MAKERERE UNIVERSITY

COLLEGE OF ENGINEERING, DESIGN, ART AND TECHNOLOGY

SCHOOL OF ENGINEERING

DEPARTMENT OF CIVIL AND ENVIRONMENTAL ENGINEERING

SECOND YEAR INTERNISHIP REPORT.

STUDENT NAME: MUGISHA HAWAH

REGISTRATION NO: 20/U/0496

STUDENT NO: 2000700496

TRAINING COMPANY: HILTON FOUNDATION IN PARTNERSHIP

WITH AQUAYA INSTITUTE

DEPARTMENT SUPERVISOR: DR. ROBERT KYEYUNE KAMBUNGU

FIELD SUPERVISOR: PROF. KABENGE ISA

 DECLARATION
I, MUGISHA HAWAH, claim that this report is original and compiled by me. It includes
activities I undertook during the training of my second- year industrial training and the
research obtained from various engineering manuals and that it has never been presented
before.

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 ACKNOWLEDGEMENT
My sincere gratitude and appreciation go to the Management of Hilton Foundation in
partnership with Aquaya Institute for giving me an opportunity to train with them.

I also appreciate and acknowledge the guidance and knowledge given to me by Prof.
Kabenge Isa, Prof Zziwa, Mr. Rawling, Dr.Nakawuka Prosper and Dr. Joel from department
of Agricultural & Biosystems Engineering, Makerere University.

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 TABLE OF CONTENTS
DECLARATION.......................................................................................................................i

ACKNOWLEDGEMENT.......................................................................................................ii

LIST OF FIGURES................................................................................................................vi

1 INTRODUCTION............................................................................................................8

1.1 HISTORY AND BACKGROUND OF HILTON FOUNDATION.......................................................8

1.2 Water Quality Overview........................................................................................................9
1.2.1 Physical characteristics................................................................................................10
1.2.2 Chemical characteristics..............................................................................................12
1.2.3 Biological characteristics..............................................................................................13
1.3 CONTAMINANTS IN WATER.................................................................................................14
1.3.1 Sewage contaminants (e.g., residential, commercial and institutional contaminants) 14
1.3.2 Industrial contaminants...............................................................................................14
1.3.3 Agricultural contaminants............................................................................................15
1.3.4 Petroleum products.....................................................................................................15
2 WATER SOURCES.......................................................................................................16

2.1 WATER SOURCE TYPES.........................................................................................................16

2.1.1 Piped water into dwelling............................................................................................17
2.1.2 Piped water into yard/plot...........................................................................................17
2.1.3 Public tap/ standpipe...................................................................................................18
2.1.4 Borehole with hand pump...........................................................................................18
2.1.5 Mechanized borehole..................................................................................................18
2.1.6 Protected dug well.......................................................................................................19
2.1.7 Unprotected Dug Well.................................................................................................19
2.1.8 Protected dug well with hand pump............................................................................19
2.1.9 Protected Spring..........................................................................................................20
3 WATER SAFETY...........................................................................................................20

3.1 Sanitary Inspections.............................................................................................................20

3.2 CALIBRATION.......................................................................................................................22
3.3 Laboratory safety practices..................................................................................................22
3.3.1 Protection in the laboratory.........................................................................................22
3.4 GENERAL HAZARDS IN A LABORATORY................................................................................23
3.4.1 SOLUTION TO THE HAZARDS........................................................................................23
3.5 Waste management in the laboratory.................................................................................24

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 3.6 Risk hazards and Risk management.....................................................................................24
3.6.1 Risk management........................................................................................................25
3.6.2 Safety Signs in the laboratory......................................................................................25
3.7 Standards and Protocols......................................................................................................27
3.7.1 Laboratory Standard....................................................................................................27
3.7.2 International laboratory quality standards..................................................................28
3.7.3 Protocols......................................................................................................................28
3.7.4 Quality assurance and Quality Control.........................................................................29
1.7 DISINFECTION......................................................................................................................30
4 MATERIALS AND METHODS...................................................................................30

2.1 EQUIPMENTS USED..............................................................................................................30

2.1.1 Aquameter box............................................................................................................30
2.1.2 Aquaprobe...................................................................................................................31
2.1.3 AP-2000 Extension cable..............................................................................................32
2.1.4 Aquameter...................................................................................................................32
2.1.5 The chlorine tester.......................................................................................................33
2.1.6 The turbidity meters....................................................................................................33
2.1.7 Hand sanitizer and Alcohol wipes................................................................................34
2.1.8 Cooler with frozen ice packs........................................................................................34
2.1.9 Whirl-Pak bags.............................................................................................................35
2.1.10 Distilled water..............................................................................................................35
2.1.11 The manifold................................................................................................................35
2.1.12 Petri dishes..................................................................................................................36
2.2 SAMPLE PRESERVATION......................................................................................................36
2.3 THE CALIBRATION OF THE MULTI-PARAMETER METER.......................................................36
2.3.1 Procedure of the calibration of the multi-parameter meter........................................37
2.4 CALIBRATION OF THE TURBIDIMETER WITH STABLCAL STANDARDS...................................38
2.4.1 Calibration Verification (Verify Cal) procedure............................................................38
2.4.2 The cleaning of spectrophotometric cells....................................................................39
2.5 PREPARE FIELD SUPPLIES.....................................................................................................39
2.6 CONDUCT WATER SOURCE SURVEYS...................................................................................39
2.7 COLLECT SAMPLE FOR MICROBIAL ANALYSIS......................................................................40
2.8 TEST SAMPLES FOR PHYSICAL-CHEMICAL PARAMETERS.....................................................42
2.9 TEST PROCEDURE FOR TURBIDITY USING THE HACH 2100Q PORTABLE TURBIDIMETER.....42
2.10 TEST PROCEDURE FOR FREE CHLORINE...............................................................................43

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 2.11 DELIVER SAMPLES TO THE FIELD LAB FOR ANALYSIS...........................................................43
2.11.1 Quality Assurance / Quality Control Measures............................................................43
2.11.2 Procedure for microbial testing...................................................................................44
5 ANALYSIS AND DISCUSSION OF RESULTS.........................................................47

3.1 READING THE SAMPLES FOR MICROBIAL ANALYSIS............................................................47

3.2 Laboratory blank samples....................................................................................................48
3.2.1 Procedures...................................................................................................................49
3.2.2 Microbial survey results from the water point sample..................................................1
3.2.3 Water point survey results from the mwater app..........................................................2
3.3 Disinfecting the compact dry plates.......................................................................................5
3.3.1 Procedures.....................................................................................................................5
6 REFERENCE....................................................................................................................6

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 LIST OF FIGURES
Figure 1: The target countries in Africa.....................................................................................9
Figure 2: Contaminated water in bottles with colored metallic ions.......................................11
Figure 3: The bottles containing suspended particles in water................................................11
Figure 4: The industrial discharge into the water source.........................................................15
Figure 5: The motorcyclists near the surface water source......................................................16
Figure 6: Piped water inside the dwelling................................................................................17
Figure 7: Piped water outside the dwelling..............................................................................18
Figure 8: A bore hole with hand pump....................................................................................18
Figure 9: The protected dug well.............................................................................................19
Figure 10: Unprotected dug well..............................................................................................19
Figure 11: Some of the sanitary inspections............................................................................21
Figure 12: The laboratory safety gears.....................................................................................23
Figure 13: Some of the prohibition signs used in the laboratory.............................................26
Figure 14: The common warning signs....................................................................................27
Figure 15: Some of the mandatory signs in the laboratory......................................................27
Figure 16: The components of the Aquameter box..................................................................31
Figure 17: The different parts of the Aquaprobe.....................................................................31
Figure 18: Connecting the extension cable onto the Aquameter probe...................................32
Figure 19: The Aquameter showing the different parameter readings....................................32
Figure 20: The chlorine tester..................................................................................................33
Figure 21: HACH 2100Q portable turbidimeter with its sample cells.....................................34
Figure 22 showing a cooler bag with whirl pak bags seated on an ice pack............................34
Figure 23 showing the Whirl Pak bag......................................................................................35
Figure 24: The single branch manifold used for membrane filtration method........................35
Figure 25: Pulling water out of the sterile funnel on the manifold..........................................47
Figure 26: The compact dry plate with colonies......................................................................48
Figure 27: The compact dry plates with the bleach solution placed in the bucket....................7

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1 INTRODUCTION.
1.1 History And and Background Of of the Hilton Foundation
The Hilton Foundation began their work in water in 1990 to help eradicate Guinea worm in
Ghana. In 2002, they created the West Africa Water Initiative, a coalition across Ghana, Mali,
Niger, and Burkina Faso. In 2006, the Foundation began work in Ethiopia and India to increase
access to basic water services for the high need areas. They also supported a watershed
management program in Mexico.
In 2011, the Foundation launched Phase I of the Safe Water Initiative. It contributed to increased
access to basic water services for nearly 2 million people. They also learned that improving
water quality at the point-of-use and making service delivery sustainable would require more
attention to governance, incentives, and other ‘software’— providing ‘hardware,’ like pipes, is
insufficient.
Phase II, launched in 2017, responded to those lessons. emphasized systems change and tested
service delivery models with the potential for replication and scale in Burkina Faso, Ethiopia,
Ghana, Mali, Niger, and Uganda. The Foundation’s 2017-2020 Safe Water Strategy sought to
demonstrate systematic ways of providing sustainable and safe water supply to multiple
countries, including fragile contexts. During the 4-year timeframe, the Foundation contributed to
595,000 people, 40 schools, and 149 health care facilities gaining access to at least basic water
services.
Phase II achievements also include:
1. the establishment of 10 district-based partnerships with accompanying government-
owned plans for water services
2. The identification of promising models such as safe water enterprises to deliver on
quality water services.
Strategy 25 (2021-2025) continues to invest in safe water services for those most in need. The
Foundation remains committed to continue working along with national government leadership,
partners, collaborators, and communities to achieve Sustainable Development Goal (SDG), 6.1,
which calls for universal access to safe water, beginning with securing safely managed water
services1 in its target districts in Ghana, Ethiopia, and Uganda as shown in Figure 1 over the
next five years. (HiltonFoundation, 2022)

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 Figure 1: The target cCountries targeted by the Hilton Foundation in Africa

In Uganda, the Hilton Foundation is in partnership with the Aquaya Institute and program is
being implemented by Department of Agricultural & Biosystems Engineering (DABE),
Makerere University. Lira and Kabarole were some of the selected districts to carry out water
quality testing across the districts. Each district was divided into different enumeration areas
established by Uganda Bureau of Statistics (UBOS). The enumeration areas are 15 in each
district, and every enumeration area consist of 10 households, over 120 water point sources, 30
schools and 15 Healthcare facilities.

1.2 Water Quality Overview

Water is the most basic need of mankind. The utilization of water for various activities depends
on its physical, chemical, and biological characteristics. This includes public water supply for
drinking and domestic purposes, industrial activities, propagation of aquatic and wildlife, water-
based recreational activities, commercial fishing, and esthetic enjoyment. And, since drinking
water is considered to be the most essential use of water for life, it must be free of health hazards
such as pathogens, toxins, and carcinogens.

Absolutely pure water is not available in nature because surface water absorbs particulates,
carbon dioxide and other gases, and mixes with silt and inorganic matters from the environment.
The problem becomes more complex when treated and untreated domestic and industrial wastes
are discharged into natural water bodies. These wastes contain different organic and inorganic

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pollutants and various pathogenic and nonpathogenic microbes. Thus, human waste, drinking
water, and communicable diseases have a direct relation. The communicable diseases mainly
transmitted by water include bacterial, viral, and protozoal infections. For this reason, drinking
water must be free of any kind of microbial, organic, and inorganic pollutant. It must be safe to
drink, esthetically pleasing to the taste, and suitable for domestic activities.

The extent of contamination in water is assessed by the level of pollutants present in water. This
could be obtained by regular analytical estimation of water samples. The analytical examination
of the quality of water and wastewater falls into three main categories;

1) Physical characteristics
2) Chemical characteristics
3) Biological characteristics.

1.2.1 Physical characteristics.

Physical characteristics are those that reflect the palatability or esthetics acceptability of water
for drinking and other domestic activities. They respond to the sense of sight, touch, taste, or
smell, thus indicating the physical state of water. Color, temperature, taste, odor, turbidity,
conductivity, and suspended solids are included in this category.

Temperature

Temperature affects the amount of dissolved oxygen in the water. The amount of oxygen that
will dissolve in water increases as temperature decreases. Temperature also affects the rate of
photosynthesis of plants, the metabolic rate of aquatic animals, rates of development, timing and
success of reproduction, mobility, migration patterns and the sensitivity of organisms to toxins,
parasites and diseases.

Color

Color is due to the presence of humic acids, fulvic acids, metallic ions, suspended matter,
plankton, weeds and industrial influents and effluents as shown in Figure 2

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 Figure 2: Contaminated water in bottles with colored metallic ions

Turbidity

Turbidity is a measure of water clarity. It describes the amount of light scattered or blocked by
suspended particles in a water sample and is caused by particles of soil, organic matter, metals,
or similar matter suspended in the water column. When turbidity is high, microbial contaminants
are often present as well. The effect of suspended particulates can be shown below in the
following bottles containing different levels of suspended particles in the Figure 3

Figure 3: The bottles containing suspended particles in water

Odor and taste

Odor and taste are often due to dissolved organic impurities, such as phenols, chlorophenols, and
sewage components. Algae can produce severe taste and odor problems. These are subjective
properties which are difficult to measure. Algae can produce severe taste and odor problems.

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Electrical conductivity

Conductivity (specific conductance) is the water's ability to conduct an electric current. It

depends on the total concentration, mobility, valence and the temperature of the solution of ions.
Electrolytes in a solution disassociate into positive (cations) and negative (anions) ions and
impart conductivity.

Total solids.

Total solids include Total Suspended Solids (TSS) and Total Dissolved Solids (TDS).
Suspended solids are the portions of solids that are retained on a filter of standard specified size
(generally 2.0 micron) and dissolved solids are solids that are in dissolved state in solution.
Waters with high dissolved solids generally are of inferior palatability.

1.2.2 Chemical characteristics.

Water is a universal solvent and so dissolves most naturally occurring substances as well as those
produced by human activities. These substances are found at a wide range of concentrations in
water depending on their abundance, solubility, and other physiochemical qualities.

Thus, chemical parameters are related to the solvent capabilities of water. Alkalinity, acidity,
PH, hardness, metallic and nonmetallic, biodegradable and nonbiodegradable organics, and
nutrients are chemical parameters of concern in water and wastewater quality management.

pH

pH is a measure of active hydrogen ions in a solution. Water that has more free hydrogen ions is
acidic, whereas water that has more free hydroxyl ions is basic. pH can be affected by chemicals
in the water; thus, pH is an important indicator of water that is changing chemically.

The pH of water determines the solubility (amount that can be dissolved in the water) and
biological availability (amount that can be utilized by aquatic life) of chemical constituents such
as nutrients (phosphorus, nitrogen, and carbon) and heavy metals (lead, copper, cadmium)

Total hardness

Total Hardness is predominantly caused by divalent cations such as calcium, magnesium,

alkaline earth metal such as iron, manganese, strontium. Hardness of water prevents lather

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formation with soap rendering the water unsuitable for bathing and washing. It forms scales in
boilers, making it unsuitable for industrial usage.

Carbonates and bicarbonates of calcium and magnesium cause temporary hardness. Sulphates
and chlorides cause permanent hardness

Nitrates (NO3- )
Nitrates are the most oxidized forms of nitrogen and the end product of the aerobic
decomposition of organic nitrogenous matter. The significant sources of nitrates are chemical
fertilizers from cultivated lands, drainage from livestock feeds, as well as domestic and industrial
sources.
Nitrates may find their way into groundwater through leaching from soil and at times by
contamination. Natural waters in their unpolluted state contain only minute quantities of nitrates.
A high concentration of nitrate in water is indicative of pollution.
Chlorine residual
Chlorine in one form or another is the principal disinfecting agent employed in most countries
against water borne diseases. Chlorine has a number of advantages as a disinfectant, including its
relative cheapness, efficacy, and ease of measurement, both in laboratories and in the field.
The absence of a chlorine residual in the distribution system may, in certain circumstances,
indicate the possibility of post treatment contamination. Exposure to strong light or agitation will
accelerate the rate of loss of free chlorine
Dissolved oxygen
Oxygen dissolved in water is a very important parameter in water analysis as it serves as an
indicator of the physical, chemical and biological activities of the water body. The two main
sources of dissolved oxygen are diffusion of oxygen from the air and photosynthetic activity
Oxygen is considered to be the major limiting factor in water bodies with organic materials
1.2.3 Biological characteristics.
This category includes the microbiological estimation of non-pathogenic and pathogenic
microbes in water. It is also related to the oxygen demand created by the ecosystem present in
water and wastewater.

Viruses

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Drinking water must be free from viruses. Sometime viruses from intestinal tract of infected
person get access to water along with feces. Some intestinal pathogenic viruses which are
transmitted through contaminated water are- Rotavirus, Poliovirus, Hepatitis A and E

Coliform bacteria

Coliforms are bacteria that are always present in the digestive tracts of animals, including
humans, and are found in their wastes. They are also found in plant and soil material. While
usually not pathogenic, the presence of coliform bacteria indicates that a sample has been
contaminated with faecal matter and, consequently, is probably contaminated with pathogens.
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.

Escherichia coli (E. coli) is the major species in the faecal coliform group. E. coli is the best
indicator of faecal pollution and the possible presence of pathogens.

1.3 CONTAMINANTS IN WATER

Contaminants are unwanted substances in water. They can be biological (e.g., pathogenic
microorganisms), chemical (e.g., salts, metals, pesticides) or physical (e.g., sediment).
Contaminants become pollutants when their concentrations in water reach magnitudes that are
harmful to humans, animals and plants.

The common water contaminants include;

1.3.1 Sewage contaminants (e.g., residential, commercial and institutional contaminants)

Sewage is the single largest source of water pollution. The sewage contains garbage, soaps,
detergents, waste food and human excreta. Pathogenic (disease causing) microorganisms
(bacteria, fungi, protozoa, algae) enter the water system through sewage making it infected.
Typhoid, cholera, gastroenteritis and dysentery are commonly caused by drinking infected water.

1.3.2 Industrial contaminants

These are as a result of discharging influent directly into the environment. This introduces highly
toxic heavy metals such as chromium, arsenic, lead, mercury, etc. along with hazardous organic
and inorganic wastes (e.g., acids, alkalis, cyanides, chlorides, etc.) into water resources.

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 Figure 4: The industrial discharge into the a water source

1.3.3 Agricultural contaminants

Manure, fertilizers, pesticides, wastes from farms, slaughterhouses, poultry farms, salts and silt
drain as runoff from agricultural lands. A water body receiving large quantities of fertilizers
(phosphates and nitrates or manures) becomes rich in nutrients (i.e., eutrophication) thus
supporting growth of algae

Excessive growth of algae (algal bloom) often results into the deterioration of water quality and
the depletion of dissolved oxygen in water body. Consumption of water rich in nitrates is bad for
human health especially because it causes “blue baby syndrome” in babies. Toxic pesticide
residues enter the human body through drinking water

1.3.4 Petroleum products

Petroleum products are widely used for fuel, lubrication, plastics manufacturing, etc. and happen
to be poisonous in nature. Besides accidental spills, oil refineries, oil exploration sites and
automobile service centers pollute different water bodies as shown in the Figure 5

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 Figure 5: The motorcyclists near the a surface water source

2 WATER SOURCES
A water source is the location from which households obtain water, such as a tap or borehole.
Samples are collected at water sources to monitor the quality of the water that people are
collecting and help with understanding how water supply systems are performing

The point of use is the place/container where households use and consume water, usually the
home. The quality of water consumed at the point of use may be very different from that at the
source because water can become contaminated during transport, storage and use

2.1 WATER SOURCE TYPES

There are many waters source types which include the following;

 Piped water into dwelling

 Piped water into yard/plot
 Public tap, standpipe

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 Mechanized borehole
 Borehole with hand pump
 Protected dug well with hand pump
 Protected dug well
 Unprotected dug well
 Protected spring
 Unprotected spring
 Rainwater collection
 Surface water (river-dam-pond-stream-canal-irrigation channels)

2.1.1 Piped water into dwelling

A water service pipe connected with in house plumbing to one or more taps and is as shown in
the Figure 6

Figure 6: Piped water inside the a dwelling

2.1.2 Piped water into yard/plot

A piped water connection to a tap in the yard or plot outside the house. It is used only by the
members of one household or dwelling (different from public taps that may be used by anyone)
as shown in the Figure 7

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 Figure 7: Piped water outside the a dwelling

2.1.3 Public tap/ standpipe

Water point from piped system with one or more taps. It is identified by use, not ownership, used
by many households and located in a publicly accessible.

2.1.4 Borehole with hand pump

A deep hole that has been driven, bored or drilled to reach groundwater It has a pump powered
manually. They are usually protected by a platform around the well as shown in the Figure 8

Figure 8: A bore hole with hand pump

2.1.5 Mechanized borehole

Borehole with a motorized pump, a tank, and tap(s) very close together. It is different from the
tap in that one can see the tank when standing at the tap and it has only 1, 2, or 3 tap stands.

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2.1.6 Protected dug well
A protected dug well is a hand-dug well that has an opening fully covered, lined with an
impermeable material to protect the well from runoff water and raised above the ground to
prevent surface runoff water entering as shown below in the Figure 9

Figure 9: The A protected dug well

2.1.7 Unprotected Dug Well

A dug well that is not covered, not lined with any material to prevent runoff from entering and
not raised to protect surface runoff from entering through the top.

Figure 10: Unprotected dug well

2.1.8 Protected dug well with hand pump

A hand pump is placed on a hand dug well instead of a borehole. This is identified by looking at
the base; a borehole has a small base, a hand-dug well has a large round base and the hand-dug
well also has an access door.

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2.1.9 Protected sSpring
A spring is where water first leaves the ground it has a constructed wall or spring box and water
comes out through a pipe.

3 WATER SAFETY
Water safety is compromised when a contaminant exceeds national or international standards, or
when problems occur in a water system that make it vulnerable to contamination. Water safety is
ensured by preventing water from being contaminated through improving infrastructure to
protect sources or by adding water treatment to piped networks.

3.1 Sanitary Inspections

A sanitary site inspection is a physical inspection that identifies the ways that a water source is
vulnerable to contamination. A sanitary site inspection is done on the water source and around
the water source

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A perimeter walk allows to systematically scan the area around a water point for potential
sources of contamination. Most contaminants (e.g., E. coli) are not visible, but sources such as
latrines or open sewers are identified.

A sanitary inspection is indispensable for the adequate interpretation of the laboratory results. No
analytical, bacteriological or chemical survey, however carefully carried out, is a substitute for
comprehensive knowledge of conditions at the water source and within the distribution system,
the adequacy of water treatment and the qualifications and performance of the operators.

The sanitary inspection provides essential information about immediate and ongoing possible
hazards associated with a community water supply, even in the absence of microbiological or
chemical evidence of contamination.

The inspection includes a checklist of the components of the water supply from the source to
distribution and incorporate all the potential points where hazards may be introduced. Any
problems identified during the inspection is highlighted so that a report maybe provided directly
to the community. Some of the defects observed at water point sources are shown I the Figure
11

Figure 11: Some of the sanitary inspections

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3.2 CALIBRATION
Calibration is defined as a set of operations that establish, under specified conditions, the
relationship between values of quantities indicated by a measuring instrument or measuring
system, or values represented by a material measure or a reference material, and the
corresponding values realized by standards.

Calibration also involves checking, by comparison with a standard, the accuracy of a measuring
instrument of any type. It may also include adjustment of the instrument to bring it into
alignment with the standard. 

3.3 Laboratory safety practices

Laboratory safety is the key to reducing injury and illness. There are many exposures in the
laboratory that pose a hazard to the health.

3.3.1 Protection in the laboratory

• Wearing the clothing and protective wear identified in the risk assessment

• Laboratory coats must be kept fastened

• Don’t wear sandals or open shoes

• Long hair must be tied back

The above gears can be shown in the Figure 12 below;

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 Figure 12: The lLaboratory safety gears

3.4 General Hazards in a Laboratory

Some of the common hazards in the laboratory include; Fire, breakage of glassware (note that
you break- u buy policy), sharps, spillages, pressure equipment & gas cylinders, extremes of heat
& cold, chemical hazards, biological hazards, radiation.

3.4.1 Solution to the hazards

Avoiding Fires

• Flammable substances

• Use minimum quantity

• Store in special storage cabinet

• Use temperature-controlled heating sources ege.g. water-bath rather than hot-plate

or Bunsen burner

• Fire Extinguishers- safety

Glassware

• Safe storage of glassware

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 • Never use glassware under pressure or vacuum unless it is designed for the job and
suitably shielded

• Dispose of chipped or broken glassware – it is a risk to you and others

• Always dispose of broken glassware in a glass bin or sharps bin and not in a general
waste bin

Spillages

• Clear up spillage promptly

• Work from outside in

• Dispose of any hazardous material as toxic waste

3.5 Waste management in the laboratory.

This is part of the risk assessment which involves determining how to dispose of waste lab
materials safely. Solvents and oils must be segregated into the correct waste bottle or drum and
special care needs to be put in place for the chemical or biological material. Precaution should be
done not to put materials down the drain or in mixed with normal waste unless authorised to do
so.

The waste management requires the following;

 Proper storage– same rules apply – making sure waste chemicals are compatible
 Proper labeling – tags are placed on bottles name of chemical
 Pre-planning – know what waste being created prior to carrying out experiments; minimize
purchases
 Record-keeping – of all waste chemicals on hand and those already picked up for disposal

3.6 Risk hazards and Risk management

Risk is a situation involving exposure to danger and possibly leading to loss or injury. Most
hazards encountered fall into three main categories; chemical, biological, or physical. Cleaning
agents and disinfectants, drugs, anesthetic gases, solvents, paints, and compressed gases are
examples of chemical hazards. Potential exposures to chemical hazards can occur both during
use and with poor storage

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3.6.1 Risk management
Risk management can minimize the chance of errors and ensure reliability of test results. Risk
management guidelines recommend that laboratories play a proactive role in minimizing the
potential for errors by developing individualized QCPs to address the specific risks encountered
with laboratory analysis.

3.6.2 Safety Signs in the laboratory

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3.6.2.1 PROHIBITION SIGNS
These mean stop/must not do. The sign is circular with a white background, black pictogram and
red border and crossbar. All writing is white on a red background. Some of the prohibition signs
can be shown in the figures below

Figure 13: Some of the prohibition signs used in the laboratory

3.6.2.2 WARNING SIGNS

These mean there is a risk of danger. The sign is triangular with a yellow background, black
pictogram and black border. All writing is black on a yellow background. Some of the warning
signs are shown below.

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 Figure 14: The common warning signs

3.6.2.3 MANDATORY SIGNS

These mean that it must be obeyed. These signs feature a blue circle with white pictogram. All
text is in white on a blue background. Some mandatory signs can be safety boots must be worn,
ear protection must be worn and eye protection must be worn.

Figure 15: Some of the mandatory signs in the laboratory

3.7 Standards and Protocols

3.7.1 Laboratory Standard
This regulation, commonly known as the Laboratory Standard, was designed to ensure that
laboratory workers are informed about the physical and chemical hazards of chemicals in their
workplace and are protected from chemical exposures exceeding allowable levels.

The purpose of establishing laboratory quality standards is to ensure the accuracy of test results,
increase the confidence of clients and communities.

All laboratory activities may be subject to errors, and studies have shown that errors in the
laboratory can occur in all the phases of analytical procedures. Examples of errors that can occur
in each phase are given below:

Pre-analytical phase

• Incorrect test request or test selection

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 • Incomplete laboratory request forms (especially when one doesn’t know which
parameters and the number of samples to pick or which site to go)

• Incorrect sample collection, labelling and

Analytical phase

• Use of faulty equipment, improper use of equipment (common with the un calibrated
ones)

• Use of substandard or expired reagents

• Incorrect reagent preparation and storage (some are influenced by light penetration)

• Incorrect technical procedures; non-adherence to standard operating procedures (SOPs)

or internal quality control (IQC)

Post- analytical phase

• Inaccurate reporting and recording

• Inaccurate calculations, computation or transcription and even the units used (mg/l, micro
segments, g/l,

• Late return of results

• Incorrect interpretation of results (reading TC instead of E. coli)

3.7.2 International laboratory quality standards

There are several internationally accepted standards applicable to laboratories and many of these
have been developed by ISO. Standards ensure desirable characteristics of products and services
such as quality, safety, reliability, efficiency and reproducibility.

ISO standards provide a technical base for health, safety and conformity assessment

3.7.3 Protocols
A lab protocol, also known as a standard operating procedure, is a list of instructions to perform
an experiment. It is a plan used to duplicate favorable results from a previous test.

27
In a research laboratory, many protocols are needed for safety, to operate analytical equipment
and to make solutions with minimal mistakes. Every protocol needs to focus specifically on one
type of experiment to be performed. This part of the protocol states the hypothesis and any
methods that can reference it.

3.7.4 Quality assurance and Quality Control

Quality Control refers to the procedures for the measures that must be included during each
examination run to verify that the test is working properly.  This is a laboratory function. These
include planned activities designed to provide a quality product.

Quality assurance refers to the overall program that ensures the final results reported by the
laboratory are correct.  These include planned activities designed to ensure that the quality
control activities are being properly implemented. Quality assurance activities are performed by
staff who are not directly involved with the analysis of water samples. This can be taking
samples to another lab or having triplicates of samples analyzed or having same sample analyzed
3 times.

In order to demonstrate that a laboratory is producing data of adequate precision, accuracy and
sensitivity, it is necessary to assess all laboratory procedures at all stages from sampling to
reporting. This is a time consuming and costly process and, for this reason, it is important to
ensure that the necessary standards of performance are clearly defined and adhered to.

In most laboratories, QA (Quality Assurance) will start with the examination and documentation
of all aspects of laboratory management. This will include clearly identifying lines of
communication and responsibility, the description and documentation of all procedures which
are carried out, and the documentation of instrumental and analytical checks.

Use of blanks

Field and laboratory blanks should be analyzed with each batch of samples. A blank consists of
reagent water, usually deionized water or distilled water. A field blank is reagent water that has
been bottled in the laboratory, transported with sample bottles to the sampling site, processed and

28
preserved as a routine sample and returned with the routine samples to the laboratory for
analysis. The analysis of a blank should not yield a value higher than that allowed by the
acceptance criteria. This procedure checks interference and the limit of detection of the assay.

Control by duplicate analysis

Since the samples are analyzsed using the same method, equipment and reagents, the same bias
should affect all results. Duplicate analyses are only useful for checking precision; they provide
no indication of the accuracy of the analyses.

1.1
1.2
1.3
1.4
1.5
1.6
1.7 DISINFECTION
Disinfection of equipment is essential for many microbiological analytical procedures, but some
of the devices used for this purpose in a laboratory are unsuitable for transporting to the field.
Some of the methods for disinfecting equipment in the lab are:

Dry heat: The flame from a liquid cigarette-lighter, for example, can be used to disinfect
forceps for manipulation of membrane filters.

Disinfecting reusable items: The two main items of reusable equipment, Petri dishes (glass or
aluminium) and bottles, may be disinfected by immersion in boiling water for a few minutes, by
dry heat sterilizsation in an oven or by heating in a pressure cooker for at least 20 minutes.

29
 4 MATERIALS AND METHODS
2

2.1 EQUIPMENTS USED

2.1.1 Aquameter box

The Aquameter consists of The Aquameter unit, quick release lanyard, set of 5 AA Alkaline
batteries, USB Cable for downloading logged data to a PC, cross-head screwdriver for fitting the
batteries and Probe maintenance, getting started card for quick reference.

The Aquameter box also consists of; protective Sleeve End Cap. Calibration bottle filled with
Rapid Cal Solution, Spare calibration / rinse bottle, One mounting nut (pre-fitted), 25mL bottle
of pH storage solution, Pot of silicone grease, Spare Galvanic DO Membrane Cap (AP-700 &
AP-800 only), 25mL bottle of Galvanic DO filling solution (AP-700 & AP-800 only). The figure
showing the contents of the Aquameter box.

Figure 16: The components of the Aquameter box

2.1.2 Aquaprobe
Aquaprobe is made primarily from hard-galvanised marine grade aluminium. It is constructed
with an aluminium sleeve surrounding the delicate sensing electrodes. The Sleeve are easily
removed by unscrewing to allow cleaning of the individual electrodes. The different parts of the
Aquaprobe are shown in the Figure 17 below.

30
Figure 17: The different parts of the Aquaprobe

2.1.3 AP-2000 Extension cable

The AP-2000 is designed to connect to the Aquameter using an AP-2000 Extension Cable. The
AP-2000 Extension Cable features high-pressure metal connectors, which incorporate several O-
ring seals at the Probe end. Prior to first connection, the seals are lubricated using the silicone
grease found in the Aquameter box.

To connect the Extension Cable to the AP-2000, we align the coloured dot on the AP-2000 with
the ◄Aquaread logo on the plug body, then press the plug into the socket and tighten the
retaining collar fully as shown in the figure below.

Figure 18: Connecting the extension cable onto the Aquameter probe

2.1.4 Aquameter
The GPS Aquameter is the original handheld water quality meter with built in GPS. The built in
GPS receiver allows the meter to record the location of every dataset. The meter is designed to
be picked up and used by anyone intuitively. It requires no boot up or warm up time and allows
to record data with a single button press.

31
Figure 19: The Aquameter showing the different parameter readings

2.1.5 The chlorine tester

The chlorine tester contains DPD tablets (diethyl-p-phenylenediamine) which are used to
determine chlorine in water and it can also be used to test bromine, iodine and other
disinfectants.

Figure 20: The chlorine tester

2.1.6 The turbidity meters

Turbidity Meter is used to determine the number of suspended particles in water. This easy-to-
use handheld instrument accurately measures turbidity by the intensity of reflected light. The

32
values are proportional to the number of suspended particles in the sample. The turbidity meter
was used when the turbidity probe was found faulty due to lack of calibration, presence of air
bubbles and contamination of the Rapid Cal solution.

Figure 21: HACH 2100Q portable turbidimeter with its sample cells.

2.1.7 Hand sanitizer and Alcohol wipes

Hand sanitizer is a handy on-the-go way to help prevent the spread of germs when soap and
water aren’t available. Alcohol-based hand sanitizers help keep the lab attendants safe and
reduce the spread of pathogens.

2.1.8 Cooler with frozen ice packs

The cooler bag with the frozen ice packs help to preserve the microbes in the water sample. It is
always necessary to prevent the extinction of the microbes during transportation as well as to
avoid the multiplication of the microbes, therefore the ice packs help to this job.

Figure 22 showing a cooler bag with whirl pak bags seated on an ice pack.

33
 2.1.9 Whirl-Pak bags
Whirl-Pak bags are sterile polyethylene laboratory sample bags used to transport samples for
biological testing and other applications. Bags should not be used at temperatures above 82°C.
The bag contains sodium thiosulphate solution used to preserve the sample.

Figure 23 showing the Whirl Pak bag

2.1.10 Distilled water

The distilled water is used to wash the equipment in the
laboratory as well as in the field after analyzing the
water sample. The distilled water is always assumed that it
does not contain any contamination, therefore, its recommended to use it as a washing reagent

2.1.11 The manifold

This is made up of stainless steel with a funnel-like shape containing a porous stony filter
membrane that is used to support the filter membrane paper. It contains holes where a pressure
vacuum is applied to suck the water sample using a syringe.

Figure 24: The single branch manifold used for membrane filtration method

2.1.12 Petri dishes.

Petri dishes are used for biological examinations and should be of adequate size and depth to
ensure constant surface area of the medium for growth and propagation of microbes . Disposable
plastic petri dishes containing the culture media were used directly without sterilization and
discarded after estimation was done.

34
2.2 SAMPLE PRESERVATION
Physical preservation of samples by cooling, i.e., by keeping the sampling container on ice
during transportation, was highly essential — especially at high atmospheric temperature.
Increase in temperature might cause an increase in the rate of biochemical processes, which
would alter the characteristics of the sample.

For microbiological examinations of water, the collection, storage, and transport of samples was
carried out under controlled conditions to assess the accurate level of microbial contamination.
To achieve this goal, the following precautions were essential:

• Every sampling whirl-Pak bag was properly labeled with a unique ID number, date and the
sampling time.

• Extra care was taken to avoid any accidental contamination during sampling through
continuous sanitization before and after sampling.

• The sample was made sure that it was not exposed to light. It was transported in an insulated
container filled with ice.

• The time for sampling, transportation, and estimation (analysis) was minimized as far as
possible. The estimation was performed within 6 h of sampling.

• If the water contained residual chlorine, an adequate quantity of sodium thiosulfate (Na2S2O3)
was used embedded in the Whirl-Pak bags to neutralize it.

2.3 THE CALIBRATION OF THE MULTI-PARAMETER METER

Calibration is a very important part of successful water quality measurement and should be
carried out regularly. A great deal of development work has been put into simplifying and
automating the calibration procedures in the Aquameter in order to allow normal field operatives
(as opposed to trained lab technicians) to achieve quick and accurate results.

As a general rule, pH and EC are calibrated as close to 25ºC as possible. In order to standardize
calibration techniques, Aquaread provide plastic calibration bottles into which the Aquaprobe
can be directly inserted. The Aquaprobe is designed to be calibrated in these calibration bottles
with the Probe Sleeve and Sleeve End Cap fitted.

35
RapidCal calibrates EC at 2570µS/cm, the pH7.00 point and the zero point of all optical
electrodes (except Turbidity) simultaneously. This procedure is carried out at the beginning of
each day the Probe is to be used.

2.3.1 Procedure of the calibration of the multi-parameter meter

The lid is removed from a fresh bottle of RapidCal solution and the storage cap is also removed
from the pH electrode if fitted.

The Probe is then washed in distilled water while ensuring the Probe Sleeve and End Cap are
fitted.

The probe is into the RapidCal solution, bang against the bottom of the bottle several times in
order to remove any air bubbles that may be clinging to the electrodes.

When the Probe is inserted, the level of the solution is ensured such that it is at least up to the
minimum insertion line scribed around the Probe sleeve. If the level is low, the EC electrodes
will not be covered and EC will not calibrate properly. If the level is low, top up with fresh
RapidCal solution.

The Aquameter is switched on and waited until all readings are completely stable. The longer we
left the probe to achieve thermal equilibrium before proceeding, the better it was.

The temperature of the solution is kept between 5ºC and 40ºC (41ºF – 104ºF). The closer to 25ºC
the better.

Then, we pressed the MENU key then select Calibration. Then select the RapidCal option. The
Meter would wait until all readings are stable, then it would send the RapidCal command to the
Probe, where the calibration takes place. During calibration, the Calibrating screen is displayed
and the progress counter counts up. If the calibration is successful, the counter will reach 100%
and the following screen will be displayed.

When calibration was complete, we would then press OK then ESC to return to normal reading
mode.

After calibrating with RapidCal, we removed the Probe from the bottle, washed in fresh water,
then Shaked off ensuring there are no droplets adhering to the DO membrane

36
Moistened piece of tissue paper with fresh water was used to wrap around the open end of the
probe ensuring all the holes were covered.

2.4 CALIBRATION OF THE TURBIDIMETER WITH STABLCAL STANDARDS

1. Push the calibration key to enter the calibration mode. Follow the instructions on the
display. Note that one should always gently invert each standard before inserting the
standard so as to agitate the standard. It should be done gently so as not to form air
bubbles in the standard.
2. Insert the 20 NTU StablCal Standard and close the lid. NOTE: the standard to be inserted
is always bordered. The sample cell was inserted in the instrument cell compartment so
the diamond or orientation mark aligns with the raised mark in front of the cell
compartment.
3. Push Read key. The display shows stabilizing and then shows the result.
4. Steps 2 and 3 were repeated with the 100 NTU and 800 NTU StablCal Standard. Push
Done to complete a 2-point calibration process.
5. Push Store to save the results. After calibration is complete, the meter automatically goes
into the Verify Cal mode.

2.4.1 Calibration Verification (Verify Cal) procedure

1. Push Verify Cal to enter the Verify menu.
2. Gently invert the standard. Insert the 10.0 NTU (or other defined value) Verification
standard and close the lid.
3. Push Read. The display shows stabilizing and then shows the result and tolerance range.
4. Push Done to return to the reading display. Repeat the calibration verification if the
verification failed.

2.4.2 The cleaning of spectrophotometric cells

It needed the following precautions:

37
  Cleaning the cells was done using soft tissue to wipe the cells dry. The cleaning
of cells was done as soon as possible after each use. DDW was used for rinsing
before and after every sample measurement
 Care was taken not to blow air with the intent to dry the cell and cells were never
cleaned with a brush or any other cleaning device, which might scratch the walls
of the cells.

2.5 PREPARE FIELD SUPPLIES

1. Calibrate the multi-parameter meter - pH, turbidity, conductivity, and temperature

2. If it was needed, we would adhere barcode stickers to the Whirl-Pak sample collection
bags for identification
3. The supplies needed for the field were;
 Calibrated multi-parameter meter
 Cooler with frozen ice packs
 Whirl-Pak sample collection bags with barcode stickers on them (take at least 5
extras)
 Hand sanitizer
 Two markers
 Paper wipes
 Charged mobile phone with surveys loaded/synched and power bank
 Field notebook and pen
 One new, unopened water bottle for days when Blank samples are collected
 Distilled water

2.6 CONDUCT WATER SOURCE SURVEYS

1. We travelled to Enumeration Area.
2. Then met with a community leader and inquired about the number and types of shared
drinking water sources in the enumeration area / community. If there were more than 10
sources, we inquired about which 10 were used by the most people living in the
enumeration area.

38
 a. This included both improved and unimproved water sources, as long as they were
used to supply drinking water.
b. This included water points that were currently not providing water because of a
breakdown, seasonal dryness, or other issue, as long as they had provided water
within the previous 12 months.
c. This also included any source that was “shared” – meaning that anyone in the
community could have access to it.
d. This did not include on-plot or private sources used by households from a single
compound, unless the source was also accessible to others on an ongoing basis.
e. This finally included any sources that were physically located outside the EA, but
were used by households located within the EA boundary.
3. We visited all selected water sources and complete the Water Source survey.
a. The respondent for the Water Source Survey would be a community leader, a
water committee member, a water point caretaker, or other community member
knowledgeable about the water source particularly women
4. The water samples would then be tested from a maximum of 10 water sources in each
enumeration area.
a. If there were <=10 water sources that met the above criteria, then water would be
tested from all of the sources with water available.
b. If there were >10 water sources that met the above criteria, then water would be
tested at the 10 sources with the highest reported number of households using the
source, according to the community leader.

2.7 COLLECT SAMPLE FOR MICROBIAL ANALYSIS

1. We began by disinfecting our hands with the hand sanitizer.
2. Then took out a Whirl-Pak sample collection bag, and ensured it had a barcode sticker on
it.
3. Scanned the barcode in the survey form and checked that the correct sample ID was
recorded (e.g., “L-139”)
a. If the scan did not work, we selected “Not Applicable” and typed the barcode
number into the next question

39
4. Wrote the date and time on the Whirl-Pak sample collection bag with a permanent
marker.
5. Then disinfected our hands with the hand sanitizer.
6. Opened the Whirl-Pak sample bag carefully by tearing off the perforated top. Then we
used the two little white tabs on the sides of the bag to open the bag.
a. If your hand accidentally touched the inside of the bag where the sample went,
even slightly, we discarded the bag and used one of the spare bags. In this case,
re-scanned the new barcode of the spare bag in the survey form and wrote the date
and time.
7. Then collected a sample by filling the Whirl-Pak bag OVER the 100mL line.
a. For improved sources such as hand pumps and tap stands:
i. We held the sample bag by the wire ends. Carefully filled the bag with
water directly from the water source, being sure not to touch the bag to the
water source outlet.
b. For improved or unimproved wells:
i. We extracted water from the well using the method used by people who
collect water there, using the container that was typically used. Carefully
poured this water into the sample bag, being sure not to touch the
container to the bag.
c. For surface water sources:
i. The sample bag was held by the wire ends, and carefully scooped water
directly from the source.
8. The two wire ends of the Whirl-Pak bag were held, and closed the bag by flipping the bag
quickly so that it flipped at least three times.
9. Sealed the bag by folding the wire ends up and twisting them together.
10. We turned the bag upside down and shook it at least 3 times to see if it was leaking. If it
was leaking, we would open the bag and reclose it. We repeated until the bag was not
leaking.
11. We placed the sample upright in the cooler with ice.

40
2.8 TEST SAMPLES FOR PHYSICAL-CHEMICAL PARAMETERS
1. Measuring the pH, conductivity, turbidity, temperature
a. The Probe sleeve was removed and gently removed the pH sensor cover.
b. We fastened back the Probe sleeve and end cap before any measurements to
ensure the sensors are protected.
c. We connected the probe to the calibrated meter.
d. Rinsed the 3-liter bucket three times with water from each source.
e. Turned the meter on using the red button.
f. We checked the GPS Position 3D was displaying.
g. We placed enough water in the bucket to submerge the sensors on the probe of the
meter.
h. We submerged the meter probe in the sample water in the bucket, and moved
gently to take the reading.
i. Ensured the water level was above the groove-ring on the probe sleeve.
j. We then waited until the reading of most parameters stabilized (a double headed
arrow shows) and recorded the displayed readings in the survey form. Using the
right and left arrow button to toggle through the other parameters.
k. We saved the point reading by placing the M+ key on the Aquameter.

2.9 TEST PROCEDURE FOR TURBIDITY USING THE HACH 2100Q PORTABLE
TURBIDIMETER.
1. Before a measurement was taken, we always made sure that the water sample was
homogenous throughout and this was achieved by gently agitating the sample.
2. Collect a representative sample in a clean sample in a clean container. Fill a sample cell
to the white line (about 15ml). Take care to handle the sample cell by the top. Cap the
cell.
3. Wipe the cell with a soft lint-free cloth to remove water spots and finger prints.
4. Apply a thin film of silicone oil. Wipe with a soft cloth to obtain even film over the entire
surface.
5. Push the power key to turn the meter on. Place the instrument on a flat sturdy surface. Do
not hold the instrument while making measurements.

41
 6. Gently invert and then insert the sample cell in the instrument cell compartment so the
diamond or orientation mark aligns with the raised orientation mark in front of the cell
compartment. Close the lid.
7. Push the Read key. The display shows stabilizing then the turbidity in NTU is shown and
stored automatically.

2.10 TEST PROCEDURE FOR FREE CHLORINE

1. The DPD tablet was added to the right-hand compartment without letting the tablet
get in contact with your fingers.
2. The sample water was added to fill the compartment and we replaced the rubber
stopper.
3. The Pool tester was shaken (for about 20 seconds) to mix and dissolve the tablet.
4. We compared the color in the sample with the chlorine scale on the Pool tester by
holding it up to a window or other bright light. You could also use a piece of white
paper as a background.
5. The color scale in the middle of the Pool tester was used for the lighter sample colors
(lower chlorine levels) or the scale at the far right if the sample color was darker
(higher chlorine levels)
6. After obtaining the corresponding colors, the results were recorded.

2.11 DELIVER SAMPLES TO THE FIELD LAB FOR ANALYSIS

1. Each sample was stored in the cooler with ice packs throughout the day while ensuring
that the cooler was kept out of direct sunlight whenever possible.
2. After all samples were collected, drove back to the field laboratory to deliver the samples.
3. The samples were arrived at the field lab no more than 6 hours after the time the first
sample was collected (within 4 hours is preferred).

2.11.1 Quality Assurance / Quality Control Measures

One time per week, we collected either a Blank or Duplicate Sample

1. On Wednesday, we were assigned to collect a blank sample, we collected the sample by

the procedures described for collecting a sample under Chapter 3 in Section 2.6 and
testing the physical-chemical parameters at the first water source of the day.
a. We followed the sample collection and testing instructions listed above, except:

42
 i. Instead of collecting water from the water source and testing the water
from the source, we collected and tested the water from a new, unopened
packaged water bottle preferably Rwenzori mineral water
ii. In addition to the date and time, we wrote “B” on the Whirl-Pak sample
bag to indicate a blank sample.
2. On Friday, we were assigned to collect a duplicate sample, we repeated the procedure for
collecting a sample at the first water source of the day.
a. We followed the sample collection instructions listed above a second time,
collecting a second sample of the same water, collected in the same way.
i. In addition to the date and time, wrote “Dup” on the sample bag implying
a duplicate sample.

2.11.2 Procedure for microbial testing

Two techniques are commonly used to detect the presence of coliforms in water. The first of
these is called the “multiple fermentation tube” or “most probable number” technique. In this
method measured portions of a water sample are placed in test-tubes containing a culture
medium. The tubes are then incubated for a standard time at a standard temperature. In the
second technique, a measured volume of sample is passed through a fine filter that retains
bacteria. The filter is then placed on culture medium and incubated. This is called the
“membrane filter” technique.

2.11.2.1 Equipment needed

 Membrane filtration manifold
 200ml syringe and rubber tubing
 Pipette with 1ml sterile pipette pipe
 100ml sterile funnel
 47mm membrane filter
 Compact dry E. coli plate
 Fine tipped permanent marker
 Hand sanitizer
 Disposable gloves
 Forceps
 Disinfectant alcohol wipes
43
  Bucket for liquid waste and disinfection
 Sample water
 Deionized water
 70% Ethanol solution in spray bottle
 Bleach
 Plastic table cloth

2.11.2.2 Preparing for the testing

 We began by cleaning the laboratory table of any extra materials and after put on the
disposable gloves
 We lay the plastic table cloth and disinfect it with 70% ethanol by spraying and using the
paper towels.
 The samples were taken out from the coolers and placed them in order from the earliest to
the latest times of the collection. The times of the sample collected, the earliest and latest
were marked on the log book sheet.
 The samples were put back into the coolers and the table disinfected again with 70%
ethanol
 We then set up the cleaned table with the following wiped down solid, non-sterile items
with ethanol;
 Filtration manifold
 200ml syringe for manifold suctioning
 Forceps
 A small batch of the membrane filters in the top of the filters box
 Pipette with the sterile pipette tips
 Sterile funnels for each sample
 Compact dry plates for each sample
 Bottle of DI water
 Hand sanitizer bottle
 Alcohol wipes
 Blank log book pages
 Then we would prepare the filtration method manifold by;
 Disinfecting all the surface of the manifold with ethanol
44
  Inserting the 200ml syringe in the manifold using the rubber tubing to connect it
 Pouring a small amount of ethanol on the stone disk and carefully flaming it until
all was burned off and then placing the stone disk into the filtration manifold

2.11.2.3 Processing samples using the membrane filtration method

 We began by putting on the gloves and then writing on the log sheet the time we began
the sample processing
 The first sample was taken out of the cooler bag and one compact dry plate taken out to
be used for the sample
 The barcode sticker was then moved from the Whirl Pak sample bag to the top of the
compact Dry plate. If the barcode sticker was not able to be moved, we would write the
sample ID with the permanent marker on the compact Dry plate
 We then sanitized our hands with hand sanitizer while the forceps were disinfect using
the alcohol wipes
 The alcohol wipes were again used to disinfect the top of the manifold and forceps were
then kept on the alcohol wipes to keep it sterile
 The manifold was opened by putting the valve in the vertical direction and pulled the air
through the manifold to make sure that the alcohol evaporated and afterwards closed
before removing the syringe.
 We then carefully opened the sterile filter package without touching the filter by the help
of the forceps to separate the filter from the blue film. The filter was then placed onto the
manifold with the gidded side up.
 The sterile funnel was then placed on the manifold and making sure the manifold was
closed that is to say in the horizontal direction
 The sample was shaken for 10 seconds and then carefully opened the sample bag by
touching the white tabs on the bag.
 The sample was then carefully poured into the funnel on the manifold and filled up to the
100ml mark. The sample bag was then closed carefully and kept aside.
 A 1ml of the sample was transferred from the funnel to the compact Dry plate using the
sterile pipette in order to rehydrate the plate.
 The manifold was then opened that is put in the vertical position

45
  Using the syringe, the sample was pulled through the filter. After filtering all the water,
the manifold valve was closed.
 The funnel was then carefully lifted off from the manifold and then disposed.
 Using the forceps, the edge of the membrane filter was grabbed and carefully placed onto
the compact dry plate. The remaining water was pulled through the manifold using the
syringe.
 The manifold valve was then closed and the table cleaned for the next water sample.
 After processing all the samples, the compact dry plates were placed upside-down (to
prevent water from condensing on the lid and droplets falling on the growth medium and
disturbing the culture) into the incubator at 37 0C
 The time when the sample went into the incubator was recorded and written in the log
book.

Figure 25: Pulling water out of the sterile funnel on the manifold

46
 5 ANALYSIS AND DISCUSSION OF RESULTS
3
3.1 READING THE SAMPLES FOR MICROBIAL ANALYSIS
The compact dry plates were read after incubating for 24 hours at 370C.

The compact dry plates were removed from the incubator and gently set down slowly in order to
avoid the vapor on the lid from falling on the filter as this would make the plates unreadable.

The results were recorded in the Microbial Results survey following the steps below;

 The barcode on the compact dry plates was scanned if the barcode mark existed
otherwise, the number would be entered manually.
 The date and time on which the sample would be read was recorded in the survey form
 The number of blue/green colonies was counted, regardless of the size and result entered
in the survey.
 If there were more than 100 colonies on the plates, the result would be recorded as
“101” meaning TNTC (too numerous to count)
 If it was impossible to interpret the results or the incubated could not be
completed, the results were recorded as “Don’t know”
 Any observations on the colonies and plates noticed could then be recorded
 A photo of the plate was taken and fed in the survey form. Below is an example of the
compact dry plate having the colonies as shown in the blue color

Figure 26: The compact dry plate with colonies

47
 3.2 Laboratory blank samples
The laboratory blank sample was processed each day as part of the quality assurance/ quality
control procedure. This blank sample was used as a measure to ensure that sample handling and
analysis in the laboratory did not breed into contamination.

3.2.1 Procedures
1. The compact dry plate was labelled with “B- “and a number. For example, B-1, B-2, etc.
2. The number used was written in the log book
3. The procedures discussed for sample processing were followed but instead of the sample
water, 100ml of deionized water was used.
4. The results would be recorded in the mwater app as shown below;

User: wqtfmak2224
Response Id: wqtfmak2224-AP835V
Submitted: Sep 15, 2022 10:05 PM
IP Address: 41.210.154.58
Status: Final
Drafted by wqtfmak2224 on Sep 15, 2022 10:01 PM
Submitted by wqtfmak2224 on Sep 15, 2022 10:05 PM

Question Answer

Enter your name Irumba Collin

Enter the date and time now September 15, 2022 10:01 PM

Is this a Laboratory Blank sample? (Labelled with a "B") Yes

Enter the blank sample ID B11

Count the number of colonies and enter the number here. 0

If there are >100 colonies, meaning "too numerous to
count", enter 101. If there are problems reading the plate,
such as the colonies being smudged or wet and not able to
be counted, then select "Don't know".

Re-enter the number of colonies 0

48
What was the volume of water that was filtered when this 100 mL
sample was processed?

Take a photograph of the Plate with the lid off

Record any notes about the sample here (optional)

49
 3.2.2 Microbial survey results from the water point sample.
The procedures followed are shown in the table under the question column.

User: wqtfmak2224
Response Id: wqtfmak2224-APXNDG
Submitted: Sep 24, 2022 8:54 PM
IP Address: 41.210.141.122
Status: Final
Drafted by wqtfmak2224 on Sep 24, 2022 8:51 PM
Submitted by wqtfmak2224 on Sep 24, 2022 8:54 PM

Question Answer

Enter your name Irumba Collin

Enter the date and time now September 24, 2022 8:51 PM

Is this a Laboratory Blank sample? (Labelled with a "B") No

Scan the sample ID barcode. If the scan does not work, or Not Applicable
there is no barcode, please choose Not Applicable.

Please manually type in the sample ID A-0371

Count the number of colonies and enter the number here. Don't Know
If there are >100 colonies, meaning "too numerous to
count", enter 101. If there are problems reading the plate,
such as the colonies being smudged or wet and not able to
be counted, then select "Don't know".

Re-enter the number of colonies Don't Know

What was the volume of water that was filtered when this 100 mL
sample was processed?

Take a photograph of the Plate with the lid off

Record any notes about the sample here (optional) Some part of the filter paper had issues; therefore, it was

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 hard to count
Table 1: A summary of the results from the survey form

3.2.3 Water point survey results from the mwater app

The procedures taken for this survey are discussed are shown in the table.
User: wqtfmak2224
Response Id: wqtfmak2224-APKB4H
Submitted: Sep 20, 2022 1:48 PM
IP Address: 41.210.154.39
Status: Final
Drafted by wqtfmak2224 on Sep 20, 2022 1:31 PM
Submitted by wqtfmak2224 on Sep 20, 2022 1:48 PM
Edited by pnakawuka on Sep 22, 2022 9:40 PM

Question Answer

Metadata - Fill out without asking the respondent

1. Enter your name Irumba Collin

2. Enter the date and time September 20, 2022 1:31 PM

3. Choose the district name Kabarole

4. Select the water point site Name: KISONGI 'A'

Type: Protected dug well

Location
GPS Location: 0.722050, 30.381814

Administrative region: KAHANGI, Hakibaale, Kabarole, Uganda

Unique ID: 408857271

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 Photos:

Attributes

Pump/lifting device: Nira

Year of construction or most recent rehabilitation (EFY): 1995

Custom fields
Import code: Aquaya11-ABMHD4

External photo location:

Implementing organization: NGO

5. Is this water point associated with a No

surveyed institution?

Question Answer

6.a. Select the enumeration area Name: KISONGI 'A'

Description: KABAROLE

Location
GPS Location: 0.715817, 30.389184

Administrative region: KAHANGI, Hakibaale, Kabarole,

Unique ID: 405352508

Import code: 214

7. [Direct Observation] Water point or water Protected dug well (shallow well) with hand pump
source type  Edited

Acceptability

8. Are there issues with the quality or No

acceptability of water from this water point -

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like what it looks or tastes like?

Reliability and Functionality

9. [Direct observation] Is water available from Yes

this water point now?

10. Are there months during the year when Yes, there are months when water is not available
water is not regularly available at this water
point because it is dry? (Or was not regularly
available when it was working in the past?)

11. In the past 6 months, (since MARCH), has No

the water point broken down because of a
mechanical problem?

Water Quality Testing

13. [Direct observation] Are you collecting a Yes

water sample at this water point? [If water is
available, a sample should be taken]

13.1. [COLLET SAMPLE] Scan the barcode L-556

on the Whirl-pak bag, write the time and date,
sanitize your hands, and collect the sample [If
the scan does not work, choose Not Applicable
and enter manually]

13.1 [COLLET SAMPLE] Type in the sample

code from the Whirl-pak bag, write the time
and date, sanitize your hands, and collect the
sample.

13.1a Re-type in the sample code from the

Whirl Pak bag.

13.2 [Measure] Enter the pH 6.45

13.3 [Measure] Enter the Conductivity (uS/cm) 44

13.4 [Measure] Enter the Temperature (degC) 21

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5
Question Answer

13.5 [Measure] Enter the Turbidity (NTU) 14.74

13.6 [Measure] Enter the Free Chlorine 0

Residual
(mg/L)

Water Quality Testing - BLANK

14. [Direct observation] Are you collecting a No

BLANK water sample at this water point?
(From an unopened packaged water bottle.) Do
this at the first water point on Wednesdays.

Water Quality Testing - Duplicate

15. [Direct observation] Are you collecting a No

DUPLICATE sample at this water point? A
duplicate should be taken at the first water
point on Fridays

Photo

16. [Photo] Take a photograph of the water

point.

17. [Direct Observation] If there is any NIRA AF 85

identification number or mark on this water
point, please enter it below [If there is no
identification number or sign, select "Not
Applicable"]

Conclusion of survey

18. Thank the respondent for their time. Note The baseplate where the pipes pass has rusted, animal graz
any observations here (optional) the water point

19. Scan the GPS coordinates 0.7220405° 30.3817731°(+/-) 6.200 m (gps)

Tue, Sep 20, 2022 1:41 PM

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3.3 Disinfecting the compact dry plates
After counting all the plates, a 1-litre of 0.05% bleach solution would be used to disinfect all
the used plates. The bleach solution was made by mixing 14ml of 3.85% bleach with 986ml
of tap water to make 1L of 0.05% bleach solution.

The plastic graduated cylinder was filled with 14ml of 3.85% bleach. The volume was
checked to ensure that it was exactly 14ml by keeping the cylinder on a flat table and then
looking eye levels at the cylinder 14ml marking.

The cylinder was topped up to 1000ml using the tap water and then, the solution was poured
into the bucket.

3.3.1 Procedures
 The protective disposable gloves were put on to avoid infection from the plates.
 We could then make sure that there was enough of 0.05% bleach solution in the
bucket to cover all the plates.
 The compact dry plates were then opened and placed both sides of the plate into the
bleach solution as shown in the Figure 27. The bucket was waited for at least 30
minutes.
 After 30 minutes, the plates and filters were disposed in the garbage and bleach
solution disposed into the sink.

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Figure 27: The compact dry plates with the bleach solution placed in the bucket

6 REFERENCES

Foundation, C., 2022. Overview - Hilton Foundation. [Online] Hilton Foundation. Available

at: <https://www.hiltonfoundation.org/about> [Accessed 29 September 2022].

Aquaya.org. 2022. [Online] Available at: <https://aquaya.org> [Accessed 29 September

2022].

(Nasco), W., 2022. Whirl-Pak® Sterile Sampling Bags (Nasco). [Online]

Weberscientific.com. Available at: <https://www.weberscientific.com/whirl-pak-sterile-
sampling-bags-nasco-2> [Accessed 30 September 2022].

https://www.Aquaread.com

https://portal.mwater.co

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