SIJIL TINGGI PERSEKOLAHAN MALAYSIA (STPM) (MALAYSIA HIGHER SCHOOL CERTIFICATE

)

Student’s Manual Practical Biology Paper 964/3 (School-based Assessment)
2010/2011 Session

Majlis Peperiksaan Malaysia
Bangunan MPM, Persiaran 1 Bandar Baru Selayang 68100 BATU CAVES Selangor Tel: 03-6136 9663 Fax: 03-6136 7329

© Majlis Peperiksaan Malaysia 2010

CONTENTS page

1.0 2.0 3.0 Experiment 1 Experiment 2 Experiment 3 Experiment 4 Experiment 5 Experiment 6 Experiment 7 Experiment 8 Experiment 9 Experiment 10 Experiment 11 Experiment 12 Experiment 13 Experiment 14 Experiment 15 Experiment 16 Experiment 17 Experiment 18 Experiment 19 Experiment 20

General Information Practical Work Assessment Guide Table of Summary of Experiments Determination of osmotic potential Use of microscope, magnification, and measurement of cell size Observation of cells (a) animal cell: cheek cell (b) plant cell: leaf epidermis cell Enzyme activity Separation of photosynthetic pigments using paper chromatography Examining slides of transverse sections of C3 and C4 leaves Use of yeast in respiratory experiment Dissection of the mammalian digestive system Dissection of the mammalian respiratory system Examining slides of transverse sections of vein, artery and capillary Dissection of the mammalian circulatory system Examining slides of liver and kidney Examining slides of Spirogyra, Funaria, Marchantia, Dryopteris and Pinus Investigating the structure of flowers, Angiospermatophyta (one monocotyledon and one dicotyledon) Monohybrid and dihybrid crosses and use of χ2 test Construction of a dichotomous key using local specimens Preservation procedure (a) Insects (b) Plants 25 insect species (up to order) 25 plant species (local name and habitat) Ecological study of a terrestrial or an aquatic area. (group work)

1 2 4 5 8 12 15 18 21 24 26 29 32 35 39 43 46 49 54 58 61 63 65

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

SCHOOL-BASED ASSESSMENT OF PRACTICAL BIOLOGY
1.0 General Information 1.1 Continuous assessment of practical work will be carried out throughout form six course. 1.2 1.3 This assessment is expected to commence in early July 2010 and end before 30 August 2011. Majlis Peperiksaan Malaysia (MPM) has determined 20 experiments to be carried out by students. Of these 20 experiments, only 13 compulsory experiments will be assessed by the teacher. (Refer to the Table of Summary of Experiments on pages 4.) The assessment of practical work will be by the carried out subject teacher during the experiment and also based on the student’s practical work report. Students should plan their practical work first before the experiment is carried out. Compulsory experiments are to be carried out by students individually, or in groups as recommended in the Table of Summary of Experiments. Students may write their practical work report in either English or Bahasa Malaysia. The report is to be submitted to the teacher on the same day of the experiment is carried out unless otherwise stated. (Refer to the Table of Summary of Experiments.) Practical work reports which are not submitted on the day of the experiment are to be awarded ‘0’ mark. Practical work report which can be completed at home is to be submitted to the teacher not later than 3 days from the date of the experiment. A penalty of 2 marks is to be imposed for the report submitted late to the teacher. Practical work report which is submitted later than 7 days from the fixed date are to be awarded ‘0’ mark. For a student who is absent from an experiment, the teacher can fix another date for the student to carry out the experiment. Practical work reports which have been submitted to the teacher can be returned to the students only after the teacher has completed assessing the reports and recording the marks of all students. However, the teacher will collect all the practical work reports before 15 October 2010 for the first year of the course and before 15 September 2011 for the second year. Students can check their Student Record to ensure that the mark for each experiment and the overall total mark awarded are correct. For a student who has transferred to another school, the previous school is to send the student’s Student Record, which is partially completed and signed by the subject teacher, to the student’s new school.

1.4 1.5 1.6

1.7

1.8 1.9

1.10 1.11

1.12

A student whose Student Record has not been sent by the school to MPM will be considered as not having carried out the practical work and not having attended paper 964/3. STUDENT’S MANUAL 2010/2011

STPM BIOLOGY 2.0 Practical Work Assessment Guide Aspects to be assessed by the teacher are as follows: 2.1

2.2

2.3

Microscope and Slide 2.1.1 Skills in handling microscope and displaying specimens (10 marks − Skill A) 2.1.2 Product (drawing/accuracy, labels, scale, and identification) (10 marks − Product B) Biochemistry and Physiology 2.2.1 Preparation of materials, procedures, including preparation of solution and experimental substances (2 marks − Skill A) 2.2.2 Manipulative skill (4 marks − Skill A) 2.2.3 Planning and execution (structuring, planning and managing, neatness, efficiency, observation,and following instructions) (6 marks − Skill A) 2.2.4 Product (observation and drawing) (8 marks − Product B) Dissection 2.3.1 Flower (i) Manipulative skills (cutting the flower into two equal halves) (10 marks − Skill A) 2.3.2 (ii) Product (observation and drawing) (10 marks − Product B) Animal (one of 5 systems) (i) Ability to follow instructions on how to dissect and display system (6 marks − Skill A) (ii) Accuracy and completeness of display (6 marks − Skill A) (iii) Neatness (2 marks − Product B) (iv) Product (drawing, labels and scale) (6 marks − Product B) Collection of insect and plant specimens, and ecological projects 2.4.1 Collection of the insect and plant specimens (i) Achieving target (number and accuracy of identification) (4 marks − Product B) (ii) Quality of specimen and preservation (6 marks − Product B) (iii) Presentation (6 marks − Product B) (iv) Diversity (family and order) (4 marks − Product B)

2.4

STPM BIOLOGY 2.4.2 Ecology

STUDENT’S MANUAL 2010/2011

2.4.2.1 Project planning (4 marks − Product B) 2.4.2.2 Collecting data and keeping records (8 marks − Product B) 2.4.2.3 Product (through assessment of report) (8 marks − Product B) (i) Data, presentation, analysis, and others (ii) Summary (iii) Creativity/Innovation (iv) Overall quality of report

STPM BIOLOGY 3.0 Table of Summary of Experiments
Experiment number 1 2 3

STUDENT’S MANUAL 2010/2011
Mode of working Individual Individual Individual

Title of experiment Determination of osmotic potential Use of microscope, magnification, and measurement of cell size Observation of cells (a) animal cell: cheek cell (b) plant cell: leaf epidermis cell Enzyme activity Separation of photosynthetic pigments using paper chromatography Examining slides of transverse sections of C3 and C4 leaves Use of yeast in respiratory experiment Dissection of the mammalian digestive system Dissection of the mammalian respiratory system Examining slides of transverse sections of vein, artery and capillary Dissection of the mammalian circulatory system Examining slides of liver and kidney Examining slides of Spirogyra, Funaria, Marchantia, Dryopteris and Pinus Investigating the structure of flowers, Angiospermatophyta (one monocotyledon and one dicotyledon) Monohybrid and dihybrid crosses and use of χ2 test Construction of a dichotomous key using local specimens Preservation procedure (a) Insects (b) Plants 25 insect species (up to order) 25 plant species (local name and habitat) Ecological study of a terrestrial or an aquatic area. (group work)

4 5 6 7 8 9 10 11 12 13 14 15 16 17

Individual Individual Individual Individual Individual Individual Individual Individual Individual Individual Individual Individual Individual Individual

18 19 20

Group Group Group

Note: 1. 2. For the experiments 1 to 17, the practical work reports are to be completed in the laboratory. Project work (that is for experiments 18, 19, and 20) must be carried out during the school holidays at the end of lower six.

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 1 (Practical No. 2(a))
Title: Determination of the osmotic potential of the potato cell sap Purpose:
1. To prepare the sucrose solutions of various molarities from a 1.0 mol dm-3 stock solution 2. To plan and conduct the experiment efficiently 3. To measure and record the length of tissues accurately 4. To tabulate the results neatly 5. To analyse and interpret experimental results based on knowledge or theory

Apparatus and materials
1.0 mol dm-3 sucrose solution (40 cm3 per candidate) Distilled water Boiling tubes placed in beakers Ruler Potato Filter paper Petri dish with cover Marker pen Knife White tile Graph paper 130 cm long round wooden stick, 0.5 cm in diameter, with a pin at one end 25 cm3 measuring cylinder Forceps or glass rods ≈ 15 cm 10 cm3 pipette

Procedure
1. In labelled boiling tubes, prepare 20 cm3 of sucrose solutions of molarities 1.0 mol dm−3, 0.2 mol dm−3, 0.3 mol dm−3, 0.4 mol dm−3, and 0.5 mol dm−3 from the stock solution using the dilution technique. The volumes of the solution and of the distilled water used should be recorded in the table below. Molarity Volume of 1.0 M sucrose solution (cm3) Volume of distilled water (cm3) 0.1 M 0.2 M 0.3 M 0.4 M 0.5 M

STPM BIOLOGY STUDENT’S MANUAL 2010/2011 2. Prepare 15 strips of potato tissues 4 cm to 6 cm long and with a cross-section of 0.4 cm × 0.4 cm. Record the average length of the potato strips. 3. Mark the initial level of 0.1 mol dm-3 sucrose solution on the boiling tube before the addition of potato strips. Take 3 potato strips, record the length of each of the strips, and place the strips into the boiling tube. Repeat this step using the sucrose solutions of molarities 0.2 mol dm−3, 0.3 mol dm−3, 0.4 mol dm−3, and 0.5 mol dm−3. 4. After 30 minutes, remove the strips with the wooden stick provided. Wipe them gently with the filter paper and record the final length of each of the potato strips. 5. Record the final level of the sucrose solution in each of the boiling tubes and note down any changes to the physical condition of the potato strips. 6. Calculate the average change in length. Table

7. On the graph paper, (i) Plot a standard graph of the osmotic potential against the molarity of the sucrose solution to determine the osmotic potential of the potato cell sap.
Molarity Osmotic potential (atm) 0.0 5 1.3 0.1 0 2.6 0.1 5 4.0 0.2 0 5.3 0.2 5 6.7 0.3 0 8.1 0.3 5 9.6 0.4 0 11. 1 0.4 5 12. 6 0.5 0 14. 3 0.55

16.0

(ii) Plot a graph of the average change in the length against the molarity of the sucrose solution.

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Questions
(a) From the two graphs, determine (i) the osmotic concentration of the potato tissues in mol dm−3 of sucrose solution, (ii) the osmotic potential in atm. ............................................................................................................................................... . ............................................................................................................................................... . (b) Explain why you have chosen the above molarity. ............................................................................................................................................... . ............................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 2 (Practical No. 3)
Title: Use of microscope, magnification, and measurement of cell size. Purpose:
1. To estimate the magnification of a drawing made with the help of a microscope 2. To estimate the actual size of several examples of microorganisms 3. To determine the size of onion scale leaf epidermal cells

Apparatus and materials
Microscope Prepared slides (Euglena, Amoeba, Hydra) Glass slide Coverslip Forceps Onion Iodine solution

Procedure A
To estimate the magnification of a drawing made with the help of a microscope. 1. Examine the slides of various types of microorganisms under a high power microscope. 2. Estimate the image size (apparent object size) of the specimen. 3. Make a labelled drawing and determine the magnification of your drawing using the following formula: Magnification = Magnification of x Magnification of x of drawing eyepiece objective lens size of drawing apperent size of object

4. Estimate the actual size of each microorganism using the following formula. Actual size of object = Apparent size Magnification of microscope

STPM BIOLOGY Drawing

STUDENT’S MANUAL 2010/2011

Data
Eye piece magnification = ………………….................………………………………....... Objective lens magnification = ...........................................………………………………. Microorganism Apperent object size Size of drawing Euglena Amoeba Hydra

Questions
1. Determine the magnification of the drawing of each microorganism. (i) Euglena .......................................................................................................................………… (ii) Amoeba ......................................................................................................................……....... (iii) Hydra .........................................................................................................................……….. 2. Estimate the actual size of each microorganism (i) Euglena ......................................................................................................................……....... (ii) Amoeba ......................................................................................................................……....... (iii) Hydra .........................................................................................................................………..

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Procedure B
To determine the size of onion scale leaf epidermal cells. 1. Using a pair of forceps, peel off the epidermal layer of the onion scale leaf. 2. Mount the epidermal layer in a drop of water on a slide. Add a coverslip. 3. Stain the specimen with iodine solution. 4. Determine the diameter of the microscope’s field of view: Place a clear plastic ruler under the microscope and focus on the millimeter scale at low power. Estimate the diameter of the field of view. Convert it to μm and tabulate your observations.

Data
To determine the diameter of a microscope’s field of view. Diameter of low power field of view = .................................................................................
Magnification of high power objective lens = ...................................................................................

The diameter of a high power field of view is 1/4 of the diameter of a low power field of view = .............................................................................................................…………….. 5. Examine the epidermal cells under the microscope using high power objective lens.

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

6. Count the number of cells across the high power field of view lengthways. Repeat three times and determine the average number of cells. 7. Repeat step 6 but this time count the number of cells across the high power field of view width ways. 8. Calculate the average length and width of a single epidermal cell. State your answer in μm.

Results
Number of cells lengthways: First count = ................................................... Average = ............................……………... Second count = ............................................... Third count = .................................................. Number of cells width ways: First count = .................................................... Average = ..................................……......... Second count = ................................................ Third count = ................................................... Average length of one epidermal cell of onion scale leaf Diameter of microscope’s field of view = Number of cells lengthways = Average width of one epidermal cell of onion scale leaf Diameter of microscope’s field of view = Number of cells widthways =

Questions
1. What is the average length of one epidermal cell of the onion scale leaf? …........................................................................................................................................... . 2. What is the average width of one epidermal cell of the onion scale leaf? …........................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 3 (Practical No. 4)
Title: Observation of cells.
(a) Animal cells: cheek cells (b) Plant cells: leaf epidermal cells

Purpose:
1. To prepare the slides of the animal cells and plant cells using the correct staining technique 2. To realise that cell is a basic unit of life

Apparatus
Microscope Toothpick Microscope slide Coverslip Dropper Forceps Methylene blue Iodine solution Onion

Procedure
(a) Observation of the animal cells 1. Using a toothpick gently scrape off a thin layer of cells from the inside of your cheek. 2. Mount the scrapings in a drop of methylene blue solution on a slide. 3. Examine the specimen under low power objective lens followed by high power objective lens. 4. Draw the cheek cells and label the following parts: nucleus, nuclear membrane, chromatin granules, cell membrane, and cytoplasm. (b) Observation of the plant cells 1. Using a pair of forceps, peel off the epidermal layer of the onion scale leaf. 2. Mount the epidermal layer in a drop of water on a slide. 3. Examine the specimen under a microscope. 4. Stain the onion scale leaf epidermis with iodine solution. Then examine the specimen under the microscope. 5. Draw the onion scale leaf epidermal cells. Label the cell wall, cell membrane, cytoplasm, nucleus, chromosome, nucleolus, and vacuole.

STPM BIOLOGY STUDENT’S MANUAL 2008/2009 13 Drawing of animal cells

Drawing of plant cells

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Questions
1. Name the type of cells which lines the inner cheek. ……....................................................................................................................................... . 2. What is the effect of the methylene blue solution on the cheek cells? ……....................................................................................................................................... . 3. The onion scale leaf cells do not contain chloroplast? Give a reason for your answer. ……....................................................................................................................................... . 4. State two differences between the animal and plant cells. ……....................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 4 (Practical No. 8)
Title: Enzyme activity Purpose:
1. To investigate the effect of temperature on the enzyme-catalysed reactions 2. To determine the optimal temperature of the enzymic reactions 3. To determine the temperature quotient, Q10, of an enzyme-controlled reaction

Apparatus
250 cm3 beakers (5 units) Thermometer Test-tubes (10 units) White tile Dropper Stopwatch 30 cm3 of 1% starch solution Iodine solution

Procedure
1. Prepare a saliva solution by spitting saliva into a clean beaker and diluting it with an equal amount of distilled water. (Remember to rinse your mouth first.) 2. Prepare five beakers of water baths at the following temperatures: 0 °C, 20 °C, 37 °C, 50 °C, and 65° C, and label the baths A to E. The temperature of the water bath must be kept constant by adding hot/cold water into it. 3. Place two test-tubes labelled X and Y into the water bath A. Put 4 cm3 starch solution into test-tube X and 3 cm3 saliva solution into test-tube Y. Leave it for 5 minutes to stabilise the temperature. Then add the saliva solution from test-tube Y to the starch solution in test-tube X to start the reaction. 4. Start the stopwatch. 5. Every minute, take out a drop of solution from test-tube X and test it with a drop of iodine on a white tile. 6. Repeat steps 3 to 5 for the water baths B, C, D and E. 7. Conduct the iodine tests at a shorter interval as and when the reaction is nearing completion. 8. The time taken for each complete hydrolysis is recorded in the table below.

STPM BIOLOGY STUDENT’S MANUAL 2010/2011

Results
Test-tubes A B C D E Temperature/° C 0 20 37 50 65 Time taken for complete hydrolysis, t/minute Rate of reaction 1/t

Questions
1. Plot a graph of the reaction rate (1t) against the temperature.

2. (i) Calculate the temperature coefficient, Q10, between 30 °C and 40 °C. (ii) What conclusion can you draw from the value of Q10 in question 2 (i)?

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

3. Describe how temperature affects the enzyme-catalysed biochemical reactions. ……....................................................................................................................................... . ……....................................................................................................................................... . ……....................................................................................................................................... . ……....................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 5 (Practical No. 9)
Title: Separation of photosynthetic pigments using paper chromatography Purpose
1. To prepare a concentrated pigment extract from leaves 2. To follow the practical instructions carefully 3. To separate the pigments based on the clear difference in pigment colours 4. To calculate Rf value.

Apparatus and materials
80% acetone (to extract pigments) Red spinach leaves or Hibiscus leaves or “cekor manis” leaves Mortar and pestle Muslin (cheese cloth) Glass tube Boiling tube with stopper Dropper with pointed tip/pin Solvent (1 part 80% acetone and 9 parts petroleum ether) Ruler Chromatography strip or Whatman no. 1 filter paper cut into strip to fit inside the boiling tube without touching the sides

Procedure
Switch off the fans while conducting the experiment. 1. Prepare the pigment extract by grinding leaves with 5 cm 3 acetone using a mortar and a pestle. Squeeze out a thick extract using a cheese cloth. 2. Prepare the chromatography strip. Cut out the end of the chromatography strip to form a pointed tip. 3. Transfer the pigment extract onto the chromatography strip using a dropper with a pointed end. Leave it to dry. Repeat the process 15 to 20 times over on the same spot. (Smaller spot gives the better result). 4. Put enough solvent into a boiling tube. Using a pin/clip hang the strip (pointed end down into the solvent). Make sure that the strip is in a perfectly vertical position. Solvent level must be 1 cm from the pigment spot. Stopper the boiling tube tightly. Do not shake or move it. Place the tube on a rack.

5. Let the solvent move until it reaches near the pin/clip. Mark the solvent front with a pencil. Quickly dry it and measure the distance from the pigment spot to the solvent front. STPM BIOLOGY STUDENT’S MANUAL 2010/2011

6. Observe and mark the positions of the separated pigments and measure the distance of the movement of each of the pigments. 7. Cover the chromatography strip with a black material to protect the pigments. 8. Warning: Lift up the chromatography strip as soon as the solvent reaches the end of the strip. 9. Calculate the Rf value using the formula: Rf = Distance moved by pigment Distance moved by solvent

10. Record all the data in the table below.

Results
(i) Table Pigment colour Distance moved by pigment /mm Distance moved by solvent /mm Rf (ii) Stick on your chromatography paper in the space provided below.

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Questions
(a) Explain the basic principles of this pigment separation method. ……....................................................................................................................................... . (b) What conclusions can you draw from your observations of the leaf pigments? ................................................................................................................................... ……..... ……....................................................................................................................................... . (c) Using the pigment colours or Rf values, identify and state each of the pigments that you have separated. ……....................................................................................................................................... . ……....................................................................................................................................... . ……....................................................................................................................................... . ……....................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 6 (Practical No. 10)
Title: Examining slides of transverse sections of the C3 and C4 leaves Purpose:
1. To reinforce the theoretical understanding of the cells which involved in the HatchSlack pathway and Calvin Cycle 2. To relate the structure of the cell to its functions

Theory
Many tropical plants such as the sugar cane and corn do not use ribulose bisphosphate to fix carbon dioxide. Instead, these plants use a 3-carbon compound (PEP) and its first product of photosynthesis is a 4-carbon compound (oxaloacetate). The first photosynthetic product of the C3 plant is phosphoglyceric acid (PGA). The process of the carbon fixation via the Hatch-Slack pathway happens in two types of cell (mesophyll and bundle sheath cells), which are spatially separated.

Apparatus and materials
Microscope Prepared slide of transverse section of Zea mays leaf Prepared slide of transverse section of Helianthus leaf

Procedure
1. Examine the slides and make labelled plan drawings of each of the leaf sections. 2. Note that in the C4 leaf, the mesophyll cells are arranged to form a ring around the bundle sheath cells. The bundle sheath cells are large and are made of two types of photosynthetic cells, these are mesophyll cells which contain chloroplasts with grana but without starch and bundle sheath cells which have chloroplasts without granum but rich in starch. The chloroplasts of the bundle sheath cells are large and prominent. 3. Note that in the C3 leaf, the mesophyll palisade exists as one basic layer. Its bundle sheath cells are small, the chloroplasts in all the mesophyll cells can carry out

photosynthesis (almost a single type of photosynthetic cells), and the spaces in between cells are larger than in the C4 leaf. 4. Make a high power labelled drawing for the cells, which you have observed. State the magnification of the drawing. STPM BIOLOGY High power drawing of the C3 leaf STUDENT’S MANUAL 2010/2011

Magnification of drawing .......................................... High power drawing of the C4 leaf

Magnification and drawing .......................................... STPM BIOLOGY STUDENT’S MANUAL 2010/2011

Questions
1. What are the compounds in the C3 and in C4 plants, which act as carbon dioxide receptors during dark reaction? ................................................................................................................................... ……..... ……………………………………………………………………………………………… 2. Name the outer layer of a leaf and describe its functions. ............................................................................................................................................... . ............................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 7 (Practical No. 11)
Title: Use of yeast in experiments on respiration Purpose: To investigate the effect of different nutrients on the anaerobic respiration of
yeast

Apparatus
Test-tubes (5 units) Fermentation tubes 50 mm × 7.5 mm (5 units) Teat pipettes Labels Incubator Distilled water 20% sucrose solution 20% glucose solution 20% lactose solution 10% starch suspension 10% yeast suspension

Procedure
1. Label five fermentation tubes A to E. 2. Using teat pipettes, place the following solutions into each tube. Tube A B C D E Solution 10 drops of distilled water 10 drops of glucose solution 10 drops of sucrose solution 10 drops of lactose solution 10 drops of starch suspension

3. Add 10 drops of yeast suspension into each tube. Add distilled water to fill up the tubes. 4. Using pencil, support the fermentation tube vertically in a test-tube as shown in Diagram I. After that, invert the fermentation tube as shown in Diagram II. Take care not to spill out the fluid in the fermentation tubes.

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

5. Record the height of the fluid in the fermentation tubes A to E. 6. Place all the tubes in the incubator at 37 °C − 40 °C for 50 minutes. 7. Remove the tubes from the incubator and measure the final height of the fluid in each of the fermentation tubes. 8. Record the difference in height between the initial and final readings for each of the tubes.

Results
Fermentation tube A B C D E Nutrient Initial height of fluid, x/mm Final height of fluid, y/mm Difference (xy)/mm

Distilled water Glucose Sucrose Lactose Starch suspension

Questions
1. Name the gas collected in the fermentation tubes. ............................................................................................................................................... . 2. (i) Which tube/tubes did not release any gas? ............................................................................................................................................... . (ii) State the reasons for your answer in 2 (i). ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... . 3. Based on the experimental results, suggest the most suitable nutrient for the yeast to carry out its activity.

............................................................................................................................................... . 4. Write an equation representing the anaerobic respiration in yeast. ............................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 8 (Practical No. 12)
Title: Dissection of the mammalian digestive system Purpose:
To clearly display 1. The digestive system and the related organs; 2. The blood vessels, which are the arteries and veins of the system.

Apparatus and materials
Animal BA 5 Dissecting board Pin A set of dissecting instruments *Note: Use of chloroform to kill rats must be done by the teacher.

Procedure
1. Pin the animal BA 5 to a dissecting board, with the ventral surface upwards. 2. Open up the abdominal cavity to display the internal organs. 3. Flip over and pin the liver to the anterior of the animal and push the rest of the viscera to the left side of the animal. Carefully separate the small intestines from the large intestines by cutting the supporting mesenteries without damaging the mesenteries holding the duodenum in its looped form. Do not cut the blood vessels of the mesenteries. 4. Make a labelled drawing showing the lobes of the liver, ducts of the liver, stomach, duodenum, pancreas, and hepatic portal vein. State the scale of your drawing.

STPM BIOLOGY Drawing

STUDENT’S MANUAL 2010/2011

Scale of drawing .................................................. 5. Pull the stomach slightly downwards to display the esophagus. Display the alimentary canal and the related organs on the left side of the animal so as to display the veins to the maximum advantage. 6. Make a large labelled drawing to display the alimentary canal, related organs, and their veins. State the scale of your drawing. Drawing

Scale of drawing .................................................. STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

7. Pin the alimentary canal and the related organs to the right side of the animal BA 5 so as to display its supply of the arteries to maximum advantage. 8. Make a large labelled drawing to display the alimentary canal, related organs, and their arteries. State the scale of your drawing. Drawing

Scale of drawing .................................................

Questions:
State the main functions of the following organs and their importance to the digestive system: (a) Liver: ...................................................................................................................……... ……………………………………………………………………………………………… (b) Pancreas: ........................................................................................................................

............................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 9 (Practical No. 15)
Title: Dissection of the mammalian respiratory system Purpose:
1. To train the students to dissect small mammals 2. To train the students how to use the dissecting instruments 3. To increase the students’ skill in displaying, drawing and labelling respiratory organs 4. To enable the students to examine the structures of the main organs involved in respiration (lungs, trachea, diaphragm, rib cage, and intercostal muscles) 5. To increase the students’ understanding of the process of gas exchange in animals

Apparatus and materials Rats
Dissecting instruments Dissecting board Hand lens × 10 Transparent plastic ruler Thread

Procedure
1. Pin the rat to the dissecting board with the ventral surface uppermost. 2. Make a mid-ventral incision through the skin and cut forward as far as the lower jaw and then backwards to the anus. 3. Holding the skin with a pair of forceps, cut away the connective tissues between the skin and the body wall as far as possible around the animal’s body and pin back the skin. 4. Cut away the ventral and lateral thoracic walls to expose the thoracic cavity. 5. Remove the thymus gland.

6. Cut away muscles and tissues of the neck to expose the trachea and larynx. 7. Cut above the larynx. Cut off the connective tissues attached to the trachea. 8. Remove the heart, lungs, trachea, esophagus, and larynx together. 9. Carefully separate the esophagus from the heart. Pin the larynx, trachea, and lungs to the board. 10. Make a large labelled drawing of the structures you have taken out.

STPM BIOLOGY Drawing

STUDENT’S MANUAL 2010/2011

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Questions
1. (a) How many pairs of ribs does this animal have? ............................................................................................................................................... . ............................................................................................................................................... . (b) How does the rib cage function during gas exchange in this animal? ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... . 2. (a) Describe the appearance and characteristic of a diaphragm. ............................................................................................................................................... . (b) What is the importance of this characteristic of the diaphragm in relation to its function during gas exchange? ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... . 3. Describe the appearance of the left and right lungs. ............................................................................................................................................... .

............................................................................................................................................... . 4. Cut a part of the lung. Examine the cut surface using a magnifying glass. Describe what you can see with regards to the texture of the lung. ............................................................................................................................................... . ............................................................................................................................................... . 5. Measure the length of the trachea to the nearest mm, from the larynx to the point where it branches out into two bronchi. ...............................................................................................................................................

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 10 (Practical No. 16)
Title: Examining slides of transverse sections of a vein, an artery, and a capillary Purpose:
1. To improve students’ skills in using the microscope 2. To enable students to draw the structures of transverse sections of a vein, an artery, and a capillary accurately according to the magnification power of the microscope 3. To differentiate the structure of vein from that of the artery 4. To enable students to relate the structures of the blood vessels (artery, vein, capillary) to their functions 5. To observe and draw mammalian red blood cells under high power magnification 6. To increase students’ understanding of the mammalian circulatory system

Apparatus and materials

Microscope Slides of transverse sections of a cat’s vein and artery Slide of human blood

Procedure
1. Examine the slide of human blood under the high power microscope. 2. Draw the shape of mammalian red blood cells. 3. Examine the slides of a vein and an artery under the high power microscope. 4. Make a labelled plan drawing to show the arrangement of tissues in the transverse section of the blood vessels. State the magnification of your drawing.

STPM BIOLOGY Plan drawing

STUDENT’S MANUAL 2010/2011

Magnification of drawing ...........................................

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Questions
1. What is the shape of a human red blood cell? ............................................................................................................................................... . 2. How is the shape of the red blood cell related to its function? ............................................................................................................................................... . ............................................................................................................................................... . 3. State five opposing structural features found in a vein and an artery. Give your answers based on your observations under the microscope. ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... .

............................................................................................................................................... . ............................................................................................................................................... . 4. How are the structural features of a vein and an artery related to their functions in a mammal? ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... . 5. How is the structure of a capillary related to its function? ............................................................................................................................................... . ............................................................................................................................................... . ...............................................................................................................................................

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 11 (Practical No. 17)
Title: Dissection of the mammalian circulatory system Purpose:

1. To follow instructions correctly so that the thoracic area can be displayed to maximum advantage 2. To identify the organs in the thoracic cavity 3. To identify the position of the main blood vessels (veins and arteries) and their branches 4. To produce labelled drawings from the display 5. To state the scale of drawing accurately

Apparatus and materials
Dissecting instruments Animal BA 1 Dissecting board Pin

Procedure
1. Pin BA 1 to the dissecting board with the ventral surface uppermost. 2. Make a mid-ventral incision through the skin and cut the skin towards the mouth and then towards the posterior. 3. Using fingers or scalpel, free the skin from the underlying body wall. Pin back the skin. 4. Open up the body wall in the abdominal region of animal BA 1. 5. Pull the xiphoid cartilage downward and fix its position with a thread pulled back and pinned to the dissecting board in between the legs. 6. Using scissors make an incision by cutting a little anterior into the diaphragm, into the thoracic cavity. Continue cutting the sidewall down towards the dorsal surface. Take care not to cut too far down. 7. Continue cutting the sidewalls towards the thoracic apex. You will be cutting the ribs, intercostal muscles, and pectoral muscles. (Make sure that the tip of the scissors is always pointing upwards to avoid damaging the internal organs). 8. Once the thoracic wall is free lift up the whole ventral thoracic wall as if you are lifting a cover. STPM BIOLOGY STUDENT’S MANUAL 2010/2011

9. Remove the ventral thoracic wall by cutting the tissues near the apex. 10. Lift up the cut pectoral muscles. Carefully separate the muscles from the thoracic wall without damaging the underlying veins. Once the deeper muscles have been removed, you will be able to see the clavicle. Cut this bone in the middle taking care not to damage nearby veins. 11. Cut and remove the neck muscles to expose the trachea and larynx. 12. Remove the thymus gland and superfluous fat from the displayed section. 13. Push and pin the heart and lungs to the right side of the animal.

14. Examine and identify the veins on either the left or the right side of the animal. 15. Make a large labelled drawing to show the veins in the thoracic region of animal BA 1. State the scale of your drawing. Drawing

Scale of drawing ...................................................... STPM BIOLOGY STUDENT’S MANUAL 2010/2011

16. Examine and identify the arteries on both sides of the thorax. Make a large labelled drawing to display the arteries on both sides of the thorax. State the scale of your drawing. Drawing

Scale of drawing .................................................. STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

17. Cut the anterior section of the larynx. Using forceps, hold the cut end of the larynx, and then loosen the larynx and trachea from the tissues underlying it by cutting off some of the connective tissues where necessary. Determine the main blood vessels, which pass through the diaphragm and cut these vessels at the region near the anterior diaphragm. Cut away tissues where necessary to free and remove the heart, lungs, trachea, and main blood vessels together. 18. Remove each lung by cutting the pulmonary arteries and veins. Cut nearest to the lungs so that the maximum length of each blood vessel can be displayed.

19. Draw a dorsal view of the heart with its blood vessels attached to it. Drawing

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 12 (Practical No. 18)
Title: Examining slides of liver and kidney Purpose:
1. To understand the structures of the liver and kidney so as to reinforce the theoretical understanding of the functions of the two organs in homeostasis

2. To understand the important structures in the liver and kidney which function in homeostasis

Theory
The liver and kidney are important homeostatic organs.

Apparatus and materials
Microscope Prepared slides of liver and kidney

Method
1. Observe the slides under low power microscope to determine the plan of general tissue distribution. Examine the detailed structures under high power microscope by observing the form of the cells and other features. 2. Make two drawings for each slide. (i) Drawing of the outline as seen under low power microscope. Do not draw any cells. (ii) Detailed drawing as seen under high power microscope showing accurate cell/ tissue characteristics. Draw a few cells only. 3. Each drawing must have a complete title which gives the following information: Name of organ, type of section, and magnification. 4. Examine the prepared slides of liver and kidney. Make a large labelled plan drawing of each tissue. 5. Note that the liver consists of many lobules. For each lobules, liver cells (liver cord) are arranged in rows in between sinusoids which are blood channels. Bile canaliculus lies in between liver cords. Central vein is a tributary of the hepatic vein. The portal area contains the bile ducts, branches of hepatic artery, and branches of hepatic portal vein. 6. Examine the kidney slide under low power microscope and identify the characteristics of the various structures found in this organ. Observe the capsule (connective tissues), pelvis, cortex and medulla. Note that the Malphigian corpuscle consists of the glomerulus and the Bowman’s capsule, the cuboid epithelial cells of the proximal convoluted tubules have brush borders while the distal convoluted tubules have bigger lumen and without brush borders. 7. State the magnification of your drawing. STPM BIOLOGY Plan drawing Liver slide STUDENT’S MANUAL 2010/2011

Magnification of drawing ............................................ Kidney slide

Magnification of drawing ............................................. STPM BIOLOGY STUDENT’S MANUAL 2010/2011 High power drawing Liver slide

Magnification of drawing .................................................. Kidney slide

Magnification of drawing ............................................ STPM BIOLOGY STUDENT’S MANUAL 2010/2011

Questions
1. What are the blood vessels, which feed the liver? ............................................................................................................................................... . 2. What is meant by emulsification?

............................................................................................................................................... . 3. State the name of the two types of bile salt. ............................................................................................................................................... . 4. Why is bile salt not considered as an enzyme? ............................................................................................................................................... . ............................................................................................................................................... . 5. Why do mammals have to get rid of their excretory products? ............................................................................................................................................... . ............................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 13 (Practical No. 22)
Title: Examining slides of Spirogyra, Funaria, Marchantia, Dryopteris, and Pinus. Purpose:
1. To identify morphological and anatomical features of plants 2. To improve one’s understanding in the field of taxonomy 3. To identify specific features for the purpose of grouping an organism into a particular phylum and class

Apparatus and materials
Microscope Spirogyra (B1) Funaria (B2) Marchantia (B3) Dryopteris (B4) Pinus (B5)

Method
Examine the specimens given under low and high magnification. Make a large labelled drawing of each of the specimens. Drawing B1 B2

B3

B4

B5

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Questions
Spirogyra (B1) slide 1. (i) Make a large labelled drawing to show a typical vegetative structure of the specimen B1.

(ii) From your observation of the vegetative structure of the organism, suggest its nutritional method. 2. (i) Under which phylum is the specimen B1 classified? (ii) Give reasons for your answer. 3. Identify the structure, which is produced from fertilisation. Funaria (B2) slide 1. (i) State the phylum and class for the specimen B2. (ii) Give reasons for your answers with reference to the observable features only. 2. (i) State the male and the female reproductive organs of B2. (ii) State the names of the male and female gametes of plant B2. 3. Suggest a possible habitat for the specimen B2.

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Marchantia (B3) slide 1. (i) Under which plant group is the specimen B3 classified? (ii) Give reasons for your answer with reference to the observable features only. 2. State the male and female reproductive organs of the specimen B3. 3. Suggest a possible habitat for the specimen B3. Slide of Dryopteris (B4) leaf section 1. (i) Identify the organ from which the specimen B4 was prepared. (ii) Give reasons for your answer based on the observable structures. 2. (i) Which plant group is the specimen B4 classified? (ii) Give reasons for your answer with reference to the observable features only. Slide of structures of the reproductive organ/Pinus cone (B5) 1. Identify the organ from which the specimen B5 was taken. 2. (i) Which plant group is the specimen B5 classified? (ii) Give reasons for your answer with reference to the observable features only.

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 14 (Practical No. 23)
Title: Investigating the structure of flowers Purpose:
A. To investigate the morphology of the Flame of the Forest (Delonix regia) flower and its relation to its functions

Apparatus and Materials
Flame of the Forest flowers (Delonix regia) (2 flowers) A sharp scalpel/razor blade Magnifying glass

Procedure
By using a sharp scalpel, cut the flower into two equal halves. Make a large labelled drawing of the dissected flower. State the scale of your drawing. Repeat this procedure for orchid flower. Drawing Flame of the Forest flower (Delonix regia)

Scale ......................................... STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

B. To investigate the morphology of the orchid flower (Dendrobium) and its relation to its function.

Apparatus and Materials
Orchid flower (Dendrobium) (2 flowers) A sharp scalpel Magnifying glass

Procedure
By using a scalpel cut the flower into two equal and opposite halves. Make a large labelled drawing of the dissected flower. State the scale of your drawing. Drawing Orchid flower (Dendrobium)

Scale .............................................. STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Questions
State the family, type of ovary, and the symmetry of both flowers being investigated. Flame of the Forest Type of ovary …………………………... Symmetry of flower ……………………. Orchid …………………………………………… ……………………………………………

Family …………………………………... ……………………………………………

Questions
What are the special features, which both flowers (Flame of the Forest and orchid) have to ensure the success of their fertilisation process? ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 15 (Practical No. 28)
Title: Monohybrid and dihybrid crosses and use of the χ2 test. Purpose:
1. To know the calculations for the ratio of heritable characters (traits) which obeys Mendel’s First and Second Laws by doing calculations on the product of monohybrid and dihybrid crosses. Using the χ2 test to determine whether or not the calculations (experimental data) obey Mendel’s Law.

Introduction
The basic rules governing inheritance were first discovered by Gregor Mendel, an Australian monk who was also a scientist. In 1866 he proposed a model conceptualised by how seven characters of the garden pea plants were inherited. Unfortunately, the importance of Mendel’s theory was not recognised by the scientific community until early 1900. After his death, researchers studied the research papers, which he published, and realised that he has discovered the principles of hereditary, which they had been looking for. Why was his discovery not recognised when it was first published? At that time, the processes of cell division and fertilisation were not yet fully understood. Furthermore, the theory of inheritance at the time was based on the “blending” hypothesis, not the “particulate” hypothesis of inheritance. In addition, Mendel’s reasonings were only based on the statistical analysis of his crossbreeding experiments and not from the observation of biological structures. For these reasons, no one took notice of the writings of this unknown monk-scientist. After the rediscovery of his writings, his theory has repeatedly been proven to be true and was finally made into a law by modern biologists. Mendel’s procedure is a perfect role model for young scientists like you. Ask the right questions, collect the necessary qualitative data, analyse them, and make a wise logical conclusion. Alleles are alternative forms of the gene for one characteristic. If only a pair of alleles (for one characteristic) is considered, this cross is known as a monohybrid cross. If two pairs of alleles (for two different characteristics) are considered, this cross is known as a dihybrid cross. Genetic constitution (all genes/alleles present) of an individual is known as genotype. Physical appearance of an individual as a product of expression of alleles is known as phenotype. When a pair of alleles exist in the heterozygous form (e.g. Yy) and one of them expresses its phenotypical effect fully and suppresses the phenotypical effect of the other allele, this allele is dominant to the other allele. The suppressed allele is the recessive allele. As an example, the allele Y produces the yellow colour of corn seeds and the allele a produces the red colour of corn seeds. In the heterozygous form, Yy, the colour of corn seed is yellow. The alleles can also exist in the homozygous form, that is YY (homozygous dominant) or yy (homozygous recessive). The recessive allele would express its

phenotypical effect only in the homozygous form (in this case, corn seed colour is white). The dominant allele will always express its phenotypical effect, either in the homozygous (YY) or heterozygous (Yy) form. STPM BIOLOGY STUDENT’S MANUAL 2010/2011 Note: (a) A dominant allele and a recessive allele are usually written with a capital letter and a lower case letter respectively. (b) Mendel’s Law of inheritance would only be obeyed if alleles exist in the recessive and dominant form. The monohybrid ratio 3:1 and dihybrid ratio 9:3:3:1 are hypothetical estimations based on the following: (1) dominance/recessive; (2) segregation; (3) independent assortment; and (4) random fertilisation. The last three processes are influenced by chance events and will be subjected to normal deviation. To assess a genetic hypothesis, we need a test, which can change the deviation from the expected value/ratio to the probability that chance alone could be responsible for the deviation. Furthermore, this test must also take into account the sample size and the number of parameters (degree of freedom). Degree of freedom is the number of parameters considered (n) minus one. For most problems in genectics, degree of freedom (df) is one less the number of classes of phenotypes. As an example, in a monohybrid cross where only the colours of corn seeds are yellow and red, is considered (n = 2), df is n − 1 = 2 − 1 = 1. In a dihybrid cross where the colours of corn seeds yellow and red, and the pattern of striped and white spotted corn seeds are considered (n = 4), df is n − 1 = 4 − 1 = 3. The χ2 test takes into account all these factors. χ2 table
n/p 1 2 3 4 5 6 7 8 9 10 .99 . 00016 .0201 .115 .297 .554 .872 1.239 1.646 2.088 2.558 .98 . 00063 .0404 .185 .429 .752 1.134 1.564 2.032 2.532 3.059 .95 .0039 .103 .352 .711 1.145 1.635 2.167 2.733 3.325 3.940 .90 .016 .211 .584 1.064 1.610 2.204 2.833 3.490 4.168 4.865 .80 .064 .446 1.005 1.649 2.343 3.070 3.822 4.594 5.380 6.179 .70 .148 .713 1.424 2.195 3.000 3.828 4.671 5.527 6.393 7.267 .50 .455 1.386 2.366 3.357 4.351 5.348 6.346 7.344 8.343 9.342 .30 1.074 2.408 3.665 4.878 6.064 7.231 8.383 9.524 10.656 11.781 .20 1.642 3.219 4.642 5.989 7.289 8.558 9.803 11.030 12.242 13.442 .10 2.706 4.605 6.251 7.779 9.236 10.645 12.017 13.362 14.684 15.987 .05 3.841 5.991 7.815 9.488 11.070 12.592 14.067 15.507 16.919 18.307 .02 5.412 7.824 9.837 11.668 13.388 15.033 16.622 18.168 19.679 21.161 .01 6.635 9.210 11.341 13.277 15.086 16.812 18.475 20.090 21.666 23.209

p − Probability n − Degree of freedom

Materials
Special type of corn

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Procedure
A. Monohybrid cross The corn provided is a special type of corn. The colours of corn seeds (kernels) are yellow and red. After carrying out a few crosses, it was found that the colour of the yellow seeds is dominant to the red. Using symbol Y for the dominant allele and y for the recessive allele, a cross can be presented as follows (assuming that students have been taught how to carry out the crosses in theory): P: YY × yy (Yellow) (Red) F1 : Yy × Yy (Yellow) (Yellow) Selfing 3/4 Y −:1/4 yy (Yellow) (Red) (expected ratio)

F2 :

1. Examine the corn. Count all the yellow and red seeds. Record your data in the table below. 2. From the total number of the corn seeds (yellow and red), calculate the expected number of each seed colour. 3. Calculate the value of the χ2 and make a conclusion as to whether or not the results that you have obtained can be accepted as having obeyed Mendel’s First Law.
MPM Phenotyp e Expecte d ratio Observatio n (o) Expected number of count (e) Divergenc e (o − e) Divergence
2

(o − e)2

Divergence2/ Expected no. of count (o − e)2/(e)

Total

Conclusions
...............................................................................................................................................

............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

B. Dihybrid cross The corn provided is a special type of corn. The colours of the seeds (kernels) are yellow and red, white striped and white spotted. After carrying out a few crosses, it was found that the colour of the yellow seeds is dominant to the red, and the white striped is dominant to the white spotted. Using symbol Y for yellow, y for red, W for white striped and w for white spotted, the cross can be presented as follows (assuming that students have been taught how to carry out the crosses in theory): P : YYWW × yyww (Yellow, striped) (Red, spotted) F1 : YyWw × YyWw (Yellow, striped) (Yellow, striped) selfing F2 : 9/16 Y-W- : 3/16 Y-ww : 3/16 yyW- : 1/16 yyww (Yellow, (Yellow, (Red, (Red, striped) spotted) striped) spotted) (expected ratio) 1. Examine the corn and count all the striped yellow, spotted yellow, striped red and spotted red seeds and record your data in the table below. 2. From the total of all the corn seeds (striped yellow, spotted yellow, striped red and spotted red) calculate the expected number of each seed colour and seed pattern. 3. Calculate the value of the χ2 and make a conclusion as to whether or not the results that you have obtained can be accepted as having obeyed Mendel’s Second Law.
MPM Phenotyp e Expected ratio Observatio n (o) Expected number of count (e) Total Divergenc e (o − e) Divergence
2

(o − e)2

Divergence2/ Expected no. of count (o − e)2/(e)

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Conclusions
............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... . ............................................................................................................................................... .

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Experiment 16 (Practical No. 29)
Title: Construction of dichotomous key using local specimens Purpose:
1. To inculcate nature-loving attitude 2. To enable students to identify the external features of an organism 3. To enable students to classify living organisms 4. To enable students to identify the specific features which enable an organism to be grouped into a phylum or a class 5. To increase students’ knowledge in the field of taxonomy

Apparatus and materials
Microscope Magnifying glass (× 10) Amoeba (slide) Hydra (slide) Ant Snail (e.g. garden snail) Shrimp Marchantia/moss Dryopteris/other ferns bearing sori on undersides Grass

Procedure
1. Examine the organisms given to you. 2. Note down the opposing features, which clearly differentiate the organisms from one another. 3. Group these organisms according to the opposing features. 4. Construct a dichotomous key for this group of organisms up to the category of phylum and class.

STPM BIOLOGY

STUDENT’S MANUAL 2010/2011

Results
A1 Organism containing chlorophyll ......................... refer to B A2 Organism without chlorophyll ..................…….... refer to D B1 Organism does not produce spores. Organism has roots, stems and leaves. .......……... Phylum Angiospermae (Grass) B2 Organism produces spores ..........................……... refer to C C1 Organism has leaves (fronds) sub-divided into pinnae and pinnules bearing sori on undersides ...................………….. Phylum Filicinophyta (Dryopteris) C2 Organism with thalloid body, has gemma cups, antheridiophores and archegoniophores as reproductive structures................………………... Phylum Bryophyta (Marchantia) D1 Unicellular organism ......................……………. Phylum Protoctista (Amoeba) D2 Multicellular organism ....................................... refer to E E1 Organism without exoskeleton but has soft cylindrical body. Body wall contains two layers of cells − ectoderm and endoderm Has tentacles and a gut cavity with a single opening called a mouth ……….....................……. Phylum Cnidaria (Hydra) E2 Organism with exoskeleton .............................…. refer to F F1 Invertebrate, has soft muscular foot, and calcareous shell ..........………………………. Class Mollusca (Snail) F2 Invertebrate with jointed legs .............…………... refer to G STPM BIOLOGY STUDENT’S MANUAL 2008/2009 56 G1 Invertebrate, body divided into head, thorax and abdomen. Has three pairs of legs, wingless ...................……….. Class Insecta (Ant) G2 Invertebrate, body divided into cephalothorax and abdomen. Has two pairs of antennae and five pairs of legs ............................…………. Class Crustacea

(Shrimp) STPM BIOLOGY STUDENT’S MANUAL 2008/2009 57

Questions
1. (a) State a possible habitat for Marchantia. ............................................................................................................................................... ..... (b) State the special adaptational features which enable Marchantia to live successfully in the habitat mentioned in (a). ............................................................................................................................................... ..... 2. List down two features, which enable biologists to group ant and shrimp into the same phylum. ............................................................................................................................................... ..... ............................................................................................................................................... ..... 3. State two structural differences, which can be observed between Marchantia and Dryopteris. Marchantia ............................................................................................................................ .... ............................................................................................................................................... ..... Dryopteris ............................................................................................................................. ..... ............................................................................................................................................... ..... 4. State two structural differences, which can be observed between ant and shrimp. Ant ......................................................................................................................................... ... ............................................................................................................................................... ..... Shrimp ……....... ........................................................................................................................

…………………………………………………………………………………………… …….. STPM BIOLOGY STUDENT’S MANUAL 2008/2009 58

Experiment 17 (Practical No. 30)
Title: Preservation procedure Purpose:
1. To learn how to preserve the insects and plants

2. To improve the students’ understanding in the field of taxonomy 3. To identify the morphological features of the animals and plants 4. To draw and label the preserved animal and plant specimens 5. To acquire the skill to determine the phylum, class, and order of the preserved animals and plants

Apparatus and materials
Materials for preparing FAA solution 90 cm3 acetone 50% or 70% 5 cm3 glacial acetic acid 5 cm3 formalin solution Naphthalene Formalin solution Small insect − house fly (alive and complete) Large insect − locust (alive and complete) – or other insects of the same size Plant − Fern leaf/Dryopteris Marchantia (liverwort)/other Bryophyta UHU glue Pin Polystyrene board Box Labels Cotton wool Newspapers Oven − for dying insects Magnifying glass × 10 STPM BIOLOGY STUDENT’S MANUAL 2008/2009 59

Procedure
1. Preservation of small insect, e.g. house fly 1.1 Kill the insect using chloroform. 1.2 Dry it in the oven. 1.3 Pierce the pin through the insect’s body. Apply a little UHU glue to fix the pin to the insect’s body. 1.4 Stand the insect on the polystyrene board. 1.5 Keep the insect in a box together with a naphthalene ball to get rid of mites. 1.6 Label the specimen. 2. Preservation of large insect, e.g. locust 2.1 Kill the locust using chloroform. 2.2 Cut the abdomen dorsally to remove the digestive system to prevent the insect from rotting and to ensure good quality product, which can be kept in good condition for a longer period of time.

2.3 Soak the insect in formalin solution for a short while. 2.4 Insert a small wad of cotton wool into the abdomen to keep its original shape. 2.5 Dry the insect in the oven. 2.6 Repeat steps 1.3 to 1.6. 3. Preservation of plant in liquid 3.1 Preparation of the FAA solution Mix 90 cm3 acetone 50% or 70%, 5 cm3 glacial acetic acid, and 5 cm3 formalin, stir the solution, and leave it to stand for 24 hours. 3.2 Soak Marchantia plant in the FAA solution. 4. Preservation of plant by drying procedure 4.1 Place Dryopteris leaf in between two pieces of newspapers and press the leaf (using heavy object) to flatten it. 4.2 Leave it to dry for a week. 4.3 Display the dried leaf on paper. STPM BIOLOGY STUDENT’S MANUAL 2008/2009 60 Questions 1. (a) State the phylum for house fly and locust. ............................................................................................................................................... ..... (b) State the observable features, which can be used to support your answers. ............................................................................................................................................... ..... ............................................................................................................................................... ..... 2. (a) State the classes for the animals. ............................................................................................................................................... ..... (b) State the observable features, which can be used to support your answers. ............................................................................................................................................... ..... ............................................................................................................................................... ..... 3. State the orders for house fly and locust. ............................................................................................................................................... ..... ............................................................................................................................................... ..... 4. Complete the classification table below for the plant specimens.

Specime n Moss Fern

Divisio n

Clas s

5. Observe the underside of a Dryopteris leaf using a magnifying glass. What is the structure that you can see and what is the importance of this structure to the plant? ............................................................................................................................................... ..... ............................................................................................................................................... ..... STPM BIOLOGY STUDENT’S MANUAL 2008/2009 61

Experiment 18 (Practical No. 31 (a))
Title: Collection of 25 species of insect (group work) Purpose:
1. To be aware of the diversity of insects in our own country 2. To increase the knowledge and interest in the subject of Biology 3. To arouse the interest in the history of nature 4. To inculcate the feeling of love towards all living things 5. To learn the insect behaviour, way of life, feeding habits, methods of reproduction, locomotion, and others 6. To strengthen the theoretical understanding of naming and classification 7. To strengthen the power of observation

Apparatus and materials
Pooter or aspirator Insect net Collecting tubes Killing jars Magnifying glass Insect tubes 4% −10% formalin

Procedure
Collection of 25 insect specimens up to the category of order 1. Some of the methods that can be used to collect insects are as follows: (i) Beat small trees and tree branches using a sweep net (sweep netting). This causes the insects to fall into the net. Remove the insects and place them on a white material and examine them.

(ii) Hold a beating tray underneath small trees or tree branches, shake or beat the trees. Insects will fall onto the tray and examine them. (iii) Butterfly net is used to catch butterflies and other flying insects. (iv) Use a light trap or span a white material vertically and also place another one at the base. Switch on a light bulb. This will attract insects, which come out at night. The insects can then be collected and examined. (v) Use an aspirator or pooter to catch small insects through suction. STPM BIOLOGY STUDENT’S MANUAL 2008/2009 62 2. Transfer the collected insects into collecting tubes or killing jar. 3. Preserve the insects by following the correct procedures according to their sizes. 4. Clearly display the collected insects. Fully label each insect as follows. Local name: ............…………………..................... Order: ……………….............................................. Location: ............……………................................. Habitat: ........................………………................... Date of collection: ...................…........................... Collector’s name: ................................……............

Reference
1. Pengumpulan, pengawetan, dan pengelasan by Dr Mohamed Salleh, DBP, Kuala Lumpur 1990. STPM BIOLOGY STUDENT’S MANUAL 2008/2009 63

Experiment 19 (Practical No. 31 (b))
Title: Collection of 25 species of plant (group work) Purpose:
1. To be aware of the diversity of plants in our own country 2. To increase the knowledge and interest in the subject of Biology 3. To arouse the interest in the history of the nature 4. To inculcate the feeling of love towards all living things 5. To strengthen the theoretical understanding of naming and classification 6. To strengthen the power of observation

Apparatus and materials
Plastic bags or vasculum Thread Old newspapers Procedure 1. Choice of specimen (i) Specimen for preservation must be normal that is (a) not abnormal specimen

(b) not too young (c) not too old 2. Preservation process (i) Specimen must be pressed immediately after collection. (ii) Only the leaf and stem (flowers if any) need to be preserved. (iii) Pieces of papers (6 in × 10 in or 13 in × 8 in) must be used to display the specimen. Use of exercise books to replace loose pieces of paper is not allowed. (iv) Fully label each prepared plants as follows: Local name: .........................................…….. Family: .........................................…………. Location: .........................................……….. Habitat: .........................................………… Date of collection: ........................................ Collector’s name: ......................................... STPM BIOLOGY STUDENT’S MANUAL 2008/2009 64

Assessment
Students must collect the plants from at least 5 different families.

Reference
1. Hsuan Keng, Orders and families of Malayan Seed Plants, University of Malaya Press, Kuala Lumpur, 1969. STPM BIOLOGY STUDENT’S MANUAL 2008/2009 65

Experiment 20 (Practical No. 33)
Title: Ecological study of a terrestrial or an aquatic area. (group work) Purpose:
1. Learning the basic principles of ecology through students’ own effort 1.1 Elements of ecosystem: biosis and abiosis 1.2 Dynamic relationship of elements and flow of energy through ecosystem 2. Using the simple apparatus and instruments in ecological studies 3. Learning the methods of collecting and analysing ecological data 4. Writing an ecological study report 5. Inculcating nature loving attitude 6. Inculcating good moral values−cooperation, independence, and self-confidence

Procedure
Students are divided into groups of 4 or 5. Each group is lead by a leader who plans study proposals they have chosen. Determine the area to be studied. Determine the objectives; rough working plan, and the techniques to be carried out. Each group will have a discussion with their teacher after completing the project.

Keep the record of each project, the product of the project, and the students’ attendance. Send in a report containing the objectives of the product and the conclusions.

Assessment
(1) Assessment of the folio for the field work emphasises the way of writing of project proposal and project report. (2) Project leader will be given 2 additional marks higher than the rest of the group members. (Total mark is 20). (3) Project leaders must hand in a confidential report on the participation of project members. (4) Members of the same group may not necessarily obtain the same mark. (5) Project report must be printed. STPM BIOLOGY STUDENT’S MANUAL 2008/2009 SOIL ANALYSIS 1. Soil sampling technique Apparatus: Metal cylinder and piston (to dig out soil) Procedure: (a) Press the metal cylinder into the soil. (b) Using the piston, remove the soil sample from the cylinder. 2. Determination of the texture of soil Apparatus: 500 cm3 measuring cylinder 100 cm3 soil sample 300 cm3 water Procedure: (a) Add the soil sample to the measuring cylinder and cover with water. (b) Shake the contents vigorously. (c) Allow the mixture to settle out, according to density and surface area of particles, for 48 hours. (d) Measure the volume of the various fractions of soil sample. Results: Calculate the percentage of stone, sand, and clay components of the soil sample. Formula: 100% sample soil ofWeight sandofWeight component sand %×= 3. Determination of water content of soil Apparatus: Aluminum foil pie dish Balance Oven Desiccator Tongs Thermometer Material: 80 gm soil Procedure: (a) Weigh an aluminum foil pie dish while still empty. Record the mass (a).

(b) Add the broken-up soil sample to the pie dish and weigh. Record the mass (b). (c) Place the pie dish containing the soil sample in the oven at 110 °C for 24 hours. (d) Remove the sample from the oven and cool in a desiccator.

66 STPM BIOLOGY STUDENT’S MANUAL 2008/2009

(e) Weigh the sample when cool, and record the mass. (f) Return the sample to the oven at 110 °C for a further 24 hours. (g) Repeat stages (d) and (e) until consistent weighings are recorded (constant mass). Record the mass (c). (h) Calculate the percentage water content as follows:
100×−−abcb

(i) Retain the soil sample in the desiccator for experiment 4. Results: Calculate the percentage water content of the soil sample. Formula: 100% soil ofWeight water ofWeight soil ofcontent water ×=% 4. Determination of organic matter content. Apparatus: Desiccators and lid Tripod Bunsen burner Asbestos mat Fireclay triangle tongs Material: Dried Soil Sample Procedure: (a) Heat the crucible and lid strongly in the Bunsen Flame to remove all traces of moisture. Place in the desiccator to cool. Weigh and record the mass (a). (b) Add the dried soil sample (kept from the previous experiment) from the desiccator and weigh. Record the mass (b). (c) Heat the soil sample in the crucible, covered with the lid, to red-heat for 1 hour to burn off all the organic matter. Allow to cool for 10 min and remove to the desiccator. (d) Weigh the crucible and sample when cool. (e) Repeat (c) and (d) until constant mass is recorded. (f) Calculate the percentage of organic content as follow:
100×−−abcb

(g) Repeat the experiment on soil samples taken from different areas to demonstrate variation of organic content.

67 STPM BIOLOGY STUDENT’S MANUAL 2008/2009

Result: Calculate the percentage of organic matter of the soil sample. Formula: 100% sample soil ofWeight matter organicofWeight component organic of %×= 5. Determination of air content of soil. Apparatus: Tin can of volume about 200 cm3 500 cm3 beaker Metal seeker

Material: Water
Procedure: (a) Place the empty can open end uppermost into the 500 cm3 beaker and fill the beaker with water above the level of the can. Mark the water level in the beaker. (b) Carefully remove the can containing the water and measure this volume of water in a measuring cylinder. Record the volume (a). The water level in the beaker will fall by an amount corresponding to the volume of water in the can. (c) Perforate the base of the can using a drill, making about eight small holes. (d) Push the open end of the can into soil from which the surface vegetation has been removed until soil begins to come through the perforations. Gently dig out the can, turn it over and remove soil from the surface until it is level with the top of can. (e) Place the can of soil, with open end uppermost, gently back into the beaker of water and loosen soil in the can with seeker to allow air to escape. (f) The water level in the beaker will be lower than the original level because water will be used to replace the air which was present in the soil. (g) Add water to the beaker from a full 100 cm3 measuring cylinder until the original level is restored. Record volume of water added (b). (h) The percentage air content of the soil sample can be determined as follows:
100×ab

(i) Repeat the experiment on soil samples from different areas.

68 STPM BIOLOGY STUDENT’S MANUAL 2008/2009

Results: Calculate the percentage volume of air in the soil sample. Formula:
sample soilin air of volume% 100% sample soil of Volumesoil) free-(air particles soil of volume- sample soil of Volume×= 100% sample soil of Volume soilin air of Volume×=

6. Determination of soil pH Apparatus: Long test-tube Test-tube rack Spatula 10 cm3 pipette Material: Universal indicator Procedure: (a) Add about 1 cm3 of soil to the test-tube and 1 cm3 of barium sulphate, which ensures flocculation of colloidal clay. (b) Add 10 cm3 of distilled water and 5 cm3 of BDH universal indicator solution. Seal the test-tube with the bung. Shake vigorously and allow contents to settle for 5 min. (c) Compare the colour of liquid in the test-tube with the colours on the BDH reference colour chart and read off the corresponding pH. (d) Repeat the experiment on soil samples from different areas. Results: State the pH value of the soil.

69 STPM BIOLOGY STUDENT’S MANUAL 2008/2009

DETERMINATION OF THE TYPES OF SOIL ORGANISMS Apparatus: Tullgren funnel Retort stand Beakers Magnifying glass Microscope, glass slide Bearmann funnel Material: 4% formalin solution Results: 1. List down the types of animal such as Nematoda, Annelida, Myriapoda, Insecta, Mollusca, and Amoeba. 2. State the name of the above animals and draw the appearance of the animals. DETERMINATION OF THE DENSITY OF PLANT SPECIES IN A HABITAT The density of plant species in a habitat can be determined using quadrats and transects. 1. Quadrat sampling technique Apparatus: Quadrats measuring 1 m2 Procedure: (i) Systematic sampling procedure−quadrats are placed at the same intervals along transects which runs across the investigated area at the same intervals. (ii) Random sampling procedure−using random number table Systematic distribution Random distribution of quadrats of quadrats

70 STPM BIOLOGY STUDENT’S MANUAL 2008/2009 71

Results: Students must write their reports as follows: Student’s name ..................................... Date ........................…………….......... Habitat .................................................. Location/Place ...................................... Type of plant ........................................ Quadrat size .......................................... Table of data for the measurement of each species cover in quadrat sampling Total species No Species cover (base/air) in Percentage cover for 10 . quadrat cover (%) quadrats 1 2 3 4 5 6 7 8
1 2 3 4 5 6 7 8 9 10

9

10

Students are required to determine the percentage of relative species cover, relative density, and relative frequency of each plant species. STPM BIOLOGY STUDENT’S MANUAL 2008/2009 72 2. Sampling technique using line transect Apparatus: Rope (15.30 meters) Procedure: (i) Determine a base line along the border of the area under investigation.

(ii) Choose a series of points along this base line either randomly or systematically. These points are used as the starting points for the transects to run across the area being investigated. (iii) Record only the plants which touch the line as seen vertically above or below the transect line. (iv) 10 − 20 lines are placed randomly in the area to provide enough samples to investigate the community. Results: Students must write their reports as follows: Student’s name …........................................... Date ..........………….................. Habitat ............................................................ Location/Place ................................................ Type of plant .................................................. Distance of each interval ................................. Total number of intervals ................................ Total length of line transect ............................. Name of No Number of interval species . 1 2 3 4 5 6 7 8 9 10
1 2 3 4 5 6 7 8 9 10 STPM BIOLOGY STUDENT’S MANUAL 2008/2009 73

(a) Calculate the frequency of a species using the following formula:

100%transect ofintervalsofnumber Total foundarespeciesthewhereintervalsofnumber TotalFrequency ×= (b) Calculate % surface cover of each species. 100% transect oflength Total speciesaoflength sectional crossTotalcover species %×= (c) Calculate the relative species cover. 100% species all oflength sectional cross Total speciesaoflength sectional crossTotalcover species Relative×= Summary of the measurements obtained by the line transect technique
No . Name of species Number of intervals where species are recorded 1 2 3 4 5 6 7 8 Percentage cover Relative cover Frequenc y

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