a variety of aquatic organisms, including fish Trivedi
(1986). The BOD content of a water body is among the most important
water quality characteristics necessary for protecting fish and aquatic life.
Low DO levels can induce fish kills and reduce reproduction rates in
aquatic biota Standard Methods (2005)
2.5.0 Chemical oxygen demand (COD)

Chemical oxygen demand (COD) is a measure of the capacity of water to
consume oxygen during the decomposition of organic matter and the
oxidation of inorganic chemicals such as ammonia and nitrite Hateras
(2000), Trivedi (1986) Iyer (2003). COD measurements are commonly
made on samples of waste waters or of natural waters contaminated by
domestic or industrial wastes. Chemical oxygen demand is measured as a
standardized laboratory assay in which a closed water sample is incubated
with a strong chemical oxidant under specific conditions
of temperature and for a particular period of time. A commonly used
oxidant in COD assays is potassium dichromate (K2Cr2O7) which is used in
combination with boiling sulfuric acid (H2SO4) Iyer (2003). Because this
chemical oxidant is not specific to oxygen-consuming chemicals that are
organic or inorganic, both of these sources of oxygen demand are measured
in a COD assay. Chemical oxygen demand is related to biochemical oxygen
demand (BOD), another standard test for assaying the oxygen-demanding
strength of waste waters Harilal (2004). However, biochemical oxygen
demand only measures the amount of oxygen consumed by microbial
oxidation and is most relevant to waters rich in organic matter. It is
important to understand that COD and BOD do not necessarily measure
the same types of oxygen consumption Appleton (2007). For example, COD
does not measure the oxygen-consuming potential associated with certain
dissolved organic compounds such as acetate. However, acetate can be
metabolized by microorganisms and would therefore be detected in an

assay of BOD APHA (1998). In contrast, the oxygen-consuming potential
of cellulose is not measured during a short-term BOD assay, but it is
measured during a COD test Hateras (2000).
2.5.1 Health effects of high chemical oxygen demand
High levels of COD in water often correlate with threats to human health
including toxic algae blooms bacteria from organic wastes and seafood
contamination WHO (2002), Trivedi (1986). According to the WHO
prescribed limits high COD levels decrease the amount of dissolved oxygen
available for aquatic organisms. Low (generally under 3 mg/L) dissolved
oxygen, or hypoxia, causes reduced cell functioning, disrupts circulatory
fluid balance in aquatic species and can result in death of individual
organisms as well as large dead zones. Hypoxic water can also release
pollutants stored in sediment.

Chapter 3
Sampling and sampling points
3.0 introduction
Sampling is the process of selecting units (e.g., water, soil) from a
population of interest so that by studying the sample we may fairly
generalize our results back to the population from which they were chosen
Harilal (2004). Water quality data are as good as the water samples from
which the measurements are made Trivedi (1986). Even the most precise
study of water analysis of water sample cannot compensate for improper or
poorly executed sampling procedure or for physical or chemical alterations
of a sample due to inappropriate sample collections, transport or storage.
ESA (2015)
3.1 Sampling order

When previous water quality data is available, the least contaminated wells
should be sampled first, and the order should proceed in such a way that
the most contaminated wells are sampled last. When contaminant
distribution is unknown, begin with wells up gradient of likely contaminant
source(s), continue with down gradient wells, and finish with wells in or
closest to suspected contaminant source(s) Harilal (2004). The samples in
this project were sampled in accordance to Harilal (2004).
3.2.0 Equipment decontamination

All sampling related equipment should be decontaminated. The need for
decontamination can be avoided by use of new disposable equipment that is
certified as clean and between each sampling point all the equipment
should be decontaminated and after cleaning, residues should be reinspected for or other substances that may survive normal cleaning. If
inspection reveals that decontamination was insufficient, implement

additional measures as needed and document APHA (1998), Harilal
(2004). Decontaminate equipment in the following manner:
Clean inside and out with a laboratory grade detergent (Liquinox or
equivalent)/clean-water solution, applied with a scrub brush where
practical.
Rinse with tap water followed by a final rinse with distilled or deionized
water. Inspect for remaining particles or surface film, and repeat cleaning
and rinse procedures if necessary.
Sampling equipment that comes in contact with free product or heavily
contaminated areas requires use of a desorbing agent (dilute solution of
water and isopropanol or methanol) followed by a thorough tap water rinse
and a final distilled or deionized water rinse.
3.3. Sample Handling and Preservation

Sample preservation is not practical because biological activity will
continue after the sample has been taken, changes may occur during
handling and storage Harilal (2004). If analysis is to be carried out within 2
hours of collection, cool storage is not necessary. If analysis cannot be
started within 2 hours of sample collection to reduce the change in sample
keep all samples at 40 c. Do not allow samples to freeze. Trivedi (1986) do
not open sample bottle before analysis. Analysis should begin within six
hours of sample collection. The following precautions should be observed
while performing the experiment: Dilution water should be prepared 3 to 5
days before all experiments and if any sign of biological growth is shown
samples should be discarded and sample should be adjusted to a pH
between 6 and 7.5, using sulphuric acid for samples with pH of alkaline side
i.e. greater than 7.5 or sodium hydroxide for sample with pH in the acidic
side i.e., less than 6.5. Eaton et al (2015). The samples were handled and
preserved according to Harilal (2004), Trivedi (1986) and Eaton et al
(2015)
3.4.0 Sampling points
The selected sampling points were not really distributed because of the lack
of boreholes around locations within Chinhoyi and also because the
mucipality of Chinhoyi denied us access to its boreholes and wells with

reasons best known to the municipality but hower some we got access to
private boreholes and wells and the project was successfully completed
3.4.1 Catholic university

Located in the vicinity of the public rank were various activities occur and
also improper sewage and waste disposal is practiced. There is likely to be
contaminants in this area.
3.4.2 Alaska
Situated near an iron smelting company which is no longer functional.
there is also some agricultural practices being done at the site and these are
animal rearing and crop production even though they are at a very small
scale they might have impact
3.4.3 Ruvimbo
Due to financial crisis in the country the residents of these area havent
received piped water from ZINWA and hence they adopted use of boreholes
and wells for their sustenance. The water sources are however crowded, the
borehole to people ration is very low and this might lead to contamination.
They also use improperly constructed Blair toilets which is of great concern.
3.4.4 Delta Beverages

Various activities like beer brewing, truck servicing and storage of

petroleum occur here and they use different types of chemical which might
be threat if improperly disposed. And ambient temperatures they use for
brewing the beer might be worthy of consideration.
3.4.5 Rusununguko White City
A small residential and business areas still growing. The borehole is located
where heavy truck wait for transportation of stones and silt for building
purposes and they happen to be serviced there.

Chapter 4
Experimental Design
4.0 Introduction
The unavailability of funds and resources limited the choice of
experimental methods used extend and this could have affected the quality
of results which were. The method of analysis were chosen based on the
resources we had.
4.1.0 Biological Oxygen Demand

Sample test procedures and pretreatment were carried out before the initial
BOD test.* the incubation bottles were thoroughly cleaned with a phosphate
free detergent and drained. The bottles were inverted in a water bath to
prevent contamination between cleaning and use. The dilution water was
prepared by adding 1 mg/l of each phosphate buffer, MgSO 4 (aq), CACl2 (aq)
and FeCl3 (aq), it was the aerated using an organic free filtered air and it was
brought to 200 C both with the sample without supersaturating it. The
sample size dilution was determined*. The samples were diluted and the
samples initial dissolved oxygen concentration in parts per million (ppm)
was measured and incubated at 200 c in a dark incubator for 5 days. After 5
days the DO of all samples was measured and the BOD5 was calculated.
Incubator and Accumate Dissolved Oxygen meter Xl40

4.2.0 Total Coliform

Samples were mixed thoroughly by inverting the sample bottles several
times. 50ml of water was added using a graduated pipette to a 6*100ml
bottle with 50ml of double strength McConckey purple broth (ds MACB)
containing an inverted Durham tube. 10ml of water was added using a
graduated pipette to each 6*5 universal bottles containing 10 ml ds MACB.
To 6*five bijou bottles containing 2ml single strength MACB an equal
amount of sample from different samples was added making sure that the
Durham tubes were completely covered with broth before incubation. All
the 65 bottles containing broth and an equal amount of sample were
incubated in an incubator at 37 0 c for 48 hrs. And after incubation they
were examined and counted each bottle each bottle which had produced
both acid (color change from purple to yellow) and gas (collection of gas
bubble in Durham tubes). Using the most probable number table (MPN)
the most probable number of total coliform bacteria in a 100ml water
sample were counted and recorded. A loopful from each positive bottle was
taken and inoculated into 6*10ml in MACB with inverted Durham tubes
and another loopful from each positive bottle and inoculate in 6*1ml bijou
bottles. These were incubated at 44 0 c for 48 hrs. in an incubator. After
incubation they were carefully examined and counted each bottle with
MACB which had produced gas and color change. Referring to the MPN
table the MPN of Faecal coliform bacteria in 100ml water sample were
determined. By adding few drops of Kovacs reagent to each peptone water
bottle and checking for indole production the presence of E.Coli was
determined. All water sample results are reported by giving the MPN of
fecal coliform bacteria and E.Coli in 100ml water samples and counts of
total coliform bacteria and fecal coliform bacteria are determined by
reference to the MPN table which is found appendix A.

Incubator
4.3.0 Chemical Oxygen Demand.

10.21g of K2Cr2O7 were added to a 500ml Erlenmeyer flask and diluted up to
the mark with distilled water. 50ml of the sample was put into a reflux
condenser and 1g of glass beads were added followed by 70ml of
concentrated H2SO4 and 25 ml of K 2Cr2O7 . The mixture was refluxed for
2hrs and cooled. The absorbance of distilled water and the dichromate was
measured at 420 and at 600nm and also for the mixture. The concentration
of the dichromate remaining after oxidation using lambda at 420nm was
calculated and using the relationship below the concentration of oxygen
was computed.
K2Cr2O7 + 7H2SO4 + 6e-
Cr2(SO4)3 + 7H2O + 2K+ .1
O2 + 4H+ + 4e...2
2H 2O
Overall equation:
2K2Cr2O7 + 14H2SO4
3 [O2 ]+ 2Cr2(SO4)3 + 8 H2O + 4K+ + 12H+

Chapter 5
Results
5.0 Total Coliform
5.1.0 Table 1: total coliform/ 100ml sample
Site
Total
Fecal
e.
coliform/100ml coliform/100ml coli/100ml
Alaska 25
Ruvimbo
borehole
Ruvimbo 9538
Delta beverages
Alaska frost
cresant
Catholic
university
Rusununguko
8
13
5
0
+
-
13
5
13
1
0
0
+
-
8
3

0
8
Fig 5.1.1 Bar graph 1 fecal coliform/100ml sample

Error bars are not shown because resources didnt allow for replicates to be
done. The bar graphs shows levels of fecal coliforms in 6 selected sampling
sites which are in the range of 3-13/100ml sample the highest being
recorded in Ruvimbo and the least being recorded in Rusununguko

Fig 5.1.2Bar graph 2 fecal coliform /100ml sample

done. The data in the bar graphs shows the level of contamination due to
human waste with areas such as Alaska, delta, catholic recording a zero
fecal contamination. Areas like Ruvimbo and White city recorded a fecal
contamination with Ruvimbo recording the highest coliform units /100ml
sample
5.2.0 Biological oxygen demand

5.2.1Table 2: BOD values for 6 sites
site
Ruvimbo borehole
Ruvimbo well
Delta beverages
Alaska frost cresant
Catholic university
Rusununguko

BOD (ppm)
1.05
3.23
1.21
1.21
1.21
1.1o
Fig 5.2.2 BOD Values For all Sampled Sites

done. Hence statistical analysis wasnt possible. The bar graph show BOD
values for all 6 sampled sites which are in the range of 1-3.5 ppm with the
highest BOD value being recorded in Ruvimbo and the lowest in Alaska. All
the BOD values were within the WHO prescribed limits(6mg/L) showing a
degree of purity in the water samples.
5.3.0chemical oxygen demand (COD)

5.3.1 Table 3 COD for all the 6 samples

Site
Ruvimbo well
Ruvimbo borehole
Alaska frost
Delta
catholic
White city
COD (ppm)
326.4
273.87
192
331.68
433.44
265.25
Fig 5.3.2
Bar
graph
for COD
test
Error
bars are
not
shown
because
resources
didnt
allow for
replicates to be done. Hence statistical analysis wasnt possible. The data in
the fig above shows BOD values in 6 selected areas for sampling. The BOD
values were very low and well within the WHO limits for drinking water
(6mg/l). The range was from 100- 350mg/l with Catholic recording the
highest and Rusunungungo. High content of biodegradable and nonbiodegradable matter mainly constitute to high COD values

Chapter 6
Discussion
6.0 total coliform
6.1 Ruvimbo well
The total coliform and fecal coliform were 13 and 5/100ml sample
respectively. E.Coli was present in the sample and this showed us recent
fecal contamination. This is possible because the well is improperly
constructed and the level of the water is too high. Also the fact that they
local residents carry out their washing around the well this allows for
leakages of the dirty which poses a serious health effect to the local
residents. This water source is contaminated and unfit for drinking
purposes.
6.1.1 Ruvimbo Borehole
E.Coli was detected which some of its strains are toxic to human health and
also the levels of total coliform and fecal coliform were 8 and 5/100ml
sample signifying the fact that the water is contaminated and unfit for
human consumption. The contamination might be because of the gradient
at which the borehole is situated, the septic tank might be leaking into the
borehole water.
6.1.2 Delta Beverages and Catholic University
No E.Coli or fecal coliform were detected in the water meaning to say the
water was not contaminated by human feces and no in any way are sources
of fecal contaminants near the sampling points. Total coliform was detected
in the water at 8 and 5mg/100ml for Delta beverages and catholic
University respectively and these are just indicator organisms which
indicates presents of other bacteria but not meaning the water is fit of
human consumption. The results also showed that they are no leakages of
septic tanks into the groundwater.
6.1.3 Alaska Frost Cresant, Alaska 25 white city

E coli was detected in the water samples which is against the World health
Organization which says E.Coli should not be detected in 100ml of every
sample hence the water is contaminated and the total coliform were 13, 8
and 3/ 100ml sample . These are just indicator organisms which shows the
presence of bacteria in water sample and shows recent contamination.
Fecal coliforms were undetected showing that there are no sources of fecal
contaminants are within the sampling points and also there are no leakages
of septic tanks. These areas also practice animal rearing and small scale
farming this could also be sources of contaminants especially when good
hygienic practices are not enhanced.
6.2.0 Chemical oxygen demand
Chemical oxygen demand is a valuable water quality parameter. COD is a
measure of the oxygen equivalent of the organic matter in a water sample
that is susceptible to oxidation by a strong chemical oxidant, such as
dichromate. It is an index of organic content of water because the most
common substance oxidized by dissolve oxygen in water is organic matter
having biological origin i.e. dead plant and animal wastes (Singh, A, 2006).
COD values convey the amount of dissolved ox disable organic matter
including the non-biodegradable matters present in it. The value of COD
was found in the range of 194mg/l to 326 mg/l. Its value is higher than the
permissible limit prescribed by WHO. The increase in COD concentration
was found in the bottom water where organic matter is more (Prasad &
Qayyum, 1976).
6.3.0 Biological oxygen demand
The cardinal point about the solubility of oxygen in water is that it has an
inverse relationship with temperature, as shown in the table below. This
could give results which doesnt reflect actual values of BOD contained in
water The consequence is that the actual concentrations of DO in a
groundwater will be lowest in summertime when it is usually the case that
the risk of damage to a water supply source or of environmental pollution is
greatest, especially in areas developed as Chinhoyi were temperatures are
high during the summertime. However the values for BOD were in the
range form 1.05mg/l to 3.23 mg/l. all of them were in the range prescribed
by who which showed that there were no activities within ground water
source which consumed the total dissolved oxygen.
Chapter 7
Conclusion
The results showed that:
Data reveals that all the 6 sampling sites are polluted to some extent
Ruvimbo site is most polluted as indicated by a very high value of COD and
total coliform. The main sources of pollution of Ruvimbo sampling site are
the human settlement which outnumbers the sources of water and their
activities. Also the uses of bush toilets and improperly constructed Blair
toilets. Data also indicates that in all the 6 sampling sites total COD values
were found to be beyond the permissible limits as prescribed by WHO but
the BOD was well within the permissible limits as prescribed by WHO
standards. To improve the quality of water of all the 6 sampling sites should
be continuous monitored of the pollution level. Good hygienic practices
should be enhanced and washing around water sources were there are no
sinks should be avoided at all costs to keep the water clean and safe for use
by mankind.
References
Wallace, R. (2000). Maxcy-Rosenau-Last Public Health and Preventive
Medicine. Stamford, CT
Appleton & Lange. Bhandari N. S. and Kapil Nayal, E-Journal of
Chemistry, 5(2), 342 (2008)
Iyer C. S., Sindhu M., Kulkarni S. G., Tambe S. S. and Kulkarni B. D., J.
Environ. Monit. 5, 324 (2003)
Mayur C. Shah, Prateek Shilpkar and Sangita Sharma, Asian J. Chem.,

19(5), 3449 (2007)
Manivaskam N., Physico-chemical examination, sewage and industrial,
Pragati Prakashan, Meerut, (1986)
Trivedi P. K. and Goel P. K, Chemical and biological methods for water
pollution studies, Env. Publication, Karad, (1986)
APHA Standard methods for examination of water and waste water;
American Public health Association, Washington, DC, (1998)
APHA Standard methods for the examination of water and waste water,
Public Health Association, 19th ed., Washington, DC, (1996)
WHO, The guideline for drinking water quality recommendations; World
Health Organization; Geneva, (2002)
Manivaskam N, Physicochemical examination of water sewage and
industrial effluent, 5th Ed, Pragati Prakashan Meerut, (2005)
Harilal C. C., Hashim A, Arun P. R. and Baji S., Journal of Ecology,

Environment and Conservation, 10(2), 187 (2004)
U.S. EPA, National Recommended Water Quality Criteria,
http://www.epa.gov/waterscience/criteria/wqctable/index.html#U (last
visited June 7, 2015).
National Science & Tech. Council, An Assessment of Coastal Hypoxia and
Eutrophication in U.S. Waters, 2 (2003) available at
http://oceanservice.noaa.gov/outreach/pdfs/coastalhypoxia.pdf.
Ambient Aquatic Life Water Quality Criteria for Dissolved Oxygen
(Saltwater): Cape Cod to Cape Hatteras, 65 Fed. Reg. 71317, 71318 (Nov. 30,
2000).
Ecological Society of America, Hypoxia
http://www.esa.org/education_diversity/pdfDocs/hypoxia.pdf (last visited

june. 11, 2015).
Ecological Society of America, supra.
STANDARD METHODS, 5-14(Andrew Eaton, et al. eds., 21st ed. 2005).
Appendix A
Sampling point
Ruvimbo borehole
White city
borehole
Ruvimbo 9538
Alaska frost
cresant
delta
Rusununguko
Initial pH
7
6.8
Adjusted pH
-
7.3
7.2
6.8
6.5
Biological
Oxygen
Demand
Sample
test
procedure
It should be made sure that the sample if free from chlorine. Chlorine is
present dechlorination must be done using sodium sulphate which should
be added prior to testing. It should also made sure that the sample pH is
within a range of 6.5-7.5 if sample is not within the range sulphuric acid or
sodium hydroxide should be used to adjust sample pH. The sample also
need to have an existing adequate microbiological population. If the
microbiological is inadequate or unknown a seed solution of bacteria
should be added along with an essential nutrient buffer solution than
ensures bacteria population vitality. Specialized 300ml BOD bottles
designed to allow full filling with no air space should provide an airtight
seal should be used. The bottles should be filled with sample to be tested or
dilution water are added to reflect different dilutions and a blank as a
control. A DO meter should be used to measure initial dissolve oxygen after
being calibrated with a 0% dissolved oxygen solution.
Sample pre-treatment
Measure pH of the sample making sure that it is within the range 6.5-7.5,
adjust it by adding 1M NaOH OR 1M H 2SO4. Do not dilute the solution with
more that 0.5% of the acid/alkali. Determine the amount of reagent added
to adjust the pH. The present of chlorine is tested by adding 100ml of well
mixed portion of sample into a 250 ml Erlenmeyer flask followed by 10ml
of acetic acid and swirling to mix. 10ml of KI is also added followed by 1ml
starch indicator and letting stand for 15 minutes. If blue color doesnt
appear there is no chlorine and sample doesnt require further treatment
prior to BOD test. If sample contains chlorine (blue color appears)
0.0025M sodium sulphite should be added till the blue color disappears
noting the amount of the sulphite used. Add the calculated volume of
sodium sulphite which is the one needed to dechlorinate the selected BOD
sample, to the sample mix thoroughly and let stand for 15 minutes and
make sure that the pH is within the range after dechlorination.
Total coliform1
Most probable number table (MPN)
Volume
of 50ml
sample
in
each bottle
Number of 1
bottles used
0
0
0
0
0
0
0
0
0
0
0
0
1
1
1
10ml
1ml
0
0
0
1
1
1
2
2
2
3
3
4
0
0
0
0
1
2
0
1
2
0
`1
2
0
1
0
0
1
2
MPN
of
coliforms in
100ml of the
original
water
0
1
2
1
2
3
4
3
5
5
1
3
4
6
3
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
0
1
1
1
1
2
2
2
2
3
3
3
3
3
4
4
4
4
4
4
5
5
5
5
5
5
3
0
1
2
3
0
1
2
3
0
1
2
3
4
O
1
2
3
4
0
1
2
3
4
5
0
4
6
3
5
9
5
7
10
12
8
11
14
18
20
13
17
20
30
35
40
25
35
50
90
160
180+
)
Chemical oxygen demand
sample
Absorbance replicate
(nm)
average
Ruvimbo 9538
Ruvimbo borehole
Alaska frost
Delta
catholic
White city
0.064
0.050
0.036
0.066
0.062
0.052
0.064
0.0505
0.0355
0.065
0.0805
0.052
Calculations
0.064
0.051
0.035
0.064
0.099
0.052
The reaction equations are as follows and to add them together equation1 is
multiplied by factor 2 and equation 2 by factor 3 so that the electrons cancel
out and also equation 2 is reversed
K2Cr2O7 + 7H2SO4 + 6eO2 + 4H+ + 4e-
Cr2(SO4)3 + 7H2O + 2K+

2H2O
2K2Cr2O7 + 14H2SO4
3 O2 + 2Cr2(SO4)3 + 8 H2O + 4K+ + 12H+
Ratio K2Cr2O7 : O2
2 :3
Number of moles of O2 = n (K2Cr2O7)
Oxygen present in the sample = mg/L
= (n Mr) 1000
Number of moles K2CrO4 used =
=
=0.03472467709 moles
Concentration = number of moles/volume (dm3)
=*10000
= 6.945 * 10-2
Appling Beer Lamberts law
A = bc
=
=
= 9.41mol-1 nm

Concentration of K2Cr2O7 remaining after oxidation of all the

samples
sample
Ruvimbo
well
Ruvimbo
borehole
Alaska
frost
Delta
catholic
White
city
[ K2Cr2O7
] before
oxidation
0.0695
[ K2Cr2O7]
after
oxidation
6.8 10-3
[02]
COD
(ppm)
0.0102
326.4
0.0695
5.367 10-3
0.008055
273.87
0.0695
3.7725 10- 0.0056587

3
5
-3
6,91 10
0.010365
-3
9.03 10
0.013545
-3
5.526 10
0.008289
0.0695
0.0695
0.0695
192
331.68
433.44
265.25
The concentrations of all the samples was calculated using beer lamberts
law
A = bc
C=
Mass of oxygen in ppm = c*32*1000


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c14123361t Project Final

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Crispen Munashe Mazodze

Munashe C Mazodze c14123361t

Special thanks to my supervisor Dr Munyuki for his dedication to the

this project feasible.

commercial and industrial purposes

2.1 Definition of Groundwater

2.2 Sources of Groundwater Contamination

chemical or microbiological material into a water source. There are two

Munashe C Mazodze c14123361t

2.3.0 Total coliform

2.3.1 Health Effects of Total Coliform

2.4.0 Biological Oxygen Demand (BOD)

to the amount of food available. This microbial metabolism creates an

2.4.1 Health effects of biological oxygen demand

2.5.0 Chemical oxygen demand (COD)

metabolized by microorganisms and would therefore be detected in an

Munashe C Mazodze c14123361t

3.1 Sampling order

3.2.0 Equipment decontamination

inspection reveals that decontamination was insufficient, implement

3.3. Sample Handling and Preservation

mucipality of Chinhoyi denied us access to its boreholes and wells with

3.4.1 Catholic university

3.4.4 Delta Beverages

Various activities like beer brewing, truck servicing and storage of

Munashe C Mazodze c14123361t

4.1.0 Biological Oxygen Demand

Munashe C Mazodze c14123361t

4.2.0 Total Coliform

Munashe C Mazodze c14123361t

4.3.0 Chemical Oxygen Demand.

Cr2(SO4)3 + 7H2O + 2K+ .1

3 [O2 ]+ 2Cr2(SO4)3 + 8 H2O + 4K+ + 12H+

Munashe C Mazodze c14123361t

Munashe C Mazodze c14123361t

Fig 5.1.1 Bar graph 1 fecal coliform/100ml sample

Munashe C Mazodze c14123361t

Fig 5.1.2Bar graph 2 fecal coliform /100ml sample

5.2.0 Biological oxygen demand

Munashe C Mazodze c14123361t

Fig 5.2.2 BOD Values For all Sampled Sites

5.3.0chemical oxygen demand (COD)

5.3.1 Table 3 COD for all the 6 samples

Munashe C Mazodze c14123361t

6.1.3 Alaska Frost Cresant, Alaska 25 white city

Mayur C. Shah, Prateek Shilpkar and Sangita Sharma, Asian J. Chem.,

Harilal C. C., Hashim A, Arun P. R. and Baji S., Journal of Ecology,

http://www.esa.org/education_diversity/pdfDocs/hypoxia.pdf (last visited

Cr2(SO4)3 + 7H2O + 2K+

3 O2 + 2Cr2(SO4)3 + 8 H2O + 4K+ + 12H+

Munashe C Mazodze c14123361t

Concentration of K2Cr2O7 remaining after oxidation of all the

3.7725 10- 0.0056587

Munashe C Mazodze c14123361t

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