BASIC MICROBIOLOGY PRACTICUM REPORT

(UNIT I LABORATORY EQUIPMENT AND STERILIZATION)

Name : Ayunda Widya Frasiska

NIM : 1814441008
Group : Group VI (Six) Session I
Class : International of Biology Education

Assistant : Agung Gunawan

DEPARTMENT OF BIOLOGY
BIOLOGY EDUCATION STUDY PROGRAM
FACULTY OF MATHEMATICS AND NATURAL SCIENCES,
STATE UNIVERSITY OF MAKASSAR
2020
 RATIFICATION PAGE

Complete report of the Basic Microbiology Practicum entitled "Laboratory

Equipment and Sterilization" compiled by:
name : Ayunda Widya Frasiska
NIM : 1814441008
class : ICP Biology Education
group : VI Session I
has been checked and corrected by the Assistant and Assistant Coordinator and
declared accepted.

Makassar, October 2020

Assistant coordinator assistant

Djumarirmanto, S.Pd Agung Gunawan

NIM. 1614142001

Knowed,
Lecturer in Charge of Practicum

Prof. Dr. Yusmina Hala, MS

NIP: 19611212 198601 2 002
 CHAPTER I
ITRODUCTION

A. Background
As science develops, the human curiosity about new things in this
experience will increase. At present, with science, humans can carry out
various studies, both research related to humans, animals, plants, and even
microorganisms that cannot be seen with the eye. This has led to the study of
the challenge of microorganisms called microbiology.

Microbiology is the study of microbes. Microbiology is a branch of

biology and requires supporting science in chemistry, physics, and
biochemistry. Microbiology is often called the practical science of
biochemistry. To see and research these microorganisms, tools are needed that
can support this research. We need to be familiar with microbiology laboratory
equipment and methods or techniques for using these tools, which consist of
glass, plastic and iron.

The tools used must be sterile. Before using microbiological tools, these
tools must be sterilized first, sterilization is the process of freeing tools or
materials from unwanted microbes. To sterilize microbiological equipment, we
must know the method or technique of sterilization.

To be able to research microorganisms in the laboratory we must be

able to grow these microorganisms. Microorganisms can develop naturally or
artificially. The substrate that humans use in developing and growing
microorganisms is called media. For this reason, it is necessary to understand
the types of nutrients signaled by bacteria and the physical environment that
provides the optimum conditions for their growth. The tools used in this
breeding must be sterilized first, so that unwanted microorganisms do not grow,
thus inhibiting the growth of microorganisms.
 There are two kinds of methods that are often used for sterilization,
namely dry heat sterilization using an oven at 170ºC for 1 hour and wet
sterilization (steam) using an autoclave at 121ºC for 15 minutes. Dry heat
sterilization is used for equipment made of metal and glass, qualitative tools
and for tools or materials that can withstand high temperatures. Meanwhile,
wet sterilization is used for equipment made of heat-resistant plastic, rubber,
metal utensil sets, or quantitative tools that are less resistant to heating.

B. Purpose

The purpose of this practicum is that the practitioner is expected to be

able to know what tools are in the microbiology laboratory and how to use
them, then the practitioner is also expected to know how to properly sterilize
techniques so that errors do not occur.
 CHAPTER II
LITERATURE REVIEW

Microbiology is a science that studies microbes such as viruses, archae,

bacteria, fungi, algae, and protozoa. The term microbial (microorganism) is used
to describe an organism that has a very small size, so that it cannot be seen with
the naked eye without using a microscope. Microbes are very simple organisms.
Generally, protozoa bacteria and some algae and fungi are single-celled microbes.
Even multi-celled microbes do not have large cell sizes.(Alimuddin Ali, 2005)

Microbiology is a science that studies very small living things (less

than 0.1 mm in diameter) which cannot be seen with the ordinary eye without the
aid of a special tool. These creatures, which are called microorganisms or
microorganisms, are ubiquitous. Among them are those that are beneficial to
human life, but many are harmful, such as those that cause various diseases.
Microbiology covers a wide variety of disciplines such as bacteriology,
immunology, virology, psychology and parasitology. These sciences have grown
in size over the years, so that they are separate and independent disciplines. This
science continues to develop continuously because microorganisms as living
things are able to adapt to their new environment.(Chatim et al., 2019)

In The etymology of the word "laboratory" comes from the Latin word
which means "place of work" and in its development the word "laboratory" retains
its original meaning, namely "place of work", but specifically for the purposes of
scientific research. Laboratory is defined as a place that can be in the form of an
open space, closed space, school garden, green house or other environment for
conducting experiments or research. Room or the room in question is a building
bordered by a wall, roof, or open space. The definition of laboratory as meant in
this research is limited to a laboratory in the form of a closed room. The
laboratory is a place or place for conducting experiments as proof of the truth of
the theories given in class, stimulating certain experiments in a guided manner, or
discovering on your own as well as increasing students' reasoning power.(Muna,
2016)

As for the functions of the science / science laboratory room, among others, as a
place for learning science / science and providing skills, the place for which it is
produced new friends, both theories and objects / tools / new technology and
skills, a display or exhibition place, a place to practice and prove the truth or not
(verification) of certain symptomatic factors, a place where it takes place Biology
learning activities in practice that require special equipment.(Muna, 2016)

At the present time it's a tool is one of the supporters of the success of a
job in the laboratory. So that for make it easy and smooth berlangsaung practicum
knowledge about the use of tools is very necessary. Introduction tools laboratory
is important for work safety when conducting research. Laboratory tools can
usually be damaged or even dangerous if their use is not in accordance with the
procedure. Importance Done the introduction of laboratory equipment is so that
we can know how to use these tools properly and correctly, so that errors in the
procedure for using the tools can be minimized as little as possible.(Andriani,
2016)

Tools laboratory microbiology such as incubator, autoklav, test rack and tube,
glass beaker, suction pipette, measuring pipette, tweezers, petri dish, sterile cotton
swab, spritus lamp, ose. Total microbial testing was carried out by using the cup
method. The palig plate count method is widely used to count the number of
microbes in food ingredients. The medium used includes, plate count agar (PCA)
medium, test tubes, petri dishes, pipettes, incubators.(Andriani, 2016)

Storage of tools made from plastic, metal glass and rubber such as measuring
cups, test tubes and so on, each grouped into one and stored according to their
respective groups. Heavy equipment is placed where it is easily accessible,
expensive or dangerous equipment is stored under lock and key. Basically, tool
storage should not be placed in a place where the tool is damaged or in a
place where the process of taking / returning
it can endanger the wearer.(Muna, 2016)

Things to watch out for when The filling of the material / equipment you
want to sterilize is that the material is packaged loosely in a chamber to facilitate
the penetration of hot steam and remove air after the sterilization process is
complete. (Andriani, 2016)

Sterilization is a process to kill all organisms contained in an object (tool

or material). The purpose of sterilization in microbiology is to kill, inhibit growth
and get rid of all the microorganisms present in tools and materials to be used in a
job to create an aseptic atmosphere. In general, sterilization can be done by 3
methods: mechanical, physical and or chemically. Mechanical sterilization
includes using a microfillter, physical is divided into 2 irradiation and heating,
while chemistry is using chemicals (disinfectants). Materials, tools and work
tables to be used in practice at The microbiology laboratory must go through the
sterilization stage first, this is so that the work is carried out aseptically or free
from unwanted polluting microbes. The sterilization that is often used in the basic
practice of microbiology is physical sterilization by heating, which is divided into
dry sterilization and wet sterilization.(Murtius, 2018)

Sterilization of pharmaceutical preparations and medical devices done in

various ways, namely by heating, gas, radiation, and filtration. Sterilization by dry
heat method is used for materials in powder form. Various attempts to sterilize
alginate have been carried out, for example using ethylene oxide gas, autoclaving,
and light radiation, but this process causes damage to the uronic acid polymer
chain in the alginate solution and gel.(Ayuning Putri et al., 2017)

The sterilization of growing media in the production of AMF aims to

minimize disturbance by unwanted microorganisms (contaminants), so as not to
experience damage which can affect the quality of inoculants. Media sterilization
can be done in three ways, namely mechanically, physically and chemically. In
this study, the growth media sterilization technique in the production of AMF was
carried out physically and mechanically, namely through the media sterilization
technique using Gamma Co-60 Irradiation, autoclaving, adding 10% NaOCl
solution, and washing the media with tap water. Rays. Sterilization technique has
an effect on increasing colonization. The method of sterilization using tap water
(control) was not significantly different from autoclaving and Gamma Co-60
irradiation in increasing colonization.(Dewi et al., 2017)
 CHAPTER III
PRACTICUM METHOD

A. Time and Date

Day / Date : Monday / 28th September 2020
Time : 10.50 to 13.50 WITA
Place : Biological Laboratory, west 2nd floor of Biology
B. Tools and Materials
1. Tools
a. Stationary
b. Glass tools
1) Petri Dishes
2) Graduated Cylinder
3) Beaker Glass
4) Test Tube
5) Preparatory Glass
6) Deck Glass
7) Stirring Rod
8) Drop Pipette
9) Erlenmeyer
10) Fial Bottle
11) Volume Pipette
12) Reagent Or Diluent Bottles
13) Glass Funnel
14) Bunsen
15) Durham Tube
16) Spread Rod Or Hock Stick
17) Thermometer
 18) Cuvet
19) Burette
20) Desiccator
c. Non Glass Tools
1) Test Tube Rack
2) Test Tube Brush
3) Haemocytometer
4) 3 Legs And Gauze
5) Spoit
6) Colony Counter
7) Laminar Air Flow
8) Enkas
9) Water bath
10) Autoclave and basket
11) Furnace
12) Incubator
13) Oven
14) Analytical scales
15) Spray flask
16) Shaker
17) Vortex
18) Centrifuge
19) Spectrophotometer
20) PCR
21) Microscope
22) pH meter
23) Refrigator
24) Chamber
25) Clamp
26) Tweezers
27) Scalpel and bisturi
 28) Scissor
29) Matches
30) Mortar and pistil
31) Razor blade
32) Mikrotube
33) Micropipette
34) Drip plate
35) Bulb
36) Inoculating loop
37) Inoculating needle
2. Materials
a. Paper
A. Work Procedures
The work procedures carried out are as follows.
1. Laboratory equipment to be observed is prepared.
2. Every laboratory equipment is observed and recorded.
 CHAPTER IV
RESULT AND DISCUSSION

A. Result
1. Glass
No Tools Observation Comparison Note
. Image Image

1. Cover
2. Container

Petri Dishes
1.

1. Mouth
2. Gauge
Graduated 3. Base
2. Cylinder

1. Mouth
2. Glass
3. Gauge
3. Beaker Glass
 1. Tube mouth
2. Tube body
Test Tube
4.

1. Object glass
5.
Object Glass

1. Stirring rod
2. Spoon
Stirring Rod
6.

1. Rubber
pipette
Drop Pipette 2. Mouth
7. pipette

1. Erlenmeyer
mouth
Erlenmeyer 2. Erlenmeyer
8. neck
3. Erlenmeyer
body

1. Cover bottle
2. Body bottle
Fial Bottle
9.
 1. Mouth
pipette
Volume 2. Goiter
10. Pipette

Reagent Or 1. Cover bottle

2. Body bottle
Diluent
11. Bottles

1. Top mouth
12. 2. Lower mouth
Glass Funnel

1. Cover
13. 2. Axis
Bunsen
3. Spiritus

1. Mouth tube
14. 2. Body tube
Durham Tube

1. Spreader
Spread Rod
2. Stick
15. Or Hock
Stick
 1. Thermometer
pipa.

16. Thermometer

1. Cuvette
Cuvette
17.

1. Burette
2. Stopcock
Burette
18.

1. Cover
19. 2. Body
Desiccator Desiccator

1. Desk glass
20. Deck Glass

2. Non Glass
No Tools Observation Comparison Note
. Image Image
 1. Tube place
2. Rack tube
Test Tube
1
Rack

1. Brush
Test Tube 2. Stick
2
Brush

1. Cover
Haemocytomet 2. Sample point
3
er 3. Cover glass

1. Buffer
3 Legs And 2. Base buffer
4
Gauze

1. Hub
2. Barrel
5 Spoit

1. Base
Colony 2. Lup
6 3. Wolffugel
Counter
disks

1. Control panel
Laminar Air 2. Workspace
7 3. Support frame
Flow
4. Base
 1. Cover
2. Entrance
8 Enkas

1. Bath/chamber
2. Control panel
9 Water Bath

1. Cover
2. Key
10 Autoclave
3. Control lamp

1. Crucible space
2. Cover
11 Furnace
3. On/off button

1. On/off button
2. Incubator door
12 Incubator

1. Oven door
2. Base
13 Oven

1. Plate scale
Analytical 2. Control panel
14
Scales
 1. Bottle hose
2. Bottle body
15 Spray Flask

1.control panel
16 Shaker

1. Standart top
2. Digital wheel
17 Vortex

1. Lid and latch

2. Control panel
18 Centrifuge

1. Control panel
Spectrophotom
19
eter

1. Heated lidad
2. Door lock
3. Touch screen
20. PCR

21 Microscope
1. Occuler Lens
2. Tube
3. Revolver
4. Objective lens
5. Table
6. Condenser
7. Diafragma
 8. Mirror
9. Base
10. Inclination
joint
11. Clamp
12. Arm
13. Fine screw
14. Coarse screw

1. PH display
2. Agile glass
22 PH Meter
probe
3. Protection cap

1. Cover
2. Base
23 Refrigerator

1. Control panel
2. Chamber door
24 Chamber

1. Clamp space
2. Opening
25 Clamp
clamp
3. Handle limb

1. Tweezers
26 Tweezers
 1. Bisturi
2. Scalpel
Scalpel And
27
Bisturi

1. Scissor tips
2. Scissor Handle
28 Scissor

1. Matches
29 Matches

1. Pistil
Mortar And 2. Mortal
30
Pistil

1. Razor blade
31 Razor Blade

1. Microtube
32 Microtube

1. Micropipette
33 Micropipette
 1. Drip plate
34 Drip Plate

1. Bulb
35 Bulb

1. Round
Inoculating inoculating
36
round 2. Handle

1. Inoculating
Inoculating needle
37
Needle

B. Discussion
Microbiology is the study of microbes. Microbiology is a branch of
biology, and requires supporting science in chemistry, physics, and
biochemistry. Microbiology is often called the practical science of
biochemistry. In basic microbiology, basic understanding is given about the
history of microbial discovery, the kinds of microbes in nature, microbial cell
structure and function, microbial metabolism in general, microbial growth and
environmental factors, applied microbiology in the environmental and
agricultural fields.

Advanced microbiology has developed into various sciences, namely

virology, bacteriology, mycology, food microbiology, soil microbiology,
industrial microbiology, and so on, which study specific microbes in more
detail or according to their benefits. With the development of science today
there are various kinds of tools used to examine microbes, each tool has a name
that shows the usefulness of the tool, the working principle or the process that
took place when the tool was used. Some of the uses of the tool can be known
based on its name, the materials or equipment used in the microbiology lab are
divided into several types, including Erlenmeyer, beaker, reaction tube, hot
plate, petri dish, ose needle, Bunsen lamp, pipette, oven, autoclave.

Bunsen lights serve to minimize other bacteria when heating. The

spetula serves to store the susfensi Micro pipette is a tool for moving liquid
with a small volume. There are many choices of capacity in micro pipettes that
can be adjusted for the volume of the intake and micro pipettes that cannot be
adjusted in volume. In its use, the micro pipette requires that tips that have
been sterilized should not be touched by hands because it will cause
contamination.

A petri dish is a round container made of plastic or glass which is used

to multiply cells. Petri dishes always come in pairs, the smaller ones for the
container and the big ones for the lids. The spread rod serves to spread the
suspense on the media. The loop needle is made of chrom wire measuring 0.5
to 0.75 mm.

Hot plate functions to homogenize a solution by stirring. The plates

contained in this tool can be heated so as to accelerate the homogenization
process. Stirring with the help of a hot plate and magnetic bar, for example, can
homogenize up to 10 L, with a very slow speed of up to 1600 rpm and can be
heated to 425oC. options based on the volume scale. When measuring the
volume of solution, it is preferable to determine the volume based on the
concave meniscus of the solution. The test tube that functions as a growth
medium and other liquid reservoirs such as solvents can also be filled with
solid media.

Ovens are tools used to dry tools before they are used and are used to
dry ingredients in a wet state. The oven is a sterilizer that uses the dry heat
principle. Ovens are used to sterilize glassware or materials such as oil that
cannot be sterilized by autoclave because they are not permeable to moisture.
This tool consists of electric heat, controls the temperature and an insulation
room which is generally equipped with a fan to circulate the air to heat evenly.
Typical sterilization conditions are 160-170 degrees within 1 hour.

Dropper pipette is a tool for moving the solution by volume known.

There are various sizes of measuring pipette capacities, including pipettes
measuring 1 ml, 5 ml and 10 ml. How to use it is that the liquid is sucked with
a measuring pipette with the help of a filler to the desired volume. The volume
transferred is removed according to the available scale (it is seen that the scale
should be immediately parallel to the fluid concave scavenging) by equalizing
the filler pressure with the ambient air. Dropper pipette (Pasteur Pippete),
functions the same as measuring pipette, but the volume transferred is
unknown. One application is to add HCl or NaOH when adjusting the pH of
the media, adding reagents to biochemical tests, and so on.

The measuring cup is used to measure solution, liquid or flour at

various volume sizes. This measuring cup is made of glass and how to use it by
looking at the volume indicated on the measuring cup. Autoclafe is used as a
high pressure steam sterilizer. The use of autoclave for sterilization, do not put
the lid carelessly and opened it because the contents of the bottle or medium
container will overflow and can only be opened when the manometer shows
the number 0 and cool down little by little. Medium containing vitamins,
gelatin or sugar, then after sterilization the medium must be cooled down
immediately. This method is to prevent the substance from breaking down. The
medium can be stored directly in the ice cup if it is certain that the medium is
sterile.

Erlenmeyer is a tool used to sterilize medium storage sizes and contain

filtering. This tool can be sterilized by first covering the top with cotton and
then sterilized using an autoclave. The beaker functions as a container for
solutions and also heats microbiological substances and takes a sample of the
correct size. The test tube serves to store microorganisms in liquid or solid
nutrient medium.

Centrifuge is a device used to separate compounds with different

molecular weights by using the centrifuge force. There are two types of
centrifuge, namely electric centrifuge and manual rotary centrifuge. Incolum
needles (loop needles) function to move the culture to be planted / grown to
new media. The inoculum needle is usually made of nichrome or platinum wire
so that it glows when exposed to heat. The shape of the needle tip can be in the
form of a loop (loop) and is called an inoculating loop / transfer loop, and the
straight one is called an inoculating needle / transfer needle. The inoculating
loop is suitable for streaking the agar surface, while the inoculating needle is
suitable for stab inoculating. This inoculum needle is very useful when splitting
agar for the preparation of Henry's Slide Culture.

Laminar air flow is a basic laboratory equipment, especially medical

laboratories. There are two Laminar air flow treatment systems, namely the
vertical and horizontal air treatment systems, each of which has advantages and
disadvantages. Laminar air flow is a systematic tool that uses the principle of
air filtration and the use of ultraviolet radiation. Laminar air flow is used as a
place to carry out laboratory activities that require sterile conditions, such as
opening sterilized instruments and preparing microbial covers.

Some examples of the sterilization process that are often carried out in
laboratory and factory-scale lab work, including Dry Sterilization Dry
sterilization or dry heat sterilization can be applied by direct heating, floating
over a flame, burning and sterilizing with hot air (oven).

Fire is used to sterilize equipment such as inoculation needles, petri

dishes, slide glasses, tweezers, culture tube mouths, spatels and others. After
being sterilized, the equipment must be cooled before being used, especially
for inoculation needles and pipettes after being annealed or heated over a fire,
then cooled in 70% alcohol solution then burned again to remove the remaining
alcohol, after that it is cooled again in the alcohol solution and left / aerated and
do not need to be burned again.

Dry sterilization is sterilization with hot air. The tool used is an oven.
This method is commonly used to sterilize glassware such as petri dishes, test
tubes, and other glass tools. The working principle of this tool is simpler, that
is, the oven door is opened and all the tools to be sterilized are neatly arranged.
After that the oven door is closed, the temperature is set at 160-180oC for 1-2
hours. The advantage of dry heating is that there is no water vapor wetting the
sterilized materials or tools. In this experiment, qualitative tools are sterilized
by the dry sterilization method.

Wet sterilization or humid heat sterilization can be applied by heating

using steam with pressure (autoclaving) at high temperatures, wet sterilization
is usually used on devices that are not heat resistant, in this wet sterilization
uses a temperature of 120C for 30 minutes on autoclave Digital in this
experiment, quantitative tools that are measuring and materials that are not
resistant to high heating are starilized by this wet heat method.
 CHAPTER V
CLOSING

A. Conclusion
On the conclusion of this practicum, we already know what
microbiology laboratory tools and their functions are and we can find out how
to sterilize tools and materials. Sterilization is a process to kill all organisms
contained in an object. The sterilization process can be divided into three types,
namely, the use of heat (annealing and hot air), filtering, the use of chemicals
(ethylene oxide, peracetic acid, formal dehide and alkaline glutaraldehyde)

B. Suggestion
 The practitioner should pay attention to all practicums and do them
carefully. Because between one practicum title and other titles have a close
relationship.
 In this experiment, Praktikan really expect instructions from the assistant
to be careful in doing practicum. Given the experimental materials that
contain microbes that can contaminate and disturb health
  We recommend that every action in carrying out the practicum is directed
directly to the practitioner so that the practitioner can understand various
techniques in the laboratory related to the title being practiced.

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ATTACHMENT