The document outlines microbiology and parasitology laboratory rules concerning the borrowing and returning of equipment, emphasizing proper handling, safety protocols, and management of hazardous materials. It includes detailed instructions on safety measures, such as treating all microorganisms as potential pathogens and stringent waste disposal procedures. The document is a comprehensive guide aimed at ensuring laboratory safety and compliance with hygiene standards.

1. Microbiology &
Parasitology Laboratory

2. Rules Concerning the
Borrowing and Returning of
Apparatus and Equipment

3. 1. Fill up borrowers’ slip issued by the Office of UNP Laboratory Services
2. Borrow and reserve apparatus needed one day before the laboratory period
3. Receive apparatus during the scheduled time of your laboratory.
4. Receive only clean and dry apparatus
5. Check if all chemicals requested and apparatus were issued.
6. Report any breakage or damage to the laboratory staff. Breakage should be
replaced or equivalent cost shall be paid at the cashier’s office. Show receipt to
the lab staff for recording and clearance of your breakage. Wait for the Official
Receipt to be recorded.
7. Return only clean and dry apparatus and equipment.
8. No apparatus and equipment should be taken out from the premises of the UNP.
*Source: UNP Laboratory Operations Manual (adopted for ISO 1900 certification)

4. General Rules and
Practices for Microbiology
Laboratory Safety

5. The following is a list of rules that is designed to ensure your
safety as well as the safety of your classmates and instructor.
Failure to follow these safety rules may result in loss of points from
the related activities. Violation of the rules may result in a referral
to the Office of Student Affairs for disciplinary action.
Although we use organisms that are not usually pathogenic, the
instructors want students to develop appropriate techniques and
safeguards against the day when they will be dealing with
infectious materials. Therefore we perform as if we are in a clinical
laboratory.

6. 1. Please see or the list of Haz waste listed in RA6969. You may be exposed to one
or more of these chemicals during this course. The RA 6969 requires hazardous
waste generators to revise and publish at least once per year the list of chemicals
known to the state to cause cancer or reproductive toxicity.
2. The effects on human pregnancy of chemical agents used in this course are
unknown, and a pregnant woman is advised to consult her physician before
taking this course. Safety data sheets are available upon request. As contact with
dyes used in staining may have potential teratogenic effects, a pregnant woman
should avoid staining procedures as directed by her physician.
3. Treat all microorganisms as potential pathogens. While the majority of
microorganisms are not pathogenic to humans and have never been shown to
cause illness, under unusual circumstances a few microorganisms that are not
normally pathogenic can act as pathogens. Treat all microorganisms—especially
unknown cultures—as if they were pathogenic. A student who has a compromised
immune system or has had a recent extended illness should talk with the
instructor before working in the microbiology laboratory.

7. 4. Please be respectful of one another and the materials used in the
microbiology lab. This includes not crowding one another for resources, careful
use and care of microscopes, replacing prepared slides in their original boxes
and other materials in the appropriate containers, and ensuring everyone’s
safety by using proper aseptic and disposal practices
5. Wear a laboratory gown or other protective clothing. Lab gowns must cover
the knees and should be long sleeved. Avoid wearing garments with loose,
hanging sleeves.(Gram stains have an affinity for your best clothing).
• Wear closed-toe shoes while working in the laboratory.
• Similarly, confine long hair. They are a fire hazard.
6. Sterilize equipment and materials. All materials, media, tubes, plates, loops,
needles, pipettes, and other items used for culturing microorganisms should be
sterilized by autoclaving. Incinerate loops, needles before and after inoculating
microorganisms. Otherwise, use commercially sterilized products. Understand
the operation and safe use of the autoclave, oven and all equipment and
materials needed for the laboratory.

8. 7. Disinfect work areas before and after use. Use a disinfectant, such as a 10%
bleach or 70% ethanol solution, to wipe down benches and work areas both
before and after working with cultures. Also be aware of the possible dangers
of the disinfectant, as 70% ethanol can catch fire around open flame or high
heat sources. Bleach, if spilled, can ruin your clothing. Either alcohol or bleach
can be dangerous if splashed in the eyes. Students should know where the
nearest eyewash station and sink are located.
• Keep working area, sink and room clean.
8. Wash your hands. Use a disinfectant soap to wash your hands before and
after working with microorganisms. Nondisinfectant soap will remove surface
bacteria and can be used if disinfectant soap is not available. Gloves may be
worn as extra protection.
9. Never pipette by mouth. Do not pipette anything by mouth at any time. Use
pipette bulbs or pipetting devices for the aspiration and dispensing of liquid
cultures. Pipette fillers will be supplied for pipetting liquids. Discard all pipettes
into proper receptacles. Use pipettes properly as instructed by the professor.

9. 10. Do not smoke, eat or drink in the lab, nor store food in areas where
microorganisms are stored. Never eat or drink in the laboratory while working
with microorganisms. Keep your fingers, pencils, gum, and other objects outout
of your mouth, and wash your hands before and after the laboratory activity.
Open cuts, lesions, and the like should be covered with a band-aid, tape (gauze
as appropriate), or gloves, when handling viable organisms.
11. Label everything clearly. All cultures, chemicals, disinfectant, and media
should be clearly and securely labeled with their names and dates. Label
properly all inoculated media to be placed in incubators: name, date, section,
name of organism. If they are hazardous, label them with proper warning and
hazardous information.
12. Always handle test tubes by the tube, not the top. Place test tubes in
supportive containers at all times; a test tube rack, beaker, or can. Circular test
supports are to be used only in a double boiler, not for transport of test tubes.
Never lay a test tube directly on a desk or cart. Test tubes roll off and spill.

10. 13. Autoclave or disinfect all waste material. All items to be discarded after a
class, such as culture tubes, culture plates, swabs, toothpicks, wipes,
disposable transfer needles, and gloves, should be placed in a bag and
autoclaved 30 to 40 minutes at 121° C at 20 pounds of pressure. If no
autoclave is available and you are not working with pathogens, the materials
can be covered with a 10% bleach solution and allowed to soak for at least 1 to
2 hours.
14. Discard all used materials into proper containers.
• a. Test tubes to be used again will be sterilized before use. Place test tubes
in designated racks as directed by your instructor.
• b. Place commercially prepared slides back in their original boxes; clean
them first by removing immersion oil with alcohol moistened lens tissue.

11. • Place used slides you have prepared in containers of disinfectant. Be sure
the disinfectant covers the slides. Place swabs in beakers of disinfectant.
Do not overfill the containers.
• d. Place Petri dishes in the appropriate Biohazard container.
• e. Discard stained gloves in the regular trash; discard only de-
contaminated gloves
15. Discard broken glass into the appropriate Broken Glass container. Use the
broken GLASS container for large items such as beakers, and the SHARPS
container for small items such as broken stained slides. Place only broken
glass in the broken glass containers—do NOT place used paper towels, used
but unbroken slides, or other materials in the broken glass container.

12. 16. Clean up spills with care. Cover any spills or broken culture tubes with a
70% ethanol or 10% bleach solution; then cover with paper towels. After
allowing the spill to sit with the disinfectant for for 10 minutes or short time,
carefully clean up and place the materials in a container apprpriate to be
autoclaved. Wash the area again with disinfectant. Never pick up glass
fragments with your fingers or stick your fingers into the culture itself; instead,
use a brush and dustpan. If working with animal or plant pathogens, keep the
area clear and notify your instructorIf a test tube containing bacteria breaks,
notify the instructor. Use the dustpan and brush to gather broken glass; do not
pick up broken glass with a gloved or naked hand.
17. All materials, cultures, and equipment must remain in the lab at all times.
Do not take cultures, test tubes, or other materials from the lab.

13. 18. Refer to your instructor in case of doubt and emergency. Notify the
instructor immediately of any injury, accident, or spill of bacteria. If you get an
irritating chemical in your eye or on your skin you may be escorted to the
eyewash station or the safety shower in the lab. Your instructor will coordinate
first aid efforts, including notification of UNP guards who will call for emergency
medical assistance if needed.
19. Conserve water and energy. Switch off lights, unplug electric stove, turn-off
gas burner if not needed. Check for safety of any gas and electric outlets.
20. Photography, videos, and recordings can only be taken with instructor
consent. They may not be given, sold, or published in print or online without the
written consent of the instructor, and can only be used for the purposes of the
class.
21. In the unlikely event that we must leave the classroom, please reassemble
in the vacant area in front of the Science building.

14. Apparatus and Equipment
Used in a Microbial
Laboratory

15. Plain Test Tube
❖ Used to hold, mix, and heat
chemical experiments.
❖ Used as homes for
microorganisms when people
want to culture (grow) them

16. Screw-capped Tube
❖ Used to grow, culture, and
identify bacteria in a
microbiology laboratory.

17. Centrifuge Tube
❖ Used to separate a substance
into its components.

18. Fermentation Tubes
❖ Used to verify and measure gas
production in fermentation
experiments

19. Durham Fermentation
Tubes
❖ Used in microbiology to detect
production of gas by
microorganisms.

20. Glass slides
❖ Used to hold objects or
specimen for examination
under a microscope.

21. Depression slides
❖ Used to examine living
microorganisms in a hanging
drop.

22. Cover slip
❖ Keep solid specimens pressed
flat, and liquid samples into a
flat layer of even thickness.

23. Watch Glass
❖ Used to hold solids during
weighing and cover beakers
during sample preparation

24. Erlenmeyer flasks
❖ Used for the preparation of
microbial cultures.

25. Beaker
❖ Used for reaction container or
to hold liquid or solid samples.

26. Pipette
❖ Used to measure a volume of
solution extremely precisely.

27. Funnel
❖ Used for filtering, filling, decanting
or transferring liquids or powder
from vessels to another.

28. Graduated cylinder
❖ Used to measure the volume of a
liquids, chemicals and solution.
Are more accurate and precise
than laboratory beaker.

29. Aspirator
❖ It serves as a vacuum source for
filling reagents through a pipette
and also help control the flow of
liquid from the dropping bottle.

30. Inoculating Loop
❖ Used for transfer and streaking
of microorganisms onto solid
culture media.

31. Inoculating Needle
❖ Used for transfer and streaking
of microorganisms onto solid
culture media.

32. Test tube rack
❖ Used to hold upright multiple
test tubes at the same time.

33. Tripod
❖ Used to support or hold the
flasks and beakers during
experiments.

34. Wash bottle
❖ Used to rinse various pieces of
laboratory glassware, such as test
tubes and round bottom flasks.

35. Alcohol lamp
❖ Used for heating, sterilization,
and combustion in a laboratory.

36. Laboratory
oven/incubator
❖ Used to grow and maintain
microbiological cultures or cell
culture.

37. Autoclave
❖ Used to decontaminate
certain biological waste and
sterilize media, instruments
and lab ware.

38. Centrifuge
❖ Used to separate particles or
macromolecules like cells,
sub-cellular components,
proteins, and nucleic acid.

39. Hot plate
❖ Used to heat substances in
containers such as beaker,
flasks or test tubes.

40. Microscope
❖ Used to observe small
objects, even cells.

41. Wire gauze
❖ Used to provide a stable, heat-
resistant surface for containers
such as flasks, beakers, and
test tubes, and helps to
distribute heat evenly.

42. Test tube brush
❖ Used to clean test tubes and
narrow mouth laboratory
glassware's.

43. Evaporating dish
❖ Used to evaporate excess
solvents-most commonly water-
to produce a concentrated
solution or a solid precipitate of
the dissolved substance.

44. Volumetric flask
❖ Used for precise dilutions and
preparation of standard
solutions.

45. Candle jar
❖ Used to grow bacteria
requiring an increased CO2
concentration.

46. Stirring rod
❖ Used for mixing liquids, or
solids and liquids.

47. Spatula
❖ Used for scraping,
transferring, or applying
powder and paste-like
chemicals or treatments.

48. Clay triangle
❖ Used to support a crucible
being heated by a Bunsen
burner or other heat source.

49. Test tube holder
❖ Used to hold test tubes
securely they are being filled,
stirred, or heated.

50. Triple beam balance
❖ Used to measure the mass of
an object that is placed on the
pan.

51. Crucible with cover
❖ Used to heat and melt
substances at high
temperature.