Laboratory Manual

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Biology

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Class XII
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FOREWORD
The National Council of Educational Research and Training (NCERT) is the
apex body concerning all aspects of refinement of School Education. It has
recently developed textual material in Biology for Higher Secondary stage
which is based on the National Curriculum Framework (NCF)–2005. The NCF
recommends that children’s experience in school education must be linked
to the life outside school so that learning experience is joyful and fills the gap

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between the experience at home and in community. It recommends to diffuse
the sharp boundaries between different subjects and discourages rote learning.

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The recent development of syllabi and textual material is an attempt to
implement this basic idea. The present Laboratory Manual will be
complementary to the textbook of Biology for Class XII. It is in continuation
to the NCERT’s efforts to improve upon comprehension of concepts and

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practical skills among students. The purpose of this manual is not only to
convey the approach and philosophy of the practical course to students and
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teachers but to provide them appropriate guidance for carrying out
experiments in the laboratory. The manual is supposed to encourage children
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to reflect on their own learning and to pursue further activities and questions.
Of course the success of this effort also depends on the initiatives to be taken
by the principals and teachers to encourage children to carry out experiments
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in the laboratory and develop their thinking and nurture creativity.

The methods adopted for performing the practicals and their
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evaluation will determine how effective this practical book will prove
to make the children’s life at school a happy experience, rather than
a source of stress and boredom. The practical book attempts to provide
space to opportunities for contemplation and wondering, discussion
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in small groups, and activities requiring hands-on experience. It is

hoped that the material provided in this manual will help students
in carrying out laboratory work effectively and will encourage teachers
to introduce some open-ended experiments at the school level.

YASH PAL
Professor and Chairperson
National Steering Committee
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New Delhi National Council of Educational

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21 May 2008 Research and Training

PREFACE
The development of the present laboratory manual is in continuation to
the NCERT’s efforts to improve upon comprehension of concepts and
practical skills among the students. The present laboratory manual will
be complementary to the textbook of Biology for Class XII.
The expansion of scientific knowledge and consequently the change
in the system of education has led to the development of new methods of

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instructions. Today the stress is laid on the enquiry approach and
discussion method instead of lecture method of teaching. Biology is now

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something more than observation of living organisms. Study of Biology
includes microscopic observations to reveal minute internal details of the
organism, biochemical testing to understand complex reactions taking
place inside the organisms, experiments with live organism to understand

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various physiological processes and even much more. In other words
experiments in Biology truly represents an interdisciplinary approach of
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learning.

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The new syllabus of Biology has been designed to cater to the needs of
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pupil who are desirous of pursuing science further. The fundamental
objective of this course is to develop scientific attitude and desired
laboratory skills required for pursuing Biology as a discipline at this level.
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A similar approach has been taken while formulating the practical syllabus
of Biology for higher secondary stage. The practical syllabus includes
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content based experiments, which help in comprehension of the concepts.

There are altogether twenty-five exercises in the present manual which
are based on Biology curriculum for Class XII. For each practical work,
principle, requirements, procedure, precautions, observations, discussion
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and the questions are given in the book. The methodology of preparation
of any reagent, if required, has been given alongwith the requirements,
for the convenience of students and teachers. The questions are aimed to
develop learner’s understanding of the related problems. However, teacher
may provide help in case the problem is found to be beyond the capability
of the learner. Precautions must be well understood by the learners before
proceeding with the experiments and projects. In addition to the core
experiments enlisted in the syllabus for Class XII emphasis has also been
given for pursuing Investigation Project Work. It is expected that these
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informations will motivate the students to take up independent project

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work on topics of their interest.

Appropriate appendices related to the observation and study of
organisms are given along with the experiment. International symbols for
units, hazards and hazard warnings are given at appropriate places in
the book. It is expected that this will make the learners more careful about
the environment and make them careful while dealing with the equipments
and chemicals in the laboratory.
 It gives me a pleasure to express my thanks to all those who have been
associated at various stages of development of this laboratory manual. It is
hoped that this practical book will improve teaching-learning process in
Biology to a great extent. The learners will be able to understand the subject
well and will be able to apply the acquired knowledge in new situations.
I acknowledge with thanks the dedicated efforts and valuable contribution
of Dr Dinesh Kumar, coordinator of this programme and other team members
who contributed and finalised the manuscript. I especially thank Professor
G. Ravindra, Director (Incharge), NCERT for his administrative support and
keen interest in the development of this laboratory manual. I am also grateful

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to the participating teachers and subject experts who participated in the
review workshop and provided their comments and suggestions which helped

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in the refinement of this manual. We warmly welcome comments and
suggestions from our readers for further improvement of this manual.

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H UKUM SINGH
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Professor and Head

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Department of Education in
Science and Mathematics
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vi
LABORATORY MANUAL DEVELOPMENT TEAM

MEMBERS
Animesh K. Mohapatra, Associate Professor, Regional Institute of Education,
NCERT, Ajmer
B.K. Tripathi, Professor, DESM, NCERT, New Delhi

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C.V. Shimray, Assistant Professor, DESM, NCERT, New Delhi

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N.V.S.R.K. Prasad, Associate Professor in Botany, Sri Venkateshwara College,
New Delhi
P.K. Durani, Professor (Retired), DESM, NCERT, New Delhi

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Sunita L. Varte, Assistant Professor, DESM, NCERT, New Delhi
S.P. Sinha, Professor of Zoology (Retired), TM Bhagalpur University, Bhagalpur
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V.V. Anand, Associate Professor, Regional Institute of Education, Mysore
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MEMBER-COORDINATOR
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Dinesh Kumar, Associate Professor, DESM, NCERT, New Delhi

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ACKNOWLEDGEMENT
The National Council of Educational Research and Training (NCERT) acknowledges the valuable
contribution of the individuals and organisations involved in the development of this laboratory
manual. The council also acknowledges the valuable contribution of the following academics for
reviewing and refining the manuscript of the laboratory manual: A.K. Sharma, Reader in Zoology,
University of Lucknow, Lucknow; Iswant Kaur, D.M. School, RIE, Bhopal; K. Muralidhar,

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Professor, Department of Zoology, University of Delhi, Delhi; K.K. Sharma, Professor Department
of Zoology, M.D.S. University, Ajmer; M.M. Chaturvedi, Professor Department of Zoology,

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University of Delhi, Delhi; Nazir Ahmad Kakpori, Department of Education, Govt of Jammu &
Kashmir, Srinagar; Reena Mohapatra, St. Stephen’s Senior Secondary School, Ajmer; Savita
Sharma, Mount Carmel School, Dwarka, New Delhi; Savithri Singh, Professor and Principal,

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Acharya Narendra Dev College, New Delhi; Shalu Dhawan, Amity International School, Saket,
New Delhi; Shivani Goswami, Mother’s International School, New Delhi; V.K. Srivastava, Reader
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in Zoology, J.N. College, Pasighat; Vijay Kumar, Delhi State Science Teacher Forum, New Delhi.
We also acknowledge the contributions of Anil Kumar and Binita Kumari, Junior Project
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Fellows, DESM, NCERT, New Delhi.
Special thanks are also due to Hukum Singh, Professor and Head, DESM, NCERT for his
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interest in the work and administrative support.

The Council also acknowledges the efforts of Surender Kumar, Narender Kumar Verma, Monika
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Rajput and Girish Goyal, DTP Operators, for helping in shaping this laboratory manual. The
contributions of Publication Department of NCERT in printing out this laboratory manual are
also duly acknowledged.
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CONTENTS

FOREWORD iii
PREFACE v

Introduction 1

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Exercise 1 : To study the reproductive parts of commonly 5
available flowers

Exercise 2 : To calculate percentage of pollen germination 12

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Exercise 3 : To study pollen tube growth on stigma 14

Exercise 4 :
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To study the discrete stages of gametogenesis in 16
mammalian testis and ovary
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Exercise 5 : To study and identify various stages of female 18

gametophyte development in the ovary of a flower
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Exercise 6 : Preparation and study of mitosis in onion root 20

tips
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Exercise 7 : Study of stages of meiosis using permanent slides 25

Exercise 8 : To study the blastula stage of embryonic 30

development in mammals, with the help of
permanent slide, chart, model or photograph

Exercise 9 : To verify Mendel's Law of Segregation 32

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Exercise 10 : To verify the Mendel’s Law of Independent 35

Assortment

Exercise 11 : Preparation and analysis of Pedigree Charts 39

Exercise 12 : To perform emasculation, bagging and tagging 45

for controlled pollination
Exercise 13 : Staining of nucleic acid by acetocarmine 47

Exercise 14 : To identify common disease-causing organisms 49

and the symptoms of the diseases

Exercise 15 : To study the texture of soil samples 53

Exercise 16 : To determine the water-holding capacity of soils 59

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Exercise 17 : To study the ecological adaptations in plants living 62
in xeric and hydric conditions

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Exercise 18 : To study the adaptations in animals living in xeric 65
and hydric conditions

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Exercise 19 : To determine the pH of different water and soil 68
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samples
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Exercise 20 : To study turbidity of water samples 71
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Exercise 21 : To analyse living organisms in water samples 75

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Exercise 22 : To determine the amount of Suspended 83

Particulate Matter (SPM) in air at different sites in
a city
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Exercise 23 : To study plant population density by quadrat 85

method

Exercise 24 : To study plant population frequency by quadrat 87

method

Exercise 25 : Study of homologous and analogous organs in 89

plants and animals
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Investigatory Project Work 92–104

Project 1 : To study the effect of pH on seed germination 100

Project 2 : Quantitative analysis of phytoplankton in a water 102

body

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Introduction
Laboratory is a place where ideas and concepts can be tested through
experiments. Biology, like any other discipline of science, is based on
experimental work and therefore practical forms an integral part of learning.
Biology laboratory provides a unique learning environment where learners
inculcate scientific temper, develop relevant skills and get exposed to realms
of techniques and methodologies of scientific investigations. Laboratory
investigations in Biology increase the reasoning abilities, bring scientific
attitude in a learner and also help in acquisition of skills of scientific processes.
Also, observation of nature and the living organisms found in it is no less
important for the understanding of many aspects of the subject especially
the diversity of the living organisms, their systematic study, their relationships
among themselves and with the environment. Knowledge in the field of
Biology can be acquired or constructed only on the basis of correct
observations and experimentally verifiable processes.
Biology laboratory thus provides the learners an environment where the
process of learning is facilitated by hands-on experiments. Biology is a
unique discipline in the sense that it does not merely deal with the study of
morphology, anatomy, physiology and reproduction of the living organisms,
rather, understanding of the subject requires understanding of a number of
interdisciplinary areas and approaches. On one hand, a biologist needs to
be sufficiently skilled in handling the enormous diversity of the living
organisms, be it plants, animals, fungi or even microscopic bacteria, while
on the other hand, a biologist should be able to understand the biochemical,
molecular, physiological, behavioural, genetic and many other phenomena
pertaining to the living organisms. The study of intricate relationship of
different types of organisms among themselves and also with its environment
is an important concern of a biologist. Thus, experiments and exercises in
Biology train a learner about skills of observations, manipulation of the
organisms for the study of internal details, biochemical as well as molecular
composition and processes, investigation of the abiotic environment and even
analysis of phenomena like inheritance and evolution.
As far as the study of the living organism is concerned, correctness of the
method is very important. Such a study may be very simple, e.g., study of
habit, habitat and external features of the plants or animals, or, it may involve
certain manipulations like dissection and section cutting of the parts of the
organisms to study the minute details. Very often observation and study of
the magnified image of the minute parts under a microscope provides a
better insight about the features of the organisms. However, microscopic
study involves certain specific skills depending on type of the organisms/
tissues/cells to be studied. It involves specific preparations (peeling, section

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 LABORATORY MANUAL: BIOLOGY

cutting, fixation, staining, dehydration, mounting, etc.) so that microscopic

examination reveals the expected details. As histological and cytological
observations give us only static pictures of the continuous processes, analysis
of biochemical, physiological and ecological aspects need certain other kinds
of skills such as preparation of chemicals and reagents, designing and
performing an experiment, observation and recording of data and ultimately
interpretation and drawing conclusions. While performing experiments,
honesty in recording of data and its correct presentation is very important
as it is not only useful in the logical interpretation but also helps in the
identification of errors.
In order to perform experiments successfully, a learner needs to go to
the Biology laboratory well prepared. This includes the following:
1. Laboratory Record Book: For maintaining all the information including
recording of data and its interpretation.
2. Dissection Box: A dissection box is required in the Biology laboratory
for various purposes like handling and manipulation of living materials,
performing experiments, preparation of slide, etc. A dissection box
should contain scissors (two pairs, one small with fine tip and one
larger), scalpels (one small and one medium sized), forceps (two, one
small with sharp fine tips and the other medium sized with blunt tips),
dissecting needles (two), razor, hand lens, dropper, fine brush, etc.
3. Laboratory Manual
4. A Laboratory Coat or Apron
5. A Hand Towel
While in the laboratory a student should be very careful and methodical.
One should listen carefully to the instructions given by the teacher/
instructor before performing an experiment. In the biology laboratory a
student has to handle a number of sharp objects and hence necessary
precaution and care should always be taken while handling objects like
scissors, forceps, needles, scalpel, razor, etc. It is also very important to
follow the safety instructions mentioned on the instruments and/or on the
label of the reagent/chemical. Students should also be aware about the
use of the First-aid Box so that in case of any accident or injury the
preliminary aid can be provided to the affected person.
While describing the experiment students are expected to follow a pattern
in which the aim of the experiment, its principle, list of the materials to be
used, procedure, observation table (if required), inference and discussion
should be given. Necessary precautions to be taken should also be
mentioned appropriately in the procedure or at the end. There are a few
experiments in which field visit is essentially required. For this all the
necessary preparations (materials, equipments, reagents and chemicals)

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IENTRODUCTION
XERCISE 1

should be made in advance. Drawing of illustrations is also an important

component of the practical in Biology. Students are expected to follow certain
fundamental rules while drawing the illustrations so that it reflects your
observations correctly.
• Make your illustrations using pencil only and always use white drawing
sheet. Illustration should be in the centre of the page.
• Drawing of an object (plant, animal or experimental set-up) should be
proportionate in size.
• Draw your illustrations keeping the object before you.
• Drawing must be clear with simple outlines.
• Appropriately label your drawing. Parts of the drawing should be
indicated by straight horizontal line or arrow. Two lines or arrow should
never cross each other. As far as possible, labelling should be done on
the right side of the drawing. An appropriate legend or heading of the
drawing should also be given below it.
About the Manual
The main objective of the manual is to introduce the students of higher
secondary stage to the fascinating world of plants, animals and microbes
and their complex biological phenomena. The manual covers a complete
description of the experiments and exercises. The suggested experiments
cover almost all the units/topics including those on diversity in living world,
plant, animal and human physiology, genetics, bio-technology and human
welfare and environment. A standard format has been used to describe each
experiment which includes
• Aim: It gives a brief title of the experiment under investigation.
• Principle: It is a very brief introduction of the experiment under
investigation and explains the biological phenomenon involved. It gives
brief but comprehensive ideas about the design of the experiment and
explains the significance of the phenomenon being studied.
• Materials required: This includes the names of plants/animals to be
used as 'samples', the type of apparatus, the type and quantity of
glasswares required, reagents, chemicals and solutions needed, their
concentration and other specifications, method of preparations of
solutions and reagents. If a particular material/chemical/glassware is
not available, sufficient alternatives have been suggested.
• Procedure: This section includes full details of experimental procedure
explained stepwise, including special precautions necessary to be taken
while the experiment is being conducted. Drawings of the samples,
apparatus and the experimental setup, wherever found necessary, have
been included to facilitate the students to perform the experiment as
accurately as possible.

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 LABORATORY MANUAL: BIOLOGY

• Observation and Results: This section deals with the recording of all
observations made during the experiment. Students are advised to
consider the entire data. Data can be represented in the form of tables,
graphs and histograms wherever possible. Use of units in which various
quantities are measured has been indicated in the manual.
• Discussion: Included in this heading is a statement of the conclusions
drawn from the experimental results and compared thesis (wherever
possible) with any comparable data from other sources. The relevance
of the conclusions drawn from the experimental results to the various
processes under investigation and to the life of plant, animal and microbes
has been prompted out.
• Precaution: This section contains all the necessary precautions to be
taken during experimentation to obtain results free of errors. However,
attempts have been made to mention required precautions along with
the procedure also.
A great emphasis has been laid on a student getting valid results and
interpreting them. It is essential that the teacher should properly explain
each experiment so that inexperienced students will be able to obtain accurate
results within a reasonable time. Teachers are also expected to help students
in identifying errors and mistakes committed during experiments and ways
for correcting them. It is possible that some of the students may undoubtedly
be capable of doing more sophisticated work than that represented in the
manual. But introductory course of this sort has been designed to help all
students for some useful and joyful experience by conducting the
experiments of their own. The manual also aims that students and teachers
not be discouraged by either incomplete experiments or experiments which
yield apparently meaningless results.
With the objectives of inculcating scientific temper among learners and
providing them an opportunity to undertake independent scientific
investigation, Investigatory Project Work has been included as an integral
part of the practical curriculum of Class XII. Such investigatory projects
are expected to provide thrill in the learning process. It is also expected to
serve the real purpose of practicals, i.e., developing an ability to hypothesise
and design experiments to address certain problems, to make observations
methodically and to draw conclusions out of the experimental data. A
comprehensive idea about undertaking investigatory project has been given
in the book with a list of a few problems on which investigatory project
work can be undertaken. However, the list is only suggestive and considering
the wider scope students can undertake any kind of investigatory project
work depending on their region, its climatic condition, availability of
resources, etc.

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Exercise 1


            









Procedure
(i) Familiarise with the terms to describe the reproductive parts of flowers
given in annexures of Exercise No. 11 of Laboratory Manual: Biology
(Class XI) and at the end of this experiment.
(ii) Observe the flower with the naked eye, hand lens or under a dissecting
microscope. Study their reproductive parts and count the number of
stamens and record their cohesive and adhesive features.
(iii) Cut L.S. of the flower and place it on a slide to observe the following
characters:
(a) Placement of anthers
(b) Position of the ovary: epigynous/perigynous/hypogynous.
(iv) Mount one stamen on a slide and study the following characters:
(a) Attachment of filament to anther
(b) Dehiscence pattern of the anther lobes for discharge of pollen.
(v) Cut T.S. of anther lobe to observe the number of pollen sacs.
(vi) Mount the pistil on a slide and study style, stigma and ovary. Record
the number of stigma and nature of pistil.
(vii) Cut T.S. of ovary, mount it on a slide and observe
(a) Number of locules in the ovary

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(b) Type of placentation

(c) Number of ovules per locule
(viii) Draw labelled figures of your preparation and observations.

Questions
1. Name the most common type of placentation observed.
2. What is the most common type of dehisence pattern in anthers?
3. Name a few unisexual flower -bearing plants studied by you.
4. “Flower is a modified shoot.” Justify the statement based on your observation.

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EXERCISE 1

Annexure 1

Description of reproductive parts of flowers

Androecium

Number of stamens The number of stamens may vary from a few to many in dif-
ferent flowers

Cohesion Stamens may be free or united. If united they can be of the

(Fig. 1.1 a–e) following type:
(i) Syngenesious: Filaments free and anthers united,
e.g., Sunflower.
(ii) Synandrous: Stamens fused all through their length,
e.g., Cucurbita
(iii) Adelphous: Anthers remain free and filaments are united.
Adelphous condition can be
(a) Monoadelphous—United to form 1 bundle, e.g.,
China rose
(b) Diadelphous—United to form 2 bundles, e.g., Pea
(c) Polyadelphous—United into more than two
bundles, e.g., Lemon

Adhesion Fusion of stamens with other parts of the flower

(Fig. 1.2) (i) Epipetalous: Stamens fused with petals,
e.g., Sunflower, Datura
(ii) Epiphyllous: Stamens fused with perianth,
e.g., Lily

Attachment of filament to (i) Basifixed: Filament attached to the base of anther,

anther e.g., Mustard
(Fig.1.3 a–d)
(ii) Adnate: Filament attached along the whole length of
anther, e.g., Michelia, Magnolia
(iii) Dorsifixed: Filament attached to the back of anther,
e.g., Passion flower
(iv) Versatile: Anther lobes attached with filament in the
middle portion with both ends free, e.g., Gramineae
family

Dehiscence pattern (i) Porous: Pollens released through pores, e.g., Brinjal,
(Fig. 1.4 a,b) Potato
(ii) Longitudinal: Pollens released through the longitudinal
slit of another lobes, e.g., China rose, Cotton

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 LABORATORY MANUAL: BIOLOGY

Gynoecium

Number of stamens The number of stamens may vary from a few to many in
different flowers

Position of ovary (i) Epigynous: Position of ovary inferior to other floral parts,
(Fig. 1.5 a–d) e.g., Mustard, China rose
(ii) Perigynous: Other floral parts are attached around the
ovary, e.g., Apple, Guava
(iii) Hypogynous: Position of ovary superior to other floral
parts, e.g., Sunflower

Cohesion If number of carpels is more than one, they may be

(Fig. 1.6 a–c) (i) Apocarpous: Carpels are free. Each carpel has its own
style and stigma, e.g., Rose
(ii) Syncarpous: Carpels are united, e.g., Lady’s finger, Tomato

Number of locules in ovary Vary from one to many

(i) Unilocular: One locule, e.g., Rose, Pea
(ii) Bilocular: Two locules, e.g., Datura
(iii) Multilocular: Many locules, e.g., Lady’s finger, China rose

Placentation (i) Marginal: The placenta forms a ridge along the ventral
(Fig. 1.7 a–e) suture of the ovary and the ovules are borne on this ridge,
e.g., Pea
(ii) Axile: The ovary is partitioned into several chambers or
locules and the placentae are borne along the septa of
the ovary, e.g., Tomato, China rose
(iii) Parietal: The ovules develop on the inner wall of the
ovary or on peripheral part. Ovary unilocular but in some
cases becomes two chambered due to formation of a false
septum, e.g., Mustard
(iv) Free central: Ovules are borne on the central axis and
septa are absent, e.g., Carnation, Chilly
(v) Basal: Placenta develops at the base of the ovary,
e.g. ,Sunflower.

21/04/2018
EXERCISE 1

(a) (b) (c)

(d)
(e)
Fig.1.1 Cohesion of stamens: (a) Syngenesious (b) Synandrous
(c) Monoadelphous (d) diadelphous (e) Polyadelphous

Fig.1.2 Adhesion of Stamens: Epipetalous/Epiphyllous

(a) (b) (c) (d)

Fig.1.3 Attachment of filament to anther: (a) Basifixed (b) Adnate

(c) Dorsifixed (d) Versatile

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 LABORATORY MANUAL: BIOLOGY

(a) (b)
Fig.1.4 Dehiscence pattern of anther: (a) Porous (b) Longitudinal

(a) (b) (c) (d)

Fig.1.5 Position of ovary: (a) Epigynous (b–c) Perigynous (d) Hypogynous

(a) (b) (c)

Fig.1.6 Cohesion of carpels: (a) Apocarpous (b–c) Syncarpous

10

21/04/2018
EXERCISE 1

(a) (b) (c)

(d) (e)

Fig.1.7 Placentation: (a) Marginal (b) Axile (c) Parietal (d) Free central (e) Basal

11

21/04/2018
Exercise 2









Procedure
(i) Prepare the pollen germination medium by dissolving 10g sucrose,
30mg calcium nitrate and 10mg boric acid in 100ml of distilled water.
Alternatively 10% sucrose solution can also be used.
(ii) Take a drop of medium or 10% sucrose solution on a cover slip and
sprinkle mature pollen grains on the drop.
(iii) Invert the cover glass on to a slide
(iv) After 10 minutes, observe the slide under microscope.
(v) Count (a) total number of pollen grains seen in the microscope field,
and (b) the number of pollen grains that have germinated.

Observation
Several pollen grains germinate and put forth pollen tubes. Count the total
number of pollen grains and the number of germinated pollen grains in 3-5
different microscope fields. Tabulate your observations and calculate the
percentage of pollen germination.
Nameoftheplantusedassourceofpollen……………………………
Number of pollen grains in a field of microscope = N
Number of germinated pollen grains in a field of microscope = n
n 100n
Percent pollen germination = ×100 or
N N

21/04/2018
EXERCISE 2

Number of Total number of pollen (N) Total number of pollen germinated (n) % pollen germination
observation n
×100
N
1.
2.
3.
4.
5.
Average

Discussion
Although pollen grains of many species germinate in this medium, the
percentage of germinations and the time taken for germination varies in
different species. Draw a germinating pollen grain and label.

Questions
1. How many pollen tubes emerge from a pollen grain?
2. What does the pollen tube carry?
3. Can you explain as to why some pollen grains fail to germinate?
4. Why do we use sucrose as the medium for pollen germination?

13

21/04/2018
Exercise 3











Procedure
(i) Place the stigmas in boiling water in a beaker
for softening the tissues for 5–10 minutes. Pollen grains
(ii) Stain with cotton blue for 3–5 minutes and
wash with water to remove excess stain.
(iii) Mount one stigma in a drop of glycerine on a Pollen tube
slide. Place a cover slip on the stigma and gently
press the cover slip on the material. Observe Style
the slide under a microscope.
(iv) If you fail to observe pollen tubes mount
another stigma.

Observation
Look for long blue-coloured tubular structures
traversing through the tissues of stigma and Fig.3.1 Growth of pollen tube
style (Fig. 3.1). in the style of a carpel

21/04/2018
EXERCISE 3

Discussion
Pollen tubes are seen amidst the stylar tissue. Many pollen tubes may be
seen. Trace the origin of pollen tubes to the pollen grains present around
the surface of the stigma.

Questions
1. Can pollen grains of one plant species germinate on stigma of other species?
Give reasons.
2. Do all pollen tubes reach the ovules?
3. Are all the pollen tubes of equal length? If not, why?

15

21/04/2018
Exercise 4








          


Procedure
(i) Clean the slide and microscope’s eye and objective lenses with the
help of lens cleaning paper using any cleaning fluid.
(ii) Place the slide on the stage of the microscope and observe first under
lower magnification and then in higher magnification. Observe various
stages of gamete development.
(iii) Record your observations in the notebook and draw labelled diagrams.

Observation
T.S. of testis
(i) You will observe a large number
of seminiferous tubules under
Seminiferous tubule
lower magnification. Observe a
complete tubule in higher
magnification and view various
stages of gamete development Spermatozoa
from periphery towards lumen
(Fig. 4.1) and identify the
following types of cells namely, Germinal
Epithelium
Germinal epithelium,
Spermatogonial cells, Primary Spermatogonia
spermatocytes, Secondary
spermatocytes, Spermatids and
Spermatozoa. Fig. 4.1 T.S. of mammalian testis

21/04/2018
EXERCISE 4

(ii) In T.S. of testis the space between tubules are filled with blood vessels
and a specific cell type called Leydig's cell or Interstitial cells.

T.S. of Ovary
(i) In the section of ovary, there is a mass Graafian Follicle
of tissue lined with germinal
epithelium. Inside that you will
Antrum
observe an ovum, which is a cell
surrounded by one to several layers
of follicular cells. As the ovum
matures, the number of surrounding
follicular cell layer increases (Fig. 4.2). Corpus luteum
(ii) In the later stage of follicular Corpus albicans
development a cavity called antrum
appears. Fig. 4.2 Section of mammalian ovary
(iii) The cavity gets further enlarged and
the follicle grows bigger. This is the
stage of Graafian follicle ready to release the ovum (ovulation).
(iv) In the next stage, you may notice a Corpus luteum, and/or Corpus
albicans, which differ from each other and also from Graafian follicle in
their features.

Discussion
Spermatogenesis is a continuous process after attainment of puberty, and
that is why gamete development and spermatozoa are observed in a single
seminiferous tubule. In case of ovary, the follicular development stages are
observed.

Questions
1. What would happen if meiosis fails to occur in gametocyte?
2. At which stage of follicular development, is ovum released?
3. Spermatogenesis is a continuous process. Justify the statement.
4. Draw a labelled diagram of T.S. of testis.
5. Draw a labelled diagram of T.S. of ovary.
6. What would happen if sperms are devoid of their tail?
7. What are the consequences of failure of ovulation?

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Exercise 5











Procedure
(i) In a V.S. of ovary we generally find several ovules. Carefully observe
each ovule and locate as many stages of female gametophyte
development as possible.
(ii) Draw the diagrams as observed under microscope.

Observation Chalaza
(i) Record the features of ovule
like number of integuments,
nucellus and micropylar
and chalazal poles. Fig 5.1
Embryo sac
shows the female
gametophyte (embryo sac)
as seen in a V.S. of an ovule.
Different stages of Outer integument
development of female Inner integument
gametophyte are shown in
Fig. 5.2. Micropyle

(ii) Observe the placement of

embryo sac close to the Funiculus
micropylar pole.
Fig. 5.1 V.S. of an ovule

21/04/2018
EXERCISE 5

Egg

Synergids

Central cell
Secondary nucleus

(a) (b) (c) (d) (e) Antipodals

Fig. 5.2 Stages of gametophyte development: (a) megaspore with 2 nucleus

(b) 4-nucleate stage (c) 8- nucleate stage (d) 8- nucleate stage
showing 3+2+3 distribution of nuclei (e) mature embryo sac.

(iii) Note the contents of embryo sac, namely, an egg apparatus

(2 synergids and egg) at micropylar end, secondary nucleus in the
center and three antipodal cells at the chalazal end (Fig. 5.2).

Questions
1. Explain the difference between gamete and a gametophyte.
2. Name two differences between synergids and egg.
3. What is the function of polar nuclei?

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Exercise 6














Procedure
Growing of root tips
Select a few medium-sized onion bulbs. Carefully remove the dry roots
present. Grow root tips by placing the bulbs on glass tubes (of about 3–4
cm. diameter) filled with water. Care should be taken so that the stem portion
of the bulb (basal part) just touches the water. A few drops of water may be
added periodically to compensate evaporation losses. New roots may take
3–6 days to grow. Cut 2–3 cm long freshly grown roots and transfer them to
freshly prepared fixative, i.e., aceto-alcohol (1:3:: glacial acetic acid : ethanol).
Keep the root tips in the fixative for 24 hours and then transfer them to 70%
ethanol (for preservation and use in future). Onion root-tip cells have a cell
cycle of approximately 24-hour duration, i.e., they divide once in 24 hours,
and this division usually takes place about two hours after sunrise. Therefore,
roots grown on water should be cut only at that time to score maximum
number of dividing cells.

Preparation of slide
Take one or two preserved roots, wash them in water on a clean and grease-
free slide. Place one drop of N/10 HCl on the root tip followed by 2–3 drops
of aceto-carmine or aceto-orcein stain on it. Leave the slide for 5–10 minutes

21/04/2018
EXERCISE 6

on a hot plate (or warm it slightly on spirit lamp). Care should be taken that
the stain is not dried up. Carefully blot the excess stain using blotting paper.
Now cut the comparatively more stained (2–3 mm) tip portion of the root
and retain it on the slide and discard the remaining portion. After
(10–20 seconds) put one or two drops of water and blot them carefully using
blotting paper. Again put a drop of water on the root tip and mount a cover
slip on it avoiding air bubbles. Place the slide in between the folds of blotting
paper using the fingers in such a way that the cover slip mounted on the
slide is properly held. Now slowly tap the cover slip using the blunt end of a
pencil so that the meristematic tissue of the root tip below the cover slip is
properly squashed and spread as a thin layer of cells. Carefully seal the
margins of the cover slip using molten paraffin wax or nail polish. This
preparation of onion root tips cells is now ready for the study of mitosis.

Study of slide
Place the slide on the stage of a good quality compound microscope. First
observe it under the lower magnification (10 X objective) to search for the
area having a few dividing cells. Examine the dividing cells under higher
magnification of the microscope to observe the detailed features of mitosis.

Observation
The stages of mitosis can be broadly categorised into two parts: karyokinesis
(division of nucleus) followed by cytokinesis (division of cytoplasm, and
ultimately of the cell). Those cells, which are not in the phases of cell division
are considered to be in interphase. You may observe that most of the cells
in a microscope field are in interphase

Interphase
The cells are mostly rectangular, oval or even circular in shape, with almost
centrally situated densely stained nucleus. The chromatic (coloured) material
of the nucleus is homogeneous and looks granular. The boundary of the
nucleus is distinct. One or few nucleoli (sing: nucleolus) can also be observed
inside the nucleus (Fig. 6.1a).

Stages of Mitosis
(a) Prophase
Intact nuclear outline is seen. The chromatin (seen as a homogeneous
material in the nucleus at interphase) appears as a network of fine threads
(chromosomes). Nucleoli may or may not be visible (Fig. 6.1b).

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 LABORATORY MANUAL: BIOLOGY

a. Interphase

b. Prophase

c. Metaphase

d. Anaphase

e. Telophase

Fig.6.1 Interphase (a) and stages of mitosis (b - e) – actual microscopic

view on left side and its diagrammatic representation on the
right hand side
If the cell under observation is in the early stage of prophase then the
chromatin fibres (chromosomes) are very thin. However, in the cells
at late prophase, comparatively thicker chromatin fibres would be
visible. Besides this, in the late prophase the nuclear membrane
may not be noticed.

(b) Metaphase
The nuclear membrane disappears. Chromosomes are thick and are seen
arranged at the equatorial plane of the cell (Fig. 6.1c). Each chromosome at

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21/04/2018
EXERCISE 6

this stage has two chromatids joined together at the centromere, which can
be seen by changing the resolution of the microscope. Nucleolus is not
observed during metaphase.
(c) Anaphase
This stage shows the separation of the chromatids of each chromosome. The
chromatids separate due to the splitting of the centromere. Each chromatid
now represents a separate chromosome as it has its own centromere. The
chromosomes are found as if they have moved towards the two poles of the
cell. The chromosomes at this stage may look like the shape of alphabets 'V',
'J' or 'I' depending upon the position of centromere in them. Different anaphase
cells show different stages of movement of chromosomes to opposite poles,
and they are designated to represent early, mid and late anaphase (Fig. 6.1d).
(d) Telophase
Chromosomes reach the opposite poles, lose their individuality, and look
like a mass of chromatin (Fig. 6.1e). Nuclear membrane appears to form the
nuclei of the two future daughter cells.

Cytokinesis
In plants, a cell plate is formed in the middle after telophase. The plate can
be seen to extend outwards to ultimately reach the margin of the cell and
divide the cell into two. Such cell plates are characteristic of plant cells
(Fig. 6.2). However, in an animal cell, the two sides of the cell show inpushings
or constrictions formed from the peripheral region in the middle of the cell,
which grow inward and meet to divide the cell into two daughter cells.
Draw labelled diagrams of all the phases of mitosis.

Fig. 6.2 Cytokinesis

Discussion
Mitotic index (MI) is defined as a ratio of the total number of dividing cells (n)
and the total number of cells (N) in a particular focus chosen randomly under
n
the microscope and is calculated as MI = N ×100 . By randomly selecting
5 to 10 such foci, one can estimate the mitotic index for a given type.

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 LABORATORY MANUAL: BIOLOGY

The effect of different samples of water (polluted or contaminated) can be

assayed on the mitotic-index (an indicative feature of somatic growth rate in
them).
Further, the impact of different types of pollutants on different phases of
mitosis can also be assayed.

Tabulate your observations in the tabular form given below

Features Interphase Karyokinesis Cytokinesis

Prophase Metaphase Anaphase Telophase

1. Cell morphology

2. Nuclear morphology

3. Chromosomes/chromatids

Questions
1. Suggest names of a few tissues, which are suitable for the study of mitosis.
2. Why is mitosis also known as equational division?
3. What shape would a metacentric and a sub-metacentric chromosome exhibit
during the anaphase stage?
4. How does cytokynesis differ in plant and animal cells?

24

21/04/2018
Exercise 7











Procedure
Place the slide on the stage of the microscope and search for the dividing
cells using lower magnification. When dividing cells are located observe them
under higher magnification.

Observation
Observe various stages of meiosis and identify them on the basis of the specific
features given in the table 7.1. A significant number of cells will be in the
Interphase. These cells have a centrally positioned densely stained nucleus.
In case of slide of animal tissue a few mitotically dividing spermatogonial
cells may also be seen.
Table 7.1 Different stages of meiosis and their features
Meiosis I
1. Prophase I Unlike the prophase of mitosis, it is a comparatively complex phase
characterised by a number of events. Five sub-phases can be
identified in it.
(a) Leptotene (leptos = slender tene = band or thread)
(i) The nuclear membrane and nucleolus are not distinctly
observable (Fig. 7.1 a).
(ii) Fine network of thin threads are seen uniformly distributed
in the nucleus.These are chromatin threads, which may be
observed as more prominent structures in the later stages.

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(a) Leptotene

(b) Zygotene

(c) Pachytene

(d) Diplotene-Diakinesis

Fig. 7.1 Sub-phase of Prophase I (a-d) – actual microscopic view on left

side and its diagrammatic representation on the right hand side

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EXERCISE 7

(b) Zygotene (Zygon = paired)

This stage is characterised by the pairing of the homologous
chromosomes, which can be seen as paired chromatin threads
(bivalents) (Fig. 7.1b).
(c) Pachytene (pachy = thick)
The chromatin threads get condensed and appear shortened and
thick. Pairs of homologous chromosomes can be seen. Each
chromosome has two chromatids and thus each bivalent consists
of four chromatids. This configuration is called tetrad (Fig. 7.1c).
(d) Diplotene (diplos = double)
The homologous chromosomes (each made up of two chromatids)
show distinct separation from each other except at few regions where
attachments are seen (Fig. 7.1d). These are chiasmata (sing.
chiasma) representing the site of exchange of the parts between
two homologous chromosomes (i.e. crossing over).
(e) Diakinesis (Dia = opposite; kinesis= separation or movement)
(i) The homologous pair of chromosomes appear more shortened,
thick and prominent (Fig. 7.1d).
(ii) Chiasmata can be still observed.
(iii) All the homologous pairs appear scattered in the cell.
2. Metaphase I Homologous chromosomes are still in pairs, and are arranged along
the equatorial plane of the cell (Fig. 7.2a). At this stage, the number
of bivalents can be counted. Chiasmata may still be seen in a few
bivalents.

3. Anaphase I The chromosome pairs appear to have moved towards the two
opposite poles of the cell. At the later stage, the anaphase - I may
show the assembly of chromosomes at two poles (Fig. 7.2b). This
results into the reduction of number of chromosomes to half.
This stage can be identified by the presence of two chromatids in
each chromosome.

4. Telophase I The chromosomes present at the two poles appear decondensed

and form two distinct nuclei (Fig. 7.2c).
Note: After the telophase I stage there may or may not be cytokinesis.
Thereafter the cell enters into the second meiotic division.

Meiosis II

1. Prophase II (i) Distinct thread- like chromatin fibres or rod- shaped chromosome
are seen.

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 LABORATORY MANUAL: BIOLOGY

(b) Anaphase I
(a) Metaphase I

(c) Telophase I

Fig. 7.2 Phases of Meosis I (a-c) – actual microscopic view on left side
and its diagrammatic representation on the right hand side.

(a) Metaphase II

(b) Anaphase II

Fig.7.3 Phases of Meosis II (a,b) – actual microscopic view on left side

and its diagrammatic representation on the right hand side.

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EXERCISE 7

2. Metaphase II This phase is similar to that of mitotic division

(i) The chromosomes having two chromatids attached at the
centromere are observed arranged at the equatorial plane of the
cell.
Note: Metaphase II of meiosis can be differentiated from metaphase-I
on the basis of the following features:
(ii) Each chromosome of metaphase II has two chromatid
(Fig. 7.3a) whereas in metaphase I these are paired
homologous chromosomes each having two chromatids thus
forming tetrad.
(iii) In the metaphase I of meiosis, a few chiasmata are observed,
where as no chiasmata are observed during metaphase II.
3. Anaphase II The two chromatids of each chromosome after separation appear
to lie at the two poles of the cell (Fig. 7.3b).
Note: Anaphase II can also be distinguished from the anaphase I of
meiotic division on the basis of chromatids: In anaphase I, each
chromosome has two distinct chromatids, but in anaphase II, each
chromosome is represented by one chromatid only.
4. Telophase II The separated chromosomes appear decondensed and form nuclei
(Fig. 7.3c).

Questions
1. What is the significance of meiosis?
2. What is synapsis and crossing over?
3. How can anaphase I and anaphase II be distinguished from each other?
4. Indicate distinguishing feature of metaphase I of meiosis and metaphase of mitosis.
5. How many daughter cells are produced at the end of meiosis?
6. The daughter cells produced at the end of meiosis are genetically different. Explain.
7. What is the significance of synapsis?

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Exercise 8










Procedure
Observe the slide under lower magnification of the microscope. In case of
chart/models/photographs, note the feature of blastula in your practical
record and draw labelled diagram.

Observation
In transverse section, the blastula appears as a sphere with a cavity, called
blastocoel within it (Fig. 8.1). Notice an outer layer of blastomeres called
trophoblasts. A cellular mass, adhered to the trophoblast is present on one
end of the blastula. It is called inner cell mass.

Fig.8.1 Blastula stage of a mammal

21/04/2018
EXERCISE 8

Questions
1. What are the differences between blastula and morula?
2. What are the main structures you observe in T.S. blastula?
3. Match the stages in column I with features in column II
Column I Column II
(a) Trophoblast (i) Dividing cells of the morula
(b) Morula (ii) Outer layer of blastula
(c) Blastocoel (iii) Solid ball of cells
(iv) Cavity

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21/04/2018
Exercise 9













Procedure
Students have to work in pairs to perform the experiment. The following
steps are to be strictly followed in the sequence mentioned below.
(i) Put 64 yellow beads in one beaker/petridish and 64 green beads in
the other to represent respectively male and female gametes. Let the
yellow bead be indicated by ‘Y’ and green bead by ‘y’.
(ii) Take a bead from each container and place them together (it represents
fertilisation) on the napkin spread before you on the table. (One student
to take out beads and to put in the hands of the other student who will
put them on the table).
(iii) Just like the previous step, continue to pick beads and arrange them
in pairs. Thus 64 pairs of beads are obtained representing the 64
heterozygous F1 progeny.
Note that all the F1 individuals are represented by one yellow and one
green bead.
(iv) Put 32 F1 progeny in one petridish and the remaining 32 in another
petridish (representing the F1 males and females).
(v) Stir the beads of each petridish with a pencil/pen for about 10 times
taking care that no bead falls off.

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EXERCISE 9

(vi) To obtain the F2 generation, one student would withdraw one bead
from one beaker labelled male and one from the other beaker labelled
female keeping his/her eyes closed (to ensure randomness), and put
them together in the stretched palm of the partner, who will put them
together on the napkin spread over the table. Continue this process till
all the beads are paired. Thus 64 offsprings of F 2 are obtained.
(vii) Note the genotype (YY or Yy or yy) of each pair, and their possible
phenotype.
(viii) Have six repeats of the experiment (steps i to vii) with partners changing
their roles. Pool all the data from the six repeats together.
(ix) Calculate the genotypic and phenotypic ratios of your pooled data.
Note that larger the sample size, more accurate is the result.

Observation
Record the result in the following table:

Generation Repeat No. Total no. of Genotype (s) Phenotype (s)

individuals YY Yy yy

F1 1.
2.
3.
4.
5.
6.
Total
F2 1.
2.
3.
4.
5.
6.
Total

Phenotypic Ratio: in F1…….

in F2…….
Genotypic Ratio: in F1…….
in F2…….

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Discussion
The results are so because each diploid individual contains two copies of
every gene - one copy on each of the two homologous chromosomes. These
two copies of the gene may be of similar type (YY or yy) or are dissimilar Yy.
The former (YY or yy) are called homozygous for that particular character,
and the Yy are called heterozygous ones. The pure lines in the above cross
are homozygous ones, which contributed only one copy of their gene (as a
result of meiosis) to their F1 progeny to restore its diploid nature with genotype
Yy (heterozygous) where only one form (allele) is expressed (dominant) and
the other form (allele) is not expressed (recessive). This is the phenomenon
of Dominance.
When the F1 individuals are crossed together to raise the F 2 generation,
each F1 individual produces two types of gametes: 50% having dominant
allele, and the remaining 50% having recessive allele. These gametes undergo
random fusion during fertilisation to produce the F2 generation. According
to simple probability of mixing of opposite sex gametes (sperms and ova),
offsprings of three genotypes are likely to appear as follows: [(half of gametes
of Y type + half of remaining gamete y type) X (half gametes of Y type + half
of remaining gamete of y type)] = One-fourth of F2 individuals of YY phenotype
+ half of F2 individual Yy type + one-fourth of F2 individul of yy type. Among
these proportion of dominant phenotype would be ¼ YY+ ½ Yy = ¾ yellow
and recessive phenotype ¼ yy i.e. green phenotypes in 3:1 or 75%:25%
ratio.
This ratio of 3:1 in the F2 suggests that in the F1 heterozygotes, the
recessive allele does not get destroyed and remains only in the recessive
(dormant) state to get an opportunity to express itself when it has separated
from the influence of the dominant allele (Y). This is called Law of Segregation
of the alleles.

Questions
1. Do you expect the same results in terms of 3:1 ratio in F2 if you had started with
smaller number of beads (say 10 beads)?

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Exercise 10











Procedure
Students are to work in pair.
The following steps are to be followed sequentially:
(i) Place 64 beads of each colour in four separate beakers.
(ii) Put the beakers containing the yellow and red beads on your left side,
and those containing the green and white beads on your right side.
The beakers on your left side represent plants bearing yellow seed and
red flower (dominant character YY, RR). Beakers on the right side
represent plants bearing green seeds and white flowers (recessive
character yy, rr). These are the two parental types having contrasting
forms of two different characters.
(iii) Stir the beads in each beaker with a pencil/pen. Each bead now
represents alleles in the male and female gametes.
(iv) Pick up one yellow, one green, one red and one white bead, and put
them together on the napkin spread on the table.
(v) Continue picking up and putting together of the beads of all colours
as mentioned in the previous step, till all the beads are utilised.
(vi) Note that in all, 64 such 4-bead clusters are obtained representing the
F1 individuals. Ascertain their genotype and phenotype.
(vii) Next step is to cross these F1 individuals to raise the F2 generation. Let
us suppose half of the 4-bead clusters (32 clusters) represent the male
parents and the remaining half (32 clusters) the female parents. Now
put the 32 red and 32 white beads together in one beaker (numbered-
I), and similarly put 32 yellow and 32 green beads together in other

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beaker (numbered-II). These two beakers represent F1 female. Similarly

put remaining 32 red + 32 white beads in beaker numbered-III, and
32 yellow and 32 green one in beaker numbered-IV to represent the F1
male. The arrangement can be presented as below

Female F1 Male F1

32 red + 32 white (Beaker I) 32 red+32 white (Beaker III)

32 yellow + 32 green (Beaker II) 32 yellow + 32 green (Beaker IV)

(viii) Stir the beads in each beaker with a pencil. In order to raise the F2
generation, pick up (with eyes closed) one bead from the beaker-I of
female and one bead from the beaker-III of the male, and put into the
palm of the partner student. Similarly, pick up one bead each from the
beaker-II of female and beaker IV of male to put in the palm of the
partner. This partner would now keep all the four beads together (to
represent the F2 individual). Continue this process till all beads are
utilised. At the end, 64 F2 individuals (each represented by a 4-bead
cluster) are obtained.
(ix) Determine the genotype and phenotype of each of the 64 F2 individuals
and write down the number of individuals of different genotypes and
phenotypes in the tabular form (given below), remembering that Y
(yellow seed colour) is dominant over y (green seed) and R (red flower)
is dominant over r (white flower).
(x) Repeat the whole procedure (steps i to ix) six times, and tabulate your
results.

Observation
Tabulate the results as follow:
Symbol (-) indicates the presence of corresponding dominant or recessive
allele e.g. Y or y and R or r.
Summarise your results (adding together the data of all the six repeats)
F1 Generation
(a) Total number of individuals: _________________________
(b) Phenotype (s) _________________________
(c) Genotype (s) _________________________

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EXERCISE 10

Generation Total No. Genotype Phenotype

& repeat of offsprings Y-R- Y-rr yyR- yyrr Yellow Yellow Green Green
No. Red White Red white

F1
1.
2.
3.
4.
5.
6.
Total

F2
1.
2.
3.
4.
5.
6.
Total

F2 Generation
(a) Total number of individuals _________________________
(b) Phenotypes _________________________
(c) Number of individuals in each phenotypic class:
Number Phenotype
__________________ _____________________
__________________ _____________________
__________________ _____________________
__________________ _____________________
(d) Phenotypic ratio _____________________
(e) Genotypic ratio _____________________

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(f) Number of individuals of each genotypic class:

Number Genotype
__________________ _____________________
__________________ _____________________
__________________ _____________________
__________________ _____________________
__________________ _____________________
(g) Genotypic Ratio ______________________________________

Discussion
The four phenotypic classes in the F2 generation are in ratio of 9:3:3:1 as
expected from the Law of Independent Assortment. The genotypic ratio
would be (1:2:4:2): (2:1):(2:1):1.
Note
1. In case six repeats of the experimental procedure are not feasible due
to time limitations, either the number of repeats be slashed down to
three or the data from single repeat of six different pair of students may
be pooled together to make the final calculations.
2. This Law of Independent Assortment was later found to be true only for
traits present on two different homologous pair of chromosomes, that
is, the two are not linked together. The linked traits do not assort
independently, rather they are inherited together (linked) except when
crossingover separates them.
3. It is quiet likely that you may not find your data exactly in the
expected ratio, instead almost approximate to it. The statistical
significance of this deviation from the exact expected ratio due to
probality can be checked using chi-square (χ2) test, about which
you will study in higher classes.

Questions
1. Linked traits fail to assort independently. Explain.
2. How is independent assortment of alleles important from the point of view of
variation?

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Exercise 11








Procedure
Select a family in which any one of the monogenic traits such as tongue
rolling, widow's peak, blood groups’, red-green colour blindness, dimple in

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 LABORATORY MANUAL: BIOLOGY

the cheek, hypertrichosis of ear, hitch-hiker's thumb, etc., is found. Ask the
person exhibiting the trait to tell in which of his/her parents, grand parents
(both maternal and paternal), their children and grand children the trait in
question is present. Among surviving individuals the trait may also be
examined. The information made available is the basis for the preparation of
pedigree chart using the appropriate symbols. A careful examination of the
pedigree chart would suggest whether the gene for the character is autosome-
linked dominant or recessive, X - chromosome linked dominant or recessive,
Y- chromosome linked or not.

Explanation
1. Autosome Linked Dominant traits: These are the traits whose
encoding gene is present on any one of the autosomes, and the wild-
type allele is recessive to its mutant allele, i.e., the mutant allele is
dominant.
The pedigree-chart can be of the undernoted pattern (Fig. 11.2), where
the female being interviewed is exhibiting the trait, and is indicated by
an arrow-mark in the chart.

The characteristic features of inheritance of such type of traits are:

(a) Transmission of traits occurs from parents of either sex.
(b) Males and females are equally affected.
(c) The pedigree is vertical, i.e., the trait is marked to be present in each of
the generations.
(d) Multiple generations are characteristically affected.
Brachydactyly, polydactyly, dimple in the cheek are some of the common
traits of this type.

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EXERCISE 11

2. Autosomal Recessive trait: These are the traits whose mutant allele is
recessive to its wild type allele.
The pedigree chart can be more or less of the pattern given below (Fig.
11.3), where the lady (marked by the arrow) is showing the trait. The bar

in the example represents the presence of corresponding dominant or

recessive allele for the specific trait.
Suppose the given trait is albinism. Denote its dominant allele as ‘A’
that produces pigments, and the recessive allele as ‘a’ that fails to synthesise
the pigment, melanin. The female (our subject in generation III) is therefore
of genotype aa. She must have received each of her ‘a’ allele from both the
parents (generation-II), who are therefore themselves normal but are definitely
of genotype Aa, and are carriers of the trait. The allele a must also have been
present in her grand parents too, of course in heterozygous condition also
to make them carriers (generation-I)
Albinism in the subject’s children (generation-IV) suggests her husband
too to be of genotype Aa, a carrier. Marriage of her albino daughter to an
albino man is bound to produce all her grand-children albino (gen-V).
The following are the salient features of the inheritance of such type of traits.
(a) Occur in equal proportions in multiple male and female siblings, whose
parents are normal but carriers;
(b) The siblings are homozygous for the defective allele, but their parents,
though some may appear normal, are obviously heterozygous, i.e.,
are merely carriers of the trait.
(c) Consanguinity (marriage between man and woman genetically related
to each other, such as cousins) occasionally results in the appearance
of such traits.

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 LABORATORY MANUAL: BIOLOGY

3. X-Linked Dominant traits: These are the traits whose encoding gene
is present on the X- chromosome, and the mutant allele of which is
dominant over its wild-type allele.
Such traits are very rare, and are almost difficult to find in the
population. One example is oral-facial-digital syndrome (Duchene
Muscular Dystrophy), which results in absence of teeth, cleft (bifid) tongue
associated with mental retardation. The pedigree chart may appear as
follows (Fig. 11.4):

The possible genotypes of the above pedigree can be written as follows

(Fig 11.5):

Fig. 11.5 Genotypes of individuals shown in Fig. 11.4

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21/04/2018
EXERCISE 11

Here, the dominant mutant allele is denoted by ‘D’, and its recessive wild
type allele is denoted by ‘d’. Remember that human females have two
X-chromosomes (XX), and the males have only one X and one Y chromosome.
Males receive their lone X-chromosome from their mother, and the
Y-chromosomes from their father, whereas females receives one of her
X-chromosome from her mother, and the other X from her father.
The characteristics of such inheritance are:
(a) The trait appears in almost all the generations, and the inheritance is
vertical.
(b) If the female is affected, then about half of her sons are affected.
(c) If the male is affected then all of his daughters would be affected, but
none of his sons are affected.
(d) In short, the pedigree resembles the pattern of inheritance of autosomal
dominants, except that there is no male-to-male transmission.
4. X-linked Recessive traits: These are the traits whose encoding gene is
present on the X-chromosome and its mutant allele is recessive to its
wild-type allele.
Red-green colour blindness and hemophilia, are some of its well known
examples. The characteristic features of such inheritance are:
(a) Females express the trait only when they are homozygous for the
mutant allele, whereas the males do so even when they are hemizygous
for it.
The pedigree chart would appear as the following one (Fig. 11.6):

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 LABORATORY MANUAL: BIOLOGY

(b) About half of the sons of the carrier (heterozygous for the trait) females
are affected. In case of homozygous females showing the trait, fifty
percent of her daughters and all of her sons are likely to be affected.
Therefore, the males are most affected in the population.
(c) Affected persons are related to one another through the maternal side
of their family.
(d) Any evidence of male-to-male transmission of the trait rules out the
X- linked inheritance.
5. Y-chromosome linked traits: These are the traits whose gene is present
on the Y-chromosome. The females do not have any Y-chromosome,
whereas all the males must have a Y-chromosome to be a male, and this
Y-chromosome they get from their father. Therefore, any trait linked to
the Y- chromosome must be present only in males, and certainly not in
any of the females. This is why these traits are also called male-sex limited
traits. All the sons of the affected male would express the trait whereas
none of his daughters would do so.
The pattern of the pedigree chart would be as follows (Fig 11.7):

Hypertrichosis of the ear (presence of hairs on pinna) is one most common

example of such traits.
Note: Students may be asked to prepare the pedigree-chart from given data and analyse the
pattern of inheritance. The work may be done as a project.

Questions
1. How will you differentiate between autosome linked dominant and sex chromosome
linked dominant pedigree chart? Explain.
2. Discuss the differences in the patterns of autosome linked recessive and sex-
chromosome linked pedigree.

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Exercise 12













Procedure
(i) Select a flower in bud condition where antheses has not occurred. Open
the bud carefully and remove the stamens (Fig. 12.1). Mark this as
female parent plant.

Fig. 12.1 Showing process of Emasculation

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(ii) Cover the emasculated flower with a plastic

bag to protect it from undesired pollen
(Bagging) (Fig. 12.2). The bag should be
held securely in place with a paper clip/
string/thread. Select the size of bag in
accordance with the flower size. Bags
must be transparent with minute pores.

Fig. 12.2 Bagging of an emasculated flower

(iii) Bring into physical contact anthers of a

desired male plant containing mature
pollen grains with the stigmatic surface
of emasculated female flower (Fig. 12.3).
Use tweezers/brush if necessary to dust
the stigmatic surface with pollen.

(iv) Cover the pollinated flower again with the

bag immediately. For identification, label
the female parent (Tagging). Each
pollinated flower should bear a label
Fig. 12.3 Showing cross pollination on an containing the name of the seed parent,
emasculated flower the letter X (to signify a cross), the name
of the pollen parent, and the date on which
the cross was effected.

Questions
1. Why is emasculation performed before anthesis?
2. What are the advantages of using a bag containing minute pores?

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Exercise 13








Procedure
(i) Peel off epidermis from the fleshy leaf of onion and put it on a slide.
Add a few drops of water over it to avoid desication.
(ii) Cut out a small piece (about 0.5 cm size) of the epidermal peel and
discard the remaining portion.
(iii) Wipe out the water with a filter paper.
(iv) Put 2 drops of acetocarmine on the epidermal peel and heat gently on
a spirit lamp.
(v) Apply a coverslip over the peel avoiding air bubbles and wrinkles of
the material.
(vi) Wipe out the excess stain with help of blotting paper.
(vii) Examine the material under low magnification of a microscope.

Observation
Record your observations with regard to shape of cell, the number of nuclei
and their position in the cell. Draw a diagram based on your preparation
and label its parts.

Discussion
Nuclei in cells are extremely rich in nucleic acid which exist in a conjugated
form with protein to form nucleoproteinous structures, called chromatin
fibres/chromosomes.

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Questions
1. What are the building blocks of the nucleic acid?
2. What is DNA and how is it different from RNA?
3. Name different nitrogenous bases present in the nucleic acid.

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Exercise 14










Procedure
Observe the preserved specimens/slides/photographs and note down the
features in the practical record book. Take care to observe all the minute
details and draw labelled diagrams of the pathogens.

Observation

A. Entamoeba
Observe the following features of the parasite in the slide or photograph:
(i) It is unicellular.
(ii) Shape of the cell is irregular due to
pseudopodia.
(iii) A single nucleus is present eccentrically in
the cell.
(iv) *In the nucleus a peripheral ring of granule
of nucleoprotein and central karyosome are
observed. Rest of the space in the nucleus
looks empty (Fig. 14.1).
(v) A few food vacuoles may be seen in the
cytoplasm. Contractile vacuoles are absent.
(vi) *Mature quadrinucleated cysts may be
present.
Fig.14.1 An Entamoeba

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Note: Entamoeba is an intestinal parasite in humans and causes amoebic

dysentery. The symptoms of the disease are frequent loose, mucus filled
watery stools, abdominal pain and spasms.

Systematic position
Phylum – Protozoa
Class – Rhizopoda
Type – Entamoeba histolytica

* Distinctive feature of the pathogen

B. Plasmodium vivax
(i) It is an intracellular endoparasite seen easily within the RBC of the
infected person.
(ii) It is unicellular.
(iii) The most diagnostic stage of the parasite is "signet ring" stage in the
erythrocytes, within which it appears as a rounded
body (Fig. 14.2).
(iv) It has a big vacuole inside, and the cytoplasm is accumulated at one
place containing the nucleus. Because of the above mentioned features,
the parasite appears as a ring.
Search the stage in the blood film slide, find the signet-ring stage, and
draw its labeled diagram.
Note: It is a protozoan parasite causing malaria in humans. When an infected
female anopheles mosquito bites a healthy person, it injects the infective
stage, sporozoite, into the peripheral blood vessels. The infective stage
undergoes several rounds of multiplication in liver and erythrocytes.
Symptoms: Intermittent high fever with chills followed by profuse sweating
at an interval of alternate days.

Systematic position
Phylum – Protozoa
Class – Sporozoa
Type – Plasmodium vivax

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EXERCISE 14

C. Ascaris
The external features of round worm are as follows: Mouth
(i) Body long (20 to 40 cm), cylindrical (5 to 6 mm
diameter) with no segmentation (Fig. 14.3).
(ii) Sexes are separate; the females are longer than
the males.
(iii) Both the ends are pointed; posterior end of male
is ventrally curved.
(iv) Mouth is situated at the anterior end, and is Female genital
surrounded by three lips, one present mid- aperture
dorsally and rest two lips are situated
ventrolaterally (for viewing these lips a magnifying
lens is needed).
Penial spicule
(v) Single longitudinal lines are present on the dorsal,
ventral and on the two lateral sides, all along the
length of the body. Out of these the lateral lines
are comparatively more distinct than the others
lines. (b)
(vi) Excretory pore is present on the ventral surface
slightly behind the anterior end.
(vii) In addition to the ventrally curved posterior tip, (a)
the male worm has a pair of penial spicules very
Fig.14.3 Ascaris (a) Female (b) Male
close to the cloacal opening.
(viii) In case of female specimen a female genital
aperture is present mid-ventrally about one third
distance from the anterior end.

Systematic position
Phylum – Aschelminthes
Class – Nematoda
Type – Ascaris lumbricoides
Note: Round worm or Ascaris is one of the common parasite found in the
intestine of human beings.
Symptoms: (a) Irregular bowel, (b) Occasional vomiting, (c) Anaemia

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Trichophyton (Ringworm fungus)

It is a fungus that feeds on keratin of the skin of human beings. The features
as observed under the microscope are:
1. Texture of hyphae is waxy, glabrous to cotton like.
2. Unstained hyphae are white, yellowish brown to reddish brown in colour.

Systematic position
Kingdom – Fungi
Class – Deuteromycetes
Type – Trichophyton rubrum

Symptoms
Ringworm is a contagious fungal infection of the skin. Infected area of skin
is itchy, red, raised, scaly patches (with sharply defined edges). It is more
red on the periphery than in the center creating a ring like appearance.

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21/04/2018
Exercise 15







     







    



   



Fig.15.1 Soil Textural Triangle

Procedure
Three methods are suggested here. Any one of these may be followed.
Method I
(i) Collect about 300–500g of soil from two different locations. Label them as sample A and B.
(ii) Dry the samples in an oven, or stove or in sun to remove the soil moisture (capillary and
bound water).
(iii) Select the 3 sieves of different mesh sizes (2mm, 0.05mm and 0.002mm). Arrange them in
a collecting chamber as shown in Fig. 15.2.
(iv) Place 200g of the soil in the Ist sieve (sieve of 2mm mesh) and close the lid. To sieve the soil,
shake the set manually for 5–10 minutes and collect the three soil fractions.

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(v) Weigh the soil fractions viz-sand, silt and clay

collected in the 3 compartments
Lid
Wt of soil sample taken – .........g
Wt of sand fraction – .........g
Wt of silt fraction – .........g
Wt of clay fraction – .........g
2mm mesh The weight of three fractions should be equal to the
total weight of sample taken for analysis.

0.05mm mesh
Observations
Calculate the percentages of the various soil fractions
0.002mm mesh
and tabulate:
Calculate the percentages of sand, silt and clay
fractions.
Collecting chamber
Use the textural triangle now. Note that the three
sides of the textural triangle represent 0 to 100% of sand,
silt and clay respectively. Note that (i) the percentage
Fig.15.2 Sieve set
lines for clay run paralled to the base line of sand, (ii)
the precentage lines of silt run parallel to the clay side of the triangle and, (iii)
perentage lines of sand run parallel to the silt silde of the triangle. In reading
the textural triangle, any two particle fractions will locate the textural class at
the point where these two intersect.

Soil sample Percentage (%) Texture class

Sand Silt Clay

Note for Teachers: The sieve sets contain a number and an abbreviation BSS/ASTM/
ISS on each sieve. In the given table (Table No. 15.1) the corresponding aperture size of
the sieves is listed. For example, BSS 30 sieve aperture size will be 500 microns.

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EXERCISE 15

Appendix 1

Table 15.1 Mesh No. and the corresponding Aperture size

Sl. No. BSS Mesh ASTM Mesh ISS Mesh Aperture

1 4 5 480 4.75 mm
2 5 6 340 3.35 mm
3 6 7 280 2.80 mm
4 7 8 240 2.36 mm
5 8 10 200 2.00 mm
6 10 12 170 1.70 mm
7 12 14 140 1.40 mm
8 14 16 120 1.18 mm
9 16 18 100 1.00 mm
10 18 20 85 850 micron
11 22 25 70 710 micron
12 25 30 60 600 micron
13 30 35 50 500 micron
14 36 40 40 425 micron
15 44 45 35 355 micron
16 52 50 30 300 micron
17 60 60 25 250 micron
18 72 70 20 212 micron
19 85 80 18 180 micron
20 100 100 15 150 micron
21 120 120 12 125 micron
22 150 140 10 106 micron
23 170 170 9 90 micron
24 200 200 8 75 micron
25 240 230 6 63 micron
26 300 270 5 53 micron
27 350 325 4 45 micron
28 400 380 3 38 micron
29 25 micron

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Method II
Texture by Feel
The texture of the soil sample can also be estimated by feeling it in the dry,
moist and wet states. Sand is coarse and gritty, silt feels smooth like flour
and clay is sticky and plastic. The smallest soil particles that one can see are
coarse silt. Feel the known texture samples first, then feel the unknown ones
and decide their textures.

Procedure
(i) Feel the dry soil first. Does it crumble easily or is it hard to break?
Hard soil samples contain a moderate amount of clay.
(ii) Take in your palm a lump of soil sample about the size of a one-rupee
coin and wet it to the consistency of modeling clay. Try to press it into
a ribbon between the thumb and forefinger. An alternate test is to form
a wire by rolling the wet soil until it is about 1/8" in diameter.
(iii) If a long wire or ribbon can be formed readily the soil is plastic and
probably contains over 40% of clay. Its texture must therefore be clay/
silty clay/or sandy clay. If a ribbon or wire can be formed easily but
also breaks easily, the soil sample is probably a clay loam/silty clay
loam/or sandy clay loam. A heavy loam/silt loam/or sandy loam
sample may form ribbon or a wire if the moisture content is just right
but these will be still weaker than the ribbons and wires formed by the
clay loam samples.
(iv) Next determine whether sand or silt is dominant. If there is a gritty feel
without the smooth floury touch of silt, choose a texture-name that
includes the word ‘sandy’. If the smooth floury feel predominates and
there is not much gritty feel, choose one of the ‘silty’ texture names.
Use the name without a prefix if neither smoothness nor grittiness
predominates (simply clay or sand or silt). Often this can best be
determined by adding more water until the soil is in a wet state.
If the soil is very sandy, you must choose between sandy loam, loamy
sand and sand. In the moist state, sandy loam samples will have some
tendency to stick together but loam sand and sand samples will not do so.
Use the wet state to determine whether a sample is sand or loamy sand.
After handling wet sand, your hands will be moist but clean loamy sand will
make the hands slightly soiled.

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EXERCISE 15

Method III
Requirements: Soil samples, balance, weights, glass rod, standard sieves of
2mm and 0.5mm mesh size, blotting sheets/old newspapers, evaporating
dish and water

Procedure
(i) Collect 200-300g of soil samples from different sites, and dry them as
suggested previously to remove the moisture.
(ii) Sieve the sample through a 2mm sieve to remove stones, pebbles,
roots etc.
(iii) Take 100-150 g of the sieved soil sample and further sieve it through a
0.05 mm sieve to separate the sand fraction (collected in the sieve) from
silt and clay (collected on a blotting sheet). Weigh the amount of sand
fraction and silt + clay fraction.
(iv) Take a large evaporating dish (a shallow clay plate, glass trough or a
shallow iron plate) and record its weight.
(v) Add the clay and silt fraction to the dish and note the weight.
(vi) Add water to the dish leaving half an inch space empty at the top and
stir the liquid thoroughly with a glass rod taking care that the contents
do not spill out. Allow it to stand for several hours. Decant off the cloudy
supernatant liquid (clay fraction). Repeat the process three to four times
until the decanted liquid is quite clear.
(vii) Dry the silt left in the evaporating dish to dryness. Cool the dish and
weigh it.

Observation
Record your observation in the following table:

A B

Weight of the soil sample taken

Weight of sand fraction

Weight of silt & clay fraction

Weight of silt fraction

Subtract the weight of silt fraction from the weight of silt + clay fraction. The
difference will be the weight of clay decanted.
Calculate the % of sand, silt and clay fraction of the soil and express the texture.

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Discussion
Correlate the texture with the plants growing in the area from which the soil
sample has been collected. Discuss how the texture of soil can affect the root
penetration, tillage, soil aeration, moisture content, water holding capacity
and other aspects related to plant growth. In sandy soil the non-capillary
pore spaces will be more and the capillary pore spaces will be less. The
condition will be reverse in case of clay soil. The pore space in turn determines
water holding capacity, percolation rate, aeration, root penetration and soil
flora and fauna. Clay particles are anionic colloides and adsorb mineral
nutrients and minimise their leaching.

Questions
1. Which type of soil is better for root-penetration and better aeration?
2. Among sandy and clay soil which one has higher water holding capacity? Explain.
3. If the clay content is high, will it affect soil fertility? Explain.
4. Which type soil has poor nutrient status and high leaching?
5. What kind of plants grow in smooth texture soil? Name two plants that grow in
heavy-textured soil.

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Exercise 16











Procedure
(i) Dig a small pit about 10cm x 10cm x 10cm, Scoop 100–300 g of soil
from the pit and collect it in a small polythene bag.
(ii) Remove the pebbles and large lumps from the soil sample.
(iii) Pass the soil through a coarse sieve to remove small lumps and dead
decaying leaves and twigs.
(iv) Spread the soil into a thin layer on a sheet of blotting paper or old
newspaper and sun dry it for 2–3 hours or dry it in a pan kept on
stove. Alternatively dry the soil sample in oven at 1080C for 1 hour.
(v) With the help of pestle and mortar grind the sample into fine powder.
(vi) Put a small disc of blotting paper at the base of the Gooch crucible.
Weigh the crucible along with the blotting paper and note its weight.
(vii) Transfer the soil sample into the crucible. Tap the rim of the crucible
gently several times with the help of glass rod so that soil is compactly
filled and forms a uniform layer at the top. Add more soil if necessary.
(viii) Weigh the crucible along with soil sample and note its weight.
(ix) Fill the petridish with water and place two small glass rods in it parallel
to and at a small distance from each other.
(x) Place the crucible on the two glass rods in such a manner that its
bottom is in contact with water.
(xi) Leave the set up undisturbed till water appears at the upper surface of
the soil. Wait till entire soil surface is wet.

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 LABORATORY MANUAL: BIOLOGY

(xii) Remove the crucible and allow all the gravitational water to flow out
from the bottom. When no more water percolates, wipe the bottom dry
with the blotting paper.
(xiii) Weigh the crucible and note its weight.

Observation
Record your observation in the following table.
Calculate the % water holding capacity of the soil as follows.
Weight of crucible + blotting paper: Ag
Weight of crucible + blotting paper
+ soil sample before experiment: Bg
Weight of dry soil: B - A= Cg
Weight of crucible + blotting paper
+ wet soil sample after experiment: Dg
Weight of wet soil after the experiment: D - A= Eg
Mass of water absorbed by soil: E - C= Ng
N
% Water holding capacity: x100
C
Tabulate your results as shown below
Sample No. Wt. of Crucible Wt. of Crucible Wt. of soil Wt. of crucible Wt. of wet Amount of % water
+ blotting + blotting paper sample + blotting paper soil (D-A) water holding
paper (A) + soil sample (B) (B-A) = (C) + wet soil (D) = E absorbed capacity
(E-C) = N N
× 100
C

A Garden
soil

B Road
side soil

C……...

D……...

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EXERCISE 16

Discussion
Compare % water holding capacity of soil collected from different habitat
conditions. The variation in water holding capacity is due to varying
proportion of sand, silt and clay in the soil of different habitats. Soil with
very high proportion of sand have very low water holding capacity due to
large pore spaces between the particles which enables the water to percolate
freely into deeper layers leaving upper layers practically dry. In clay soil,
due to very small size of the pore spaces (fine capillaries) the water is retained
in the capillary spaces as capillary water. In these soil the water does not
percolate freely. Soil with more or less equal proportion of sand, silt and clay
(loam soil) combines the properties of sand and clay and therefore has
optimum water holding capacity and optimum soil-air for root growth.

Questions
1. What are heavy soil and light soil?
2. Give examples of a plant seen in heavy soil and light soil.
3. How does pore space determine the % water holding capacity of soil?
4. Why is clay soil often referred to as physiologically dry soil?
5. Which type of soil is suitable for cultivation of crop plants?
6. How can water-holding capacity of soil be improved?
7. Dead decomposed organic matter is usually added in the fields before the
cultivation of crops. Apart from providing the mineral nutrients, what additional
role does organic matter play in the cultivation of crop plants?

61

21/04/2018
Exercise 17













 


 

 










Procedure
Prepare temporary stained transverse sections of leaf, stem and root of the
specimens. Study the morphological and anatomical features of the plants

21/04/2018
EXERCISE 17

collected and look for the following adaptations. Write the name of the plant
in which a particular adaptation is observed.

Observations
Record your observation in the given tables:

Xerophytes

Adaptations Modifications Examples (from the

(Morphological/Anatomical) specimen collected)

1. Conservation of Water a. Leaves few or absent or represented ------

by spines only
b. Petiole modified into leaf like
structure
c. Stem reduced, branching sparse
d. In some cases stem flattened, leaf
like, green, photosynthetic in
nature

2. Storage of Water Thick, fleshy and succulent --------

leaves as well as stem
3. Prevention of loss of a. Intercellular spaces reduced ---------
water by transpiration b. Spongy parenchyma/ palisade
parenchyma present
c. Stomata on lower surface,
sunken in stomatal pits
d. Leaves needle like
e. Thick cuticle on leaf surface

4. Prevention of excessive a. Leaves covered with dense hairs; -------------

heat b. Leaf surfaces shiny or glaborous
c. Leaf blade remains rolled during
the day

5. Efficient mechanism of a. Long and profusely branched roots -----------------

water absorption b. Dense root hairs
c. Well developed xylem

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 LABORATORY MANUAL: BIOLOGY

Hydrophytes

Adaptations Modifications Examples

(Morphological/Anatomical)

1. Buoyancy and resistance a. Leaves long and cylindrical

to currents of water b. Petioles flexible to withstand
currents of water and to carry the
leaf blade on the surface of water
c. Petioles are modified into air
pockets
d. Leaf blade pale green in colour,
finely dissected
e. Leaf blade waxy with thin cuticle

2. Transpiration a. Stomata absent

b. Stomata present on upper surface
of leaves
3. Absorption of water
a. Poorly developed roots
b. Root hairs absent
c. Roots with air pocket to help in
buoyancy
4. Gaseous circulation and
Parenchymatous tissue of stem, roots,
storage of air
petioles and leaves modified into
aeranchyma in the form of air channels
in
a. Root
b. Stem
c. Petiole
d. Leaf
5. Mechanical tissues
a. Poorly developed xylem
b. Poorly developed sclerenchyma
c. Sclerides present

Questions
1. Give three adaptive features of water hyacinth suitable to aquatic life.
2. What are the features present in plants of xeric habitat for the prevention of
loss of water?
3. What is the importance of succulent leaves and stem for a xerophytic plant?
4. Why is air stored between tissues in aquatic plants?

64

21/04/2018
Exercise 18












Procedure
Observe the animals provided and note down their adaptive features in the
observation table with example.

Observations
Hydric adaptations
Features Adaptations Example
(For students)

Body colour (a) On dorsal surface

(b) On ventral surface

Body contour (a) Streamlined

(b) Disappearance of neck constriction
(c) Tail enlargement
(d) Position of external nostrils
(e) Loss of external ears
(f) Position of eyes
(g) Presence of eye protecting membrane

Locomotory (a) Fins or fin-like expansions of the body wall

(b) Loss of limbs
(c) Webbed feet

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 LABORATORY MANUAL: BIOLOGY

Presence of dermal/epidermal derivatives

(a) Scales
Integument (b) Hairs
(c) Mucous glands
(d) Oil glands

(a) Position
Mouth (b) Presence of teeth
a. Upper jaw
b. Lower jaw

(a) Gills/lungs
Respiratory organs
(b) Cutaneous

Xeric adaptations
Features Adaptations

Moisture getting (a) Preference for juices as food

(b) Hygroscopic skin

Moisture Conservations (a) Storage of water in body

(b) Avoidance of evaporation (non-perspiring)

Body colour (a) Protective mimicry

(b) Predating mimicry

Body contour (a) Position of

a. Nostrils directly upward
b. Reduction to pin-head size
(b) Position of eyes
a. Covering of eyes
b. Size

Skin (a) Hard

(b) Spiny
(c) Poison glands

Limbs (a) Speed

(b) Long slender
(c) Padded feet

Scrotum Present or Absent

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EXERCISE 18

Discussion
You may have noticed many features in the body of aquatic animals
which support their life. As the different aquatic bodies vary to a
great extent, there are many other adaptive features you may notice.
For example the aquatic organism in ponds, lakes, river and sea.

Questions
1. Name the features that helps a frog for aquatic life.
2. What are the adaptations present in xeric animals for conservation of water?

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Exercise 19














Procedure
(i) Weigh 10 g of the soil sample A. Add 50 mL of distilled water to soil
sample to make a soil solution.
(ii) Filter the soil solution through a filter paper and collect the filtrate in a
beaker. Label it as soil solution -A.
(iii) Take a clean dry porcelain cavity tile. Place 5 drops of soil solution A
in three cavities of the tile as shown in Fig. 19.1.
Soil - A Soil - B Soil - C

Universal pH indicator
solution

pH indicator paper
(Broad range)

pH indicator paper
(Narrow range)

Fig. 19.1 Porcelain cavity tile

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EXERCISE 19

(iv) To the 5 drops of soil solution present in one cavity add 5 drops of
universal pH indicator solution. Note the colour developed and compare
it with the colour chart given on the universal pH indicator
solution bottle.
(v) To the soil solution present in the second cavity, dip a small strip of broad
range pH indicator paper (pH 2-11). Note the colour and compare with
the colour chart given on the broad range indicator paper and get a rough
estimate of pH of the sample solution.
(vi) Choose a suitable narrow range pH indicator paper (for e.g. If the pH of
soil is determined by you as 8.0, choose a narrow range 7.0 to 9.0) and
dip a small strip of it in the soil solution present in the third cavity. Note
the colour developed and determine the pH to the nearest possible value
with the help of the colour chart.
Repeat the same steps for determining the pH of sample B and C. Follow the
same procedure for water samples collected from different sites.

Observation
Record your observations in the given table.
Table: Measurement of pH of soil samples A, B and C

pH value as determined by Soil Samples

A B C

Universal indicator solution

Broad range indicator paper

Narrow range indicator paper

Discussion
Based on the pH values obtained, categorise the samples into acidic, basic,
neutral type.
Record the plant species present in the site from which the samples are collected.

Note for teachers: The colour developed should be noted against direct sun
light. Also, sometimes the soil solution colour may interfere with the readings.
Thus one has to be careful while making the observations.

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Questions
1. What will be the pH of chalk (calcareous) soil?
2. pH measurement with indicator paper is not very accurate. Comment.
3. Water logged soils are acidic. Comment.
4. Why are soil around mineral mining areas acidic?

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Exercise 20








 

 


 







Procedure
(i) Visit a nearby pond.
(ii) Reach to the center of the pond in a small boat.
(iii) Slowly immerse the Secchi disc into water vertically holding
the rope tightly in the hand till the black and white segments
of the disc just begin to disappear. On reaching to a
particular depth, the disc becomes completely invisible.
Mark the length of the rope when the disc just disappears
(say A cm).
Fig. 20.1 A Secchi’s Disc

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 LABORATORY MANUAL: BIOLOGY

(iv) Slowly pull up the disc and find out the length of the rope where the
black and white segments of the disc just reappear (say B cm).
(v) Find out the mean length (X) of the rope by the following method.
A+B
X= cm
2
(vi) Repeat the process at different sites of the pond.

Tabulate the results in the given table

Water body Depth at which Disc Depth at which Depth of Photic

A+B
disappears (A cm) disc reappears (B cm) zone X=
2

Pond Site 1
Site 2
Site 3

Observations
The value X represents the depth of the photic zone upto which sunlight
penetrates in the water body and photosynthesis takes place.

Discussion
Greater the value of 'X' less turbid is the water. In crystal clear deep lakes, the
value of 'X' will be very high indicating, thereby, that the water body does not
have large quantities of flocculating silt or organic matter residues. This may
be due to no discharge of effluents or domestic sewage into the water body.
The high clarity of water is also an indication of very less density of phyto and
zooplanktons. These water bodies are called as non-productive or oligotrophic,
while highly turbid water bodies are eutrophic in nature.

Precautions
Students are advised to perform this experiment under the strict supervision
of teacher to prevent incidents due to drowning.

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EXERCISE 20

II. Measurement of turbidity using measuring cylinder

Requirements: Water samples from different sources, three measuring
cylinders (500mL) of the same height.

Procedure
(i) Collect about 2 liters each water samples from different sources.
(ii) Transfer 500ml of water sample in the measuring cylinders of same
volume and height.
(iii) Mark the three cylinders A, B and C and leave them undisturbed
overnight.

Observations
Observe the amount of sediment settled at the bottom of each cylinder and
also note whether the water above the sediment is still turbid.

Record your observations in the following table:

Water sample Thickness of sediment Clarity of water—turbid/

semiturbid/clear

'A'

'B'

'C'

Discussion
 Do all the samples show same amount of sediments?
 Which sample shows maximum sedimentation and correlate it with the
source of the sample?
 Find out whether in all the cylinders, water above the sediment is clear or
turbid. Explain with reasons.
 Draw conclusions on the basis of the observations.

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Questions
1. Is turbid water fit for drinking? Explain.
2. Why is the penetration of sunlight in any water body important?
3. Green plants are seen only in photic zone. Comment.
4. It is a common practice to use alum for clearing turbid waters. Explain.
5. Turbidity of water body varies with season. Comment.

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Exercise 21


















Procedure
(i) Collect about a liter of water sample from nearby water body (pond lake,
reservoir, river etc).
(ii) Add about 5 ml of FAA to fix and preserve the living organisms present in
each sample at the place of collection.
(iii) In the laboratory, transfer the water sample into a measuring cylinder of
one litre capacity. Label each water sample to indicate the site from which
the water sample has been collected.
(iv) Leave the water samples undisturbed for 48-72 hours.
(v) Decant off the clear water, leaving concentrated sediment at the bottom.
(vi) Transfer the sediment into a vial or a small test tube. Cork and label each
vial for future use.
(vii) With the help of a dropper, transfer a few drops of sediment liquid from a
vial into a watch glass. Dilute the sediment with water if the sediment is
highly concentrated.

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 LABORATORY MANUAL: BIOLOGY

(viii) With the help of a dropper transfer a drop of water from the watch glass
on the center of a slide and mount it. Blot the excess water using blotting
paper.
(ix) Prepare a few more slides of each water sample in the same way.
(x) Observe each slide, first under lower magnification and then under higher
magnification.

Observations
1. Record the different types of organisms present.
2. Count the number of organisms under each field of microscope.
3. Some of the commonly found organisms of water bodies are given in
Annexure 2.

Discussion
Prepare a list of organisms observed in each water sample and make an
assessment of type and density of different organisms in each water sample.
Polluted waters may contain very few types of organisms but in very high
density. The non-polluted waters will have large variety of organisms in
low density.

Questions
1. Why do you find few organisms in polluted water? Explain.
2. Why is FAA (Formaline Aceto Alcohol) added after collecting the water sample?
3. Name at least one phytoplankton and zooplankton commonly found in polluted
water.

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EXERCISE 21

Annexure 2

Cosmarium

Desmickum

Stigeoelomium

Spirogyra

Draparnaldiopsis

Nitella

Dinobryon Euglena Moubeotia

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 LABORATORY MANUAL: BIOLOGY

Ceratium
Gymnodinum Peridinium

Calothrix

Rivularia

Cyanomonas Cylindespermum

Anabaena Gleotrichia

Scytonema Fischerella

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EXERCISE 21

Chlamydomonas Volvox

Coelastrum

Closterium
Golenkinia

Pedrastrum
Hydrodictyon Chlorella

Scenedesmus Staurastrum
Ankistrodesmus

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 LABORATORY MANUAL: BIOLOGY

Synechococcus
Gloeocapsa
Microcystis

Phormidium Oscillatoria
Synechocystis

Spirulina

Lyngbya Nostoc

Melosira
Chaetoceros

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EXERCISE 21

Rhizoselina

Amphora

Biddulphia
Bacillaria

Synedra

Centronella
Nitzschia

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 LABORATORY MANUAL: BIOLOGY

Navicula

Somphonema
Fragilaria
Pleurosigma

Coscinodiseus
Skeletonema

Lauderia

Cocconeis
Asterionella

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Exercise 22














Procedure
This experiment is an outdoor activity and may be
conducted by assigning 2–3 students into a group.
(i) Collect a few locally available broad leaves from a
nearby tree plant (Canna, Peepal, etc.).
(ii) Wash the leaves gently in running water to remove
any dust settled on their surfaces.
(iii) Blot dry the surface area of the leaves. To calculate
the area of the leaf, trace the outline of the leaf on
graph paper (Fig 22.1). Within the traced area
calculate the total number of full squares, 1/2, 1/3
and 2/3 squares and individual small squares. Add
all the squares to get the total leaf area. Multiply
their value with two to obtain total area of both the
surfaces.
(iv) Take 8–10 feet long twine thread and tie five leaves
leaving a foot distance in between. Apply an
extremely thin layer of vaseline on both surfaces of Fig. 22.1 Calculating the area of a
each leaf. Make a bundle of these leaves and pack leaf on a graph paper

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 LABORATORY MANUAL: BIOLOGY

them in polythene bags. Ensure that the outer surface of polythene

bag does not have any vaseline sticking on it.
(v) Make three such bundles of smeared leaves, each bundle containing 5
leaves.
(vi) Mark bundles as A, B and C and carefully weigh each bundle of leaves
along with the polythene bags.
(vii) Select three spots (X, Y and Z) near by your school. Spots selected
should be in a manner that spot 'X' has very heavy vehicular traffic,
the spot Y has moderate traffic and spot 'Z' has little or no vehicular
traffic. At spot 'X' expose each leaf of bundle 'A' by stretching the
attached thread and tie the two ends to two poles or branches of trees
preferably at 10 feet height above ground. Keep leaves exposed for
about two hours.
(viii) After exposure at spot 'X', collect the leaves and carefully re-bundle
exposed leaves and place them along with the string in the polythene
cover 'A'.

Record your findings in the following table:

Site Leaf bundle Weight of leaves (g) Weight of Total leaf area
sample suspended (cm2) of five
particle (W2- W1) leaves
Before After
exposure (W1) exposure (W2)

X 'A'

Y 'B'

Z 'C'

(ix) Repeat the same process at spot 'Y' and 'Z' exposing leaves of 'B' and
'C' bundles respectively.
(x) At the end of the experiment, return back to the laboratory. Reweigh
each bundle of exposed leaves along with their respective polythene
cover.
- Calculate the amount of suspended particles deposited in mg cm2 of
leaf at each spot.
- Compare the results of three different spots and interpret.
Since the weight of suspended particles will be in milligrams or even less
it is advised to use a very sensitive laboratory balance.

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Exercise 23
















Procedure
(i) In the selected site of study, make a 1 m X 1 m quadrat with the help of
nails and thread. Hammer the nails firmly and make sure that the
vegetation is not damaged while laying the quadrat.
(ii) List the names of the plant species seen in the quadrat (if the name is
not known mark these as species A or B etc., and the same species if
seen in other quadrats assign the same alphabet).
(iii) Count the number of individuals of each species present in the quadrat
and record the data as shown in the table.
(iv) Similarly make nine more quadrats randomly in the site of study and
record the names and number of individuals of each species.

Observations
Record the total number of species seen in the ten quadrats. This will give
an idea about the composition of the vegetation.
There will be difference in the species composition in the quadrats made in
shady areas, exposed areas with bright sunlight, dry or wet areas etc.

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 LABORATORY MANUAL: BIOLOGY

Table 23.1: Density studies of the given vegetation

Plant Quadrats employed in study & no. of Total No. of Total no. Density (D)
Species individuals in each quadrat individuals (S) of Quadrats
studied (Q)
I II III IV V VI VII VIII IX X

A 2 5 7 10 3 27 10 27/10 = 2.7

Z 1 2 4 8 3 2 20 10 20/10 = 2.0

Discussion
Plants growing together exhibit mutual relationships among themselves and
also with the environment. Such a group of plants in an area represent a
community. The number of individuals of a species varies from place to
place, making it necessary to take many random sample areas for reliable
results. Density values are significant because they show relative importance
of each species. With increasing density the competition stress increases
and the same is reflected in poor growth and lower reproductive capacity of
the species. Data on population density are often very essential in measuring
the effects of reseeding, burning, spraying and successional changes.
Discuss the vegetation composition of the area (herbs/shrubs) and
comment on the dominant component species.

Questions
1. What factors influence the population density?
2. What is the significance of quadrat method?
3. What conclusion can be drawn if density of a plant species is low?

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Exercise 24








 

 



Procedure
(i) In the selected site of study, make a 1 m X 1 m quadrat with the help of
nails and thread. Hammer the nails firmly and make sure that the
vegetation is not damaged while laying the quadrat.
(ii) List the names of the plant species seen in the quadrat (if the name is not
known mark these as species A or B etc. and if the same species is seen in
other quadrats assign the same alphabet)
(iii) Similarly lay nine more quadrats randomly in the site of study and record
the names of individuals of each species.
(iv) Calculate the percentage frequency of occurrence using the formula given.

Observations
Record the total number of species seen in the ten quadrats. This will give an
idea about the composition of the vegetation.
There will be difference in the species composition in the quadrats made in
shady areas, exposed areas with bright sunlight, dry or wet areas etc.
Observe that the frequency of occurrence is not the same for all species.

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Table 24.1: Frequency studies for the given vegetation

Plant Number of quadrats employed No. of quadrats Percentage

Species in the study (Q) in which the of frequency
species is present (N) F=N/Q X 100
I II III IV V VI VII VIII IX X

A √ √ √ √ √ 5 5/10 × 100 = 50%

B √ 1 1/10 × 100 = 10%

C √ √ √ √ 4 4/10 × 100 = 40%

Discussion
Variation in distribution of a species is caused by factors like soil conditions,
quantity and dispersal of gemmules, vegetative propagation, grazing, predation,
diseases and other biotic activities. Also frequency values differ in different
communities. They are influenced by micro-habitat conditions, topography,
soil and many other environmental characteristics. Thus unless frequency is
not correlated with other characters such as density, frequency alone does not
give correct idea of the distribution of a species.
Frequency determinations by means of sample areas are often needed in
order to check general impressions about the relative values of species. Many
species having low cover or population density also rate low in frequency, but
some may have high frequency because of their uniform distribution. Usually
if the cover and population density are high, the frequency will be high. The
plants with high frequency are wide in distribution.

Questions
1. If frequency of a plant is high, what will be your interpretation?
2. Can many micro-habitat in an area affect frequency of a species? Comment.

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Exercise 25


















Observations

1. Homologous Organs in Plants

(i) Tendrils of passion flower and thorns of pomegranate

Tendrils of passion
fruit and thorns of
pomegranate are Tendril
structurally and
functionally different
but they have similar
origin i.e. they arise
from axillary bud Thorn
(Fig. 25.1a & b).
(a) (b)

Fig. 25.1 (a) Tendrils of passion fruit (b) Thorns of pomegranate

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 LABORATORY MANUAL: BIOLOGY

Tendril Thorns (ii) Tendrils of Vitis and thorns of

Carissa
Tendrils of Vitis and thorns of
Carissa originate from the terminal
bud, but they are functionally
different (Fig. 25.2 a & b).

(a) (b)
Fig. 25.2 (a) Tendrils of Vitis (b) Thorns of Carissa
(iii) Tendrils of baloon vine
Tendril (Cardiospermum) and bulbils of
Agave.
Both are modifications of floral bud,
but they perform different functions.
Tendrils help in climbing but bulbils
are meant for reproduction
(Fig. 25.3 a & b).
(a) (b)
Fig. 25.3 (a) Tendrils of baloon vine (b) Bulbils of Agave
(iv) Scale leaves of onion and spines
of prickly pear (Opuntia)
Both the scale leaves and spines are
Spines modifications of leaves but are
structurally and functionally
different. Scale leaves of onion are
thick and fleshy and store food. On
(a) (b)
the other hand spines of cactus are
Fig. 25.4 (a) Scale leaves of onion (b) Spines of cactus defensive organs (Fig. 25.4 a & b).

2. Analogous Organs in Plants

Tendril
(i) Stem tendrils and leaf tendrils
All tendrils are analogous with one
another, being structurally and
functionally similar, irrespective of
their origin.
Example: Tendrils of pea and
tendrils of Vitis. Tendrils of pea are
Tendril modification of leaf and in Vitis it is
(a) (b) the modification of terminal bud
Fig. 25.5 (a) Tendrils of pea (b) Tendrils of Vitis (Fig. 25.5 a & b).

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21/04/2018
EXERCISE 25

(ii) Thorns and spines

Thorns and spines are analogous structures being
defensive in function. Thorns are modifications of
axillary or terminal buds, and spines are (a) (b)
modifications of leaves. Fig. 25.6 (a) Modified root of carrot
e.g: Thorns of pomegranate and spines of (b) Rhizome of ginger
prickly pear.
(iii) Modified underground stems and modified roots Spine
Modified stems (rhizome, corm, tuber) are
analogous to modified roots (carrot, radish) as they
perform similar function of storage of food but their
origin is different. Rhizome of ginger, potato tuber,
(a) (b)
Colocasia are stems and beetroot, radish etc. are
Fig. 25.7 (a) Phylloclade (b) Cladode of
roots. (Fig. 25.6 a & b)
ruscus
(iv) Phylloclade, cladode and leaves
They perform the same function i.e. they
photosynthesise but phylloclade and cladode are
modifications of stem. Phylloclade of Opuntia,
Parkinsonia, Asparagus and leaves of any local
plant like mango are analogous organs.
(Fig. 25.7 a & b) (a) (b)
3. Homologous Organs in Animals Fig. 25.8 Fore limb of (a) human (b) bat

(i) Wings of birds, and forelimb of mammals/reptiles/

frog: All have the same bony elements (humerus radio-
ulna, carpals, metacarpals and phalanges), but
perform different (flying in birds, for holding or walking
etc. in other) functions. (Fig. 25.8 a & b) (a) (b)
4. Analogous Organs in Animals Fig. 25.9 Wing of (a) dragonfly (b) bird
(i) Wings of dragonfly/cockroach/butterfly and of
birds. (Fig. 25.9 a & b)
(ii) Mandible of cockroach and mandible (lower jaw) of
a vertebrate. (Fig. 25.10 c & d)
(a) (b)
Note: Students and teachers are suggested to discuss
Fig. 25.10 Mandible of (a) cockroach
more examples.
(b) rabbit

Questions
1. Suggest examples of homologous and analogous organs other than what are
given in the manual.
2. Why are stem and leaf tendrils considered as analogous organs?

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Investigatory Project Work



















Choosing an Investigatory Project

You may be guided by your teacher for your choice of topic. The more original
or new the project is, the better it would be. But it must be realistic in terms of
the time available and at a level attained in the higher secondary biology.
You must review the available literature to find out what type of work has
been done. This will help you to reject some of the alternatives, and possibly
cause you to modify others. It may also be the source of new ideas.
By doing these investigatory projects you will gain experience of research
besides providing opportunity for learning skills such as photography,
electronics, etc.

Identifying the Objectives of the Project

Having identified a possible project, you should be able to identify and list the
tentative objectives you hope to attain by completing that investigation. For example,

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¨ Suppose your project involves studying the biodiversity of birds in your

district/state, examine the data in the light of some questions (say, how do
the birds in Rajasthan differ from those in Assam or Bihar?) your
investigation might attempt to answer.
¨ Suppose your project involves investigating leaf mosaics revealing the
complexity of the growth correlations that lead to efficient light interception,
suggest also the factors that might affect this type of study.
Keep the aim of your project simple. Investigate only one factor at a time
and never allow yourself to be side-tracked. Remember that time may be too
short for follow-up and any fascinating secondary aspects that you may come
across.

Designing Projects
Having established the objectives of your chosen project, you must have an
experimental design. This will allow you to collect the data you need in a
scientific way to test the hypothesis. For example, if your project involves
investigating the hypothesis that stale milk contains more bacteria than fresh
milk, devise the procedure you would adopt to carry out your investigation.

Planning Investigations
Having decided your topic for scientific investigation, you should give careful
thought to the plan of your investigation in some detail. These may include
¨ What hypothesis can you make?
¨ How can you ensure that the experimental tests and measurement you
carry out are accurate and reliable?
¨ What controls do you need?
• How many variables are you investigating? Correctly identify key variables
as independent and dependent.
¨ Are your variables discrete or continuous?
¨ Identify appropriate control variable for fair test.
¨ How many repeat observations or samples will you require?
¨ What instruments/equipment or techniques will you use to obtain relevant
information? Identify suitable materials and equipment to be used.
¨ If your investigation requires the use of a questionnaire, design and
standardise before implementation.
¨ Is your intended procedure safe and ethically permitted, i.e., taking care of
the distress or suffering of living organisms and damage to the environment?
¨ How will you collect your data?

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¨ How do you plan to analyse your results? Would you employ statistical or
other methods? Are scale range, interval, number of values chosen are
adequate and reasonable ?

Executing the Project

Following planning, a brief description of the expected procedures has to be
approved in advance by the teacher. Having decided what controls you need
to use, list the components of your experiment and decide what quantities of
substances to use, how to set the experiment. You should also decide what
type of readings or measurements you are going to make, how often and how
many. Note the source of error, if any, that you come across.
¨ Handle instruments and equipments appropriately to give accuracy.
¨ Repeat measurement.
¨ Keep proper controls and the variables constant.

Reporting/Writing of Project
A format, such as given below, can be followed.
(i) Title of the investigatory project: Write the title of the project, for
example, ‘Inheritance pattern of eye colour’.
(ii) Objectives: Express as clearly as possible the effect of one variable that
the experiment is designed to investigate.
(iii) Materials needed: This might be just a list, or a diagram if a particular
piece of apparatus was used.
(iv) Method: Describe the procedure stepwise including the precautions
taken, if any.
(v) Result: A suitable chart or table for recording and organising your
readings or measurements should be made out before you start the
experiment.
(vi) Analysis and interpretation: Observation data are factual, and may
not be as expected by you.
(vii) Discussion: Discuss briefly the implication of your results and suggest
extensions of any kind that can be undertaken.
(viii) Conclusion: In view of the results obtained and related work done on
the topic of the project, write conclusion briefly.
(ix) References: Any work related to the project which you have come across
through books/articles or any other source should be written as reference,
for example: Michael Michalco (2001), Cracking Creativity, Berkeley, Ten
Speed Press.
This write up is meant to train the students in scientific methods. In other
words, it accentuates the spirit of enquiry and investigation in young minds.

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The operational aspects of doing a project include choosing a hypothesis or

problem to be investigated, collecting data in a designed manner, analysing
the data in a scientific way, drawing conclusions which are justified and
discussing the results in the light of known knowledge and bringing out its
importance. Finally it includes the scientific way of communicating the findings.
While your discovery during the investigatory project may not merit a Nobel
Prize it may help you discover something, a fact or relationship that was
unknown to you and that was not recorded in any book available to you.
Scientists refer to this as an independent discovery. Your investigation will
certainly give a sample of the thrill of discovery.
Following are pages on procedural guideline about a few suggestive
investigatory project work.

1. Investigating the pH of a water sample

Background information
Monitoring the physico-chemical properties of water is of vital importance.
Normal maximum permissible limit of pH for our life and health is 6.5–8.5.
Abnormal levels of pH and their consequences are given below:
pH 3 to 5 is too acidic for most organisms to survive, when the pH of water falls
below 4.5 most of the fishes die, leaving only a small number of acid-tolerant
insects such as water boatman and whirligig. These insects (beetles) can survive
and multiply even at pH 3.5. Similarly, pH>8.5 is too basic for most organisms
to survive.

Materials needed
¨ Universal indicator test paper (broad range, narrow range PH 2–11)
¨ Water sample

2. Investigating the biochemical (also called biological oxygen

demand [BOD]) of a water sample as pollution indicator.
Background information
A dissolved oxygen (DO) test measures the current status of oxygen in a water
body. This is a useful starting point.
However, DO content can vary considerably from day-to-day as affected by
many factors like temperature, wind velocity, eutrophication, pollution, etc.
The unpolluted water is characteristically rich in DO and low in BOD.
Higher the BOD, lower would be DO. Conversely, the polluted water has
high values of BOD. Water for drinking should have a BOD less than 1. Typical
BOD value for raw sewage run from 200–400 mg of oxygen/litre. The maximum

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permissible limit of BOD followed by Central Pollution Control Board (India)

for national water quality monitoring purposes is less than or equal to 3mg/L.
For example, in a study the prevalence of some organisms were done at
two different sites in a water body. The result can be tabulated on the basis of
following facts:
A = Abundant; R = Rare; X = Very few

Concentration Indicator organism Site 1 Site 2

Organisms in order of Red sledge worm (Tubifex worm) A A

tendency to disappear Larvae of midge (Chironomus) R A
as degree of pollution
Blood Worm X X
increases
Leech (Hirudinea) X X
Water louse, water skaters X X
(Asellus) X X
Fresh water shrimp (Gammarus) X X
Water boatman X X
Diving beetle (Dytiscus) X X
Caddisfly larva (Ochrotricha) X X
Damselfly larva (Lestes) X X
Stonyfly nymph (Isoperla) X X
Mayfly nymph (Stenonema) X X
Snail (Lymnaea) none X
Clams (Corbicula) none X
Fungi X X
Bacteria (anaerobic) X X
Utricularia X X
Chara X X
Water fern (Salvinia) X X
Water velvet (Azolla) X X
Water meal (Wolffia) X X
Lesser duckweed (Lemna minor) X X
Greater duckweed (Spirodela) X X
Diatom X X

3. Population density of plants

(i) Identify any 5 weeds from your locality.
(ii) Collect information about them from various sources. Focus on their
economic importance especially their medicinal importance (collect
samples for herbarium preparation).

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(iii) Study their distribution in different localities by quadrat method.

(iv) Register their data and draw comparisons of their distribution by
histograms.
(v) Try to analyse the differences in distribution and density.
(vi) Correlate their presence with their habitat/adaptability.

4. To make an inventory of local tree, shrub and herb

Informations can be listed in following categories:
(i) Avenue trees
(ii) Wind Breakers
(iii) Road dividers
(iv) Sound barriers
(v) Medicinal and other uses

5. Agrochemicals and their effects

The project may be carried out in a survey mode with a questionnaire prepared
with the help of the teacher to cover the following aspects.
(i) List of pesticides used, amount used/hectare or acre, periodicity of spray,
name of the crop plant grown, recommended dose and the dose employed,
known effects on pest, whether the chemical pesticide is biodegradable
or not, alternate ecofriendly biocides.
(ii) List of fertilisers used, cost incurred/acre/year, recommended dosage/
time of use and the dosage used, known effects on fertility of soil, any
decrease in crop productivity, use of ecofriendly biofertilisers (VAM fungi,
leaf mold, green manure, dung, etc.).

6. Ecological role of some animals observed in a local area

Record the various plant species growing in the area under study—trees,
shrubs, annual/perennial herbs, etc.
Note the season when flower/seed is formed.
Note the various types of insects, birds, reptiles, amphibians, mammals, etc.,
and record their role as a/an
(i) Herbivore
(ii) Pollinator
(iii) Agent in seed dispersal
(iv) Prey

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(v) Predator
(vi) Vectors for transmission of diseases
(vii) Any other

07. Study the effect of a local industry on environment

(i) Select an industry of your choice.
(ii) Note the source of energy used, product formed, raw materials used
(locally available or imported) mode of transport used to move the final
product.
(iii) Possible types of pollutants released by the industry (air/water/
soil).
(iv) Measures undertaken by the management to comply with the standard
set by Central Pollution Control Board (CPCB), PCBs, etc.
(v) Awareness about ISO 2000.
(vi) Impact assessment carried out or not.

08. Study of the effect of chemicals and pollutants on the Mitotic

Index of the mitotically dividing onion root tip cells
This study may include
(i) Growing of onion root tip cells in the solution of pollutant/chemical
and also in normal water as control.
(ii) Preparation and observation of slide for the study of mitotic index both
in experimental and control set-ups.
(iii) Analysis of the effect of pollutant/chemical by comparing the data of
mitotic index between experimental and control variable.

09. Study of the genetic markers in the human population

In this investigation a few selected inherited traits can be investigated in the
family members of a small population in the locality. The compilation and
analysis of data will provide an about prevalence of trait in the said
population.

10. Inventory of weeds in aquatic bodies/agricultural fields

11. Inventory of birds in your locality, their ecological role as

scavangers, pollinators, etc.

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12. Impact of local industry on the environment and the remedial

measures taken by the industry
The guidelines and a brief outline of a few projects have been given with a
purpose to design and perform such investigation. Students and teachers can
think and design investigatory projects based on almost all concepts about
which experimental protocol have been given in the manual. However, a small
list of suggestive projects are also given below. Please note that these are only
suggestive and it is expected that students and teachers will take up many
more types of investigatory projects depending on the specificity of their area,
need and problems.

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











 

 
 
 

 

 
 
 
 

Procedure
(i) Prepare a range of pH buffers using Na2HPO4 and KH2PO4.
(ii) Wash the seeds with water and blot them dry.
(iii) Select an appropriate place in the laboratory where there is sufficient
light. Arrange petridishes in three horizontal lines, with 5 dishes in each
line. Arrange petridishes horizontally in three rows A, B and C with seven
dishes in each row.

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INVESTIGATORY PROJECT 1

(iv) Place one blotting paper disc in each petridish.

(v) Wet the blotting papers with small quantities of buffer solution of 4.0
pH for petridish No. 1, 5.0 pH for petridish No. 2 and so on till all the
blotting papers in petridishes No. 1–5 in row 'A' are wet with appropriate
buffer solution.
(vi) Repeat the process for rows B and C also.
(vii) Spread selected seeds in each of the five petridishes in such a way that
each petridish contains 25 seeds in row 'A'.
(viii) Repeat the process for rows B and C using two other types of seeds.
(ix) Cover the petridishes and record your observation after every 24 hours
for 7 days. Tabulate your results as given.

Observation
Observe the emergence of radicle as an indicator of germination and record in
the table. Calculate the percentage of germination every day.

Table: Percentage of ................ seed germination

No and % of seed germination Total % of seed
germination
pH 1 2 3 4 5 6 7 A B C
4
5
6
7
8

Use the data of the table for graphic presentation.

Inferences and conclusion

The inferences and conclusion may be drawn on the basis of observations and the points given
below
• Find out at what pH range seeds of different species had maximum % of germination.
• Find out at what pH seeds failed to germinate or showed minimum % of germination.
• Is there any general pattern of seed germination with regard to pH ranges?
• What are the common features exhibited by the various types of seeds under varied pH
ranges? For example, did all the type of seeds show maximum germination in acidic range or
alkaline range or did pH preference varied between acidic and alkaline ranges?
• Did you observe any relationship among time period, seed germination and pH range?

101

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Investigatory Project 2



















 
 
 
 




Procedure
(i) Plankton net resembles the butterfly net in several aspects. Plankton
net, however, is prepared from bolting silk cloth which is readily available
at shops dealing in scientific equipments and chemicals. Procure about
one metre of bolting silk cloth of 40 mesh size and stich out of it a 40 cm

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INVESTIGATORY PROJECT 2

long cone with a diameter of about 20 cm at the mouth and a diameter

of 3–4 cm at the other end (both the sides open). Fasten to the mouth of
the cone a circular iron ring (with handle) of about 20 cm diameter with
the help of twine thread. Fasten a small steel bucket (plankton bucket)
or glass specimen tube of 50 ml capacity at the lower end.
(ii) Visit a nearby pond, pool or river bank carrying along with you the
plankton net fitted with the plankton bucket, graduated plastic bucket
and 5–10 ml of F.A.A.
(iii) Since this is a group activity, ask your friend to hold the plankton net
firmly a few centimetres above the water surface.
(iv) Immerse and fill the plastic bucket with water completely upto 15–litre –
graduated mark and filter the water through the plankton net. Repeat
the process several times (say10 times).
(v) At the end calculate the quantity of water in litres (X) filtered by
multiplying the amount of water in one bucket and number of buckets
of water filtered.
(vii) During this process of filtering, the planktons are collected in plankton
bucket. Only the water free of planktons escapes through the mesh of
the net.
Splash a few buckets of water against the net from outside taking care that
no water enters into the cone from the mouth. This will wash all the planktons
sticking against the inside wall of the net into the plankton bucket.
Detach the plankton bucket from the net and add a few drops (1–2 ml) of
5% F.A.A. to the plankton concentrate. Transfer the concentrate collected into
a suitable specimen tube and cork it. Note the volume of the concentrate (Y).

In the laboratory
1. With the help of 1 ml pipette, draw 1 ml of concentrate and transfer it
dropwise into the watch glass. Count the total number of drops that make
1 ml of concentrate (A).
2. Transfer one drop of plankton concentrate from the watch glass on a clean
slide. Cover it with square shaped cover slip. (For convenience divide the
area of the cover slip into parts with the help of lines drawn by Indian ink).

Observation
Observe the slide under microscope and count the number of total organisms
(B) by moving the slide from one corner of the cover slip to another horizontally
as well as vertically till the entire sample under the cover slip is completed.
With the help of following calculations find out the total number of different
organisms per litre of water.

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1000×(ABY)
Unit cells/L = Unit/L =
x
Where A = number of drops in 1ml concentrate
B = number of organisms counted in 1 drop of concentrate
X = total amount of water filtered
Y = total volume of concentrate after filtration

Note: Another alternate method is the use of haemocytometer to calculate the

density of organisms under the guidance of teacher.

Inferences and conclusion

Find out the density and composition of organisms in different water samples
(polluted/non-polluted).

Note the common organisms in both the water samples and those specific
to each sample.

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Notes

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Notes

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