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UNIVERSITY OF THE WEST INDIES (MONA)

DEPARTMENT OF LIFE SCIENCES

PRELIMINARY BIOLOGY (BIOL0011/BLO5A)


LABORATORY MANUAL

SEMESTER I
Biological Chemistry, Cells, Variety of Life
BIOL0011 Preliminary Biology Laboratory Manual Page 2

BIOL 0 0 1 1 Pre l i m i n a ry Bi o l o g y I. C o u rse Ti m e ta b l e Se p t-N o v, 2 0 0 9


Wk D a te s L EC TU R ES PR AC TIC AL S
1 Chemical & Physical Phenomena 0 Introductions, Group Assignment
1 Sep 07-11 2 Water 1 Collection & Preservation of Data
3 Functional Groups
4 Carbohydrates 2 Physical & Chemical Phenomena - I
2 Sep 14-18 5 Lipids 3 Physical & Chemical Phenomena - II
6 Proteins
7 Nucleic Acids, Protein Synthesis 4 Characteristic reactions of compounds
3 Sep 21-25 8 Enzymes 5 Enzymes
9 Enzymes
10 Cell Structure 6 Light Microscopy
Sep 28-
4 11 Cell Membrane Transport 7 Cell Structure
Oct 02
TEST # 1 Th e o ry l e ctu re 1 -9 [6 % ]
12 Cell Cycle, Mitosis, Meiosis 8 Mitosis
5 Oct 05-09 13 Cell Cycle, Mitosis, Meiosis 9 Meiosis
14 Mutation
15 Human Reproduction 10 Mammalian Reproduction
6 Oct 12-16 16 Human Reproduction 11 Angiosperm Reproduction
17 Plant Reproduction
18 Plant Reproduction Heroes Day (Oct 19)
7 Oct 19-23 19 Mendelian Inheritance TEST # 2 Pra cti ca l 1 - 1 1 [1 2 % ]

20 Mendelian Inheritance
Linkage & Chi-Square Test 12 Monohybrid & Dihybrid Crosses; Chi-
21 Square Test
8 Oct 26-30
22 Natural Selection, Speciation 13 Natural Selection & Evolution

23 Natural Selection, Speciation


14 Kingdom Protoctista: Algae, Protozoa;
24 Taxonomy, Viruses Kingdom Fungi
9 Nov 02-06 15 Kingdom Plantae: Bryophytes,
25 Bacteria, Fungi, Protoctista
26 Plants
27 Plants 16 Kingdom Animalia: Porifera, Cnidaria
28 Animal Features 17 Kingdom Animalia: Platyhelminthes,
10 Nov 09-24
Nematoda, Annelida and Mullusca
29 Porifera, Cnidaria, Platyhelminthes
30 Nematoda, Annelida 18 Kingdom Animalia: Arthropoda
Nov 16- 31 Arthropoda & Mollusca 19 Kingdom Animalia: Echinodermata and
11
27 Chordata
32 Echinodermata, Protochordata
33 Chordates (Vertebrates) 20 Kingdom Animalia: Chordata (Vertebrata)
12 Nov 23-27 34 Chordates (Vertebrates) TEST # 3 Pra cti ca l 1 2 -2 0 [1 2 % ]
35 Biodiversity & Conservation
13 R e vi e w
BIOL0011 Preliminary Biology Laboratory Manual Page 3

BIOL0011 / BL05A— PRELIMINARY BIOLOGY I - COURSE OUTLINE

Level 0; Semester 1; Credits: 6 P

Aim: To equip students with a basic knowledge of biological principles and


processes.

Objectives: The course will enable students to:

• describe the chemical and biological foundation of life


• show understanding of the basic principles of evolution
• show variations in plant and animal life

Pre-requisites: CSEC Biology or equivalent

Syllabus: Biological techniques, biological chemistry, chemicals of life,


enzymes, cells and tissues, cell division, genetics, evolution, mechanisms
of speciation and variety of life: bacteria, protist, fungi, plants and animals.

I. CHEMICAL & PHYSICAL PHENOMENA


Matter
ƒ Atoms, molecules, ions, isotopes
ƒ Elements & compounds, mixtures
ƒ States of matter
Bonds (ionic, covalent, polar)
Dispersion systems
ƒ Homogenous & heterogeneous dispersions
Types of dispersions (Solutions, Colloids, Suspensions)
II. WATER
Structure of water
Properties of water
ƒ Solvent
ƒ pH properties
ƒ Temperature Regulation (Heat capacity, Heat of fusion, Heat of vaporization)
ƒ Surface Tension
ƒ Capillarity
III. FUNCTIONAL GROUPS
Inorganic versus organic molecules
Importance of the carbon in organic molecules
Characteristics & chemistry of Carbon
Functional Groups: -OH, -COH (aldehydes & ketones), -COOH, -NH2, -SH, -PO4.
IV. CARBOHYDRATES (STRUCTURE AND FUNCTION)
Structure and levels of organization
ƒ Monosaccharides e.g., glucose
ƒ Disaccharides e.g., sucrose
ƒ Polysaccharides e.g., starch, cellulose, glycogen
Isomerism – structural & stereoisomerism
Role in energy transfer, structural frameworks, storage
V. LIPIDS (STRUCTURE AND FUNCTION)
Structure (components) & properties
ƒ Saturated and unsaturated fatty acids
ƒ Phospholipids, micelles
ƒ Steroids eg. cholesterol
BIOL0011 Preliminary Biology Laboratory Manual Page 4

VI. PROTEINS (STRUCTURE AND FUNCTION)


Major roles in physiology & structural frameworks
Structure
ƒ Amino acid
ƒ Primary, Secondary, Tertiary and Quaternary levels of organization
Relation of structure to properties
Major classes of proteins
VII. ENZYMES
Function as Biocatalysts
Classification
Mode of action (Lock & key hypothesis, Induced fit)
Properties (relate to protein structure): effect of pH, temperature; concentration (enzyme &
substrate). Interpretation of related graphs
Competitive & non-competitive inhibition
VIII. NUCLEI ACIDS, GENES, PROTEIN SYNTHESIS
Nucleic acids Structure (components, H-bonding, base pairing)
Functions of DNA & RNA
DNA replication
Gene concept
Protein Synthesis
IX. CELL STRUCTURE
Cell Theory, endosymbiont theory
Generalized cell structure
ƒ Prokaryote cells
ƒ Eukaryote cells (plant and animal)
Structure of organelles
The plasma membrane
X. TRANSMEMBRANE TRANSPORT
Mechanisms of transport
ƒ Passive (Diffusion, Facilitated Diffusion, Osmosis)
ƒ Active (Carrier Protein mechanisms, Coupled channels Na/K pump)
ƒ Bulk Transport (Endocytosis, Exocytosis)
XI. CELL CYCLE, CELL DIVISION
Stages of the cell cycle
Mitosis cell division
ƒ Principle (Conserve chromosome number and genetic variation)
ƒ Purpose (Growth, Maintenance, Repair, Asexual Reproduction)
ƒ Process of mitosis: stages
Meiosis Cell Division
ƒ Principle (haploid/diploid, conserve chromosome number, increase genetic
variation)
ƒ Purpose (Gametogenesis, Genetic recombination)
ƒ Process of meiosis (stages, Crossing over, Independent Assortment)
Genetic variation further enhanced by random fertilization during sexual reproduction
XII. MUTATION
Gene Mutation: types
Chromosomal Mutation: types
XIII. HUMAN REPRODUCTION
Male & female reproductive systems
Gametogenesis (oogenesis, spermatogenesis)
Structure & function of gamete
Reproductive Endocrinology
ƒ Hormonal regulation of oogenesis and spermatogenesis
ƒ The principle of negative feedback
Pregnancy & Development
ƒ Fertilization
ƒ Placenta: structure & function
ƒ Foetal development & factors affecting it
BIOL0011 Preliminary Biology Laboratory Manual Page 5

XIV. PLANT REPRODUCTION


Sexual & Asexual reproduction
Flower structure & gametogenesis
Pollination mechanisms
Fertilization: self & cross pollination mechanisms
Seed & fruit formation
XV.MENDELIAN GENETICS
Monohybrid Inheritance
Dominance, Codominance, Incomplete Dominance
Sex determination, sex linkage
Dihybrid inheritance
XVI. LINKAGE & CHI-SQUARE TESTING
Accounting for disruptions in expected Mendelian ratios
Application of the Chi-square test
XVII. THE THEORY OF EVOLUTION
Natural selection
Evidence of evolution (Geographical distribution of animals, Comparative anatomy,
Comparative biochemistry, Embryology, Paleontology)
Variation
Species Concepts (Morphological, Phylogenetic, Biological)
Isolating mechanisms
Speciation methods (Parapatric, Allopatric, Sympatric, Hybridization, Phyletic)
XVIII. TAXONOMY (and DIVERSITY OF ORGANISMS)
Introduction to Taxonomy & its importance
Hierarchical nature of classification systems
Construction & use of a dichotomous key
Classification of organisms
ƒ Viruses
ƒ Domains, Kingdoms (features)
XIX. PROKARYOTES
Bacteria – Eukaryotic features
ƒ Eubacteria,
ƒ Archaeobacteria
XX. KINGDOM PROTOCTISTA
General features and structure
ƒ Algae (Chlorophyta, Phaetophyta, Rhodophyta)
ƒ Protozoa (Rhizopoda, Mastigophora, Cilliophora)
ƒ Slime Moulds
XXI. KINGDOM FUNGI
Structure and general features including nutrition and biological importance
ƒ Higher fungi
ƒ Lower fungi
ƒ Lichens
ƒ Mycorrhizae
XXII. KINGDOM PLANTAE
Structure and general features
Classification
ƒ Nonvascular Plants: Bryophyta (Hepaticae, Musci)
ƒ Vascular Plants
o Ferns
o Seed Plants
ƒ Conifers
ƒ Flowering Plants
BIOL0011 Preliminary Biology Laboratory Manual Page 6

XXIII. KINGDOM ANIMALIA


General features of animals
Multicellularity (advantages)
Animal Body Plan
• Body Symmetry (asymmetry, radial & bilateral symmetry)
• Body layers (diploblastic, triploblastic)
• Body Cavities (acoelomate, pseudocoelomate, coelomate)
• Body Segmentation (advantages)
• Embryonic development (protosomes, deuterosomes)
Classification and structure of animals
• Porifera
o Histology
o Types (asconoid, leuconoid, synconoid)
• Cnidaria
o Body layers, Nutrition
o Classification (Hydrozoa, Scphozoa, Anthozoa)
• Platyhelminthes
o Symmetry, body layers, nutrition, habitats, locomotion
o Classification (Turbellaria, Trematoda, Cestoda)
• Nematoda
o Symmetry, body layers, body cavity, nutrition, locomotion
o Classification
• Annelida
o Symmetry, body layers, body cavity, segmentation, locomotion
o Classification (Polychaeta, Oligochaeta, Hurudinea)
• Mollusca
o Generalized Molluscs
o Body Plan, symmetry, body layers, body cavity, segmentation
o Classification ((Gastropoda, Cephalopoda, Bivalvia)
• Arthropoda
o Symmetry, body layers, body cavity, segmentation
o Classification (Subphylum Crustacea, Chelicerata, Uniramia [Class
Hexapoda, Chilopoda, Diplopoda]
• Echinodermata
o Symmetry, body layers, body cavity, segmentation, locomotion, water
vascular system
o Classification (Asteroidea, Ophiuroidea, Echinoidea)
• Chordata
o Symmetry, body layers, body cavity
o Four characteristic features
o Invertebrate Chordates (Protochordata, Urochordata, Cephalochordata):
Classification and features
o Vertebrate Chordates (Agnatha, Gnathostomata): Classification and
features
ƒ Chondrichthyes
ƒ Osteichthyes
ƒ Amphibia – habitats, three chambered heart & double circulation,
integument, pentadactyl limb
ƒ Reptilia – amniote condition, the heart & circulation, integument,
internal fertilization
ƒ Aves– ectothermy, flight adaptations, care of young
ƒ Mammalia – endothermy, increased cerebral capacity, diversity of
form, increased range/habitats, care of young
XXIV. BIODIVERSITY
Definition and Value of Biodiversity
Species Status (endemic, endangered, threatened, extinct)
Resources (renewable, non-renewable)
Human Influence (hunting, habitat destruction, alien introductions) and conflicts of interest
Preservation (methods, organizations)
BIOL0011 Preliminary Biology Laboratory Manual Page 7

Practical Work:

• Experiments demonstrating biochemical and biological processes


and principles.

• Studies of fresh (living) and preserved organisms to demonstrate


external variations in different life forms.

Evaluation:
• Course work — 40% consisting of:
o Two in-course practical tests — 24%
o One in-course theory test — 6%
o Laboratory reports — 10%
• Final examination – 60% consisting of :
o One 2-hour theory paper (I) — 30%
o One 2-hour comprehensive paper — 30%

Note:
ƒ The theory paper has essay questions.
ƒ The comprehensive paper has a combination of multiple choice, fill in, matching, true
or false, short answer and labelling questions.
BIOL0011 Preliminary Biology Laboratory Manual Page 8

BIOL0011/ BLO5A TUTORIAL QUESTIONS


SECTION A
1. State the importance of the properties of water in life processes.
2. Compare hydrophobic and hydrophilic colloidal dispersions.
3. Describe the structure of proteins [lipids, carbohydrates]
4. Describe three roles played by lipids [proteins, carbohydrates] in the bodies
of living organisms.
5. Using examples, describe the characteristics of enzymes.
6. Describe how an enzyme works
7. Outline the steps involved in the replication of DNA.
8. State the functions of the three types of RNA.
9. Distinguish between DNA replication, transcription and translation.
10. Explain why the structure of the plasma membrane is well suited for its many
and varied roles in living organisms.
11. Describe the generalized eukaryotic [prokaryotic] cell.
12. Using examples, explain what is meant by active transport. [diffusion,
osmosis]
13. Describe the process of mitosis [meiosis].
14. Describe how genetic diversity is obtained in sexually reproducing organisms.
15. Define the term mutation and explain how gene [chromosomal] mutations
occur.
16. Describe the structure of the mammalian ovary [testis] and outline the
process of oogenesis [spermatogenesis].
17. State the functions of the hormones involved in gametogenesis.
18. Outline the process of fertilization in Angiosperms.
19. Explain the laws of segregation and independent assortment.
20. In TV bugs, Cable (A) is dominant to rabbit ears (a) and Colour (B) is
dominant to black & white (b). Work Aholic crosses a completely
homozygous Cable, black & white bug to a completely homozygous rabbit
ear, Colour bug. Determine the F1 genotype and phenotype. When he
crossed the F1 to get an F2 Work Aholic obtained 85 Cable Colour bugs, 32
Cable black & white, 26 rabbit ears Colour and 8 rabbit ears black &
white progeny for a total of 8000 TV bugs. Determine if these results are
significantly different from that expected.

SECTION B
21. Outline the theory of evolution by natural selection.
22. Outline the evidence that supports the modern theory of evolution.
BIOL0011 Preliminary Biology Laboratory Manual Page 9

23. Define the three concepts of species.


24. Outline the ways by which speciation can occur.
25. Explain what is meant by the binomial system of nomenclature.
26. Explain what is meant by the hierarchical taxa by which organisms are
classified.
27. Describe the characteristics that might lead you to the conclusion that an
organism is to be classified as a bacterium [virus, protist, fungi, plant,
animal]. (Include how you distinguish it from the other groups).
28. What are the three main groups of Protoctists (Protists)? [any phyla] Using
examples, explain why Protists are so difficult to classify.
29. State the biological importance of lichens and mycorrhizae.
30. Compare the morphology of algae and bryophytes. [any other groups]
31. Compare the ferns and the gymnosperms. [any other plant groups]
32. Using appropriate examples, distinguish between diploblastic and triplobastic
animals.
33. Illustrate the types of symmetry found in animals.
34. Describe the types of coelomic cavities. What is the function of the coelom.
35. Animals are distinguished by the ways in which the early embryo develops.
Explain how the Protostomes and Deuterostomes differ.
36. Describe the main features of the Porifera. [any other animal phyla]
37. Using examples from the Cnidarians, explain what is meant by
polymorphism.
38. Compare the methods of locomotion found in worms.
39. Comment on features possessed by the Arthropods which contribute to their
success in colonising a wide range of habitats.
40. Describe the ways in which amphibians are adapted to life on land, and in
what ways are they still restricted to a watery/moist environment.
41. Describe the adaptations of reptiles which allow them to live on dry, often
desert environments.
42. State the advantages of a coelomic cavity; complete digestive tract, bilateral
symmetry and cephalisation.
43. Describe the features that are unique to the class Mammalia.
44. Describe the human threats to biodiversity.
BIOL0011 Preliminary Biology Laboratory Manual Page 10

INTRODUCTION

These exercises are designed to introduce you to cells, bio-molecules,


reproduction, patterns of inheritance and principles of classification in
animals and plants.

These sessions will be scheduled opportunities for you to examine live and
preserved specimens as well as prepared slides in order to gain a better
understanding of the Biological concepts dealt with in lecture and tutorials.

Take full advantage of these practical classes in order to thoroughly


familiarize yourself with the material available as well as acquire training in
practical skills and the thought processes associated with the study of
Science.

Come prepared: Read this manual and lecture notes before you arrive for
your laboratory session. Always take this manual, your notes and
text books to help with your labeling, comments and discussion.
Be punctual: Any additional information regarding the class or modifications
of protocol will be announced at the beginning of the session. It is in
your best interest not to be late!
Observe carefully: A Greek scholar once said, ‘It is more difficult to observe
than to invent.’ Precise observation is essential in biology and an
important aid to observation is the preparation of large, clear
accurately labelled drawings.
Record accurately: Only the precise, up-to-date and detailed record book
will be useful for reference and revision. It is essential, therefore,
that drawings be made at the time of observation; copies of textbook
illustrations are of little value to you and will not impress your
examiners!
Think: You are the investigator. Try to solve problems and identify
structures yourself before consulting the lecturer, teaching assistant
or demonstrator. You may, however, request help when you are in
difficulty.
Be tidy: Leave the benches clear after each lab. Replace the microscope
and ensure it is covered. Turn off the electric lamps. Throw
discarded material into the appropriate receptacles provided. Do not
throw non-liquids substances into the sinks.
adapted from the Introductory Level 1 Biology Laboratory Manual
BIOL0011 Preliminary Biology Laboratory Manual Page 11

LABORATORY REPORTS

The cover page for each report should include the following information:
Name:
ID No.:
BIOL0011 Stream No.:
Lab No. & Title:

A typical experimental report is expected to have the following headings:


Title:
Aim1:
Materials and Methods2:
Results3:
Comments and conclusion4:

1 You should formulate an appropriate aim after reading through the experiment.
2You may or may not be required to record the methods and materials. If not, you may
write “As in lab manual”. If required, the methods should be listed in past tense. Diagrams
of apparatus may help your revision.
3Your results may include diagrams and tables; all should have appropriate titles.
4Explain your results; Refer to known theories and indicate sources of errors.

A typical page of illustration is expected to be as follows:

Title: Half-flower drawing of Cassia sp.

Kingdom:
Phylum & Sub- phylum:
Class:
Genus & Species:

Exercises should be placed in sequence, stapled together and submitted at


the end of each laboratory session.
BIOL0011 Preliminary Biology Laboratory Manual Page 12

Notes on Scientific Illustrations

Observe the specimen carefully before starting to draw. Use the textbook, notes
and picture in your lab manual to identify the main features. The aim of your
drawing is to illustrate the main features of the specimen.

Do not copy from books or other students. Develop the essential scientific honesty
which is required in higher studies where your observations could become a
source of knowledge for others. An accurate personal record will help you to
remember features you observed.

Illustrations
A large, clear diagram which shows the specimen being studied in its proper form
and proportion is required. Use plain paper. Draw on one side of the paper only.
Do not make more than two drawings on a page.

Your illustration should show the proper form and proportion of the specimen. The
lines should be clear, continuous and of even thickness. Use dotted lines to show
borders lying beneath structures. Do not shade or use inks. Use the stippling
style, i.e. making small dots with the tip of the pencil at right angles to the paper.
The density of dots should indicate darker or lighter areas.

Titles, Labels, Annotations


Each drawing should have an appropriate title, e.g. “T.S. of Rat Testis”.

As far as possible, all the labels should be to the right of the drawing.
The lines from the specimen to the label should be narrow and straight, should
never cross each other and should end at the beginning of the label.
Labels should form a vertical column and be parallel to each other, in small print
so they do not detract from the drawing.

Annotate your drawing. To annotate means to add a brief note to a label i.e. a
simple phrase, not a sentence. The annotation should provide more information
on the structure, or give the function, depending on the purpose of the drawing.

Magnification
Always state the magnification/ reduction of your drawing. Magnification = size of
drawing/ size of actual specimen. For microscopic observations, also state the
magnification at which the specimen was viewed. E.g. ‘T.S. of the Root of a
Dicotyledonous Plant viewed at x100’. This is obtained by multiplying the power of
the objective by the power of the eyepiece.

A scale bar should also be placed on the drawing to show the actual
measurement of an important region of the specimen.
BIOL0011 Preliminary Biology Laboratory Manual Page 13

PRACTICAL 1 COLLECTION AND PRESERVATION OF DATA

One aspect of science is to collect and organize facts. These facts are then
analyzed, conclusions are drawn, generalizations are made and predictions
are tested.

Information collected from the observations or experiments may be


qualitative or quantitative. Qualitative data are those which do not lend
themselves to precise numerical expression, while quantitative data are the
result of measurement and can be expressed in some definite and precise
form.

1. Collection and Preservation of Numerical Data

As a means of extracting as much information as possible from quantitative


data, these may be arranged in tables or graphs and subjected to statistical
analysis. The data, arranged in a table, yields limited information. A further
step is to represent the results in a graphical form, either to compare the
size of different quantities or to represent functional relationships, or, in
many cases, to estimate an unmeasured quantity based on the trend
shown by the graph (extrapolation procedures).

One way to graph data is to construct a histogram. This is constructed from


a frequency distribution table where the data on the variable is sorted into
classes. The height of each bar on the plot represents the number of
individuals in each class. If a line is drawn through the top of the bars on a
histogram connecting the midpoints of each category, the data can be
shown as a line graph. Typically, graphs are used to illustrate the
correlation between two variables. Here the graph is usually constructed by
plotting relevant points from the data and either joining them by straight
lines or drawing a smooth curve through them.

When constructing graphs always bear the following points in mind:


- The given or selected quantity (independent or basic variable) is always
shown on the horizontal axis (X-axis). While the observed or calculated
quantity (dependent or derived variable) is shown on the vertical axis
(Y-axis).
- Note the range of values of the quantities to be shown and select
appropriate scales for each axis. Scales should be as large as possible
and linear in nature.
- The intervals between values on the scales should be equal,
representing the same number of units. (Only when graphs are drawn
on an exponential scale should the intervals represent the logarithm of
the number rather than the number itself).
BIOL0011 Preliminary Biology Laboratory Manual Page 14

- Write along each axis the name of the quantity represented and the
units of measurement.
- Plot the points precisely according to the table of values.
- Join the points with a straight line or a smooth curve according to the
nature of the data. A line of best fit may have to be used for data which
in theory should generate a straight line but in reality gives a ‘near
straight’ relationship.
- Write a title clearly at the top to signify what the graph represents.
- Where several relationships are illustrated on the same graph use a
different plotting pattern for each relationship. A key should denote what
each pattern represents.

1A. Hand Span of Students on Campus

Materials: Metric rulers, people, graph paper or graphing software

Procedure: Collect hand-span data from at least 25 persons


¾ Spread the hand flat, stretching out the distance from the thumb to the
little finger as far as possible the measure the distance from the tip of
the thumb to the tip of the little finger.
¾ Record hand span measurements (to the nearest mm) and gender.

¾ Present the data in ascending numerical sequence.


¾ Present frequency tables for all students, males only and females only.
¾ Present a histogram to represent the hand spans of all students.
¾ Present another histogram which compares male & female hand spans.

o A frequency distribution table shows how often group of data points


appears in a given data set. Divide the numbers over which the data
ranges into intervals of equal length and tally how many data points
(hand spans) fall into each interval.
o Mean = Sum (frequency X midpoint value) / total sample – 1.
o Mode =midpoint with highest frequency.
o Median = middle midpoint /class (if there are an odd number of classes)
or the average of the two middle midpoints (if there are an even number
of classes). You may also use the raw data for the calculations.

i. Identify sources of error and means of improving the data collection.


ii. Is the hand span measurement discrete or continuous?
iii. What is the mean, mode and median hand span for all students, for
males only and for females only?
iv. What information does the histogram communicate?
BIOL0011 Preliminary Biology Laboratory Manual Page 15

1B. Class data on Reasons Students Study Preliminary Biology

A pie chart is a circular graph showing the relative frequencies or percents


of each class. Like the histogram, it is used for one variable. It allows
comparison of one class with the whole.

¾ Represent the data collected from the survey (to be done in class) as a
pie chart

FPAS entry requirement ________


Environmental/ Conservation concerns ______
Hope to study Zoology ________
Hope to study Botany ________
Hope to study Veterinary Medicine _______
Hope to study Human Medicine ________
Other _______

Determine the percentage of each class from the total number of students.

i. What advantage is obtained by presenting the data as a pie chart (over


presenting the list)?

2. Collection of Morphological Data

¾ Create and complete a data table on the external morphological


features of the organism provided. Include diagrams, measurements
and written descriptions of various body parts.

Use abbreviated writing, e.g. “skin covered with bony scales; body length
122mm”, instead of, “the skin is covered with scales which are bony and its
body is 122mm long”.

3. Collection of Ecological Data

¾ Write a description of the site indicated. Record site location, types of


organisms present, abundance and any interactions observed.
BIOL0011 Preliminary Biology Laboratory Manual Page 16

PRACTICAL 2 PHYSICAL AND CHEMICAL PHENOMENA - PART 1

The physical sciences are widely used in experimental work on living


organisms. Knowledge of certain fundamental facts of Physics and
Chemistry are essential for the broad understanding of physiological
processes. To understand the macroscopic or microscopic structures of
living organisms and the processes responsible for the formation,
modification or destruction of these structures, it is necessary to consider
the nature and properties of those particles of which all matter is
composed.

Living organisms are made up of numerous different compounds, each of


which is composed of molecules of specific chemical composition. These
compounds owe their properties both to the kinds of molecules of which
they are composed as well as to the arrangement of these molecules.

Diffusion In order to understand more clearly what is happening when a


substance diffuses from one area to another, imagine a large auditorium
that contains a great number of balls bouncing around in all directions. A
ball’s direction of movement will change only upon collision into a wall or
another ball. The result is a zig-zag fluctuating movement and the room
becomes filled with flying rubber balls. Similarly, any chamber becomes
filled with gas molecules because of their inherent motion, and the average
speed of the entire group will remain constant as long as the temperature
and pressure do not change. Now if the wall between this auditorium and
the adjacent one were removed, some of the balls would pass into the
empty room because they would no longer be impeded. The random
movement finally results in an equal number of balls in each room. At this
point the number moving from the first room to the second and vice versa
will be equal, a condition termed dynamic equilibrium. NB: Movement takes
place in both directions and the concentrations in two areas are equal.

Net Diffusion: Direction of movement of the greatest number of


molecules, i.e. net diffusion of rubber balls was from the first room to the
second. Net diffusion ceased at equilibrium, while diffusion continues even
at equilibrium.

Osmosis: One of the most important materials that enter cells is water.
The term osmosis refers to the movement of water through a differentially
permeable membrane from a region of high free energy to a region of low
free energy of water. In a strict sense, osmosis refers to the movement of
a solvent through a selectively permeable membrane, however, the only
solvent involved in living cells is water, so the more restricted definition
tends to be used. Osmosis is a special example of net diffusion.
BIOL0011 Preliminary Biology Laboratory Manual Page 17

Capillarity: If the tip of a tube of small diameter is immersed in water,


the level of water in the tube will rise higher than the surface into which the
tube was placed. Such a rise in tubes, termed capillary action, depends
upon the cohesion of water molecules to each other and the adhesion of
water molecules to the tube walls. The smaller the diameter of the tube,
the higher will be the capillary rise.

Cohesion: refers to the attraction of similar molecules to each other (e.g.


water molecules).

Adhesion: refers to the attraction between dissimilar molecules (e.g.


water molecules to the tube wall).

Carry out the following experiments and try to explain the results obtained.

1. Diffusion

1A. Half fill a test tube with water and stand it in a beaker of water. Leave
for 10 minutes for temperature equilibration. Drop a small crystal of copper
sulphate into the test tube. Leave the apparatus standing without any
disturbance and examine it at half-hour intervals.
¾ Record your aim, your observations and explanation.

1B. You are provided with three Petri dishes each containing a film of agar.
During its preparation the agar was mixed with a few drops of potassium
ferrocyanide (K4Fe(CN)6) solution.
¾ Use a cork borer to punch a hole in the center of the agar in each of
the three dishes.
¾ Fill the holes (don’t overflow) with 0.05, 0.25 or 1.25% FeCl3 (ferric
chloride) solution. Label the petri dishes accordingly.
¾ Record the times at which FeCl3 was added and, at half hour
intervals, measure the progress of the blue colour (prussian blue)
around the hole in each dish.
¾ Record your aim, your results (tabulate) and explanation.

4 FeCl3 + 3 K4Fe(CN)6 = Fe4[Fe(CN)6]3 + 12 KCl


Ferric chloride + Potassium ferrocyanide = Prussian blue + Potassium chloride
BIOL0011 Preliminary Biology Laboratory Manual Page 18

2. Osmosis

2A. Examine the osmometer and comment on your observations.


¾ Explain why coloured fluid rose up the tube.

2B. Solanum tuberosum (potato) slices were immersed in sucrose solutions


for two hours. The initial and final weights are recorded in the table.

Table: Change in weight of potato slices placed in varying concentrations


of sucrose.
Concentration of Initial Weight of Final Weight of
Sucrose (M) potato slices (g) potato slices (g)
0.15 3.45 3.98
0.20 3.25 3.73
0.25 3.20 3.60
0.30 3.10 3.40
0.35 3.02 3.21
0.40 2.80 2.92
0.45 3.30 3.23
0.50 3.25 3.04
0.55 3.18 2.83
0.60 3.21 2.82

¾ Plot the difference between initial and final weight against


concentration of sucrose.
¾ Explain the data in terms of the concentration of water and the
concentration of dissolved chemicals in potato slices. Also explain
the significance of the point at which the line crosses the x-axis?
Use terminology: hyper/hypo/isotonic, hyper/hypo/osmotic

2C. You are provided with a live slug (Phylum Mollusca, Class Gastropoda)
¾ Sprinkle the slug with a small amount of NaCl (sodium chloride).
¾ Observe what happens and explain.

3. Capillarity.
Select three capillary tubes with bores of different sizes. Stand them in a
beaker half filled with water. Observe the extent of the rise of water in the
three tubes.
¾ Explain why water rises in the capillary tubes?
Use terminology: Hydrogen bonds, adhesion, and cohesion.
¾ Explain the relationship between the diameter of the bore and the
height to which the water rises?
BIOL0011 Preliminary Biology Laboratory Manual Page 19

PRACTICAL 3 PHYSICAL AND CHEMICAL PHENOMENA - PART II

4. Surface tension of water.


Half fill a small beaker with water. Carefully lower a razor blade (flatly) on to
the surface of the water. Observe the razor.
Repeat the process using soapy water (prepared with soap powder).
¾ Record your observations and explain them in relation to surface
tension forces. Refer to properties of soap and water.
Use terminology: hydrogen bonds, cohesion, amphiphatic.

5. Colloids.
Half fill two 10ml test tubes with distilled water. Add about a teaspoonful of
clean sand to one tube and the same amount of clay (break up any clumps)
to the other tube. Shake the tubes well and let them stand for about five
minutes. Observe the level of sedimentation.
Divide the supernatant liquid from the tube containing the clay into two
equal parts. Use one part as control; while to the other add about 0.5g
magnesium sulphate or solid calcium chloride. Shake both tubes and let
them stand for about three minutes.
¾ Comment on your observations of the extent of sedimentation.

6. Emulsions
Place 2ml of water in a test tube and add five drops of vegetable oil. Shake
well and let stand. Observe. Add five drops of 10% NaOH and two drops of
oleic acid. Again shake well and observe.
¾ Explain your results. Use terminology: hydrophobic colloids,
saponification.

7. Adsorption.
Half fill two 10ml test tubes with a dilute solution of methylene blue. To one
test tube add a small amount of activated charcoal and shake well. Filter
both solutions separately through filter papers.
¾ Compare the colour of both filtrates and explain.

8. Coagulation and Denaturation.


To about 1ml of 30% egg albumen solution add an equal volume of 95%
ethyl alcohol and shake gently by inverting the tube repeatedly. Observe.
Take another 1ml of the egg albumen solution in a clean test tube and heat
in a water bath for 5-10 minutes. Observe.
¾ Comment on the effects of heat and alcohol on proteins.
BIOL0011 Preliminary Biology Laboratory Manual Page 20

PRACTICAL 4 CHARACTERISTIC REACTIONS OF


MACROMOLECULES

Carbohydrates, fats, proteins and nucleic acids are the major


macromolecules that occur in cells.

Carbohydrates range from relatively simple molecules called sugars to the


complex molecules of starches and cellulose. The simplest class of
carbohydrates is represented by the monosaccharides, e.g. glucose and
fructose. Disaccharides are formed by combining two monosaccharides.
One of the most important disaccharides is sucrose, made by linking
glucose with fructose. Polysaccharides such as starch and cellulose are
made up of a large number of monosaccharides joined in long chains.

The fats or lipids are a group of organic substances originating in living


cells. They are composed of two main groups, fatty acid and glycerol.

Protein molecules are of gigantic size made up of a large number of amino


acids units connected one to another by peptide bonds. A significant
portion of the protoplasm of plant and animal cells is made up of proteins.

The nucleic acids are of two types: DNA (deoxyribonucleic acid) and RNA
(ribonucleic acid). DNA is the hereditary material of the cell, while RNA is
important in protein synthesis.

Carry out the following tests which demonstrate some of the means
by which the above mentioned compounds can be identified.

1. Detection of Carbohydrates (reducing sugars)

1A. Add 5ml of Benedict’s solution to 5ml of the each of the carbohydrate
solutions provided. Immerse the tube in a boiling water bath for 5 minutes.
Record the colour change and the relative amount of precipitate formed for
each of carbohydrate tested. Explain the results.

1B. Label three test tubes.


To tube 1 add about 5ml of 10% sucrose solution and then 2 drops dilute
hydrochloric acid (HCI). Place in water bath for about 2 minutes then
add 5ml of Benedict’s solution.
To tube 2 add about 5ml of 10% sucrose solution then 5ml of Benedict’s
solution
To tube 3 add 5ml water and 2 drops of dilute hydrochloric acid (HCI)
Place all the tubes in a boiling water bath for 15 minutes then observe.
BIOL0011 Preliminary Biology Laboratory Manual Page 21

i. What was the point of heating Benedict’s solution with dilute HCI?
ii. If a food sample contains sucrose as the only sugar, what two tests
would have to be carried out to show the presence of this sugar?

1C. Place about 1ml of the carbohydrate solution into a test tube and add a
drop of iodine in potassium iodide solution (I2 in KI). Which of the
carbohydrates gave a blue colour? Immerse the blue coloured solution in a
beaker of boiling water and heat for 3 minutes then remove from the heat
and cool under the tap. Record and explain the results.

2. Detection of Lipids
To about 2mls of water in a test tube add 2mls of coconut oil followed by a
few drops of Sudan III or Sudan IV solution. Shake the tube well then allow
it to stand for 1 minute. Note the formation of two layers. Which layer is
coloured red? Why?

3. Detection of Proteins
The Biuret Test : In separate test tubes place 3mls of 1% egg albumen
solution or gelatin, 1% starch solution, 10% glucose solution and water.
Add 1ml 10% Sodium Hydroxide (NaOH) solution. (CAUSTIC). Mix
thoroughly then slowly add drop by drop 1% copper sulphate, (CuSO4)
solution. Note the colour changes. With which of the sample did the Biuret
test give a purple colour?
BIOL0011 Preliminary Biology Laboratory Manual Page 22

PRACTICAL 5 ENZYMES

Enzymes are complex proteins which act as biological catalysts that


regulate chemical reactions in the cell. Many of the characteristic reactions
of cellular activity, when carried out in the test tube, will proceed only at
temperatures, pressure and pH values that are incompatible with life. In
living organisms, complex metabolic reactions proceed rapidly under
physiological conditions since the organisms produce enzymes.

Enzymes are effective in minute amounts and are usually unchanged by


the chemical reactions they promote. Many enzymes are very specific in
the reactions they effect. Enzyme activity is usually expressed in terms of
the rate of the reaction catalyzed by the enzyme. The following
experiments will help in obtaining information of some of the factors that
affect enzyme-catalyzed reactions.

1. Effect of pH on enzyme activity.


Diastase is an enzyme (or mixture of enzymes) that catalyzes the
hydrolysis of starch with the production of dextrins, maltose and glucose.
The disappearance of starch from the reaction mixture could be
ascertained by the iodine test.

¾ Collect a petri dish containing starch agar and mark crossed lines on
the underside with wax pencil to divide the plate into four quadrants, A,
B, C & D. Use a cork borer to make a central well in each quadrant.
To well A add two drops of buffer solution pH4; well B, pH5; well C,
pH6 and well D two drops pH7 buffer solution.
Add two drops of diastase to each well; cover the dish and leave for 2
hours. Stain the agar with dilute iodine for two minutes. Pour off the
excess iodine and measure the diameter of the clear zone around each
well. Tabulate your results. Comment on the effect on pH on enzymes.

2. Effect of enzyme concentration on enzyme activity.


Dehydrogenases are widely distributed throughout the plant and animal
kingdoms. They transfer hydrogen from donor to acceptor molecules. In
certain enzymatic reactions coenzyme I and II act as hydrogen carriers for
the hydrogen atoms removed by the dehydrogenase. Under aerobic
conditions oxygen will act as the final hydrogen acceptor causing the
formation of water. In anaerobic conditions, other materials can serve as
hydrogen acceptors from reduced coenzyme. Methylene blue can act as
hydrogen acceptor becoming leucomethylene blue (colourless) when
reduced. It can therefore be used as an indicator of the enzyme’s activity.
BIOL0011 Preliminary Biology Laboratory Manual Page 23

2A. Place 0.5mls of 0.02% methylene blue in three 5mls snap cap vials. Fill
the first vial with fresh bottled milk; the second with bottled milk that has
been left unrefridgerated for at least three days; and the third with milk that
was left unrefrigerated then boiled. Close the tubes and leave at room
temperature. Examine at twenty minute intervals for two hours. Tabulate
your results.

i. Which milk sample became decolourised first? Which didn’t? Why?.


ii. What is the source of the enzyme?
iii. Uncork the tubes in which the milk has decolourised and blow gently
into it. Explain your observations.

2B. A methylene blue test was historically used to determine the quality of
milk in terms of its bacterial population.
¾ Repeat the test in 2A with the sample of box milk provided,
this time incubating at 37°C. Record the time taken for complete
decolourization of your sample. Rate the sample.

Time Taken For Milk Of Varying Quality To Decolorize Methylene Blue


Time to Decolourize Methylene Blue Rating of Milk
Less than 20 minutes Highly contaminated
20 minutes to 2 hours Poor
2 to 5 ½ hours Fair
2½ to 8 hours Good
More than 8 hours Excellent

3. Effect of temperature of enzyme activity.

Tyrosinase is an enzyme which occurs widely in plants and animals alike. It


is responsible for the production of most of the dark brown and black
pigments of animals. A crude extract of tyrosinase is prepared from potato
tubers by breaking down potato slices in a blender then filtering the juice
through a coarse filter.

Tyrosinase is an oxidase. It catalyzes the removal of two hydrogen atoms


and electrons from molecules of such compounds as catechol (colourless)
to form benzoquinone (coloured). The hydrogen atoms are then passed on
to oxygen to form water.
BIOL0011 Preliminary Biology Laboratory Manual Page 24

¾ In each of three test tubes add 5ml of water and ten drops of the potato
extract.
Place the first tube in an ice-water bath, the second in a beaker of water
at room temperature and the third in a hot-water bath. Allow five minutes
for temperature equilibration.
Add ten drops of catechol (POISON!) to each tube, shake the tube then
replace it in the appropriate incubator.
At five minute intervals, for the next 30 minutes, examine the tubes.
(Ensure the temperatures are kept relatively constant throughout the 30
minutes).
Compare the relative colour intensities. Present your results in a table.
Comment on your results and explain the effect of temperature on
enzymes.
BIOL0011 Preliminary Biology Laboratory Manual Page 25

PRACTICAL 6 USE AND CARE OF THE COMPOUND


MICROSCOPE

A microscope extends our vision beyond the limits of the eye. It allows us
to see cells. It consists of two main parts: a stand, comprising the foot and
limb and a tube, containing the optical system. Objects to be examined are
commonly mounted on a glass slide and retained on the stage by a clamp.
The object is illuminated from below the stage by light reflected from the
mirror. The microscope is fitted with a sub-stage condenser, which focuses
the parallel light rays reflected into the condenser so that the quality of light
illuminating the object can be adjusted. The tube is a hollow cylinder with
an eyepiece (ocular) lens system. At the base is a revolving nosepiece to
which objectives of different magnifications are attached. Objects are
brought into focus by the movement of the tube, controlled by the coarse
adjustment and the fine adjustment knobs.

To obtain the best results use the following procedure:


1. Place the microscope on the bench with the tube in a vertical position
facing a good light source (artificial or diffused daylight but not direct
sunlight)
2. Move the condenser right up (in older models the condenser slides in a
sleeve mounting) and using the low power (LP) objective obtain a full
illumination of the field of view by adjustment of the mirror.
3. Mount the piece of material to be examined on a slide with a small drop
of water or dilute glycerol (the latter will dry out much more slowly).
Then using a needle, cautiously lower a cover slip on to the specimen
so as to exclude air bubbles.
4. Place the slide on the stage with the specimen directly below the LP
objective and bring it into focus by using the coarse adjustment. The
focal length of the LP objective (the distance between lens and the
objective when in focus) is about 9mm.
5. Re-adjust the condenser so that an object (e.g. a pencil) placed
between the lamp and the mirror also appears in focus.
6. Adjust the iris diaphragm so that a satisfactory compromise is obtained
between brightness and contrast.
7. To view an object under the high power (HP) objective, place it in the
centre of the LP field and swing the nosepiece to bring the HP objective
into position. The distance between the objective and the coverslip is
very small, so bring it into focus using the fine adjustment only. The
condenser will still be in focus on the specimen and will not require
adjustment; the iris diaphragm will probably require some readjustment.

Develop the habit of cleaning the ocular and objective lenses after each
use of the microscope. Use only lens tissue, never filter paper, towels or
BIOL0011 Preliminary Biology Laboratory Manual Page 26

any other material that may scratch the lens. Remember that the
microscope is an expensive precision instrument: always handle with care.

Magnification
Eyepieces – usually x 6, x10 and x15
Objectives – LP magnifies X10; HP magnifies X40 or X45; Oil immersion
magnifies X100 approximately
Total magnification of the microscope is obtained by multiplying together
the magnification for eyepiece and objective lenses.

Measuring Microscopic Objects


It is often important for the biologist to know the dimensions of the object he
is observing under the microscope. To do this you will need to know the
diameter of your microscope’s field of view at different magnifications.

Determination of the diameter of the field of view


1. Place a short plastic scale (or strip of 1 mm graph paper) on the centre
stage so that the scale is visible through the eyepiece.
2. Line up an intersection of one of the vertical lines and a horizontal line
so that the point at which they meet is just visible at the left side of the
circular field of view. The horizontal line should run across the diameter
of your field of view.
3. Count the number of units (intersecting lines) included from one side of
the field to the opposite side (i.e. left to right). If the right side of the field
does not coincide with one of the lines, you will have to estimate the
fraction of a unit.
4. Write down this value for the diameter of the LP (in micrometers) of our
microscope and note the number of the microscope.
5. As the diameter of the HP field is less than 1 mm, it is more accurate to
obtain the diameter of the HP from the following formula:

Diameter of HP field = diameter of LP field÷ magnification of HP objective


magnification of LP objective

Write down this value for the diameter of both LP and HP (in micrometers)
of your microscope and note the number of the microscope. You will need
this information every time you use the microscope.

Microscope #: _____ LP field diameter ________ HP field diameter _______

To find the length or width of an object under the microscope, estimate it in


relation to the diameter of the field in view. This method is not accurate, but
it will enable you to obtain estimates of the size of microscopic objects.
BIOL0011 Preliminary Biology Laboratory Manual Page 27

PRACTICAL 7 CELL STRUCTURE

You will be using the microscope to observe the structures of cells.


Illustrate the structures by drawing 2-3 cells (refer to page 12 – scientific
illustrations).

1A. Strip a small sheet of inner epidermis from the fleshy leaf of an onion
(Allium cepa) bulb, and place in a drop of water on a clean side. Gently
spread out or cut off any folds in the sheet and cover with a clean cover slip
(lower the cover slip from one edge in order to exclude air bubbles).
Illustrate the cell to show the cell walls, cytoplasm, nucleus and nucleolus.
You may need to irrigate the mount with iodine solution to see the nucleus.
¾ Prepare a fresh sheet of onion epidermis but irrigate with 0.4M
sucrose solution. Illustrate. Annotate your drawing to show the
differences between these cells and those mounted in water.

2. Mount a young leaf of Elodea sp. in distilled water. Examine and draw a
group of cells near the edge of the leaf.

3. Examine and draw cells from very thin sections of the inside of a potato
(Solanum tuberosum) tuber. Irrigate with a dilute solution of iodine. What
effect does the iodine have on the cells? Illustrate.

4. Gently scrape the inside of your cheek with the broad end of a toothpick.
Add a drop of water to a clean slide, and then spread the scraping from the
toothpick in the water. Cover the preparation and examine with the
microscope. Notice that the cells are either square or diamond–shaped
(squamous epithelial cells). You may irrigate with methylene blue to
observe more cellular details.

5. Examine a thin longitudinal section of an onions root tip. From LP draw a


plan diagram to show the root cap, the region of cell division (meristem)
and region of elongation. Illustrate and compare the cells from each of
these regions.

¾ Prepare a table comparing the cells from the onion bulb, onion
elodea leaf, potato tuber and human cheek.
BIOL0011 Preliminary Biology Laboratory Manual Page 28

PRACTICAL 8 CELL DIVISION - PART 1: MITOSIS

The cell cycle consists of interphase and mitosis.

Interphase – Cell outline box-like, nucleus is round, and situated within a


relatively small amount of cytoplasm. The nucleus is bounded by a
membrane (the nuclear membrane). The granular material enclosed by the
nuclear membrane is termed chromatin. The nucleus may also contain a
small dense body (or bodies) known as the nucleolus (nucleoli).

Mitosis is the basic form of cell division in somatic eukaryotic cells. Cell
division actually involves two processes: karyokinesis (division of the
chromosomes) and cytokinesis (division of the cytoplasm). The result is
daughter cells with the same number of chromosomes as the parent. The
stages which may be observed are:
ƒ Prophase – This is the initial stage of cell division. At this stage, the
chromatin appears as diffuse thread-like structures, the chromosomes.
The chromosomes have become visible because each chromatin fiber
has undergone a coiling process which shortens its length while
increasing its diameter. Each chromosome in prophase is constructed
of two identical chromatids, held together at a region called the
centromere. In some cells the individual chromatids are not visible at
this stage.
ƒ Metaphase – The nuclear membrane has disappeared and the
chromosomes are now lined up in a row across the middle of the cell.
The imaginary line on which the chromosomes are arranged is the
equatorial plate. Spindle fibres radiate away from the equatorial plate
towards the opposite ends (poles) of the cell.
ƒ Anaphase – The centromeres holding pairs of chromatids together have
divided and one chromatid now called a daughter chromosome, of each
pair is now moving toward each pole. A spindle fibre appears to be
attached to the centromere of each chromosome.
ƒ Telophase – The chromosomes are now grouped at opposite poles of
the cell and a thin partition, the cell plate may be seen cutting across
the mitotic spindle along the equatorial plate. At this stage the
chromosomes uncoil and appear as chromatin and the nuclear
membrane and nucleolus re-appear.
ƒ Cytokinesis – This final stage in the separation of the daughter cells is
produced by completion of the cell plate in plant cells and cytoplasmic
furrowing in animal cells.
BIOL0011 Preliminary Biology Laboratory Manual Page 29

1. Examine the meristematic zone of an onion root-tip (longitudinal section)


under HP. Identify and illustrate cells which are in Interphase, Prophase,
Metaphase, Anaphase and Telophase.

2. You are provided with the following pairs of chromosomes. The letters
represent genes. Illustrate the chromosome as they would appear in a cell:
a. after replication in the S phase of Interphase
Mitosis
b. Prophase
c. Metaphase (remember that homologous chromosomes do not pair up in
mitosis so they attach to separate spindles).
d. Anaphase
e. Telophase and
f. at the end of Cytokinesis.
BIOL0011 Preliminary Biology Laboratory Manual Page 30

PRACTICAL 9 CELL DIVISION - PART II: MEIOSIS

Meiosis (reduction division) occurs only during the formation of gametes.


There are two divisions. The first results in two daughter cells which each
contain half of the chromosomal material of the parent cell. The second
division usually results in the production of either four spermatozoa or an
ovum and three polar bodies.

Meiosis 1 begins with Prophase 1 in which the members of homologous


pairs of chromosomes physically join. Crossing-over takes place. This is a
process of genetic recombination during which homologous (non-sister)
chromatids exchange segments of DNA strands. Homologous
chromosomes appear on the spindle in Meiosis 1 as a pair. They are
separated in Anaphase 1 so at the end of Meiosis 1 each nucleus contains
the haploid number chromosomes each consisting of two chromatids.
Chromatids of each chromosome are separated in Anaphase II so at the
end of Meiosis II there is one chromatid in each daughter cell.

1. Examine prepared slides of Lilium anthers showing stages of meiosis in


pollen mother cells. Make drawings to show as many stages of the process
as possible.

2. Examine prepared slides of grasshopper testis. Illustrate chiasma.

3. You are provided with the following pairs of chromosomes. The letters
represent genes. Illustrate the chromosome as they would appear in a cell:
a. after chromosome replication
Meiosis
b. when the homologues pair up (prophase I),
c. when a chiasma is formed (prophase I),
d. at the end of crossing over (prophase I),
e. Metaphase I,
f. Anaphase I,
g. Telophase I,
h. Metaphase II,
i. Anaphase II,
j. Telophase II, and
k. at the end of Cytokinesis.
BIOL0011 Preliminary Biology Laboratory Manual Page 31

PRACTICAL I0 MAMMALIAN REPRODUCTION – TESTIS & OVARY

The survival of the species depends upon a consistent succession of new


individuals. This is accomplished by a process called reproduction in which
the organism produces a group of new offspring. Among animals, a variety
of reproductive methods (sexual and asexual) and organs exist. Mammals
utilize sexual reproduction.

The Male Reproductive System.

The male reproductive system comprises the testes, the ducts of the
testes, the auxiliary glands associated with them (seminal vesicles,
prostate gland) and the penis. The primary function of the male
reproductive system is the production of spermatozoa.

The testis is a double gland since functionally it is both exocrine and


endocrine. It is suspended within the scrotum and is surrounded by the
testicular capsule which is composed of three layers; the outer tunica
vaginalis, the middle tunica albuginea and the innermost tunica vasculosa.

The testes contain seminiferous tubules in which the process of


spermatogenesis (sperm production) occurs. It starts in the germinal
epithelium which is composed of a mobile set of proliferating and
differentiating spermatogenic cells and a permanent set of supporting
Sertoli’s cells.

The main endocrine secretion of the testis is testosterone produced by the


interstitial cells of Leydig after stimulation by Interstitial Cell Stimulating
Hormone (ICSH) or Luteinizing hormone (LH).

¾ Make a drawing from HP to show the details of a seminiferous tubule.

The Female Reproductive System

The female reproductive system consist of the ovaries, a system of genital


ducts, (the uterine tubes, uterus, and vagina) and the external genitalia.

The ovaries like the testes are classified as double glands since they
produce both exocrine and endocrine secretions. They are slightly flattened
ovoid bodies (about 4cm x 2cm x 1cm) lying on each side of uterus in the
lateral wall of the pelvic cavity. In sections of the ovary two zones may be
distinguished as an outer cortex and inner medulla.
BIOL0011 Preliminnary Biologyy Laboratoryy Manual Pagee 32

The corttex consistss of a comp


pact cellularr stroma tha
at contains the ovariann
follicles which may be seen inn all stages of development. The progressive e
development of folllicles occurrs after puberty and is characterizzed by growwth
and diffeerentiation of
o the ovum
m. The appe earance of the ovariann cortex
dependss upon the age of the individual a and the stagge of the ovvarian cycle
e.

dulla consissts of loose


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ontaining
numerou
us blood ve
essels, lymp phatics andd nerves.

e a LP plan
¾ Make n of the ova
ary
¾ Make
e a HP draw
wing to shoow details o
of a graafian
n follicle.

T.S. of Seminiferou
S us Tubule frrom Mamm
malian Testis
BIOL0011 Preliminaary Biology Laboratory Manual Page 33

ammalian Ovary
L.S. of Ma O

ature Graaffian follicle from Mamm


L.S. of Ma malian Ova
ary
BIOL0011 Preliminary Biology Laboratory Manual Page 34

PRACTICAL 11 ANGIOSPERM REPRODUCTION

Among plants, reproduction may take place in a variety of different ways.


Angiosperms, however, most commonly use sexual reproduction; the
reproductive organs being on the flower.

1a. Examine and record the features of the flower provided.

The outer most whorl of floral parts make up the perianth. In many flowers
the perianth is distinguishable into the calyx and the corolla. Typically, the
calyx (sepal) is green and protective, although it may be coloured. The
corolla (petal) lies inside the sepals and are often brightly coloured and
serve to attract insects to pollinate the flowers. The two whorls of perianth
usually alternate with one another.

¾ Determine if the parts of each whorl are free from each other
(polypetalous, polysepalous) or fused together (gamosepalous,
gamopetalous.) Count the number of petals and sepals.

¾ Note the positions of the perianth parts on the enlarged end of the
flower stalk, the receptacle.

Using forceps, gently pull off the perianth parts. Find the stamens that
surround the pistil. The stamens collectively form the androecium. Observe
that the stamen is composed of two parts: the filament that is terminated by
a sac like anther. The pistil (gynoecium) has an enlarged ovary and a stalk
like style terminated by one or more stigmas.

¾ Determine if the stamens grow directly from the receptacle, or from the
petals (epipetalous). Determine if they are free or fused to one another.

¾ Determine whether the flower is hypogynous (perianth below, ovary


superior), epigynous (ovary below perianth) or perigynous (ovary and
perianth on the same level).

1b. Mount an anther in a drop of water, crush it with a needle, cover and
observe the contents microscopically. A large number of pollen grains
should be apparent. Draw a few pollen grains to show their characteristic
shape.

1c. Use a razor blade to cut a transverse section of ovary and examine it
microscopically. Note the immature seeds (ovules) and observe that they
grow from swellings (placentas) on the inside of the ovary. Make a low
power diagram.
BIOL0011 Preliminary Biology Laboratory Manual Page 35

1d. A Half Flower Drawing is a conventionalized longitudinal section


through the flower. The flower should be cut through the vertical plane of
symmetry.
Represent the cut edges of the flower by double lines and draw, with single
lines, the background of the uncut portions of the half flower. Use your
magnifying glass or dissecting microscope to observe the longitudinal
section of the ovary. Note carefully the shapes of the receptacle, ovules
and pedicel.

¾ Make a half-flower drawing of the flower provided.

Figure: Half-flower diagram

2. Examine the prepared slide of (a) T. S. unripe and (b) T. S. dehisced


anthers. Make low power diagrams to show the structure of the anther and
to illustrate the method of dehiscence.

3. Examine the flowers provided. In each case, state the possible method
of pollination, giving at least two reasons for your conclusion.
BIOL0011 Preliminary Biology Laboratory Manual Page 36

PRACTICAL 12 MONOHYBRID & DIHYBRID CROSSES

1. Monohybrid Cross

This involves two different alleles at a single locus. One locus may govern
seed colour, another seed shape. Alleles are genes that govern variations
of the same character (e.g. green versus yellow seed colour) and occupy
corresponding loci on homologous chromosomes. Coloured beads will be
randomly selected to simulate monohybrid inheritance. Beads drawn
represent the genotype of the new progeny. A chi squared test will then be
used to compare the difference between the observed and expected values
to determine whether or not the difference is significant.

¾ Collect two bags. To each bag add 20 red beads and 20 blue beads.
Shake the bags, and then randomly select two beads, one from each
bag. Record whether you picked two red beads (RR), a red and blue
(RB) or two blue beads (BB). Replace the beads in their respective
bags and repeat the process 99 times.

¾ Calculate the chi-square value to determine whether the difference is


significant at 0.05 probability value.
• Record the null hypothesis.
• Total the observed (O) number of RR, RB, and BB beads.
• On the basis of expected Mendelian ratios and probability, calculate
the expected (E) values. Then, complete the chi squared table.

X2=∑ (o-e)2 Phenotype O E O-E (O-E)2 (O-E)2/E


e Red Red
Red Blue
Blue Blue
∑ 100 ∑

The number of degrees of freedom = (number of classes -1) represents the number of
independent classes that contribute to the calculated value of x2.
The 0.05 probability value is used as the significance level by statistical convention.

At two degrees of freedom, if the calculated chi-square value is less than


the table value of 5.991, (see Chi squared table) then the deviation is not
significant and the difference could have arisen by chance. If the calculated
x2 is equal to or greater than the table value then the deviation is
considered significant and the likelihood of such a large discrepancy arising
by chance is too low.
• Conclude whether or not the null hypothesis is accepted.
BIOL0011 Preliminary Biology Laboratory Manual Page 37

2. Dihybrid Cross

This involves different alleles at two loci. A homozygous black, short haired
guinea pig (BBSS) and a homozygous brown long haired guinea pig (bbss)
are mated.

¾ Determine the parental gametes and the first filial genotype and
phenotype.

Parental genotype BBSS bbss


F1 generation Gametes
F1 genotype
F1 phenotype

Record in the Punnet square the gametes produced in the F1. Remember
that gametes are formed by segregation and independent assortment of
alleles. The four kinds of gametes are produced with equal probability.

¾ Determine the genotype and phenotype of the progeny. Also record the
genotypic and phenotypic ratio.

F2 generation
F1 gametes

3. In TV bugs, Cable (A) is dominant to rabbit ears (a) and Colour (B) is
dominant to black & white (b). Work Aholic found two Cable Colour bugs
and did test cross (cross with the homozygous recessive) to determine their
genotypes.
¾ For Cable Colour bug #1 he obtained 30 bugs, all Cable Colour.
Explain why he obtained these results.

¾ For Cable Colour bug #2 he obtained 11 Cable Colour bugs, 8 Cable


black & white, 8 rabbit ears Colour and 6 rabbit ears black &
white progeny for a total of 33 TV bugs. Explain why he obtained
these results. Determine if these results are significantly different
from those expected.
BIOL0011 Preliminary Biology Laboratory Manual Page 38

PRACTICAL 13 NATURAL SELECTION & EVOLUTION

The aim of the biological sciences is to understand life. Any coherent


theory of biology must account for overarching features of the natural
world.
1. The unity of life
2. The diversity of life
3. The existence of biological adaptations
4. The dynamic history of life on Earth

The theory of evolution is the cornerstone of modern biology that seeks to


explain these. It attempts to explain why millions of species exist. As a
principle, evolution states simply that species change through time. New
species of organisms emerge and older forms of life disappear.

Charles Darwin was the person who brought the evidence together and by
1838 he had proposed a mechanism, natural selection, for evolution. His
ideas are summarized in four points.
1. There is variation among individuals of most populations.
2. Some proportion of that variation is inherited.
3. Populations tend to produce more offspring than the environment can
support.
4. Those individuals whose traits are best adapted to the prevailing
environment will survive better and leave more offspring than those with
less adaptive traits.

The weakest part of Darwin’s theory was his inability to explain how genetic
variation arose and how it was maintained in populations, both of which
were poorly understood at the time. With the entry of Gregor Mendel’s work
into the mainstream of biological thinking in the early twentieth century, the
broad principles of the relationships between genes and natural selection
were elucidated.

The combination of Darwin’s theory of evolution with the principles of


Medelian genetics is known as the neo-Darwinian synthesis of the synthetic
theory of evolution. During the past fifty years, the synthetic theory has
dominated scientific thinking about the process of evolution and stimulated
a great deal of investigation. Current controversies revolve primarily around
the rate of macroevolutionary change and the role played by chance in
determining the direction of evolution. They do not affect the basic
principles of synthetic theory. The debate, however, promises to give us a
more complete understanding of evolutionary mechanisms than we have at
present.
BIOL0011 Preliminary Biology Laboratory Manual Page 39

VARIATIONS IN ANIMAL POPULATIONS

1. Sources of variation: Variations in a population arises from two primary


sources, mutation and genetic recombination Mutations add new genes or
alleles to the gene pool and thus supply the genetic foundation on which
evolutionary forces operate. You are already familiar with the variation
produced by sexual reproduction from your studies of genetics and your
experience with families.

1. Researchers have estimated that there are one or two mutations per
gamete but that these are usually inconsequential as sources of variation
within a population. However, in asexual species, such as bacteria,
mutation can be an adequate source of variation. Can you suggest one
reason for this?

2. Variation is a fundamental attribute of the individuals of a population and


the raw material for evolution. Variation includes behavioural and
biochemical traits as well as the common visible (morphological) traits.
Such variation must be genetically based to be significant to evolution.

¾ Examine the sample of arthropod or mollusk individuals taken from a


single population.

2a. Identify at least five morphological trait in which the individuals vary.
Present your results in table which shows the feature and the number of
individuals with each variation.

2b. Think about the variation in the population. Decide whether each
morphological trait varies continuously or discontinuously and whether it is
likely to be determined solely by genetic factors or also influenced by
environmental factors. Suggest advantages and disadvantages that may be
associated with different variations of each trait.

3. Comparison of Cytochrome C from Man and Seven Other Animals: An


Exercise Using the Genetic Code

Cytochrome c, or cyt c is a small, water soluble heme protein associated


with the inner membrane of the mitochondrion. Each cytochrome c carries
one electron between two different electron transport complexes (III and IV)
embedded in the inner membrane. In doing this, cytochrome c repetitively
undergoes either oxidation or reduction, but it does not bind oxygen. This
electron transport chain drives the production of ATP.
BIOL0011 Preliminary Biology Laboratory Manual Page 40

Cytochrome c has been studied extensively because its small size (about
100 amino acids; molecular weight 12,000 daltons) and its water solubility
permit researchers to isolate it from other mitochondrial proteins, which
tend to be not only larger than cytochrome c but also fat soluble and
embedded in the membrane. The amino acid sequences for the
cytochrome c occurring in many organisms from yeast to humans have
been determined. It is found universally in aerobic organisms.

Comparison of amino acid sequences of the molecule in diverse species


shows a great deal of similarity among organisms. This protein is highly
conserved across the spectrum of species and so is useful in studies of
evolutionary relatedness. Similarities in the amino acid sequences suggest
a common ancestor that must have been using this protein even before
basic divergences between plants and animals arose. The degree of
similarities correlates closely with the apparent degree of relatedness. The
sequences from monkeys and cattle are more similar than the sequences
from monkeys and fish. Chickens and turkeys have the identical molecule
(amino acid for amino acid) within their mitochondria, whereas ducks
possess molecules differing by one amino acid. Similarly, both humans and
chimpanzees have the identical molecule, while rhesus monkeys possess
cytochromes differing by one amino acid.

A sequence of thirteen amino acids, near the N-terminal end of


cytochrome C from human tissue is:
gly-asp-val-glu-lys-gly-lys-lys-ileu-phe-ileu-met-lys

The sequence, derived from the identical region of peptide chain of the
cytchrome C of a number of other organisms (a-g) is as follows:
a. gly-asp-val-ala-lys-gly-lys-lys-thr-phe-val-glu-lys
b. gly-asp-pro-asp-ala-gly-ala-lys-ileu-phe-lys-thr-lys
c. gly-asp-val-glu-lys-gly-lys-lys-ileu-phe-ileu-met-lys
d. gly-asp-ileu-glu-lys-gly-lys-lys-ileu-phe-val-glu-lys
e. gly-asp-ala-glu-asp-gly-lys-lys-ileu-phe-val-glu-arg
f. gly-asp-val-glu-lys-gly-lys-lys-ileu-phe-val-glu-lys
g. gly-ser-ala-lys-lys-gly-ala-thr-leu-phe-lys-thr-arg

¾ For each of the sequences, calculate the least number of mutations that
would be necessary if the sequence is to be changed to that found in
man. (The least number of mutations as this is more likely to occur than
a large number of mutations).
o You will need to compare each amino acid in the sequence with
that in man’s sequence and, using the genetic code, determine
BIOL0011 Preliminaary Biology Laboratory Manual Page 41
4

the numbe er of base p es required for each am


pair change mino acid in
n
the animal’s sequencce to be cha
anged to th
hat in man’ss seguence
e.
urpose of th
o For the pu he exercise, one mutattion = one change
c in a
base of the code for amino acids).

¾ Arrang
ge the sequ
uences in order
o of incrreasing num
mbers of mu
utations.

etic code
The gene

Exam
mple: Sequence A to man.
1. Determine the number of amiino acid changes in the sequencee.
2. Determine
D the least number of mutations
m for each amino acid chaange.
3. Determinne the number off mutations for thhe sequence.

4 amino acid channges requireed. Ala →Gluu; Thr→ Ileuu; Val →Ileuu; Glu →Meet
To change Ala→Glu:
A
Alaa genetic code: GCU or GCG oor GCA or GCG → Glu genetic code: GAA oro GAG.
GCA (Ala) → GAAG (Glu) by one base changge C → A therrefore 1 mutattion.
To change Thr → Ileu:
Thr geenetic code: ACCU or ACC or AACA or ACG→ Ileu genetic coode: AUU or AUUC or AUA
ACU (Thr) → AUU (Ileu) by one base channge C→U therrefore 1 mutattion
To change Val
V →Ileu:
Val gennetic code: GUU or GUC or GUA or GUG → Ileu genetic ccode: AUU or AUC
A or AUA
GUC→AUC by one baase change G→ →A therefore 1 mutation
To change Glu
G → Met:
Glu genetic code: GAA or GAG → Met: genetic code AUG
GAG→AUG
G byy two base chaanges G →A and
a A→U therrefore 2 mutattions
Total number
n of mmutations foor sequence A to man = 5
BIOL0011 Preliminary Biology Laboratory Manual Page 42

¾ Rank the following organisms in order of increasing ‘evolutionary


distance’ from man: Silk moth, Tuna fish, Chicken, Rhesus monkey,
Yeast, Wheat, Horse

¾ Suggest which of the sequences a-g could belong to each organism.

Sequence No. mutations Organism Organism description

4A. Some persons find weak solutions of the chemical phenylthiourea


(PTC) bitter and distasteful. Others are unable to taste it.

¾ Place a piece of PTC paper on your tongue. Can you detect a bitter
taste? Calculate the percentage of tasters and non-tasters in the class.

i. PTC is a harmless chemical but can you suggest a situation in the


evolutionary history of humans in which the inability to taste a chemical
might have had evolutionary consequences?

4B. Animals vary not only in their appearance but in their behaviour. For
some behavioural variations, it is easy to suggest evolutionary or adaptive
significance; for others, possible adaptive value is much harder to predict.

¾ Check yourself for thumb and arm crossing and record the class data.

Thumb crossing: Interlock fingers and check if left thumb is over or below
the right thumb. The left thumb over is due to a dominant gene.

Arm Crossing: Cross arms and check if left arm is over or below right arm.

i. Can you suggest any adaptive significance for left or right dominance in
thumb or arm crossing?
BIOL0011 Preliminary Biology Laboratory Manual Page 43

PRACTICAL14 PROKARYOTES; KINGDOM PROTISTA; KINGDOM


FUNGI

Nb. Refer to page 7 for hints on preparing illustrations. Remember to


include the classification of each organism illustrated.

1. PROKARYOTES: Examine prepared slides to observe various bacterial


shapes.

2. EUKARYOTES
KINGDOM PROTISTA

2a. Algae
Examine and draw, from fresh specimens and/or prepared slides, the
following range of Algae.

¾ Chlorophyta (green algae)


i. Single celled eg.
ii. Colonial eg.
iii. Filamentous eg.
iv. Thalloid eg.
¾ Phaeophyta (brown algae) eg.
¾ Rhodophyta (red algae) eg.

Ulva sp Sargassum sp.

2b. Protozoa
Examine and draw specimens from prepared slides. Annotate the labels
relating to feeding and locomotory structures.

¾ Amoebozoa/ Rhizopoda eg. Amoeba sp. – Locomotion is by means of


pseudopodia.
¾ Euglenophyta/ Mastigophora eg. Euglena sp. – Locomotion is by one or
two anterior flagella (often difficult to see). This species also possesses
BIOL0011 Preliminary Biology Laboratory Manual Page 44

chloroplasts, and the cell surface is covered with a pellicle. There is no


cellulose cell wall.
¾ Euglenophyta. Volvox sp. is colonial. Colonial protist species are
common (16 groups of protists contain colony-forming species) and
colonies are often very difficult to distinguish from multicellular animals.
Coloniality is favoured by biologist as the means by which multicellular
animals evolved. Cells of Volvox are not differentiated into tissues and
organs but show how the preliminary stages to such an evolutionary
sequence may have occurred. In the slide you will be able to see the
regular spacing of each cell, as well as the formation of new daughter
colonies.
¾ Ciliophora eg. Paramecium sp. – Characterized by the presence of cilia
over the surface of the cell.
¾ Place a drop of pond water on a slide with a coverslip and examine it
under the microscope. Add a few drops of methyl cellulose to slow
down the organisms if they move too quickly. Draw examples of the
protists present. Identify each as amoeboid, flagellate, or ciliate.

Amoebozoa Ciliophora

Euglenophyta Euglenophyta (colonial)


BIOL0011 Preliminary Biology Laboratory Manual Page 45

3. KINGDOM FUNGI

Examine and draw from fresh specimens, preserved specimens and


prepared slides, the following range of fungi.

¾ Lower fungi: Hyphae are coencytic, sporangia contain indefinite number


of spores. Identify hyphae, sporangiophore, sporangium, columella,
sporangiospores and zygospores
Eg. Rhizopus sp. and Mucor sp.

¾ Higher Fungi: Hyphae are septae.


Eg. Aspergillus sp. and Penicillum sp.

Examine and draw fruiting bodies: mushrooms and bracket fungi.

Lower fungi Higher fungi

Mushroom
BIOL0011 Preliminary Biology Laboratory Manual Page 46

PRACTICAL 15 KINGDOM PLANTAE (BRYOPHYTES,


PTERIDOPHYTES & GYMNOSPERMS)

1. PHYLUM BRYOPHYTA

¾ Examine and illustrate entire liverwort and moss plants, identifying the
reproductive and vegetative portions.

Class Hepaticae (Liverworts). Eg. Marchantia sp.


Note apical notch and gemma cup (asexual reproductive structures) on
dorsal surface, antheridia and archegonia (sexual reproductive structures).

Class Musci (Mosses). Eg. Polytrichum sp.


Note the male plant with stem (axis), leaves and artheridial cup; the female
plant with archegonial rosette and female plant with sporophyte

Marchantia (female & male) Polytrichum (gametophyte with sporophyte)

2. PHYLUM PTERIDOPHYTA

¾ Examine and draw the habit of heterosporous and homosporous


peteridophytes.

Heterosporous Eg. Selaginella sp.


Note different size leaves, fertile leaves forming cones. Draw spores from a
squash of fertile leaves

Homosporous eg. Eg. Nephrolepis sp.


Illustrate the gametophyte. Illustrate the sporophyte noting circinnate
vernation. On the leaf underside note the sori and indusium (covering sori).
BIOL0011 Preliminaary Biology Laboratory Manual Page 47
4

Fern sporop
phyte Fern
n gametophyte
e Selaginella strobilus
s

UM GYMNO
3. PHYLU OSPERMAT
TOPHYTA

¾ Exxamine the reproductivve and vege


etative featu
ure of Gym
mnosperms.

Class Cyccadopsida Eg. Cycas sp.


Illustrate a leaf; a maale cone wiith microspo w the microsporangia
orophylls with a
containing g the microospores andd a megasp porophyll wiith megasp
porangia
externallyy borne.

Class Pin nopsida Eg.. Pinus sp.


Illustrate a male con
ne; a femalee with ovule
es between papery sca
ale and
woody sccale cone; stem
s with le
eaves (needdles) cone sscale.

Pine (male and female cone)


c

s scale
Ovuliferous cas (megaspo
Cyc orophyll &male cone)
BIOL0011 Preliminary Biology Laboratory Manual Page 48

HOW TO CONSTRUCT AND USE DICOTOMOUS KEYS

Biologists use taxonomic or identification keys to assist them in identifying


organisms. Many identification keys are dichotomous; ie made up so that
there are two choices. Each couplet had two leads. Choice of a lead
directs the user to the possible identity of the organism or to another
couplet.

As a simple example to help you understand how an identification key


works, let us examine a simple key to certain familiar animals: earthworm,
grasshopper, dog, octopus, fish, snail, lobster, clam, alligator, bird and frog.
As you proceed, especially note how the animals are grouped according to
their similarities. Start at No. 1, examine your animal and select lead 1a or
1b. Follow to the next number as you are instructed and again select a or b.
Proceed until you come to the name of the animal.

Identification Key to Selected Animals.


Couplet Characteristic Next move/Id
1a Animal with bony endoskeleton 2
1b Animal with exoskeleton or no skeleton 6
2a Fins present Fish
2b Fins absent 3
3a Oviparous 4
3b Viviparous Dog
4a Eggs laid on land 5
4b Eggs without shells laid in water Frog
5a Wings present Bird
5b Wings absent Alligator
6a Body soft, without external skeleton 7
6b Body covered with exoskeleton or shell 8
7a Body with eight arms (tentacles) Octopus
7b Body without arms Earthworm
8a Jointed exoskeleton present 9
8b Shell presented, not jointed 10
9a Six legs present Grasshopper
9b More than six legs present Lobster
10a Shell coiled Snail
10b Shell double and hinged Clam

Preparing a dichotomous key.

¾ You are provided with a collection of plants or animals. Prepare a


dichotomous key to distinguish them. (See guide overleaf).
In addition to your observations, use your notes and text to obtain
information on the important features of the organisms.
BIOL0011 Preliminary Biology Laboratory Manual Page 49

NB. The key should be strictly dichotomous throughout.


The leads in each couplet should be contrasting so that one fits the situation at hand while
the other does not. For example:
a. Leaves opposite………………….
b. Leaves alternate………………….
The leads are usually in phrase form (not complete sentences), with each feature
uniformly separated from one another by a semicolon. For example:
a. Leaves opposite; flowers blue; sepals 4……….
b. Leaves alternate; flowers red; sepals 5………
Attempt to phrase leads to read as positive statements, especially the initial lead of the
couplet. So, for example, use
a. Inflorescence an umbel…………
b. Inflorescence a panicle………..
rather than
a. Inflorescence an umbel……….
b. Inflorescence not an umbel………
To avoid unnecessary confusion the initial word of each lead of the couplet should be
identical. So, for example, use
a. Flowers blue; sepals 5……….
b. Flowers red; sepals 4………...
rather than
a. Flowers blue; sepals 5
b. Corolla red; calyx of 4 sepals…
Also, for clarity, two consecutive couplets should not each commence with the same word.
So, for example, use
1a. Petals blue……………………..
1b. Petals red………………………
2a. Corolla actinomorphic (radially symmetrical)………
2b. Corolla zygomorphic (bilateral or asymmetrical)……….
rather than
1a. Flowers blue……………………..
1b. Flowers red…………………
2a. Flowers actinomorphic………
2b. Flowers zygomorphic………..
Avoid the use of generalities or relative terms in contrasting leads of the couplet. So, for
example, use
a. Petioles 2-3cm long; flowers 5-8 cm. in diameter……
b. Petioles 6-7cm long; flowers 0.5-1cm. in diameter…..
rather than
a. Petioles short; flowers large and showy………….
b. Petioles long; flowers smaller and less conspicuous…….
Avoid using overlapping limits of variation. The following example is in poor form. Here,
petiole length and flower color should be avoided. Other, more contrasting characters
should be sought and embodied in the key.
a. Petioles 2-6cm long; flowers pink to red………
b. Petioles 3-8cm long; flowers red to purple……..

Use clearly discernible morphological characters whenever possible. Keys to dioecious


plants should take into account both staminate and pistillate material. If vegetative parts do
not display diagnostic features use both flowers and fruit. Within a single key, avoid
switching from flower characters to fruit characters unless fruit and flowers are commonly
present together – or unless sufficient supplemental characters are given to identify
material in either condition.
BIOL0011 Preliminary Biology Laboratory Manual Page 50

PRACTICAL 16 KINGDOM ANIMALIA, PHYLA: PORIFERA AND


CNIDARIA

1. PHYLUM PORIFERA
The ciliate protists demonstrate the greatest complexity that has evolved in
a unicellular organism. Further elaboration demands an increase in size
and for this a multicellular structure has proved necessary. Sponges are
the simplest multicellular animals. They have neither true tissues nor
organs and their cells retain a considerable degree of independence.
However unlike protists the various kinds of cells do not act entirely as
individuals. The cells show a definite social organization with individual
cells specialized for the functions of feeding, support or reproduction.

Body Development Plan of Phylum Porifera: diploblastic, asymmetrical.

¾ Examine and illustrate each type of sponge. Try to determine which


are the inhalant and exhalent (osculum) openings.

¾ Cut a small section of the sponge and treat it with bleach. Examine a
drop of the digested material with the aid of a microscope. Observe and
draw the spicules.

¾ Illustrate the sponge Grantia from a prepared slide. Identify the


spongeocoels (flagellated chambers) and the choanocyte cells.

Asconoid, Leuconoid and Synconoid sponges. Sponge spicules

2. PHYLUM CNIDARIA

Body development plan of Phylum Cnidaria: diploblastic, radially


symmetrical.

The Cindaria show two advances in complexity over the sponges.


They have an internal space for digestion, the gastrovascular cavity, which
has a single extensible opening, the mouth. This allows them to feed on a
BIOL0011 Preliminary Biology Laboratory Manual Page 51

much greater range of food sizes than is possible for either the protists or
the sponges. The body wall is composed of tissue layers, an outer
ectoderm or epidermis, the middle non–cellular layer mesogloea and the
inner endoderm or gastrodermis. Cnidarians are polymorphic, ie they have
more than one body form. The “medusa” form is adapted for pelagic
existence. It has the mouth and tentacles facing downwards. The “polyp”
form is adapted for benthic existence with the mouth and tentacles facing
upward. Cindaria are classified based on whether polyps or medusae are
the dominant body form during the life cycle.

Hydrozoa: polyps are generally more conspicuous than medusae.


¾ Place a small section of the hydroid colony on a slide, with a cover slip,
and examine it using the microscope. Look for the following features;
hydranth, hydrotheca, tentacles, hypostome, mouth, gonotheca,
blastostyle and stolon. Illustrate.
¾ Observe live specimens in the lab if they are available. How does it
feed? (include the role of cnidocytes)

Scyphozoa: the medusa is more conspicuous than the polyp.


¾ Examine a preserved specimen of Aurelia (a jellyfish) using a hand
lens. Illustrate the specimen. Label the oral arms, gastric pouch,
gonads, circular canal, radial canals and tentacles.

Anthozoa: the dominant form is the polyp and there is apparently no


medusae. They may be solitary or colonial.
¾ Examine and illustrate a live specimen of Aiptasia (a local sea
anemone).
¾ Touch one of the tentacles with a T-pin and note the animal’s reaction.
¾ Present your anemone with small pieces of oyster flesh. Observe and
describe its feeding behaviour.

Corals are colonies of polyps that feed by photosynthesis using mutualistic


algae during the day. They also use their tentacles to catch small
planktonic organisms at night. They have a calcium carbonate exoskeleton
¾ Examine and Illustrate colonies of corals.

Polyp and Medusa form Cnidarians.


BIOL0011 Preliminary Biology Laboratory Manual Page 52

Practical 17 KINGDOM ANIMALIA, PHYLA:


PLATHYHELMINTHES, NEMATODA, ANNELIDA AND MULLUSCA

3. PHYLUM PLATYHELMINTHES (Flatworms)

Body Development Plan of Phylum Platyhelminthes: bilateral symmetry,


triploblastic, acoelomate.

Flatworms show true bilateral symmetry; they have roughly mirror-image


right and left halves, a head at the anterior end, a posterior tail end, an
upper dorsal surface, and a lower ventral surface. They are triploblastic – in
addition to the ectoderm (forms the epidermis) and endoderm (forms the
gastrodermis) seen in the Cnidarians there is a third layer of tissue, the
mesoderm. Flatworms are, however, acoelomate; the only body cavity is
the gut which has a single opening. Platyhelminths also have their bodies
dorso-ventrally flattened due to the dependence on diffusion for distribution
of nutrients, gases and excretory products to and from tissues. They also
have true organs - tissues are grouped into functional units.

¾ Examine and illustrate Planaria sp. from a prepared whole mount slide
or live Dugesia sp. Note the organs involved in feeding (muscular
pharynx, central mouth and the intricately branched intestine).

Platyhelminthes: Dorsal of Dugesia sp.

4. PHYLUM NEMATODA (Roundworms)


Roundworms show a major evolutionary advance over the flatworms: they
possess a separate mouth and anus. This arrangement allows a more
efficient, one-way passage of food through the digestive system. They have
complex cuticle and their body walls contain only longitudinal muscles.
BIOL0011 Preliminaary Biology Laboratory Manual Page 53
5

Body Devvelopment PlanP of Phyylum Nematoda: Bilate


eral symme
etry,
triploblasttic, pseudco
oelomate; p
protostome
e, non-segm
mented.

¾ Exammine and illu


ustrate male and fema ens of Asca
ale specime aris sp.
noting the obvio
ous sexual d
dimorphismm.

Nematoda: Female and male Ascariss limbricoides


s

UM ANNEL
5. PHYLU LIDA (Segm
mented worm
ms)

Several characterist
c ics distingu
uish Annelid
ds from the phyla prevviously
studied. They
T have a well-deve eloped metaamerised bo ody; segmeentation is
externally
y as well ass internally obvious.
o An
nnelid bodyy walls have
e an outer
layer of circular musscle and an inner layerr of longitud
dinal muscle
es.

Body Devvelopment PlanP of Phyylum Anneliida: Bilatera


ally symme
etrical,
triploblasttic, coeloma
ate; protosttome, segm
mented.

Annelida: Anterior-
A ventrral of the eartthworm Lumb
bricus terrestri
ris
BIOL0011 Preliminnary Biologyy Laboratoryy Manual Pagee 54

¾ E
Examine and illustrate the externa
al features a preservedd specimen n
of Lumb
bricus terresstris or fresh specimen
ns of Phereetina sp. No
ote the
segmentation, prosstomium, mouth,
m anus, clitellum, and chaeta
ae (setae).

LUM MOLLUSCA
6. PHYL

Body De evelopmentt Plan of Phhylum Mollu


usca: bilateral symmettry,
triploblastic, coelom
mate, protos
stome and non-segme ented.

aracteristic features
The cha f are
e:
• “HHead-foot” region (withh sensory, feeding, an nd locomoto ory organs),
• Visceral
V masss (with dig
gestive, circculatory, excretory andd respiratoryy
organs)
• Mantle
M (whicch secretess the shell aand enclose es the visce
eral mass).

s are classiified as Gasstropoda (ssnails & slug


Molluscs gs), Bivalvia (oysters,
clams, scallops)
s an
nd Cephalopoda (squid d, octopus)).

¾ Exaamine and illustrate the of the snail removed frrom its shelll.
e features o
Notee eyes at end
e of anten nna and head-foot regions.
¾ Exaamine the vaariety of Ga
astropod shhells; make quick sketcches to sho ow
variation in sha
ape.
¾ Obsserve live sp
pecimens ofo the Bivalvve Perna viridis
vi . Note the byssuss
thre
eads, siphonns (modifica
ation of ma
antle) and mantle.
m

Mollusca: External feattures of a terrrestrial gastro


opod
BIOL0011 Preliminary Biology Laboratory Manual Page 55

PRACTICAL 8 KINGDOM ANIMALIA: PHYLUM ARTHROPODA

7. PHYLUM ARTHROPODA

Arthropods are the dominant animals on earth, both in terms on number of


individuals and species. They have the following characteristics:
Exoskeleton - an external skeleton that encloses the body like a suit of
armour. In places it is thin and flexible to allow movement of
appendages.
Appendages - jointed to provide added flexibility.
Segmentation - considered evidence of annelid ancestry; segments tend to
be reduced in number, fused, and for specialized functions.
Cephalization – arthropods have well developed heads and sense organs.

Body development plan of Phylum Arthropoda: Bilaterally symmetrical,


triploblastic, coelomate, protostome, segmented.

Extant arthropods are classified into Crustacea, Uniramia or Chelicerata on


the basis of the anatomy of their limbs, particularly those used for cutting
and preparing food.

Crustacea
Crustacean appendages are all basically biramous (branched into two),
with both sections normally being of different size and shape, often bearing
further secondary branches. The appendages of the head comprise two
pairs of antennae, one pair of mandibles and two pairs of maxillae.

¾ Examine and illustrate the external features of the mangrove crab,


Aratus sp. or the blue crab, Callinectes sp. (Class Malacostracea,
Order Decapoda). Make one composite diagram, the left half showing
the features of the dorsal surface and the right half showing the
features of the ventral surface.

Note that the body consists of a large cephalothorax, with a smaller


abdominal region folded underneath. Identify the various appendages,
including the chelipeds (claws), walking legs, antennae, mouthparts and
telson. Note the compound eyes on the stalks.
BIOL0011 Preliminnary Biologyy Laboratoryy Manual Pagee 56

Crusta
acea: Dorsal and View of Caline
a Ventral V ectes sp

Uniramia
Uniramia a are by farr the most diverse
d andd abundant arthropod sub-phylum m.
Their apppendages are uniramous (no bra anching); th hose of the head
comprising one paiir each of antennae,
a m
mandibles anda maxillae e. Some
groups have
h a secoond pair of maxillae. U
Uniramia inc cludes the cclass
Hexapod da (insects), Chilopoda (centiped des) and the e Diplopoda a
(milliped
des). The in
nsects have e three pair of legs, usually supple emented byy
two pairs of wings, giving them m the capaccity for fligh
ht. This ability to flight
distinguiishes them from otherr invertebrates.

¾ amine and illustrate external featu


Exa ures of Blabberus sp. (oorder
Blattteria). Rem
move the wings of the ccockroach fromf one siide of the
body. Make on ne diagram,, the left half showing the features of the
dorssal surface and right half
h showing g the featurres of the ve
entral
surfface.
BIOL0011 Preliminaary Biology Laboratory Manual Page 57
5

Note the three


t main body segmments; the head,
h thoraxx (with thre
ee
segmentss; prothoraxx, mesothorrax, metath
horax) and ssegmented abdomen.
Note the compound eyes, antennae, wing gs, anal cercci. Note the
e
prothoraccic, mesotho
oracic and metathoraccic legs.

Uniramia
a, Hexapoda:: Dorsal and Ventral
V of Perriplaneta sp.
BIOL0011 Preliminary Biology Laboratory Manual Page 58

Chelicerata
The bodies of chelicerates are divided into an interior prosoma and a
posterior opisthosoma. All have eight walking legs attached to the
prosoma. Chelicerates include the spiders, ticks and scorpions. The
spiders (class Arachnida, order Aranae) are the most numerous
chelicerates. Their appendages are uniramous, the prosomal appendages
comprising one pair of chelicerae, one pair of pedipalps and four pairs of
walking legs, all attached near to the ventral mid line.

¾ Examine and illustrate the external features of Argiope sp. Make one
diagram, the left half showing the features of the dorsal surface and the
right half showing the features of the ventral surface.
Note, on the ventral surface, the chelicerae, pedipalps, eyes and legs. On
the dorsum, note the genital pore, spinnerets and openings to the book
lungs.

Chelicerata: Dorsal and Ventral of Argiope sp.


BIOL0011 Preliminary Biology Laboratory Manual Page 59

PRACTICAL 19 KINGDOM ANIMALIA: PHYLUM ECHINODERMATA


AND CHORDATA

8. PHYLUM ECHINODERMATA

Body Development Plan of Phylum Echinodermata: pentaradial symmetry,


triploblastic, coelomate, deuterostomes, non-segmented.

The major classes are Asteroidea (starfishes), Ophiuroidea (brittlestars) &


Echinoidea (sea urchins).

Asteroidea
The spiny aboral (upper) surface has the madreporite (the external opening
of the water vascular system) and the inconspicuous anus. The oral
(lower) surface has the mouth and tube feet (located in the ambulacral
grooves, and used in locomotion). The arms are not conspicuously
separated from the central disc.

¾ Examine and illustrate both surfaces of the starfish.

Ophiuroidea
These are the most highly mobile of all echinoderms. The arms are long
and distinctly separate from the central disc. Tube feet are not located in
ambulacral grooves. Ophiuroids do not use the tube feet as the principal
method of locomotion, but instead use the arms directly, pulling with the
anterior arms and pushing with the posterior arms. The oral surface has the
mouth and madreporite.

¾ Examine and illustrate both surfaces of the brittlestar.

Echinoidea
Sea urchins do not possess arms. The ossicles have become fused into a
shell or test. Tube feet are located in ambulacral grooves. They have
moveable spines, which are arranged in rows parallel to the ambulacral
grooves. The oral surface has the mouth, with the Aristotle’s lantern
(scraping organ used to remove algae from the substrate) inside.

¾ Examine and illustrate the sea urchin. Pry the mouth open to view the
Aristotle’s lantern.
BIOL0011 Preliminnary Biologyy Laboratoryy Manual Pagee 60

Asteroidea: aboral an
nd oral surface
es

Ech
hinoidea: aboral and oral ssurfaces of a sea
s urchin
BIOL0011 Preliminary Biology Laboratory Manual Page 61

9. PHYLUM CHORDATA

Body Development Plan of Phylum Chordata: bilaterally symmetrical,


triploblastic, coelomate, deuterostome, segmented.

The Chordata, classified into three major subphyla, the Cephalochordata,


Urochordata and Vertebrata, is extremely diverse but is characterized by
four features, which all chordates possess at some stage of their lives.
Notochord: a stiff but flexible rod running the length of the body and
providing an attachment site for muscles.
Dorsal nerve cord: lying dorsal to the digestive tract, this hollow, nervous
structure develops a thickening at its anterior end which becomes
the brain.
Pharyngeal gill grooves: located in the pharynx, these may form functional
openings or may appear only as grooves during an early stage of
development.
Post-anal tail: an extension of the body past the anus.

Subphylum Cephalochordata:
These chordates are acraniate but have a head and segmented muscle. All
the typical chordate features are present in the adult organism.

¾ Examine and illustrate the lancet Branchiostoma sp, a small fish-like


animal that lives half-buried in sand on the sea floor. It is also called
Amphioxus. Note all the chordate features in the adult.
¾ View the demonstration of the transverse section of the lancet.

Cephalochordata: lateral view and T.S. of Branchiostoma sp.


BIOL0011 Preliminary Biology Laboratory Manual Page 62

Subphylum Urochordata:
The chordates are also acraniate. No head or segmentation is present. The
larvae have all the chordate features but adults have only the pharynx;
other features degenerate during metamorphosis.

Adult sea squirts (tunicates) are sessile, filter-feeding and vase-shaped.


Their ability to move is limited to a forceful contraction of the saclike body.
The larvae are active swimmers that possess all the chordate features.

¾ Examine and illustrate the tunicate provided. Place the specimen on a


black tile to increase visibility of internal organs.
Note the inhalant and exhalent siphons, the perforated pharynx, stomach
and heart.

Urochordata: features of a sea squirt

Subphylum Vertebrata
In vertebrates, the embryonic notochord is replaced during development by
a vertebral column, composed of cartilage or bone. This structure provides
support for the body, a site of attachment for muscles and protection for the
delicate nerve cord and brain. The vertebral column is part of a living
endoskeleton, capable of growth and self-repair. It has allowed vertebrates
to achieve great size and mobility. Examine the skeletons of various
vertebrate organisms displayed in the laboratory. They will also be present
at the next practical session.

The subphylum vertebrata contains the superclasses Agnatha and


Gnathostomata (with classes Chondrichthytes, Osteichthyes, Amphibia
Reptilia, Aves and Mammalia). The Agnathans are jawless while the
Gnathostomathans have jaws. The Agnathans, Chondrichthyes and
Osteichthyes are fishes and they have a two chambered heart.
BIOL0011 Preliminary Biology Laboratory Manual Page 63

Superclass Agnatha (jawless fishes)


These are regarded as the most primitive living vertebrates. Superficially
they are fishlike but without scales, jaws and paired fins.

¾ Examine and illustrate the lamprey. Note the dorso-ventral fins, gill
openings, sucker and epidermal “teeth”. How does the lamprey feed?

Agnatha: External features and mouthparts of a lamprey


BIOL0011 Preliminary Biology Laboratory Manual Page 64

PRACTICAL 20 KINGDOM ANIMALIA, PHYLUM CHORDATA II

Superclass Gnathostomata

Class Chondrichthyes (cartilaginous fishes)


This class includes the sharks, skates and rays. They have skeletons
formed entirely of cartilage. The body is protected by a leathery skin with
tiny tooth-like, placoderm scales.

¾ Examine and illustrate a shark. Note the presence of paired pelvic and
pectoral fins, the heterocercal tail fin and the rigid dorsal fin.

Class Osteichthyes (bony fishes)


This class has fishes with skeletons made of bone. They have a swim
bladder that allows the fish to float effortlessly at any depth. Osteichthyes
includes sardines, tuna, eels luminescent deep-sea forms. The group is
enormously successful and has spread to nearly every possible watery
habitat, both freshwater and marine. They have paired fins and some
groups have modified fleshy fins that could be used (in an emergency) as
legs, dragging the fish from a drying puddle to a deeper pool.

¾ Examine and illustrate a bony fish. Note the fins, lateral line, eyes,
teeth and scales.

Class Amphibia
The organisms in this class straddle the boundary between aquatic and
terrestrial existence. They include the frogs, toads and salamanders. The
limbs show varying degrees of adaptation to movement on land, from the
crawl of salamanders to the efficient leap of frogs. Lungs replace gills in
most adult amphibians, but the skin must remain moist, since it serves as
an additional respiratory organ. A three chambered heart circulates blood
more efficiently than in fishes.

¾ Examine and illustrate the external features of either the toad, Bufo
marinus (Order Anura) or a salamander (Order Urodela).

Class Reptillia
This class includes the lizards, snakes, turtles, crocodiles and the extinct
dinosaurs. Some reptiles live in an aquatic environment and lay eggs on
land while others have achieved complete independence from their aquatic
origin. Adaptations for terrestrial life include:
ƒ presence of a dry, tough, scaly skin that resists water loss.
ƒ internal fertilization.
BIOL0011 Preliminary Biology Laboratory Manual Page 65

ƒ shelled amniotic egg (which can be buried in sand or dirt, far from
water) with an internal membrane, the amnion, which encloses the
embryo in the watery environment all animals require.
ƒ efficient lungs.
ƒ three-chambered heart with a septum in the ventricle to improve
separation of oxygenated and deoxygenated blood
ƒ modified limbs and skeleton to provide better support and more efficient
movement on land.

¾ Examine and illustrate the external features of the crocodile (Order


Crocodila) or lizard (Order Squamata).

Class Aves
This class contains the birds. The important features of this group are the
presence of feathers on the skin and wings which allow flight. Adaptations
for flight include:
ƒ elevated body temperature to allow both muscles and metabolic
processes to operate at peak efficiency, supplying power and energy.
ƒ four chambered heart that completely separates oxygenated and
deoxygenated blood.
ƒ respiratory system supplemented by air sacs that supply oxygenated air
to the lungs even as the bird exhales.
ƒ feathers to protect and insulate the body; to form lightweight extensions
to the wings and tail for lift-off and control
ƒ hollow bones to keep the skeleton light weight.

¾ Examine and illustrate the external features of a bird. Note


Chordate and Avian features.

Class Mammalia
This class contains all animals with mammary glands and hair/fur on their
skin, including rats, goats, cats, dogs, monkeys, bats, elephants, seals and
whales. Mammals are homeotherms, with four-chambered hearts and high
metabolic rates. They show great diversity of form and live in a wide range
of habitats, aerial, terrestrial (on or below ground) and aquatic.

¾ Examine and illustrate the external features of an egg laying


mammal (Sub-class Protheria), the platypus (Order Monotremata).
¾ Examine and illustrate the external features of a pouch mammal
(Subclass Metatheria), the kangaroo (Order Diprotodontia).
¾ Examine and illustrate the external features of a placental mammal
(Subclass Eutheria), the monkey (Order Primates).
In each case note Chordate and Mammalian and subclass features.
BIOL0011 Preliminary Biology Laboratory Manual Page 66

Chondrichthyes: External features of a shark

Osteichthyes: External features and skeleton of a bony fish


BIOL0011 Preliminaary Biology Laboratory Manual Page 67

Amphibia: Skeleton
S of a Toad

keleton of a Lizard
Reptilia: Sk
BIOL0011 Preliminary Biology Laboratory Manual Page 68

Aves: External features and skeleton of a bird.


BIOL0011 Preliminaary Biology Laboratory Manual Page 69

Mammaliia: Skeleton
n of a man