Mekdela Amba University

Course: Biological Laboratory and Field Techniques (Biol 2021)

 Course credit: 2 cr.hr (3 hours practical/week
 Target group: Year II ,semester I Biology students
 Course status: Compulsory

By
Kasaye Teshome (MSc. in Ecology and Systematic zoology )

March 2014E.C/ 2022G.C

1
Tuluawlia, Ethiopia
 1. Introduction

• Scientists investigate natural events to try to find out exactly why and how they
happen.

• To arrive at an answer, they need conclusive evidence that a certain factor causes the
event.

• Very often, this kind of evidence can only be obtained by carrying out experiments.

• You will learn how to proceed from identifying the problem to planning and carrying
out an investigation in such a way that the results will enable you to conclude that
the factor you are investigating does (or does not) cause the event to happen.

2
 Introduction cont.
• Along the way, you will see how some of the greatest biologists
have used this scientific method in their investigations.

• You will also learn how to write a report on a scientific

investigation in such a way that scientists all over the world would
be able to instantly recognize the stages in your investigation and
carry it out for themselves if they wanted to check your results.
3
 1.1. The Scientific Methods

The Scientific Method involves a series of steps that are used to investigate a natural occurrence.

• What are the main steps of the scientific method?

1. Observation / Asking a Question/

2. Do background research

3. Form a Hypothesis

4. Design and carry out an experiment to test the hypothesis

5. Record and Analyze Results

6. Accept or reject the hypothesis

7. Draw Conclusions

8. Report results 4
 1. Observation / Asking a Question/

 Develop a question or problem that can be solved with an experiment.

 A problem or a question must first be identified.

Examples of questions

1. We know that tomato seeds germinate when they are planted. But, why don’t
tomato seeds grow inside tomatoes?

2. How much water can a root hair absorb?

3. Why does a plant stem bend toward the light?

4. What effect does temperature have on heart rate?

5
 2. Do background research
• Before we start trying to do the whole investigation ourselves, we will
first check scientific magazines and the internet to see if anyone else
has looked into the problem, or into a similar problem.

6
 3. Form ( Construct) a Hypothesis

• Hypothesis is a possible and suggested explanation to the question

or problem.

• It is simply a prediction and has not yet been proven or disproven.

• It must be stated in a way that is testable.

• A statement is considered “testable” if evidence can be collected

that either does or does not support it.

7
 4. Design and carry out an experiment to test the hypothesis

Experiment: A procedure to test the hypothesis.

 It include a detailed materials list. The outcome must be measurable (quantifiable).

4.1. Designing a Controlled Experiment

1. The factors in an experiment that can be changed are called variables.

Some example of variables would be: changing the temperature, the amount of light

present, time, concentration of solutions used.

2. A controlled experiment works with one variable at a time. If several variables were

changed at the same time, the scientist would not know which variable was responsible
8
for the observed results.
 Design and carry out an experiment to test the hypothesis cont.

3. In a “controlled experiment” only one variable is changed at a time.

All other variables should be unchanged or “controlled”.

4. An experiment is based on the comparison between a control group with an

experimental group. These two groups are identical except for one factor.

The control group serves as the comparison. It is the same as the experiment

group, except that the one variable that is being tested is removed.

The experimental group shows the effect of the variable that is being tested.
9
 Types of variables in an experiment
1. The independent variable is the variable that is deliberately changed by the scientist.

Example: Tomato seed germination in tap water.

2. The dependent variable is the one observed during the experiment. The dependent variable is the data we
collect during the experiment. This data is collected as a result of changing the independent variable.

Example: Tomato seed germination in tomato juice.

• Occasionally, there is a variable that might influence the results that you can’t control.

• Such a variable is a confounding variable.

This is because it ‘confounds’ the interpretation of the results. You couldn’t be certain that it was the IV

producing the changes in the DV because of the presence of the confounding variable.

For example, if you measure the carbon dioxide uptake by wheat plants as the light intensity changes over the

day, you cannot control the effect of change in temperature. It could be a confounding variable. 10
 5. Recording and Analyzing Results

The data that has been collected must be organized and analyzed:

1. To determine whether the data are reliable.

2. Does the data support or not support the hypothesis?

11
 6. Accept or reject the hypothesis

• Rejected Hypothesis - An explanation that has been ruled out through

experimentation.

• Accepted Hypothesis - An explanation that has not been ruled out through
excessive experimentation and makes verifiable predictions that are true.

Examples:

1. If soil temperatures rise, then plant growth will increase.

2. There are chemicals in tomatoes that stop the seeds from growing whilst
they are still in the tomatoes themselves .
12
 7. Draw Conclusions
• A summary of what was learned during the experiment.

• Experiments must be repeated over and over.

• When repeated, the results should always be the same before a valid
conclusion can be reached.
8. Report results

• Communicate the Results: Share the results with others.

13
 Reading Assignment
• People often confuse these by the following terms , but they are really quite
separate notions and all are important to how well an experiment is received
by other scientists.

• So you should have to read and understand each term and ideas
Prediction

Fair test

 Accuracy

Validity

Reliability 14
 Reading Assignment
Prediction an educated guess as to how the biologist thinks his/ her
experiment will turn out.
Fair test an experiment in which the only difference between different repeats
of the experiment is the different values of the independent variable, all other
factors that could affect the outcome have been kept constant (they have been
controlled)
Accuracy how precisely something has been measured or counted.

15
 Cont.
Validity This is about whether or not our experiment measures what it
says it is measuring.
In the tomato seed experiment, we said that our results were due to the
presence or absence of tomato juice.
 For our experiment to be valid, we must be certain that our results were
only due to the changes in the independent variable and nothing else.
 So had we not controlled all the other variables, our experiment would
not have been valid.
16
 Reading Assignment
Reliability is a measure of how dependable our results are.

If we were to repeat the investigation, would we get more or less the same
results?
 There are several things we can do to increase the reliability of our
experiments.

• We can standardise all our procedures, so that we always do exactly the same
thing.

• This makes it much more likely that we will be able to repeat our results.
17
 Cont.
This allows us to see, hopefully, a general pattern. It also allows us to: –
spot any anomalous results and, if it is justified, to exclude these –
calculate an average result, which is likely to be more representative
than any individual result

• We can try not to use personal judgement.

For example, if in a given experiment we have to wait until a solution

turns a certain shade of red, one person’s judgement will almost
certainly differ from the next person’s. There are ways around this:
18
 1.2. Laboratory safety rules
Introduction

• Science is a hands-on laboratory class.

• You will be doing many laboratory activities, which require the use of
hazardous chemicals and expensive lab equipment.

• Safety in the science classroom is the 1st priority.

• To ensure a safe science classroom, a list of rules has been developed

and provided to you in your safety contract.

• These rules must be followed at all times.

19
 General lab rules
• For the chemicals you are working with, you should be familiar with:
the Standard Operating Procedure for using that chemical in your lab.
the hazards associated with that chemical
the personal protective equipment (PPE) required for using that
chemical
storage requirements
waste disposal procedures
the procedures to be followed in the event of an emergency
20
 General lab rules cont.
• Avoid working alone in the laboratory.

• If you must work after hours or on weekends:

 make arrangements with others in the building to check in with you
periodically.
Let someone know you are working alone, and make arrangements to call
and check in periodically.
 avoid conducting hazardous experiments during this time.
Do the most hazardous aspects of your work during regular work hours
when there are others present. 21
 Eye Safety

• Wear safety goggles when working with chemicals, flames,

or heating devices or if possibility of flying debris

• If you wear contact lenses let your teacher know

• If a chemical gets in your eye, flush in water for 15 minutes

and notify the teacher.

• Flush in water for 15 mins. and notify the teacher

22
 Eye Safety cont.
 Chemical Goggles
 Laser
 Welding
 Chemical goggles protect your eyes, eye sockets, and the facial
area immediately surrounding the eyes from impact, dust, and
splashes.
 Chemical goggles are generally stronger than safety glasses and
are used for higher impact, particle and chemical splash
protection.
 Laser and Welding goggles protect the eyes from harmful light.
 All eye protection must be ANSI Z87 approved.
23
Eye Safety cont.
Goggles
Chemical
Laser
Welding
(Metal work or joining
metallic parts by heat )

24
 Eye Safety cont.
 In case of emergency in which a chemical goes into
one’s eye, use the eyewash station.
 Flush in water for 15 mins. and notify the teacher

25
 Hand Safety
 If a chemical spills on your skin, notify the teacher and
rinse with water for 15 minutes
Wash hands after every laboratory.
Handle glassware, sharp tools and heated containers
carefully.

26
 Sharp Objects
• Always cut away from fingers and body
• Always carry sharp objects
with points and tips facing
down and away
• Never try to catch falling
sharp instruments
• Grasp sharp instruments only by the
handles

27
 Sharp Objects
• Notify teacher if you get cut
• Broken glass and sharp objects do not
go in trash cans
• Teacher will clean up broken glass

28
 Electrical Safety
• Only electrical plugs are to be placed
into an electrical outlet
• Unplug electrical equipment after use

• Keep all electrical cords, wires, and

appliances away from water
29
 Heating Safety
• Tie back hair and loose clothes when
working
with open flames
• Never look into a container as you
are heating it
• Never point the end of a test tube
being heated at yourself or others
• Never heat in a closed container

30
 Heating Safety

• Never leave a heat source

unattended
• Heated metal and glass looks
cool, use tongs or gloves
before handling
• Do not place hot glassware
directly on lab desk or in cold
water 31
Chemical Safety
• Read all labels twice before removing
a chemical from the container
• Only use the type and
amount of chemical
instructed to use
• Never touch, taste, or
smell a chemical unless instructed by the
teacher
• Never mix chemicals
unless instructed to do so 32
Chemical Safety cont.
Transfer chemicals carefully!
• Keep lids on chemical containers when not in
use
• When diluting an acid, pour the
acid into water
• Consider all chemicals dangerous

33
 Animal Safety

 Only handle living organisms with teacher

permission
 Always treat living organisms humanely
 Wash your hands after handling animals
Treatment of Specimen
 Respect the life of all laboratory specimen.
 They gave their life for your education

34
 Plant Safety
 Do not eat any plants in lab

 Wash your hands after handling plants

 Tell your teacher of any plant allergies

 Like any organism, plants should be
considered possibly harmful

35
 General danger

You Should Never…

Enter store room unless given permission

Take any chemicals from lab or store room

Touch any equipment, chemicals, or other

materials until instructed to do so

36
 General danger cont.
You Should Never…
Eat or drink in the lab
Use lab glass-ware to eat or drink out of

37
 General danger cont.
You Should Never…
Engage in….
– practical jokes

– horse play
– rough house

38
In case of an
emergency…
• Know the locations of:
• fire extinguisher
• fire blanket
• body shower
• eyewash station
• first aid kit

• If you spill a harmful

chemical on yourself or
in your eyes, start
rinsing immediately
and send your partner 39
to get teacher’s help
 Remember to…
Stay at your work station

Maintain a clean work area

Read and follow all directions

Report any spills, accidents,

or injury to the teacher immediately
Clean and put away all equipment at the end of the
lab period
Dispose of waste products according to instruction 40
 1.2.2. Personal Protective Equipment
(PPE)
Personal Protective Equipment (PPE) is any safety equipment workers wear to
prevent injury in the laboratory workplace controls

Head
Eyes
Face
Hands
Feet
Body
Hearing
Respiratory
41
 Eye PPE
Needed when an learners work presents the potential of causing eye injury from physical,
chemical, or radiation agents.
Examples of hazards:
 Machines
 Lasers (device that produce narrow and powerful beam of light)
 Heat
 Tools
 Flying Particles / Dust
 Electrical work
 Chemical handling

42
 Types of Eye Protection
 Non-Prescription safety glasses.

Prescription safety glasses.

43
 Types of Eye Protection
Goggles
 Chemical
 Laser
 Welding
Chemical goggles protect your eyes, eye sockets, and the facial
area immediately surrounding the eyes from impact, dust, and
splashes.
Chemical goggles are generally stronger than safety glasses and
are used for higher impact, particle and chemical splash protection.
Laser and Welding goggles protect the eyes from harmful light.
44
Care and Maintenance
Check prior to each use for cracks or
damage.

Replace as necessary.

Store in a clean area.

45
 Face PPE
Needed when learners work presents the potential of causing
facial injury from physical, chemical, or radiation agents.
Examples of hazards:
 Contents under pressure
 Splash hazard
 Flying objects / particles
 Electrical work
46
 Types of Face Protection

Face Shield
Welding Shield

47
 Donning Face PPE
Safety goggles or goggles must always be worn under a
face shield.

Once goggles are in place, position face shield over face

and secure on brow with
 headband.

Adjust to fit comfortably.

48
 Hand PPE
 Needed when work presents the potential of causing hand injury
from physical, chemical, or radiation agents.
 Examples of hazards:
 Absorbing harmful substances
 Sharp objects capable of causing cuts, abrasions, or punctures
 Chemical or thermal burns
 Electrical work
 High/Low temperatures 49
 Types of Hand Protection

Puncture / cut / abrasion Resistant

Those with a latex allergy can use vinyl, nitrile, etc.

Chemical Resistant
Voltage Rated
Temperature Resistant
Infectious Agent / Biohazard Resistant – Latex, Vinyl,
Nitrile, etc) 50
 Feet PPE
Needed when work presents hazards that have potential to cause a
foot injury:
Examples of hazards:
 Falling objects
 Rolling objects
 Piercing/cutting injuries
 Electrical work
51

 Types of Foot Protection

Steel toed
Electrical resistant
Chemical resistant

52
 Body PPE

Needed when work presents a potential for contamination or injury to

other parts of the body such as legs, arms, back, chest.
Examples of hazards:
Heat
Splashes
Hot/cold metals and liquids
Sharp objects
Chemicals
Electrical work
Radiation
Types of Body Protection
Lab coats
Aprons (front body coat)
Chemical resistant sleeves ( arm cover)
Coveralls (cloth over other clothes) 53
Body PPE Removal

54
 Hearing PPE
 Needed when the average (over an 8 hour period) noise level of an area reaches 90
decibels.
Examples of high noise areas can be:
 Mechanical rooms

 Shops

 Construction Sites

 When working with machinery/power tools

Types of Hearing Protection

Ear Plugs
Ear Muffs
Canal Caps 55
 Ear Plug Fit Check

Check the fit when you're all done. Most of the foam
body of the earplug should be within the ear canal.
Try cupping your hands tightly over your ears. If
sounds are much more muffled with your hands in
place, the earplug may not be sealing properly. Take the
earplug out and try again.

56
 Respiratory PPE
Needed when work presents an inhalation hazard.

Examples of hazards:
Working with uncontained chemicals.
Working with highly toxic chemicals.
Working in dusty environment.
Painting.
Welding. 57
 Types of Respiratory
Protection
Dust Mask

½ mask

Full Mask

Powered Air Purifying respirator (PAPR)

Supplied Airline Respirator

Self Contained Breathing Apparatus (SCBA)

58
 PPE care, Maintenance and
Repair
Do not use PPE if it is damaged and in need of repair.

It is the responsibility of the user to make their supervisor

aware as soon as PPE becomes damaged so that new
PPE can be obtained.

59
 1.2.2. Handling common laboratory equipment, chemicals,
reagents, media, cultures and other materials

All lab materials must be stored in the appropriate locations at the end of
each session and gloves and other disposables placed in biohazard bags.
Tubes and racks should be placed in the appropriate location for
autoclaving.
All spills should be reported IMMEDIATELY to the lab instructor for
proper cleanup reported spills can result in biohazardous conditions.

60
 Cont.
Storage and handling of chemicals
The following rules should be strictly observed in storing and handling of chemicals.

After using a chemical, the container should be tightly closed and returned to its
original place.

 Care should be taken to see if all the containers have the appropriate labels stuck
to them.

 No bottle should be left unlabeled.

 Corrosive chemicals should be stored in corrosion- resistant chambers.

 Some chemicals especially concentrated acids, produce lot of fumes on exposing

to the atmosphere and therefore, it is advisable to handle these inside a fume-
hood.
61
 Cont.

Flammable solvents like benzene, alcohol, ether and carbon

disulphide should never be handled without turning off

burners, heaters, etc.

Some of the organic solvents like ether, because of their high

volatility, build-up a pressure in the containers. Such

containers should be cooled in ice and then opened.

 Flammable liquids should not be heated in an open vessel

over a free flame. They should be heated in a flask fitted

with a reflux condenser 62
 Cont.
Handling of acids
Store strong acids separately and away from volatile organic chemicals.

Wear acid resistant chemical gloves and aprons, when working with acids.

Emergency flood showers or eye wash foundation must be available.

Dilute acids by stirring the concentrated acid slowly into the water.

Do not pour water in to acid.

When using acids, make available suitable neutralizing agents for use in
the event of spills.
Acids should be neutralized with weak bases such as sodium carbonate or
bicarbonate.
63
 1.2.3. Laboratory organization
Laboratory as the place for experiments new processes and phenomena
always face with unknown dangerous experiments and in many cases
unknown risks and hazards.

Regardless of nature of the laboratories, risk is the inescapable subject

of the laboratories activities.

Developing laboratories with using of various modern technologies,

chemical reagents, biological agents make great contribution to progress
and development in science and technology is needed.

But it has sever and serious hazard consequences in some case which
may have been arisen acute (immediately), chronic (middle or long
term) hazards which have been shown in the long term and very late. 64
What Should the Laboratory Look Like?

General Ventilation
Supply air diffusers and room air exhausts should be
located so as to avoid intake of contaminated air

Windows should be operable

Local Ventilation
Fume hoods used for operations that give off:
 Noxious Odors
 Flammable or Poisonous Vapors

65
Safety Showers and Eyewashes
Must be available in all lab areas that use or store
chemicals which are corrosive or an irritant to the eyes or
skin
Combination eye wash and drench hose units at the sink
are now available

66
Match the Extinguisher to the Risk!
Fire Extinguishers Must Be:
Clearly labeled to indicate the types of fire they are
designed to extinguish.
Visibly inspected monthly and maintained annually.
 Class ABC Extinguishers should be located:
– At the Laboratory exit
– Within 50 feet of any point in the lab.

 Class D Extinguishers are required for combustible metals.

67
 Means of Egress/Exit
Two or more well- marked and unobstructed
evacuation exits are recommended in a lab.

It’s Shocking

There should be no accessible live, exposed electrical wiring.

Consideration should be given to installing ground-fault circuit

interrupters on electrical circuits within 6 feet of water sources .

68
 Chemical Storage in the lab.
Safe Storage of Chemicals is a Necessity in Every School
Laboratory!
Minimizes exposure to students and staff to corrosive and
toxic chemicals
Lessens the risk of fire

Prevents the mixing of incompatibles & the creation of an

emergency situation

69
The “Don’ts” of Chemical Storage in the lab!
Avoid storing any chemical above eye level.

Don’t store incompatible chemicals together.

Don’t store chemicals near sources of heat or sunlight.

 Don’t store chemicals in the hoods or acids on metal shelves.

Avoid storing anything on the floor, especially glass bottles.

70
 Proper Incidental Spill Control Equipment Includes

Spill control materials such as

Spill Control Pillows,
Pads
Booms
 Scoops
Brooms
 Pails
 Bags
 Absorbent – such as Diatomaceous Earth for filter
 Neutralizers – for Acids & Alkalis
 Mercury Spill Control Kit

71
 Waste chemical disposal
• Requires:
• Proper storage– same rules apply – make sure waste
chemicals are compatible
• Proper labeling – tags should be placed on bottles
name of chemical
• Pre-planning – know what waste you’re creating prior
to carrying out experiments; minimize purchases
• Record-keeping – of all waste chemicals on hand and
those already picked up for disposal 72
1.3. Introduction to basic biological laboratory equipment
Beakers

Beakers are used for holding

various chemicals.

Not for measuring precisely.

Sizes vary.

73
Graduated Cylinder
Used to precisely measure the
volume of liquids or run
experiments.
Read from the meniscus at eye
level.
Plastic ring always on top if
applicable.
Sizes vary.
74
 Erlenmeyer Flask

Used to approximately

measure the volume

various liquids.

Erlenmeyer
Useful for mixing by

swirling

Sizes vary.
75
Florence Flask

Used to boil liquids.

Also used to collect gases, if applicable.

Sizes vary.

76
Volumetric Flask
Used to prepare precise standard

solutions.

They are only good for 1 specific

volume.

Comes in many sizes

77
Reagent Bottle

Used to store, transport, or view

reagents such as acids or bases.

Rubber Stoppers
Used to close flasks and test tubes.

The holes allow the insertion of

glass tubing, probes, or

thermometers as needed by the
experiment.
78
 Test Tubes and Rack
 Used to hold chemicals/tubes while

experimenting.
 Not for measuring precisely.

 Waft!

 Aim away from faces.

 Sizes vary.

 Label tubes.

79
 Buret and Buret Clamp
Buret: Used for precisely measuring
dispensed liquids
Buret
Buret Clamp: Holds buret to
ring stand.

Double buret clamp

Single buret clamp
80
Ring Stand and Ring Clamps

Base/Pole of set-up for

experimenting.
Holds glassware in place for
heating or evaporating.
Test Tube Brushes
Cleaning.
• You must clean tubes
before and after you use.

81
Test Tube Holder
Used for carrying or holding hot
test tubes.

Thermometer
Measuring temperature.
Use metric!!
82
 Hot Plate
Used to heat substances.
Bunsen Burner

Used to heat substances

quickly or if > 400oC is
needed.
Do not use with
flammable substances.

83
Rubber Tubing

• Used for a variety of things.

• Example:
• Connecting Bunsen burner to gas
valve stem.
• Connecting glass tubing together.

84
Clay Triangle
Used to hold a crucible in place
on a ring stand.

Also helps absorb and spread

heat of flame.

Part of ring stand set-up.

Crucible and Cover

Used for heating substances.

Can withstand high direct

heat.
85
 Crucible Tongs

Used to carry crucible.

Beaker Tongs
Used to carry beakers.

Mortar and Pestle

Used to grind substances into
powder or slurry.
86
Dropper and Bottle

Used to measure out small

amounts of liquids for
experiments.

87
 Triple Beam Balance
Measures the mass of an object.
Make certain the balance is calibrated correctly before use.

Double Pan Balance

Used to compare the masses of two substances.
Digital Balance
Used to accurately measure mass.
Only up to 200g in our labs.
88
 Pipet, Pump, Bulb

Used to precisely measure the volume of liquids in

small amounts.

89
 Berol Pipet
 Disposable pipets used to transfer small amounts of
chemicals.
 Graduated pipets can precisely measure small amounts
of chemicals.

90
Tools of biologist
Microscope
1. Optical –
use beam
of light
2. Electron
use
electric

91
Tools of biologist
Petri dish/agar plate/
Petri dishes are often used to make
agar plates for microbiology studies. The
dish is partially filled with warm liquid
containing agar and a mixture of specific
ingredients that may include nutrients,
blood, salts, carbohydrates, dyes, indicators,
amino acids or antibiotics. Once the agar
cools and solidifies, the dish is ready to be
inoculated ("plated") with a microbe-laden
sample. Virus or phage cultures require a
two-stage inoculation: after the agar
preparation, bacteria are grown in the dish
to provide hosts for the viral inoculum.

92
 Centrifuge

• A laboratory centrifuge is a piece of

laboratory equipment, driven by a motor,
which spins liquid samples at high speed.
There are various types of centrifuges,
depending on the size and the sample
capacity.[1]

• Like all other centrifuges, laboratory

centrifuges work by the
sedimentation principle, where the
centripetal acceleration is used to separate
substances of greater and lesser density. 93
Crucible tongs
to hold hot crucibles

Dropper pipet or
disposable pipet for
drawing in a liquid and expelling it
in drops

Evaporating dish
liquids are heated over a flame so that they
evaporate, leaving a solid residue

94
 Tools of biologist

Filter paper
special paper used to separate solids from
liquids

Forceps
used to pick up or hold small items

Funnel
for pouring liquid or other substance through
a small opening

95
 Tools of biologist
Goggles
Protects eyes from
chemical splashes

Graduated cylinder
accurately measures
liquid volumes

Hot plate / stir plate

used to heat and stir
substances
96
 Tools of biologist
Spatula
small scoop used to transfer powder
and crystal chemicals

Spot plate
a flat plate with multiple "wells" used
as small test tubes

Stirring rod used for stirring

97
 Tools of biologist
Striker
used to light Bunsen burner

Test tube
open tube used to hold liquids

Test tube clamp

clamp used to hold hot test-tube

98
 Tools of biologist

Watch glass
to hold solids while being
weighed, or as a cover for a
beaker
wire gauze
used to support a
container (such as a
beaker or flask) during
heating

99
• Quadrat
• A quadrat is a frame,

traditionally square, used in

ecology and geography to

isolate a standard unit of area

for study of the distribution of

an item over a large area.

Modern quadrats can for

example be rectangular,

circular, or irregular. The

quadrat is suitable for sampling

plants, slow-moving animals,

and some aquatic organisms.

100
• The pitfall trap is an
adaptation by the ecologist
of a common hunting
technique: the use of a pit in
the ground into which an
animal falls and cannot
escape. The ecologist's
pitfall trap consists basically
of a glass, plastic or metal
container, sunk into the soil
so that the mouth is level
with the soil surface. Many
ground dwelling animals fall
into the trap and are unable
to escape.
101
Trapped animals in pit

102
 Dry pitfall traps

• Used to collect reptiles or frogs and generally consisting

"of jars, tins or drums which are buried in the ground
with their lips flush with the ground's surface. The
openings are covered by a slightly raised lid or stone, or
other object to keep out predators and prevent trapped
animals from being overheated (during the day) or
drowned (when it rains)
Wet Pitfall Trap
• A wet pitfall trap is defined as a dry pitfall trap containing
a solution designed to trap, kill and preserve an animal or
animals. Aqueous solutions used in these traps include;
formalin (10% formaldehyde), alcohol, methylated
spirits, trisodium phosphate and picric acid.

103
Data logger

is electronic device that

records data over time
or in relation to location
by the help of sensor
Data can be Temptature,
Weather rain…..

104
Tools of biologist
• PH kit

105
Flow meter and Field microscope

106
• Theodolite

107
G.P.S. • Binocular is device
global positioning system
having two telescopes
and can magnify far
objects/ matter/

108
 1.3.1. Dissecting kits
• Dissecting Pan
• Used to hold specimens in place
• To use:
• Lay the specimen on the tray and pin to the rubber
bottom

109
 Dissecting Needle (Probe)

Multi-purpose tools to move parts of the specimen

Scalpel
Use: to make precision cuts and/or take
samples

110
Forceps
• Use: to grasp and manipulate parts of a specimen

111
1.3.2. Sterilization equipments

• Freeing of an environment from all living

microorganisms includes bacteria and their spores, fungi,
parasites and viruses.

• Sterilization means germ free objects.

• Sterilization occurs by:

• Physical methods.

• Chemical methods.

112
 Sterilization
by
Physical
methods

Heat Radiation Filteration

113
Physical methods
• Heat:
• Exposure of the objects to heat will kills microbes by
coagulation of protein, denaturation of enzymes and
oxidation.
• Filtration:
• Sterilization through removing of microbes from fluids by
exposing to small size filter. Used for heat sensitive fluids
like serum, antibiotic, suger, and urea.
• Radiation:
• Exposure to irradiation causes denaturation of proteins and
enzymes.

114
sterilization by heat

Heat

Moist
Dry heat
heat
115
Dry heat

Dry heat

Hot air
Red hot Flaming Incineration
oven

116
Red hot

• Exposure of wires and forceps to the Bunsen flame until it becomes

red hot, then cool down and use.

• Used for loop, forceps, and metal rods.

117
Flaming

• Slowly passing of an objects to the Bunsen flame will reduce the

number of microorganisms.

• Used for sterilization of the mouth of bottle, flasks, containers and

test tubes, smear slides etc,,,.

118
Flaming

119
Hot air oven

• Instruments consist of heater, oven.

• Used for sand, powder, metal, glass.

• Thermal death point and Thermal death time:

• 160C for 60 min.

• 180C for 30 min.

120
Incineration

• Is treating of an objects to heating over 250C until become black.

• Done for used equipment.

121
Moist heat

Moist heat

Less than Above

At 100C
100C 100C

122
Less than 100C
• Pasteurization of milk:
• Holding method (65C for 30 min)

• Flash method (72C for 20 sec)

• Preparation of vaccine:
• By heating at 56C for 30-60 min.

123
At 100 C
• Steaming:

• Single exposure of the microbe to steam at 100C for 90

min.

• Boiling:

• Boiling water is the most common form of application of

moist heat but is not capable of killing endospores or killing
all viruses
• At 100º C for 30 min.

124
 Above 100C (Autoclaving)
• Depends on steam and pressure.

• Steam is a hot air able to penetrate through things.

• Pressure will rise the temperature from 100C to 121C.

• Moist heat is more effective than dry heat at a given temperature or

length of exposure. also more penetrating than dry heat

• Make complete killing of bacteria, their spores, fungi and their spores,
parasites and viruses including Envelop and non Envelop virus.

• 121C for 15 - 20 min.

• Flash autoclaving at 134C for 4-5min.

125
Radiation
• Sterilization by radiation kills microbes by causing
mutation to the cellular protein and disrupting
cellular elements.
Types of radiation:
1) UV ( not a good sterilizing method).
2) Ionizing , most medical disposables ( syringes,
needles).

126
Filtration
• Sterilization by mechanical removal of pathogenic
microorganism by passing through membrane filter. Unable to
filter viruses according to their small size.

• The pore size is less than 0.45 µm(bacteria size 100-1 µm).

• Used for sterilization of heat sensitive fluids like serum,

glucose, urea, and Amino acids.

127
 Dyes:

• Crystal violet and Eosine are very effective antiseptic.

Surface active agents:

Soap and other detergent make mechanical remove of

microbe by scrubbing of dead tissue so reduce their
number.

Alcohols:

• E.g.: Anti-bacterial, sanitizer.

• They able to act and Evaporate, short period of time..

128
1.4. Laboratory report writing methods

A title

 theory

Aim or objective

Chemicals and apparatus used procedures

Results/in graphs, charts, tables/

Conclusion

Evaluation/ assessment of the limitations that has been

uses/
Acknowledgment/ person’s work/ 129
When you write the report

1) Write complete sentences in passive voice

2) When using graphics, label and describe them

3) Do not “cut and paste” from another source, even the

lab manual

4) Print and proofread before you upload

5) Hand in reports on time

130
2.Microscope

131
 Microscope cont.

 A microscope is an optical instrument that uses a lens or a

combination of lenses to magnify and resolve the fine details of an

object.

 The magnified image seen by looking through a lens is known as a

virtual image, whereas an image viewed directly is known as a real

image.
 Virtual image- an image that cannot be seen directly. It can only be seen by a viewer by

looking through a lens.

 Ex. –Looking through a microscope.

 Real image- an image formed by the actual convergence of light rays on a screen. Ex.

Movie projected on a screen. 132

2.1. Types of microscopes

1. Simple Microscope:
It is called simple because it has only one lens.
Hand lens
Hooke’s Microscope

133
2. Compound, Light Microscope:
It is called compound because it is composed of two lens systems.
It uses light to view the object.

134
• A---------------------------------------------
• B---------------------------------------------
• C---------------------------------------------
• D---------------------------------------------
• E---------------------------------------------
• F---------------------------------------------
• G---------------------------------------------
• H---------------------------------------------
• I----------------------------------------------
• G---------------------------------------------
• K---------------------------------------------
• L--------------------------------------------
• M--------------------------------------------

135
3. Comparison Microscope

 Provides a side by side comparison

 of specimens

 Structure consists of 2 compound microscopes

combined into one

 unit.

 Designed to use a bridge incorporating a series of

mirrors and lenses to join 2 independent objective
lenses into a singular binocular unit.

 Viewer will see a circular field divided into 2 equal

parts by a thin line.

 The specimen on the left will be seen

 in the left half of the field of view and

 the specimen on the right will be seen

136

4. Electron Microscope:

• Uses electrons to gain data and view

objects at extremely high magnifications.
• The SEM shows very detailed 3-
dimensional images created without
light waves.

Microscopes and

137
Transmission Electron Microscopes

 Invented in the 1950’s

 Magnifies up to 350,000x

 Allows scientists to observe viruses

and substructures

 Aims a beam at a thin section of

specimen, beam is transmitted

through the specimen.

138
Stereoscopic Microscope
The most commonly used microscope in crime labs
Two separate monocular microscopes each with its own
set of lenses
Produce a three-dimensional image with a right-side-up,
frontward orientation

Stereo Microscope

Compound Microscope
139
2.2. Parts and functions of the light microscope

140
 Magnification power of each lens

Types of lenses
Eye piece Objective lenses
/ocular/
lenses Low power Middle High Oil emersion
power
power
Magnification power

10x 4x 10x 40x 97-100x

141
2.3. Handling of the microscope

142
2.3. Handling of the microscope

The microscope is an extremely valuable instrument of the biologist

and it may be seriously damaged by careless use you should observe
the following rules in handling a microscope:
When carrying a microscope grasp the arm (Limb) of the
microscope firmly with one hand and support the base (foot) with
the other hand, carrying a microscope with one hand only and the
lamp or any other object with the other hand is strictly forbidden.
The table on which we place the microscope should be always flat.

Do not put the microscope at the edge of the laboratory table to
avoid falling and breakage.
143
Handling of the microscope cont.

 It should be put on the table the arm facing you and the base away from you

 Always carry the microscope in an upright position. If tilted, the ocular (the lens on

the top) may fall and be damaged.

 Place the microscope near, but not on the edge of your lab bench with its arm facing

you

 The magnifying lenses of the microscope can easily be scratched. Therefore they

should be cleaned only with special lens paper. In the absence of the latter, tissue or
very soft toilet paper may serve as a substitute.

 When using a microscope do not incline it . to have an easy look into the microscope,

pick a sufficiently high stool, or in case your seat is adjustable, adjust it to the level
that allows an easy look into the microscope without bending of your neck.

144
Handling of the microscope cont.
Before you return your microscope to its box see that no slide is left on
the stage and also make sure that the low power objective (the shortest
objective) is in exact position i.e. right above the stage opening.
Place the microscope in its box with its arm facing you.

The objective lenses may be seriously damaged and slides may be broken
on the stage if the correct procedure of focusing is nor followed.
 Therefore when you are focusing with your microscope you are expected
to follow the focusing steps given in another page of this manual strictly.
In summary treat your microscope respectfully.

It is a more precise instrument than those used by workers who made
many of the basic discoveries in biology. 145
2.4. Setting the microscope for use (adjusting the light)

• In order to study your mounted dot or any other object under the microscope first you
have to set up your microscope for use.

1. Place the microscope near the edge, not on the edge, of your laboratory bench with
its arm facing you

2. Arrange your lab stool such that it allows you an easy look through the ocular of
your microscope without inclining it.

3. Make the diaphragm wide open

4. If the condenser is in a lowered position, raise it as near the stage as it will go.

5. Turn on your microscope through and while looking into the microscope without
the ocular adjust the concave surface of the mirror until the field of vision (the circular
area you see in the microscope) is uniformly illuminated, Note that once you have
adjusted the light you should not move the microscope or the lamp otherwise you may
need to readjust the light. 146
 Practical 3 : Magnification and resolution
1. Magnification- the power of the microscope to
enlarge the image of an object
2. Resolution- the power of the microscope to
show detail clearly.
It means the ability to separate fine details.

For example, if the distance between two dots is less than

0.1 mm, the two dots would appear as a single dot to the
unaided human eye.
The resolving power of the human eye is about 0.1mm.
147
Magnification and resolution cont.
The microscope magnifies
and resolves.
The resolving power of a
microscope depends upon
the kind of illumination used
because,
The resolving power is ½ x
the wave length of the
illumination used.
148
Magnification and resolution cont.
• White light having an average wave length of 5500
angstroms, the objective cannot resolve anything less than
2750 Angstroms (i.e. ½ x 5500) or 0.275 micron since many
parts of the cell have dimensions less than 0.275 microns (or
275 mill microns) their presence remained undetected until a
moans of greater resolution was found.

149
Magnification and resolution cont.

The electron microscope uses beam of electrons instead of light.

When electrons are propelled by a charge of about 50,000 volts
they have a wave length of about 0.05 A (angstrom).

• Therefore, theoretically, an electron microscope can resolve objects

with a diameter of about 0.025 A (i.e. ½ x 0.05 A).

1 mm = 1000 microns or;

1 microns = 1000 mill microns,

1 mill micron= 10 Angstroms.

150
Mounting (dry, wet and permanent mount)

Mounting: - is preparing a specimen for observation under a

microscope.
Put a dot on a piece of paper and lay it on a microscope slide, using a dropper
put a small drop of water on the dot (wet mounting).

 Cover the dot with a clean cover slip.

To do this, first make the cover slip stand at right angle to the glass slide and
slowly lower it until it covers the object.

 If this is done carefully-there should the glass slide and slowly lower it until
it covers the object.

If this is done carefully-there should be no air bubbles imprisoned in the

water between the slide and the cover slip.

 You may practice this procedure of MOUNTING until you have perfected it. 151
CONT.
 Put the slide on the stage of the microscope, and move the
slide until the object (the dot) comes in the centre of the stage
opening i.e., on the path of the light rays coming from below.
 Wet mounting preparing an object for study under the
microscope.
Hold a glass slide by the edge between your fore fingers and
thumb and clean it with a piece of soft paper.
 Place the piece of paper bearing a dot in the center of the
slide with the dot facing upwards.
152
 CONT.
 Place a drop of water on the piece of paper.

 Holding a cover slip by the edge between the forefinger and

thumb clean it with soft paper.

 Take care, or else it can easily break.

 Holding the cover slip between the forefinger and thumb, stand it
at one end of the paper and lower it to about 45 degrees.

 Supporting with a needle or a pencil or a needle lower the cover

slip gently until it covers the paper.
 If the above steps are carefully done no air bubbles get entrapped.
153
Focusing with Low Power Objective

If the low power objectives are not in position (i.e. if it is not right
above the stage opening) hold any two objectives between your thumb
and the forefinger and turn the nose piece until the objective clicks into
position.

Looking from: the side, lower the body tube with the coarse adjustment
to within 5 mm of the slide.

The body tube of some types of the microscopes cannot be

lowered to the extent the low power objective touches the
slide. Thus, if you cannot lower it further, do not apply force.
In some types of the microscopes it is the stage that is raised
154
up towards the objective instead.
Focusing with Low Power Objective
Looking into the microscope about 2cm away from the eye piece,
slowly raise the body tube with the coarse adjustment until the
image comes into view.
Do not plant your eye right on the ocular for if you see anything
you will only see a reflection of your eye and eyelashes. Keep
both eyes open while looking into the microscope.
Spectacles used to correct near or far-sightedness need not be
worn when using the microscope.
Only those who are very near-sighted may need spectacles to
avoid headaches.
If the image viewed is blurred, focus it with the fine adjustment
until it becomes clear.
If the image is off the centre of the field of vision, move the slide
very gently, while looking into the microscope, until the image is
in the centre of the field.
Adjust (increase or decrease) the diaphragm opening as necessary 155
Focusing with Middle Power

In order to focus the object (dot) under the medium power goes to

step 5 without changing the focus under the lower power.

 If you have followed the above steps, your object is in focus under

the low power. Now, holding any two of the objectives between the

thumb and the forefingers rotate the nose-piece until the middle

power objective clicks (i.e. you hear a clicking sound) into position.

 Focus with the fine adjustment until you get a clear image

 Increase the diaphragm opening if more light is needed 156

Focusing with the High power

Now you can directly (i.e. without changing the focusing under the
medium power), go to step 10 in order to focus the object under the
high power.

Your object is already in focus under the middle power, taking hold of
any two objectives rotate the nosepiece until the high power objective
clicks into position care should be taken so that the objectives do not
hit against the clips

Focus with the fine adjustment until you get a clear image. Never use
the coarse adjustment when focusing with the high power objective.

 If more illumination is needed the opening of the diaphragm may be

increased you can practice focusing until you perfect it. 157
Some Important Points of Caution in Focusing

 Never focus downwards without looking at the objectives from the side if

your microscope is the type that is focused by lowering or raising the body

tube.

 Never focus upwards without looking at the objectives from the side if your

microscope is the type that is focused by raising or lowering of the stage.

 Never focus the high power objective or an oil immersion objective, if your

microscope has one, with coarse adjustment. Always use the fine adjustment

otherwise you may break the slide and possibly damage the microscope

itself. 158
Some Important Points of Caution in Focusing

Do not try to focus an object directly with the high

power objective or with the oil immersion objectives.

First locate or focus the object with the low power, then

shift to the medium power and then to the high power.

Alternatively, you can focus it with low power and them

high power without focusing it with the middle power.

159
4. Observing cellular structures

Onion Epidermal cells (plant cells):

An onion bulb is made up of several layers of modified leaves.
Each leaf has delicate epidermal layers which can easily be peeled
off.

 The epidermal layer consists of cells that can be readily seen under
the microscope.

Mounting onion epidermal cells:

The thin whitish transparent epidermis is removed from piece of

onion leaf and is mounted in a drop of water.

160
 4.1. Steps on Mounting onion epidermal cells:

1. Peel the inner epidermis from one of the leaves.

2. Place the piece of the epidemic on a glass slide, add a drop of

water and then a cover slip (Note that when you cover the cover

slip, try to avoid air bubbles that may mistake you with cell

organelles, your instructor shows you how to avoid air bubbles).

3. Examine it under the middle power. Is the nucleus clearly

visible? Is the cytoplasm thick in the center or in the periphery

of the cell? 161

Steps on Mounting onion epidermal cells cont.

4. Remove the slide from the stage and place a drop of iodine solution at one

end, hold a place of absorbent paper at the opposite end of the cover slip. Wait

for about 5 minutes till the cells get stained. (N.B. if your mount is not good

you can mount fresh place of epidermis directly in a drop of iodine).

a. Examine under the middle power and draw two or more adjacent cells and

label the parts.

b. Which part (s) of the cell became more deeply stained?

c. How do you account for the difference in staining ?

162
4.2. Cheek Cells (Animal Cells).

1. Gently scrape the inside of your check or the inside of your

lower lip with the broad end of clean toothpick (See figure 3.4 for
procedures)

2. Stir the scraping into a drop of water on a clean glass slide.

3. Place a cover slip on it and examine it under the middle power.

a. Are the parts of the cell clearly visible?

b. What is the color of the cells?

4. Place a drop of methylene blue at one end and hold a piece of

absorbent paper at the opposite ends of the cover slip.
163
5. Wait for about 5 minutes until the cells get stained.
4.2. Cheek Cells (Animal Cells).

5. Examine your preparation under the middle power (or

you can use the high power with good care).
a. Can you now distinguish the nucleus and the cytoplasm?

b. Compare the position of the nucleus in cheek cell (animal cells)

with that of onion cell (plant cell).

c. Draw two or more adjacent cells and label the parts.

6 Cell structure of Unicellular Organisms: A culture of unicellular

organisms is placed in the lab. Mount a drop of the culture and examine it
under the middle (or high power) With the help of the charts, identify,
draw, and label at least on representative unicellular organism from the
culture (Note: since some of them are fast. 164
Practical 5: Diffusion and osmosis

Brownian movement is random movement of microscopic particles

suspended in a liquids or gases resulting from the impact of molecules of
the surrounding media.

The random movement of molecules from a region of higher

concentration to a region of lower concentration is known as diffusion.

 Osmosis can be defined as the diffusion of water molecules across the

cell membrane from a region of their higher concentration to region of
their lower concentration.
The net movement of water into or out of the cell depends up on the
relative concentration of the protoplasm and the solution surrounding
the cell. This can be checked by experiments by using different
chemicals.
165
Practical 6: Testing for biological important molecules

Test for carbohydrates

166
 Molisch, s test
Principle.

This general test is used to identify the carbohydrates from other

macromolecules proteins and lipids.

Sulfuric acid which is the test reagent dehydrates pentose to form

fufural and hexose to form 5- hydroxylmethylfufural.

 The furfural and 5-hydroxymethylfurfural further react with α-

naphthol present in the test reagent to produce violet compound
appears as a ring at the junction between the two layers.

167
Practical 6: Testing for biological important molecules

Procedure.

1. In clean dry test tube add two ml of 5% glucose solution or any other sugar.
2. Add for the sugar solution in the test tube two drops of (10% αnaphthol in
absolute ethanol) and mix well.

3. Add for the mixture by dropping 1-2 ml of concentrated sulfuric acid on the
internal wall of the tube in a slant manner without mixing as shown below.

4. Observe the appearance of violet ring between the two layers of the mixture
solution.

168
Benedict’s test cont.
• This test is used to distinguish between reducing and non reducing sugars.

 All monosaccharides are reducing sugars. Some disaccharides including lactose and
maltose are also reducing sugars (sugars with a free aldehyde or ketone group).

 Other disaccharides such as sucrose are non-reducing sugars and will not react with
Benedict's solution. Starches are also non-reducing sugars.

• Benedict test is most commonly used to test for the presence of glucose in urine.

• The presence of glucose in urine is an indication of diabetes mellitus.

• Reducing sugars reduce cupric ions (Cu+2) to cuprous form (Cu+1) which is responsible
for the color change to brick-red.

• The color appearance and the amount of the precipitate formed (Cu2O) depends on the
amount of reducing sugar that present.

• RCHO + Cu(citrate)2 RCOO- + Cu2O

• Aldose Benedict reagent carboxylate ion brick-red ppt. 169

Benedict’s test cont.
Procedure. 1.
In clean dry test tube add 1 ml of 5% glucose solution (reducing sugar).

2. In the second test tube add 1 ml of 5% sucrose solution (non reducing
sugar).
3. For each tube add 2 ml of Benedict's reagent and mix well.

4. Keep both tubes in boiling water bath for five minutes.

5. Observe the formation of brick-red precipitate for glucose solution

indicating that the sugar is reducing (positive result). Change in colour due
to red ppt. of Cu2O
And no color change occurs with solution of sucrose indicating that the
170
sugar is non-reducing (negative result)
Benedict’s test cont.

171
Iodine test
The Iodine test is used to test for the presence of starch.

Iodine solution – Iodine is dissolved in an aqueous solution of

potassium iodide - reacts with starch producing a deep blue-black
color.Iodine forms colored complexes with polysaccharides.

 Starch is a polysaccharide with coiled structure which turns blue

Black when bound to Iodine.

172
Iodine test Cont.

• To test for starch you add iodine solution.

• If starch is present the reddish

brown iodine solution changes to a blue black
colour.

• To test for cellulose you add Schulze's reagent.

If cellulose is present it will turn a purple colour.

173
Acid hydrolysis of disaccharides and polysaccharides

• Acid hydrolysis of disaccharides and polysaccharides produces

monosaccharides by breaking the glycosidic links (ether bonds)

between monomer units in the structure of the molecule.

• Human beings cannot digest the polysaccharide known as cellulose

because we lack an appropriate enzyme to hydrolyse cellulose.

174
Practical 7: Testing for biological important molecules
Testing for fats and oils
Grease spot test
• In the test, some oil and some water are smeared onto a piece of paper.
• Some time later, the water smear would become not translucent.
• But the smear of oil would keep translucent for a long time.
• This is known as the grease spot test.

Working principle
The working principle is that most grease or fat have a high boiling point.
• So, they are non-volatile.
• In room temperature, the spot of water can absorb enough heat from the air
and evaporized.
• But the spot of grease can never absorb enough heat to evaporized.
• When the liquid is inside the sheet of paper, it diffracts light.
• So, light can pass from one side of the paper to another side. This gives the
phenomenon of "translucent".
• When there is no liquid in the paper, there is no diffraction.
175
• So, light cannot pass it through.
Emulsion Test
The emulsion test is a method to determine the presence of lipids
using wet chemistry.
The procedure is for the sample to be suspended in ethanol,
allowing lipids present to dissolve (lipids are soluble in alcohols).
The liquid (alcohol with dissolved fat) is then decanted into water.

 Since lipids do not dissolve in water while ethanol does, when the
ethanol is diluted, it falls out of the solution to give a cloudy
white emulsion. Any lipid in the filtrate will not dissolve in the
water. It will form an emulsion that makes the liquid appear milky
white.
176
Test with dyes
Sudan III Test

Sudan III dye is a special dye used for identifying

lipids.
Because lipids dissolve in nonpolar compounds and does
not dissolve in water, and Sudan III also dissolves in
nonpolar compounds, but not water, when 2-3 grains of
Sudan III are added to a food sample, if the dye is soluble
lipids are present in the substance and if it is insoluble
they are not.
177
Testing for proteins

• Biuret structure:
Principle:

• The biuret reagent (copper sulfate in a

strong base) reacts with peptide bonds in

proteins to form a blue to violet complex

known as the “biuret complex”.

• This color change is dependent on

the number of peptide bonds in the
solution, so the more protein, the more
178
intense the change.
Biuret structure cont.
• The NaOH is there to raise the pH of the solution
to alkaline levels; the crucial component is the copper
II ion (Cu2+) from the CuSO4.

• When peptide bonds are present in this alkaline solution,

the Cu2+ions will form a coordination complex with
4 nitrogen atoms from peptide bonds.

• N.B. Two peptide bonds at least are required for the formation of
this complex.

179
 The Biuret Reagent is made of
sodium hydroxide and copper
sulfate.
The blue reagent turns violet in the
presence of proteins, and the darker
the purple color, the more protein is
present.
Biuret’s Reagent is unstable, but can
be mixed on the spot using NaOH &
Benedicts
180
 Xanthoproteic test
Objective:
• to differentiate between aromatic amino acids which give positive results
[yellow color] and other amino acids.
Principle:
• Concentrated nitric acid react with aromatic nucleus present in the amino acid
side chain [nitration reaction] giving the solution yellow color.

Some amino acids contain aromatic groups that are derivatives of

benzene.

These aromatic groups can undergo reactions one such reaction is the
nitration of a benzene ring with nitric acid.

The amino acids that have activated benzene ring can readily undergo
nitration.

In the presence of activated benzene ring, forms yellow product. 181
 Xanthoproteic test Cont.

• Amino acids tyrosine and tryptophan contain

activated benzene rings [aromatic nucleus]

which are easily nitrated to yellow colored

compounds.

• The aromatic ring of phenyl alanine dose not

react readily with nitric acid despite it

contains a benzene ring, but it is not activated,

182
therefore it will not react
Xanthoproteic test Cont.
• Procedure

• Add 2 mL amino acid solution in a

boiling test tube, add equal volume of
concentrated HNO3.

• Heat over a flame for 2 min and observe

the color.

• Now COOL THOROUGHLY and

CAUTIOSLY run in sufficient 3ml
NaOH (why)

• Observe the color of the nitro

183
derivativities of aromatic nucleus.
Million’s test
Objective:

• This test is specific for tyrosine. Because it is the only amino acid
containing a phenol group.

• Note: phenol group, a hydroxyl group attached to benzene ring.

Millon’s reagent contains mercury and HNO3 and is very toxic,

corrosive a strong oxidant, an irritant, and can cause burns.
Phenolic amino acids such as Tyrosine and its derivatives respond to
this test.
Compounds with a hydroxybenzene radical react with Millon’s reagent
to form a red colored complex.
184

Million’s test cont.

• Principle:

• The phenol group of tyrosine is first nitrated by

nitric acid in the test solution. Then the nitrated
tyrosine complexes mercury ions in the solution to
form a brick-red , appearance of red color is
positive test.

Note: all phenols (compound having benzene ring

and OH attached to it) give positive results in
Millon’s test
185
Million’s test cont.
Procedure
Add 2 ml of protein solution in a test tube,

add 3 drops of Millon’s reagent.

 Mix well and heat directly on a small

flame. boil with boiler for 5 min

 A white precipitation is formed with

albumin and casein (but not gelatin);

the precipitation gradually turns into brick

186
red.
Practical 8: Introduction to Field Technique

Purpose of biological field trips

 The term “field trip” is usually used when a person or group of persons
undertake tour of places where they expect change from normal daily life.

The purpose of the trip is usually observation for education, non-

experimental research or

 To provide students with experiences outside their everyday

activities, such as going camping with teachers and their classmates.

major source of providing knowledge to the students by giving

opportunity for self-experiences and observations and self-

long-lasting learning. 187

Purpose of biological field trips Cont.
 When educational field trips are undertaken by students of an educational
institution the main aim is not only recreation and pleasure but also gain
additional knowledge through direct experiences.

 It is easier to grasp the lesson when students do hands on activities because

remembering actions is easier than remembering words.

Myers and Jones (2009) describe that educational field trips should
be designed around specific educational objectives.
If a field trip not planned well in advance will end in confusion and
will be a waste of time and money.
So field trip should be planned as a cooperative activity involving full
pupil participation under the teacher’s supervision. 188
Materials and equipment used in field trips

Alarm Clock Fuel Pajamas

Alcohol/Formaldehyde Gloves Pens, Pencils and Eraser
Blotter/ News paper Glue Pocket Knife
Camera,& Extra Batteries Ground Cloth Pruning Shears/ Scissors
Canteen or Water Bottle Hand lens Quadrats
Climbing Equipment Hat Raincoat
Coat or Jacket Insect Repellent Knife Rope
Collecting Permit Language Dictionary Ruler
Cook Stove Large Plastic Bags Shaver
Deodorant Lighter or Matches Shoes
Entry Permit Lunch, Candy Snake Antivenom
Extra Collecting Bags Map or GPS Soap
Feminine Needs Medicines (antibiotics, Spare Glasses
Field Book malaria pills, etc.) Sweaters
Field Press Money Swim Suit
Field Shirts Mosquito Net Toilet Paper
Field Socks Newspaper Toothbrush
Flash Small Plastic Bags (for Underwear
Flashlights small specimens) Watch
Forceps Notebook Water Filter
Pajamas Wooden press
189
Planning field trips (organization)

 Teachers must consider the needs of the student and the requirements of the particular
field trip site.

 In addition, it is imperative to consider the field trip site location.

 Is the site near enough so that the student will not get overly tired?

 How will you get to the site? Are there fees involved?

 Who is responsible for transportation?

 Teachers must consider the entire day's schedule, from beginning to end.

 For example, when must the group arrive at the site?

 Who will arrange for meals? Successful field trips planner must consider safety issues.

 Are there any hazards involved?

 Do you have enough parents or volunteers?

 Have arrangements been made for student with special needs? 190
Planning the Field Trip

Step1 –Selecting the Site

Step 2 –Field Trip Transportation

Step 3 –Health & Safety Instructions

Step4 –Emergency Planning

Step 5 Evaluating the Trip

191
Step 1 ‐ Selecting the Site

Identify the rationale, objectives and plan of evaluation for the field
trip.

Select the area where the field trip is to be held.

Contact the educational coordinator for the site and arrange the date and
time.

Obtain the pre-trip information package if one is available.

Record addresses, directions, contact persons, phone numbers, email

addresses, etc.

Conduct a pre-visit to familiarize yourself with the major features of the

field trip.
192
Take digital photographs to share with students prior to the visit.
Step 2 Field Trip Transportation

• Have the students meet you at the site and take transportation.

• For instances where chartered/contract/ transportation service

(e.g., bus rental) is warranted for a field trip, the field trip is then
recommended to begin and end on campus.

• Use only chartered transportation services contracted through

Procurement and Support Services.

193
Step 3 Health & Safety Instructions

 Develop health and safety instructions for all participants.

 These instructions will differ according to the specifics of your field trip activity and
location.

 Consider the need for special clothing or equipment that may be required due to site

conditions. Plan for hazards that could be encountered and mitigation

procedures (e.g, Sun Exposure‐ sunscreen, hat/clothing to avoid sunburn,

water, Chap Stick, etc.). Assess Personal Protective Equipment (PPE) that is
available/required and how and when it is to be used. Physical demands that
may be required (e.g., long walks, climbing hills, embankments) where this is
likely to be excessive or beyond the capacity of some participants.
Forbidden/restricted items (e.g., firearms, alcohol, etc.) and forbidden/
restricted behaviours (e.g., rock climbing with ropes, illegal specimen
194
collection, etc.).
 Step 4 ‐Emergency Planning
The type of field trip also dictates /focus/ the level of emergency
planning needed.
For example, if the trip location is remote, you need to consider the
availability of a first aid kit, availability of individuals with first aid
skills, and a cell phone or appropriate means of communication in
the event emergency aid is needed.
Confirmation that cell phones will operate from the field trip site
should be made in advance so that alternative arrangements can be
made if needed.
Compile and take with you a list of emergency contacts, including
local police.
Provide these numbers to field trip participants, along with your
emergency contact number.
 In addition to local emergency responders, notify the campus when
an emergency occurs during a field trip by contacting
195
Step -5 Evaluating the Trip

Complete a "Journey" regarding the field trip.

This will provide a good reference for future field trips.

 What was of unique educational value in this field trip?

 Did the students meet the objectives/expectations?

 Was there adequate time?

 Was there adequate staff and adult supervision?

 What might be done differently to make this an even better experience in

the future?
 What special points should be emphasized next time?

 What special problems should be addressed in the future?

 What would improve a visit to this site in the future? 196

Practical 9: Methods of sample collection and preservation
Basic equipment that is readily available (or can be purchased easily) will enable the
collection of plant specimens to be much more effective and less wasteful of the specimens
obtained. Such equipment would include:
• Notebook - to record information in the
field • Plant press and newspaper - for
• Secateurs - for cutting specimens pressing plants.
• Tags - for numbering specimens • Sleeping bag, torch light, warm
• Plastic bags - large (for plant specimens) cloths and personal kits,
and small (seeds etc.) • Rain coat and shoes,
• Water - to moisten specimens once • Alcohol or rectified spirit,
placed in bags;
• Rubber bands - to seal plastic bags
• Digital camera with extra batteries
• Hand lens - to examine specimens in the • Old Newspaper, inch tapes and
field field press,
• Pen/pencil - for recording information on • Markers, pen, pencils and eraser,
tags and in notebook
• Lighter. Spirits, soaps and towels,
• Ruler - for measuring variation of leaves
etc. • Permission letter, identity proof
• paper. The specimen is deposited and and tour programme.
maintained in a herbarium in order to be • GPS, compass and field lens, 197
accessible for future studies.
Steps to collect the representative samples of plants

1. Permits
Before going on to private land you must request permission from
the owner to access and traverse their land.
Collecting specimens in National Parks and protected forests is
illegal unless you have a permit.
2. Safety /Protective equipment /
• It is advisable to take personal protective equipment such as
sunscreen, a hat, long-sleeved shirt and long trousers, sturdy/strong/
shoes, water and food on any collecting trip.
3. Safe travel procedures
• Always let someone know where you are, and when you expect to
return. For prolonged journeys, details of your intended route and
destination, call-in procedure and expected time of return should be
left with someone who can raise help if necessary. 198
Steps to collect the representative samples of plants cont.

4.Selecting the plant material

A good specimen includes stems, leaves, flowers and fruits.

 Basal parts of grasses, sedges, ferns and bulbous plants are essential for
identification.

Underground parts e.g. tubers, rhizomes are important for some plant
groups.

 The plant material should be fertile i.e. in flower or fruit (both if

possible), as these characteristics are often vital for identification.

This might entail returning to the site when the plant is in flower/fruit.

Spend time looking at a number of individuals, and choosing one with a

199
number of flowers or more mature fruits.
5. Size of the specimen

A specimen should ideally be 25–40 cm long and up to 26 cm wide, allowing

it to fit on a standard herbarium mounting sheet which measures 42 x 27 cm.

This is also the approximate size of tabloid newspapers.

 Plant parts that are too large for a single sheet may be cut into sections
pressed on a series of sheets, for example a palm or cycad frond.

 Long and narrow specimens such as grasses and sedges can be folded once,
twice or even three times at the time of pressing.

 In this way a plant of up to 1.6 metres high may be pressed onto a single
sheet.

For very small plants, a number of individuals may be placed on each sheet.

200
6. Features of the plant

When collecting from trees or large shrubs, distinctive or notable

features should be recorded, for example branching habit, height
and width of the plant and details of the bark.

You may need to collect more than one specimen to show the range
of variation that is present, for example mature and immature parts,
juvenile and adult leaves, coppice shoots.

201
7. Handling plants during collection

• For best results, specimens should be pressed within a few minutes of

being removed from the plant.

• Many plants wilt and fade soon after collection.

• A day press is convenient for short trips taken from the vehicle.

• If specimens cannot be pressed at the point of collection, for example if

it is raining or on steep terrain, they may be stored in large plastic
bags.

• The bags should be kept moist, and the specimens not jammed in too
tightly.

• Make sure that each bag is correctly labelled, using one bag per
202
collection site.
 8. Data to be recorded in the field

•A field notebook should contain the following information to

ensure that all the relevant field notes are taken.

 field name
 Scientific name:
 Family:
 Collector:
 Collection number:
 Date:
 Location:
 Latitude: Longitude: Altitude:
 How common: (e.g. dominant, localized, occasional, rare)
 Habit: (e.g. tree, shrub, herb or climber) 203
Plant presses
Plant presses are designed to dry and flatten leaves and flowers.

Presses were used by early explorers for identification and

preservation of new species they encountered on their journey.

204
 Plant presses cont.

Steps in pressing plants

1. Bottom of press is one of the boards.

2. Place one piece of cardboard on top of bottom board.

3. Place a couple of pieces of newspaper on top of the cardboard.

4. Place your collected plants to be pressed on top of newspaper.

(Note: take time to position plants as you wish for them to be
pressed; once dry they are brittle. Succulent plants (large &
chunky) do not press well.

5. Use plants/flowers that are flatter in nature, such as violets).

5. Place a couple of pieces of newspaper on top of the plants. 205

 Plant presses cont.
6. Place a piece of cardboard on top of newspaper.
7. Continue layers (newspaper, specimen, cardboard and repeat).

8. Once you have filled your press, place the other board on top.

9. Wrap the belt around the press and cinch it tight (use two belts if
you are using a large press). After 24 hours, cinch belts.

10. Place the press in a dry, well‐ventilated area and do not open for
at least 48 hours. (More time would be better).

11. After 48 hours, you may remove the plants and mount them on
paper using glue or tape.
206
 Mounting plant specimens
 Mounting of specimens decreases fragmenting of most fragile material and
prevents specimens becoming separated from their labels.
 If the plant collection is a long-term project, specimens should be mounted on
sheets of archival (permanent) cardboard or paper that are usually about 42 cm
long x 27 cm wide.
 Specimens should be attached to the sheets with archival-quality tape or thread.

 One disadvantage of mounting specimens is that it can make parts of the

specimen inaccessible for examination, so it is essential that this be borne in
mind during specimen arrangement and mounting.
 For example, easily reversible mounting media should be used, specimens
should be strapped to the sheet, rather than glued all over, and the specimen
should be carefully arranged before it is attached so that it shows all features. 207
Several points should be kept in mind when mounting specimens:

Do not tape over flowers or small fruits.

Remove any loose soil from the underground organs.

Leave sufficient space on the lower right-hand corner for the specimen
label.
Arrange the specimen to display as many features as possible, for example
upper and lower leaf surfaces, and inner and outer aspects of flowers.
Arrange larger specimens diagonally across the sheet, or place them in a
V, N or M form if they have been pressed in that particular shape.
 For small plants it is usual to mount more than one on a sheet.

If so, distribute them evenly on the sheet with the largest towards the base.

208
Collection, preservation and labeling of animal specimens
(terrestrial and aquatic)

Most invertebrates can be preserved by Kill and preserve in jars

of 10% formalin solution or 70% alcohol (ethyl or isopropyl; make
label with India ink and place inside jar.

 Most insects except butterflies, moths and dragonflies: Kill in

killing jar or freezer; pin properly; allow to dry; label should be no
larger than 1"x1/2" and placed on pin below insect.

 Butterflies, moths and dragonflies: Kill in killing jar or freezer;

pin and arrange wings on spreading board or piece of styrofoam;
allow to dry; label should be no larger than 1"x1/2" and placed on
pin below insect 209
 Collection, preservation and labeling of animal specimens
(terrestrial and aquatic) cont.
Vertebrates except for birds and mammals (e.g.: fish, frogs,
salamanders, lizards, snakes, turtles) kill in freezer; fix and
preserve in jar of 10% formalin solution; use syringe to
inject formalin solution into the body cavity in several
places; make label with India ink and place inside jar
Birds and Mammals: Hunt following all applicable state and
federal laws; prepare a "study skin" using the techniques
outlined in the 'ZACT' manual mentioned above; a label no
larger than 1"x3" is attached by string to rear leg.
210
Practical 9 : Population and density estimates
 Population is a group of organisms belonging to the same species occupying a particular
space at a particular time.

 Individual members of a population are interbreeding and live in a particular place, in the
same time and interact to one another as a society.

To be considered as a population it should fulfill the following points:

 members must belong to the same kind of species

 all the members must occupy the same place

 members must live in the same time and

 members must interact with one another

 Population is the basic unit of a community

 Population ecology, hence, deals with the distribution and abundance of a

211
population and the factors governing them
 Methods of Measuring Population Density
Census Vs sampling
Total counting e.g. Census

- Involves encountering (counting) each and every individual of a

population in a given area or volume

 Have higher precision as compared to the sampling method

 Could be aided by aerial photography and sophisticated radio

and infrared technologies.

 may demand much resources and time.

As a result, ecologists are inclined to using the sampling methods 212

 Sampling

Is a process of selecting representative part of a population, and

drawing conclusions about the larger population using the
information gained.

Samples are individuals that fully represent the larger population.

Sampling involves counting a small proportion of a population and

estimating the total.

213
Sampling cont.

 The two major ways of sampling employed in ecological

studies are quadrat method and trapping techniques.
 Quadrat Technique is a sampling area of any shape (usually
rectangular or triangular frames) with limited area, made either
from wood, metal, strings or any materials.
 The quadrat is simply a delineation used to demarcate the
sampling area
 A simple square quadrat could be constructed from four
sticks that have equal length. It is used to sample less or non-
mobile organisms such as plants, amphibians and the like 214
 Sampling cont.
Quadrat sampling follows the following procedure:
Selection of appropriate sampling strategy to survey the study area.
You can prefer random, systematic or stratified sampling
depending on the nature of the environment.
– Random sampling is used if individuals are distributed
randomly in space, and there is equal probability of finding any
individual at any place.
 Decide the quadrat size. There are different techniques of
determining the quadrat size. One of the famous techniques is the
Nested Plot Analysis. We usually use 20m by 20m for natural
forestlands, 4m by 4m for shrub lands, 2m by 2m for herb lands
and 1m by 1m or less for managed pasturelands.
 Determine the sampling size (number of samples) you should take
 Estimate the number of individuals in each quadrat.
Take the average of each quadrat and extrapolate to the larger study
area.
215
Sampling cont.
Image of quadrat

216
 Sampling cont.
Image of quadrat

217
 Sampling cont.

You can measure how common an

organism is in two or more sampled
areas of a habitat using a small
quadrat and comparing the distribution
numbers of species of plants or animals
in each location in a much larger area.

For a plant in the same habitat (e.g.

same field) you might choose
dry/damp areas or bright light/shaded
areas or any permutation of conditions
(here 4 possibilities, yes?).
218
 Sampling cont.
Quadrat sampling follows the following procedure:
 Suppose you are surveying a field, you can place
the 1 m2 quadrat in specific locations or choose
some places at random over a wide area.
 The frame of the quadrat can be made of wood or
metal.
 Illustrated is 1 m x 1 m quadrat and wire strung
across at 10 cm intervals.
 In this case there 100 10x10 cm square possibilities
for sampling, each has x,y coordinates of 1-10,1-10.
 You do NOT count all 100 mini-squares, instead
you can use a random number function on your
calculator to select e.g. 10 of them.
 The square with x,y co-ordinates of 7,4 is shown
on the quadrat diagram. This 'mesh' size is ok for
very small organisms e.g. tiny flowers.
 219
Sampling cont.

After placing the quadrat at selected

locations you e.g. count the flowers in
each 10 x 10 cm2 square or the total
in the whole 1 m2 of the quadrat - the
whole quadrat is 1 m x 1m.
Here the yellow flowers are quite
large and best counted per 1 m2,
giving you quantitative data
e.g. species of flower/m2.
To count the population using 10 x 10
cm squares it needs to be a very small
flower or insect. 220
Sampling cont.
Example of quadrat calculations based on sampling
Example 1. Calculating a
data
population density

Suppose you did a count of

some very small species of
flower in 10 of 10 cm2 mini-quadrats
(10 cm x 10 cm) of a 1 m2 quadrat
placed in a sunny location. The mini-
quadrats can be selected using the
random number generator.

Data counts 1-10: 7, 8, 12, 9, 9, 10,

11, 10, 9, and 8 flowers

Total count = 93 flowers

Average per 10 cm2 = 93/10

= 9.3 flowers/mini-quadrat

Now there are 100 10 cm2 squares in the full 1 m2 quadrat.

Therefore total in 1 m2 quadrat = 9.3 x 100 = 930 flowers.

The 'flower density' = 930 per m2

If you repeated the measurements in a more shaded spot, you might find a
much lower population density of the same flower.

If you know the total area, call it A m2, you just multiply the 930 x A = total
population in that area (see next example). This is just a scaling up exercise
from several small sample areas chose at random. 221
 Suppose you counted the abundance of a relatively rare flower using
Example 2 Calculating a population size (abundance)

a 1 m2 quadrat placed 8 times at random across a piece of land (its

habitat) measuring 80 m x 120 m.
Flower data counts 1-8: 2, 5, 0, 1, 2, 0, 1 and 4
(a) Calculate the average density of the rare flower per metre2
 Total population
Total flower count = 15
Flower density = 15/8 = 1.875/m2 (no need to round up at this
stage)
(b) Calculate the whole population size of the flower in this
particular habitat
Total area of habitat = 80 x 120 = 9600 m2
Total population = density x total area
Population size = 1.875 x 9600 = 18,000 flowers
(maybe its not that rare in this made-up calculation!)
222
Surveying: (2) Surveying using transects

A belt transect is a path/gradient along which one counts and

records occurrences of the species of study.

You might wish to study how the distribution of organisms changes

by sampling across a transect.

A transect is used to survey a wider area in a more systematic way

than just doing a few quadrats.

e.g you can use a sequence of quadrats along a transect to find out
how organisms are distributed across a change in habitat due to an
abiotic factor - bright light to shade, damp to dry ground, change in
soil composition (due to underlying differences in geology e.g.
223
.Total count of large animals
A complete count, or total count, counts every member of a
population.

Where populations of large species occur in open areas, such as

waterfowl on lakes, seals on breeding beaches, or pronghorns on
shortgrass prairie

Aerial counts of most individuals are possible, especially with the

aid of photography.

Sometimes, wildlife managers can count deer in enclosed

populations using a drive approach: a large group of people crosses
the enclosure in a line, counting all deer that pass in each direction. 224
 Mark-recapture methods

• These methods are used extensively to estimate populations of fish,

game animals, and many non-game animals.

• Their method involves capturing a number of animals, marking

them, releasing them back into the population, and then determining
the ratio of marked to unmarked animals in the population.

• The population (P) is estimated by the formula:

• P = M*C2nd /R, where M is the number of animals marked in the

first trapping session, C is the number of animals captured in a
second trapping session, and R is the number of marked animals
recaptured in the second trapping session.
225
 Mark-recapture methods

For example, suppose you took 200 mice out of a forest

having an unknown number of mice, put leg bands on
them, return them to the forest and let them mix
thoroughly.
If you then take 250 mice from the forest and find 50 of
them to be have leg bands, then M = 200, T = 250, R = 50,
and the unknown total number of mice (N) could be
estimated as:

 P = M*C2nd /R = (200*250)/50 = 1000 mice 226

 Summary on population estimation
1. Direct Observational Mode

Technique Target Groups of Species References

Quadrats; fixed- Sessile or relatively immobile organisms Bonham 1989
area plots

Avian point Bird species that sing or call on territories Ralph et al. 1995
counts

Spot mapping & Territorial bird species Ralph et al. 1993

nest searches

Line transect Large mammals, birds Anderson et al. 1979

Call playback Wolves, ground squirrels, raptors, Ogutu and Dublin 1998
response woodpeckers

Standardized Large herbivores, Cook and Jacobsen 1979

visual searches

Census Cave-dwelling bats; large herbivores Thomas and West 1989

227
 Summary on population estimation cont.
2. Animal sign, pellets, and scat counts

Technique Target Groups of Species References

Foot track surveys Medium-large mammals Wilson and Delahay 2001

Food cache searches Large carnivores Easter-Pilcher 1990

Arboreal mammals; fossorial

Structures (e.g., dens, nests) Healy and Welsh 1992
mammals; bears

228
Summary on population estimation cont.
3. Remote Sensing Photo and video stations

Technique Target Groups of Species References

Track plates Medium-large mammals Wilson and Delahay 2001

Ultrasonic detectors Bats Thomas and West 1989

Audio monitoring Frogs Crouch and Paton 2002

Hair traps Small-medium mammals, large carnivores McDaniel et al. 2000

Radio telemetry Limited by animal body size (>20 g) ? USGS 1997

GPS telemetry Limited by animal body ( >2000 g) ? Girard et al. 2002

Marine radar Bats, migrating birds Harmata et al. 1999

Harmonic radar Bats, amphibians, reptiles Pellet et al. 2006

229
4. Passive Capture

Technique Target Groups of Species References

Salamanders, lizards, small Enge 2001, Mengak and

Pitfalls
mammals Guynn 1987

Snap traps Small mammals Mengak and Guynn 1987

Box traps Small-medium mammals Powell and Proulx 2003

Funnel-type traps Snakes, turtles Enge 2001

Leg-hold & snares Mist

nets Large mammals Bookhout 1994

230
 5. Active Capture
Technique Target Groups of Species References

Medium-large mammals deCalesta and Witmer

Drives to an enclosure with predictable flight
response 1990

Cannon nets Medium-large mammals Bookhout 1994

Immobilizing agents Large mammals Bookhout 1994

Kolozsvary and Swihart.

Hand capture Salamanders
1999

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 6. Capture
Technique Target Groups of Species References

Mutilation/kill/ Small mammals Wood and Slade 1990

Lemen and Freeman

Pigments Small mammals
1985

Collars &
Birds/mammals Nietfeld et al. 1994
Bands/rope/

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Brief introduction of animal behavior
ETHOLOGY is the study of animal behavior with emphasis on the
behavioral patterns that occur in natural environments
Animal behavior means an animal does and How an animal
does it!
Types of animal behavior

1. innate behaviors
• automatic, fixed, “built-in”, no “learning curve”
• despite different environments,
all individuals exhibit the behavior
• ex. early survival, reproduction, kinesis, taxis
2. learned behaviors
• modified by experience
• variable, changeable
• flexible with a complex & changing environment
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There are different types of innate or stereotyped behaviour

1. Taxisis
The simplest type of innate or stereotyped behaviour.

It is an orientation of an animal (directed either towards or away)

in response to the source of stimulus.

If the orientation is towards the stimulus it is called as positive

taxis and if it is away from the stimulus it is known as negative
taxis

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 2. Kinesis
• Kinesis is a type of locomotory behaviour in relation to the source of stimulus.

• There are two types of kinesis .

a. Orthokinesis Ortho kinesis is a response that involves changes in the speed of

movement of the whole body in response to stimuli like humidity, preserve and
diffused light. Eg:Wood louse, Porcellerio scaber, a small crustacean that live in
damp areas, and they tend to lose water from their body fairly rapidly when
exposed to low humidity.

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b. Klinokinesis

In Klinokinesis the speed of locomotion remains constant

but the rate at which the animal changes direction depends on
the intensity of the stimulus.

 Eg: A planaria changes its direction ever so often as it crawls.

 If the light intensity above the animal is increased it changes

direction more frequently but moves at the same speed .

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 3. Reflexes

A simple movement of a part of the animal in response to a stimulus is

called reflex.
It is a quick, innate and immediate response of a part of the body to an
external or internal stimulus, which has great adaptive and survival value
to the organism.
Reflexes are inherited and unlearned behaviour found in all members of
the species. The knee-jerk, constrict of pupil of eye in the bright light,
blinking of eye, peristalsis, coughing etc. in man, flight in birds, web
spinning in spiders are all examples of reflexes. A reflex action requires a
reflex arc which consists of a sensory organ (receptor), a sensory nerve
(afferent nerve), the spinal cord or brain, an intermediate motor or
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efferent nerve and a motor organ or effector.
 Advantages of reflex

 Enables the animal to respond immediately to harmful

stimuli hence it has great adaptive and survival value
 Since many of the reflex actions are controlled by the spinal
cord, it relieves the brain from too much work.

4. Instinct is the most complex type of stereotyped behviour

which is unlearned, predictable, genetically controlled and
species specific and it is in response of a sign or releaser
stimuli. Of all the stereotyped behaviour instincts are the most
fascinating to study. Eg: Building of nest by birds, singing to
attract males, territoriality, migration, parental care etc. 238
 2. Learned behaviour/ acquired behaviour/

It is acquired during the life time of an individual.

Associative learning
learning to associate a stimulus with a
consequence

1. Operant Conditioning – Trial and Error Learning

B.F. Skinner

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 Trial and Error Learning by Edward Lee Thondike
• The doors of these puzzle boxes could be
open by pushing a lever or button pulling
a string.

• The cat confined in such a puzzle box

tries to escape by moving about randomly
or restlessly inside the cage and in this
process accidently it pushed the lever or
pulled the string and the door opened
which resulted in obtaining its food.

• This process was repeated for the next

few times till the cat learned to associate
the pushing of the lever or pulling of the
string resulted in obtaining the reward ie,
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food.
Classical Conditioning

Ivan Pavlov

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Spatial Learning

• Associative learning:

• Animals associate attributes of a location (landmarks) with the

reward it gains by being able to identify and return to that
location.
Nikolass Tinbergen- observed wasps used pine cones as markers to
locate their nest.
When Tinbergen removed the pine cones the wasps were unable to
locate their nest.

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 Observational Learning

• Animals copy the behavior of another animal without

having experienced any prior positive reinforcement with
the behavior.
Survival Responses Learning
• Fight or flight response
• Triggered by stress
• Adrenaline & cortisol is produced which dilates the blood vessels, increases
heart rate, increases the release of sugar from the liver, slow digestion to
conserve energy…

• Avoidance response
• Avoid stressful situations
• EX: areas where predators can hide or areas with little camouflage

• Alarm Response
• Triggered when presence of a predator or other animal that’s a
threat is detected.
• Warning is given for other in their group. 243
 Foraging Behaviors
• Most in a group and Individuals in the group can trade off jobs
(foraging and watching for predators)
• Can mob their predator and protect their young

• Social Behavior Interactions between individuals

• Develop as evolutionary adaptations

• Communication/language/sign / or by other.

• Agonistic Behaviors

• Dominance Hierarchy

• Cooperation

• Altruistic Behavior
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 Agonistic
Threatening & submissive rituals
-symbolic, usually no harm done

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 Dominance hierarchy
Indicate power and status
relationships among individuals in a
group.
-minimizes fighting for food
and mates

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Methods used to collect animal behavioral data

• Your assignment

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End
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 End
Stay Safe!!!

THANK YOU!!!

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