DB024 Lab Manual Updated: 26/04/2022 26 Figure 8.3 Male cone of Pinus sp. (l.s.) Source: https://www.vcbio.science.ru.nl Figure 8.4 Female cone of Pinus sp. (l.s.) Source: https://www.vcbio.science.ru.nl microsporangium microspore microsporophyll Cone axis Cone axis Ovule megasporoph yll
DB024 Lab Manual Updated: 26/04/2022 27 Questions: 1. State the unique characteristics of gymnosperms. 2. In Pinus sp., male cone matured earlier than female cone. Describe how this species ensures the success of pollination? Experiment 8.2: Diversity of Angiosperms Apparatus: Dissecting microscope Razor blade Tile Materials: Fresh specimen: Hibiscus rosa-sinensis Procedures and Observation: 1. Observe the morphology of the flower. Draw and label the stamen and pistil. 2. Dissect longitudinal section of the flower. Draw and label the ovary and ovule. Figure 8.5 Flower structure of Hibiscus rosa-sinensis Source: http://www.natureloveyou.sg/Plant%20Story/Plant%20Story%20- %20Hibiscus.html
DB024 Lab Manual Updated: 26/04/2022 28 Figure 8.6 Longitudinal section of the flower, Hibiscus rosa-sinensis Source: https://www.life.illinois.edu/help/digitalflowers/Malvaceae/6.htm Questions: 1. State the unique characteristics of angiosperms. 2. Which parts of the plants produce male and female gametophytes? 3. What are the differences between the cones and flowers? Ovary Ovule Stamen tube Style
DB024 Lab Manual Updated: 26/04/2022 29 EXPERIMENT 9: ANIMAL DIVERSITY – INVERTEBRATES AND VERTEBRATES Objective: At the end of this lesson, students should be able to identify unique characteristics of invertebrates and vertebrates. Introduction: Animals are successful multicellular eukaryotes belong to Kingdom Animalia. Animals are divided into two groups, whether or not they possess a backbone (vertebral column); invertebrates and vertebrates. More than 90% of animal species found in the world belonged to invertebrates. Invertebrates Invertebrates are multicellular animals without vertebral column. Many invertebrates such as jellyfish or worm have a fluid-filled hydrostatic skeleton that function as a support mechanism. Other invertebrates such as insects and crustaceans possess hard outer shell for protection. Eight phyla of invertebrates are Porifera, Cnidaria, Platyhelminthes, Nematoda, Mollusca, Annelida, Arthropoda and Echinodermata. Vertebrates Vertebrates are classified in the phylum Chordata. Vertebrates are the animals from the groups of jawless fishes, bony fishes, sharks and rays, amphibians, reptiles, mammals, and birds. All vertebrates have the following characteristics during the development stage: i. notochord ii. dorsal hollow nerve cord iii. pharyngeal slits iv. post anal tail v. myotomes The above characteristics may undergo changes or diminish during the embryonic development.
DB024 Lab Manual Updated: 26/04/2022 30 Experiment 9.1: Invertebrates Materials: Preserved/ live specimens / diagrams: Phylum Porifera: Sponges (Leucosolenia sp.) Phylum Cnidaria: Sea fur (Obelia sp.) Phylum Platyhelminthes: Tapeworm (Taenia sp.) Phylum Nematoda: Roundworm (Ascaris sp.) Phylum Annelida: Earthworm (Pheretima sp.) Phylum Mollusca: Garden snail (Achatina sp.) Phylum Arthropoda: Grasshopper (Valanga sp.) Phylum Echinodermata Starfish (Asterias sp.) Procedures and Observation: 1. Observe the morphological characteristics of the given specimens. Experiment 9.2: Vertebrates Material: Diagram: Phylum Chordata: Lancelet (Amphioxus sp.) Procedures and Observation: 1. Identify the unique characteristics of Amphioxus sp. by referring to the diagram. 2. Draw and label the unique characteristics of Amphioxus sp. Question: 1. State the unique characteristics of vertebrates. 2. Draw and label the specimens to show their unique characteristics. 3. Refer to Appendix 9.1 to assist your investigation. Question: 1. State the unique characteristics of invertebrates.
DB024 Lab Manual Updated: 26/04/2022 31 Appendix 9.1: Characteristics of selected phyla in invertebrates Phylum Porifera Example: Leucosolenia sp. (sponges) Characteristics: Parazoa animals (the cells are arranged loosely without real tissue); have simple body structure (saclike bodies perforated by many pores); have asymmetry body; the body consists of two layers of cells (pinacoderm on the external surface and choanoderm on the internal); the choanoderm consists of flagellated collar cells, folded body walls and numerous pores; the body walls are supported by a skeleton from calcium, spicules from silica or spongin fibers; the body cavity is called spongocoel; there is a large body opening (osculum) at one end; undifferentiated nervous system. Phylum Cnidaria Example: Obelia sp. Characteristics: Eumetazoa animals (form tissues); radial symmetry; diploblastic (two germ layers); nervous system consists of network of nerve cells, have tentacles with specialized stinging cells, cnidocytes around the manubrium (mouth), each cnidocyte contain nematocyst; alternation of generation between polyp and medusa stage. Phylum Platyhelminthes Example: Taenia sp. (tapeworm) Characteristics: Eumetazoa animals (form organs); bilateral symmetry; triploblastic (three germ layers); acoelomate (no body cavity); most are parasitic lacks both respiratory and circulatory systems, simple digestive system or none at all; have flame cells (specialised for excretion); have mouth on the ventral side; have eyes on the dorsal side. Phylum Nematoda Example: Ascaris sp. (roundworm) Characteristics: Triploblastic; pseudocoelomates (false body cavity); bilateral symmetry; unsegmented body; have complete digestive tract; the muscles of nematode are all longitudinal; body covered with tough cuticles.
DB024 Lab Manual Updated: 26/04/2022 32 Phylum Annelida Example: Pheretima sp. (earthworm) Characteristics: Triploblastic; coelomate (true body cavity); bilateral symmetry; segmented body; the segments are normally separated by compartments which are called septum; the chaetae on the body are used to help in locomotion. Phylum Mollusca Example: Achatina sp. (garden snail) Characteristics: Triploblastic; coelomate; unsegmented body; has a large muscular body; visceral organ present; has radula; has mantle (may secrete shell). Phylum Arthropoda Example: Valanga sp. (grasshopper) Characteristics: Triploblastic; coelomate; bilateral symmetry body; segmented body; mostly terrestrial; exoskeleton from chitin; paired jointed appendages; respiratory system consists of the tracheal system which opens through spiracles; aquatic arthropods respire through external gills; the sensory and nervous systems are concentrated at the head region. Phylum Echinodermata Example: Asterias sp. (starfish) Characteristics: Triploblastic; coelomate; locomotion using tube feet; has endoskeleton from calcium carbonate plates; simple digestive system; part of the coelom is modified to become water vascular system.
DB024 Lab Manual Updated: 26/04/2022 33 EXPERIMENT 10: CHLOROPLAST IN AQUATIC PLANT (Elodea sp.) Objectives: At the end of this lesson, students should be able to: i. Prepare the slide of an Elodea sp. Leaf; and ii. Examine the internal structure of an Elodea sp. leaf: the cell membrane, cell wall and chloroplasts Introduction: Elodea sp. is an excellent plant for our studies of photosynthesis and cells because the leaves are only a few cells thick so they will be easy for us to observe under the microscope to look at cells and cell parts. Figure 10.1 Elodea sp. View of Elodea sp. leaf
DB024 Lab Manual Updated: 26/04/2022 34 Figure 10.2 Cells of Elodea sp. Figure 10.2 shows leaf cells of an Elodea sp. These living plant cells are viewed by light microscopy. With a microscope we can see the chloroplasts move around inside living cells. Chloroplasts and the other living parts of the cells are found near the cell wall, but not usually in the centre of the cell. Since they contain chlorophyll, which are green, chloroplasts can be seen without staining and are clearly visible within living plant cells. However, viewing the internal structure of a chloroplast requires the magnification of an electron microscope. Apparatus: Microscope Slide Cover slip Toothpick/dissecting pin Forceps Dropper Material: A sprig of Elodea sp.
DB024 Lab Manual Updated: 26/04/2022 35 Procedures and Observation: 1. Obtain an Elodea sp. leaf from the main plant and place it on a clean slide. 2. Place one drop of fresh water onto the leaf, and carefully place the cover slip on top of the leaf. (Caution: make sure there is no formation of air bubbles under the cover slip) 3. Place the slide under the microscope and observe under low power and medium power. 4. Turn the microscope to high power and draw what you see. Label the chloroplasts (the green objects), the cell wall and the cell membrane. Questions: 1. Why would we use an aquatic plant (Elodea sp.) for this experiment instead of a terrestrial plant? 2. Where does aquatic plant get the CO2 for photosynthesis? 3. If a plant were under water and was photosynthesizing, what gas would be visibly bubbling from the plant? 4. Why does chloroplast move in Elodea sp.?
DB024 Lab Manual Updated: 26/04/2022 36 EXPERIMENT 11: HOMEOSTASIS Objective: At the end of this lesson, students should be able to detect the effects of exercise on the body temperature and pulse rate in a human body system. Introduction: Homeostasis is to maintain constant internal environment of the living organism through physiological regulation. Various organs are responsible for regulating different environmental variables. In human, heart rate and temperature need to be regulated within certain set ranges. Hypothalamus for example plays important roles in regulating body temperature through negative feedback mechanism. When body temperature increases, thermoreceptors will detect this changes and cause physiological adjustment to bring the temperature back to normal range. Medulla oblongata likewise plays the similar functions to regulate body pulses. Apparatus: Clinical thermometer Digital pulse monitor Height scale Stopwatch Weighing scale BMI chart Precaution to students: Do not attempt this activity if you have a health problem or if you are recovering from an illness.
DB024 Lab Manual Updated: 26/04/2022 37 Procedures and Observation: 1. Students will be divided into groups to conduct this exercise. Before undertaking this exercise, students should fill vital parameters in Table 11.1. Use Appendix 11.1 to assist you in calculating of body mass index (BMI). Table 11.1 Vital parameters taken before exercise for each student Student’s Name Gender Height (m) Body weight (kg) BMI Average initial pulse rate Average initial body temperature 2. Sit down comfortably on a chair and take 5 minutes to settle, locate your pulse, and count the initial number of pulses per minute. Take three readings and record the average in Table 11.1. 3. Measure your initial body temperature. Take three readings and record the average in Table 11.1. 4. For the first exercise, each student will walk around in class for 5 minutes. Immediately measure the average pulse rate and average body temperature and record in Table 11.2. 5. For the second exercise, each student will run/ jumping jack/ high knees on the spot for 10 minutes (intermediate intensity). Immediately measure the average pulse rate and average body temperature and record in Table 11.2. 6. While resting, take the average pulse rate and average body temperature for every 10 minutes interval in half an hour (three times) and record in Table 11.2. 7. Analyse on class basis the changes in pulse rate and temperature after different exercises according to the gender and BMI and record them in Tables 11.3 and 11.4.
DB024 Lab Manual Updated: 26/04/2022 38 Table 11.2 Vital parameters taken after exercises for each student Pulse rate (beats/ minute) Body temperature (°C) After walking (First exercise) After running/ jumping jack/ high knees (Second exercise) Recovery period After 10 minutes After 20 minutes After 30 minutes Initial (From Table 11.1)
DB024 Lab Manual Updated: 26/04/2022 39 Table 11.3 Results of the effect of exercises on the pulse rate (for the whole class) Gender BMI No. of students Pulse rate (beats/ minute) Initial After walking (First exercise) After running / jumping jack/ high knees (Second exercise) Recovery period After 10 minutes After 20 minutes After 30 minutes Male < 18.5 18.5 – 24.9 25 – 29.9 > 30 Female < 18.5 18.5 – 24.9 25 – 29.9 > 30
DB024 Lab Manual Updated: 26/04/2022 40 Table 11.4 Results of the effect of exercises on the body temperature (for the whole class) Gender BMI No. of students Body temperature (°C) Initial After walking (First exercise) After running / jumping jack/ high knees (Second exercise) Recovery period After 10 minutes After 20 minutes After 30 minutes Male < 18.5 18.5 – 24.9 25 – 29.9 > 30 Female < 18.5 18.5 – 24.9 25 – 29.9 > 30
DB024 Lab Manual Updated: 26/04/2022 41 Questions: 1. What is the significance of homeostasis in human body? 2. What are the normal ranges of human body temperature and pulse rate? 3. Name two other examples of vital parameters in human that are regulated by negative feedback mechanism. Appendix 11.1 Body mass index (BMI) = Body weight (kg) (Height)2 (m) BMI Categories (for adult over 20 years) BMI Nutritional Status Below 18.5 Underweight 18.5 – 24.9 Normal weight 25.0 – 29.9 Pre–obesity 30.0 – 34.9 Obesity class I 35.0 – 39.9 Obesity class II Above 40 Obesity class III Source: WHO/ Europe 2021
DB024 Lab Manual Updated: 26/04/2022 42 Normal Heart Rate by Age Age range Heart Rate (bpm) New-born 100 – 160 0 – 5 months 90 – 150 6 – 12 months 80 – 140 1 – 3 years 80 – 130 3 – 5 years 80 – 120 6 – 10 years 70 – 110 11 – 14 years 60 – 105 15 years and older 60 – 100 Source: emedicine 2021 Healthy adult at rest Healthy adult at rest Normal value Heart rate (pulse) 60 – 100 bpm Respiratory rate 16 – 20 breaths per minute Blood pressure 120/80 mm Hg Temperature 36.6oC – 37oC Source: emedicine 2021
DB024 Lab Manual Updated: 26/04/2022 43 EXPERIMENT 12: COORDINATION Objectives: At the end of this lesson, students should be able to display reflex action of the nervous system. Introduction: Living organisms depend on the information obtained from the environment for their survival. Most of the information about the environment is detected by the special sense organs called the receptor organ. These receptors receive stimuli such as light, sound, temperature, taste and smell in the environment. Although stimuli and sensory organs are different, the sequences of events that occur in the receiving of stimuli are the same (see diagram). External stimuli are received by the sense organs such as ears, eyes, nose, skin and tongue. From sensory organs, the external stimuli are received by one or more sensory neurons that become impulses. Sensory neurons have many of the same character, but different from one organ to another organ that is sensitive to a specific stimulus. Example: there are sensory neurons sensitive to light and sensory neurons that are sensitive to vibration. Central nervous system will translate these impulses and the information obtained is used by the organism to react to the stimulus. Stimulus → Receptor organ → Neuron → Impulses transmit → CNS In humans there are two types of reflexes, which are: tendon reflex and organ reflex. Tendon reflex consists of patella, achilles and plantar reflex. Reflex organs consist of pupil accommodation, ciliospinal and swallow reflex. All the reflexes are to coordinate our body protection. Experiment 12.1: Reflex All experiments must be done in pairs. Apparatus: Torch light Wooden Ruler/ Reflex hammer
DB024 Lab Manual Updated: 26/04/2022 44 Procedures and Observation: Tendon Reflex A. Patella reflex 1. Sit on the table and let your legs freely hang and dangle over the edge of the table. 2. Close your eyes, your partner will knock on your knee tendon by using a wooden ruler/reflex hammer from the side of the leg. 3. Observe knee jerking. (Figure 12.1A) 4. Record your observations in Table 12.1A. Figure 12.1A Tendon Reflex Source: https://surgeryreference.aofoundation.org/spine/trauma/subaxialcervical/further-reading/patient-examination-neurological-evaluation#insci )
DB024 Lab Manual Updated: 26/04/2022 45 B. Achilles reflex 1. Ask your partner to stand on the knee down on one knee on a stool. 2. One of the legs should be bent downward stretching the gastrocnemius muscle. 3. Your partner surprisingly knocks on your Achilles tendon using a wooden ruler/reflex hammer from the side of the leg. 4. Observe the other foot jerking (Figure 12.1B) 5. Record your observations in Table 12.1A. Position of Gastrocnemius muscle and Achilles tendon. Figure 12.1B Achilles reflex Source: https://www.hothonda.top/products.aspx?cname=soleus+running&cid=153
DB024 Lab Manual Updated: 26/04/2022 46 Table 12.1A Observation of tendon reflex Types of reflexes Observation Patella Achilles Organ Reflex C. Pupil accommodation reflex 1. Ask your partner to focus an object at a distance. 2. Observe your partner’s pupil. 3. Ask your partner to focus on a very close object. 4. Observe your partner’s pupil. 5. Flash your partner’s eyes starting from the side of the eyes using a torch light. 6. Observe your partner’s pupil (Figure 12.1C) 7. Record your observations in Table 12.1B.
DB024 Lab Manual Updated: 26/04/2022 47 Figure 12.1C Pupil accommodation reflex when exposed to light Source: https://pmgbiology.files.wordpress.com/2014/05/pupil_reflex_med.jpg Table 12.1B Observation of organ reflex Types of reflexes Observation Pupil a. Distance object: _____________________________ b. Close object: ________________________________ c. Exposure to light i. With flashlight: ___________________________ ii. Without flashlight: _________________________ Questions: 1. What are the functions of reflex? 2. Explain the flow of the reflection process. 3. List the types of neurons that are involved in this experiment.
48 REFERENCES Campbell, N. A., Reece, J. B., Urry, L. A., Cain, M. L., Wassermen, S. A., Minorsky, P. V. & Jackson, R. B. (2018). Biology. (11th Ed.). Pearson Benjamin Cummings. USA. Lawrence, E. (2016). Henderson’s Dictionary of Biological Terms (16th Ed.), Prentice Hall. Solomon, E. P., Berg, L. R. & Martin, D. W. (2018). Biology. (11th Ed.). Nelson Education, Ltd, Canada. Morgan J. G & Carter M. E. B & Stout (2015). Investigating Biology: Laboratory Manual (8th Edition), Pearson Education Limited. www.bio.miami.edu www.crochetspot.com www.k-state.edu www.math.arizona.edu www.news.makemeheal.com www.pc.maricopa.edu www.quia.com www.sci.waikato.ac.nz www.sfsu.edu www.sharewhy.com www.sols.unlv.edu www.stolaf.edu www.users.rowan.edu www.vcbio.science.ru.nl www.wikispace.psu.edu
49 ACKNOWLEDGEMENTS The Matriculation Division, Ministry of Education Malaysia wish to thank everyone who has contributed in shaping and writing this BIOLOGY LABORATORY MANUAL (5 th Edition) for the Two Semesters Matriculation Programme. Special thanks go to those for their many valuable suggestions and conscientiousness in completing this manual. Dr. Hajah Rosnarizah binti Abdul Halim Director of Matriculation Division Haji Mohd. Yusof bin Samad Deputy Director of Matriculation Division (Academic) Mohd Junaidi bin Abd Aziz Senior Principal Assistant Director Ruslan bin Achok Assistant Director Reviewers for the 5 th Edition ● Prof. Ts. Dr. Shahrul Hisham bin Zainal Ariffin, UKM ● Prof Madya. Dr. Yahya bin Mat Arip, USM ● Abdul Aziz bin Abd.Kadir, KMM ● Siti Mahayan binti Hashim, KMM ● Zulkefli bin Shahruddin, KMNS ● Wan Noor Meeraisha binti Wan Jamaluddin, KMNS ● Siti Noor binti Daud@Othman, KMPP ● Siti Aliza binti Shaik Ali, KMPP ● Mohamad Masrizan bin Abdul Patah, KML ● Mohd Badruddin bin Abdullah, KML ● Elham bin Ismail, KMJ ● Azlina binti Ruhani, KMJ ● Norhaizan binti Mohamed Idris, KMPk ● Noorlizawati binti Ishak, KMPk ● Roslan bin Abu Bakar, KMK ● Hjh Intan Zahrah binti Mohd Borkhan, KMK ● Siti Hajar Rifqah binti Md. Khairuddin Pang, KMPh ● Norulhuda binti Che Ismail, KMPh ● Siti Nadira binti Nordin , KMS ● Aniza binti Abu Bakar, KMS ● Rosmah binti Yahya, KMKt ● Nor Maizura binti Harun, KMKt Cover designed by Syed Nassir bin Syed Ahmad, KML
DB014 & DB024