Chapter 3

3.1 Designing and Methodologies

A research design is a scheme of action or frame work for answering questions needed in the

data collection. It is also regarded as blue print of a project that enables the researcher to come

up with steps of solutions to a problem . Thus it is a detailed outline that shows how the project

is going to be designed ,ways of data collection and the tools to be used and the tools to be

used and the intended means of analysing data.

The scientific approach is the one to be employed ,in this project a set of experiments is going to be
used.

Design

A design is a plan ,a structure of any scientific work intended to give direction and systematizes

the project. In this project the researcher is going to design experiments on how ultraviolet light

can be used to purify water.An artificial source of ultraviolet light is going to be used.The impure

water is going to be exposed to ultraviolet light for at least 12 hours and the exposure time is

also going to be increased to observe the effect of increase time on purity of the water.The

researcher is also going to compare the purifying power of ultraviolet light and that of boiling the water.

3.2 Methodologies

Experiments

Materials:

Nutrient agar,petri dishes ,stirring rods,500 ml PET bottles,clamp stand ,connecting wires,ultra

violet light,electric power source ,droppers,gloves ,water source ,beakers,burner ,distilled water

Preparing the agar plates

Materials:

Agarvpowder,distilled water,flask or beaker,glass stir rod,laboratory thermometer,sterile petri

dishes( plastic or glass),flame or boiling mixture,heat resistant hand protection

Procedure

1. Measure agar and distilled water into clean flask or beaker.

Recipe: Agar + Distilled Water = Yield

Agar distilled water yield

23g 1000ml 50 plates

11.5g 500ml 25 plates

9.2g 400ml 20 plates

4.6g 200ml 10 plates

2. Flame, sterilize a clean glass rod to stir the medium as it mealts

3. While wearing heat resistant hand protection ,hold the flask or beaker over the flame .

Swish or stir the mixture constantly while heating

4. Boil the mixture for 1 minute .Remove from heat.

5. Place a sterile lab thermometer.In the mixture and monitor the temperature until it falls to

approximately 45- 50 degrees celsius or is a lab thermometer is not available ,cover and let stand few
minutes.

6. Pour enough melted agar into each sterile plastic petri dish to cover the bottom about 0,5cm deep.
Replace the lid immediately.

7. Place agar plates on a counter top to cool and set.Agar medium will set like stiff gelatine at room
temperature.

8. The agar medium is now ready for storage or use.

Storage: stack agar plates upside down is to prevent condensation from dripping down into the agar
surface which could then facilitate movement of organisms between colonies.

Preparing the Plates

1. If plates have been refrigerated ,set them out and allow them to warm to room temperature

2. Sterilize the loop

Allow the loop to cool for 3- 5 seconds before touching the collection area

Desterilize the loop after each inoculation

Do not allow the loop to touch any surface other than the collection area and the agar

3.Uncover each agar plate just long enough to inoculate the medium .Hold the petri dish lid

directly over the petri dish( or tilt the just enough to allow the loop inside) while inoculating the

medium to help prevent contamination from ai - borne particles.Do not allow loop to touch the petri
dish.

4. Do not dip the loop in the agar ,let it glide over the surface

5. Make a pattern of inoculation lines( parallel lines,tic- tac- toe,zigzag,initials,etc) to help

determine that what is growing is what you put there and not an air borne contaminant.

Preparing the Setup

6. Place the cover back on the plate immediately.

Incubation

1.Turn the plates upside down and put them in a warm place.The ideal temperature for

incubating is 32 degrees celsius or 90 degrees celsius F. Bacterial growth should start to

become visible in about 2- 3 days.

1.The first step is to sterilize all of your tools prior to collecting the water sample.

Sterilization kills all of the bacteria on the tools. Using sterilized tools prevents you from adding bacteria
to the samples.

a. Preheat the oven to 150 degrees celsius

b. Cover the cookie sheet completely with aluminium foil

c. Fill the pot with tap water. Put all of the medicine droppers ,the glass rods ,and the jar and lid into the
pot.
d. Place the pot on the stove and bring the water to a boil. Boil all of the tools for 5 minutes and then
turn off the heat.

2 Using the metal tongs ,lift each item from the pot and place it into the cookie sheet .Once all

of the items are on the sheet ,place it into the pre heated oven to dry. Let the items dry

completely for 10- 15 minutes.Carefully remove the cookie sheet from the oven with an oven mitt when
the time is up.

3.Use the tongs to place each medicine dropper into the glass jar. Put the lid on the jar so that

the medicine droppers remain clean. Completely wrap all of the glass rods in a sheet of

aluminium foil to keep them clean for the duration of the project. Store all of the sterilized tools in an
area that will not be disturbed.

4. Go to your local creek or stream with the clean ,plastic 1- gal jug and a pair of disposable

gloves. Place the jug into the water and fill it up. Avoid trapping any large particles or foreign

objects in the jug. Once the jug is full,replace the cap. Take the jug back to where you are conducting
your testing.

5. Now put the clamp - lamp and the Daylight Blue Reptile Bulb together,following all

instructions that came with the lamp. Find a quiet location near an electrical outlet. Clip the

lamp onto something ,such as the bottom of a cabinet door over a counter,allowing the lamp to face
downward.

Preparing the solar water disinfection Samples

Note: it is important that you observe the following agar plates at the same time.You will apply

the boiled water and untreated water samples to the agar plates while the solar water

disinfection samples are still under the ultraviolet lamp. You must keep track the time when you

start and stop tests so that the observations can be compared.

1. While the solar water disinfection samples continue resting under the ultraviolet

light,measure 1 cup of the creek or stream water in the liquid measuring cup and pour it into the

1- gal pot. Place the pot onto the stove to and bring the water to a rolling boil. Boil the sample

for 5 minutes. Remove the pot from the burner ,cover the pot ,and let the water cool to room
temperature.
2. Put on a pair of disposable gloves.Get a clean medicine dropper from the glass jar and attach a bulb to
the top. Get out three agar plates and a glass stirring rod from your aluminium foil package. Suck some
of boiled ( but now room temperature) water into the medicine dropper.

3. Remove the cover of one of the plates. Apply three drops of the boiled water to the agar.

Use the glass rod and smear the water drops in a zigzag pattern on the surface of the

agar,starting in the center.Smear the water sample from the center of the plate to the edge of the plate.
Replace the cover.

4. Repeat steps 2-3 with the two other agar plates. Reuse the same medicine dropper and bulb.

Using the permanent marker,note down the time ,the date ,and the treatment process on the

bottom of the plates( in this set of trials,it is boiling ; for the next set of trials it will be untreated).

Discard the medicine dropper and the bulb. You can reuse the glass rod later,but you will have to

sterilize it again. If you dont sterilize right away,then keep it separate from the clean glass rods

and use a clean glass rod for the next trials with the untreated water.

5. Keep your agar plates in a warm location in your house that will not be disturbed.

6. Repeat steps 2-5 with untreated water from 1- gal jug. Use a fresh medical dropper for the

untreated water sample. You can reuse the same dropper for all three agar plates. You should have six
agar plates.

7. Let the boiled water and untreated water plates sit undisturbed for 24 hours. Do you observe

any growth on the plates? Record time ,date and your observations in your lab notebook. Check

again in 48 hours ,72 hours and 96 hours. Record your observations each time you check. If you

see any growth ,count the number of bacterial colonies and record the number in you lab

3.3 Observing and Recording

Observations are going to be made from the experiment designed above and the data collected is going
to be recorded and analysed.

3.4 Conclusion

This chapter examined the experimental procedure and methodologies used to conduct the

experiments to be used to investigate the effectiveness of ultraviolet rays on water purification.