Activity 2 - Determination of Bacterial Motility

Amarillo, Rachelle
Bade, Angelica
Victoria, Celine
HUB35

1. Activity 2:DeterMInation of microbial motility Amarillo, Rachelle Louiese Bade, Angelica Mae Victoria, Celine

2. introduction Many but not all bacteria exhibit motility Flagella, Cilia, Pseudopods (Eukaryotic Cells) Gliding Motility, Spirochaetal Movement (Prokaryotic Cells)

3. Introduction Importance of Bacterial Motility Chemotactic behavior and survival Ability to change direction (moving away or towards repellants or attractants), avoids unfavorable conditions of habitat and choose favorable environment Pathogenesis For attachment and colonization of cell wall of host cell Microbiology Nutrition , Water Expulsion

4. introduction Types of Bacterial Motility 1. Brownian Motion – movement exhibited by particles suspended in liquids due to the bombardment of water molecules example: Saccharomyces cerevisiae , Staphylococcus aureus 2. True Motility – independent movement brought by different mechanisms of self propulsion example: Pond water microorganism, E. coli

5. https:// upload.wikimedia.org / wikipedia /commons/thumb/1/15/ Flagellum_base_diagram_en.svg /1148px-Flagellum_base_diagram_en.svg.png https:// en.wikipedia.org /wiki/ File:Brownian_motion_large.gif

6. INtroduction Bacterial Motility Test A. Non-Pathogenic Wet mount – simplest way to determine motility Hanging drop Method – useful in observing the general shape of living bacteria and the arrangement of bacterial cells when they associate together

7. INtroduction Bacterial Motility Test (cont.) B. Pathogenic Soft Agar Stabbing (Culture Method) Negative result – growth in distinct zone directly along the stab Positive result – indicated by diffuse or cloudy growth mostly at the top or bottom of the stab

8. objectives 1. Observe the motility of the microorganisms using different techniques 2. Recognize Brownian motion from true motility (self – propulsion) based on the movement of particles 3. Identify the observed microorganisms whether they are motile or non- motile 4. Differentiate the movement of microorganisms under wet mount from hanging drop technique

9. Materials Binocular Compound Microscope Alcohol lamp Cultures of Proteus vulgaris Staphylococcus aureus Micrococcus luteus Saccharomyces cerevisiae Pond water Glass slides Petroleum jelly Applicator sticks Cover slips Inoculating loop/ needle Disinfectant Motility Medium

10. Procedure A. Wet Mount Preparations 1. Place small drop of pond water on a clean slide using inoculating loop. Cover with cover slip. 2. Observe under HPO. * repeat steps using Saccharomyces cerevisiae B. Hanging Drop Preparation 1. Place four small drops of petroleum jelly on the edge corner of the coverslip 2. Place two loopfuls of Micrococcus suspension at the center. 3. Invert the coverslip and place it in the depression slide. The drop must hang into the depression but should not touch the bottom of the depression slide. 4. View under HPO.

11. procedure C. Culture Method ( Proteus vulgaris, Staphylococcus aureus, Micrococcus luteus ) Aseptic Technique Touch the bacterial suspension with the cooled needle. Stab in the center of the motility medium. Aseptic Technique Incubate the medium at 37˚C for 48 hours.

12. Asceptic technique ASEPTIC TECHNIQUE

13. RESULTS AND DISCUSSION Microorganisms in pond water (HPO)

14. RESULTS AND DISCUSSION Saccharomyces cerevisiae (HPO)

15. RESULTS AND DISCUSSION What type of motion was exhibited by the microorganisms in pond water and by the Saccharomyces cerevisiae ? Microorganisms in pond water – true motility Saccharomyces cerevisiae – brownian motion

16. RESULTS AND DISCUSSION Differentiate between Brownian motion from true motility. Brownian Motion – the random movement exhibited by particles suspended in liquids due to the bombardment of water molecules. True Motility – an independent movement brought about by different mechanisms of self-propulsion (e.g. flagella, cilia).

17. RESULTS AND DISCUSSION Differentiate the motility of the microorganisms observed in wet mount from hanging drop mount based on the rate of movement. The motility of microorganisms observed in wet mount is slower or eventually won’t have any movement at all in comparison to the hanging drop mount, which, in this case, has an observable original movement exhibited by the microorganisms.

18. Wet Mount Hanging Drop Mount The rate of movement slows down because the wet mount slide dries due to the heat from the light of the microscope. The rate of movement is constant because the petroleum jelly keeps the hanging drop mount hydrated. The microorganisms’ movement is limited because they are pressed in between the flat surface of the slide and the cover slip. The microorganisms can move freely because of the curvature on the slide which provides them space to move.

19. RESULTS AND DISCUSSION Show the dispersion of the cultures in the tube inoculations.

20. Micrococcus luteus Proteus vulgaris Staphyloccocus aureus

21. RESULTS AND DISCUSSION Motility of the Examined Organisms Microorganisms Motility Proteus vulgaris + Staphylococcus aureus - Saccharomyces cerevisiae - Micrococcus luteus -

22. objectives 1. Observe the motility of the microorganisms using different techniques 2. Recognize Brownian motion from true motility (self – propulsion) based on the movement of particles 3. Identify the observed microorganisms whether they are motile or non- motile 4. Differentiate the movement of microorganisms under wet mount from hanging drop technique

23. References https:// en.wikipedia.org/wiki/Brownian_motion http:// www.cellsalive.com/animabug.htm http://www.microbehunter.com/making-a-wet-mount-microscope-slide /