I.

PENGENALAN ALAT, BEKERJA SECARA ASEPTIK, STERILISASI DAN PEMBUATAN MEDIA

A. Introduction 1. Latar Be a!an" Microbiology is a branch of the science of biology that studies micro-sized creatures or microorganisms. In the microbiology work we often can not be separated from the tools that are in the laboratory. It is necessary for an understanding of the functions and properties of the tools used. Equipment used in microbiology laboratories similar to the equipment that is commonly used in chemistry labs. Besides knowing the equipment used is needed also how to work in the laboratory asepticall. According to ram !"##$ aseptic technique is necessary to pre%ent microorganisms from contaminants that can inhibit the growth of microbes . Aseptic technique is used throughout the acti%ity takes place $ both the tools $ materials $ en%ironment and praktikannya . &or equipment and lab materials can be applied to the method of sterility . Mastery of aseptic technique is %ery necessary to the success of the microbiology laboratory $ and it is the beginning of one of the methods are studied by microbiologists . Aseptically work $ not in spite of sterilization materials used while working in the laboratory . 'terilization is generally useful for materials used in the laboratory free from undesirable microorganisms or free from contaminants . 'terilization is necessary in creating the media . (he media ser%es to culti%ate microbes $ isolation $ multiply the number $ e)amine the attributes and calculating the amount of microbial physiology $ where the manufacturing process must be sterilized and aseptic methods to apply in

order to a%oid contamination of the media . (he media is a place where the de%elopment of the organism occurred . #. Pur$o%e After doing this practicum students are e)pected to be able to*
a. +now the types of equipment used in microbiology laboratories and

how to use them.

b. ,a%e the basic skills to work aseptically.

c. +nowing the ways of sterilization equipment.

d. +nowing how to manufacture medium and function of each media.

&. Date and P ace o' Practice -raktikum dilaksanakan pada hari 'elasa tanggal ." 2ni%ersitas 'ebelas Maret 'urakarta. B. Too %, Materia %, and Met(od% ktober !"#!

pukul #/."" 0IB di 1aboratorium Biologi (anah &akultas -ertanian

. #. (ools *
a. analytical scale b. stirrer c. erlenmeyer d. glass beaker e. petri dish f. gas sto%e 3 hot plate stirrer.

!. Materials a. 4utrient Agar 54A6 * #6 Beef e)tract . g$ !6 -eptone / g$ .6 Agar #/ g$ 76 aquades up to #.""" ml$ /6 4a8l / g. b. -otato 9e)trose Agar 5-9A6 * #6 -otato . g$ !6 -eptone / g$ .6 Agar #/ g$ 76 Aquades up to #""" ml. .. Methods
a. Making 4utrient Agar 54A6

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1) 8hop the potatoes into small parts . 2) 0eigh the medium components using analytical scale

.6 -repare #"" ml of distilled water which is di%ided into two parts $ one part to dissol%e Beef e)tract and peptone and partly to dissol%e the order . It is better to water in order to dissol%e more . 76 9issol%e agar on some of the water with constant stirring and heat gi%en . 8an use a gas sto%e or hot plate stirrer 5 not to o%erheat $ as will be formed so that the e)panding foam and spill 6 /6 Meanwhile$ partially distilled water used to dissol%e the peptone and beef e)tract $ simply by stirring . :6 nce both are soluble $ the solution was poured into a solution of agar and stirred until homogeneous . ;6 nce the media is inserted into the erlemenyer flask and sterilized by autocla%ing <6 -ouring sterile medium into sterile petri dishes in aseptic . If the media wants to point upright or tilted to / $ the media immediately poured into tubes and then sterilized . b. Making Potato Dextrose Agar 5-9A6
1. 8hop the potatoes into small pieces.

!. 0eigh the media components using analytical scales.

3. Boil the potatoes in the earlier part distilled water for #-. hours until tender$

then e)tract the oil by filtering using filter paper and squeeze it and hold on the new glass beaker.
4. 9issol%es =agars= with stirrer in /" ml of distilled water and then after

dissol%ed can be added de)trose and homogenized again.

5. After all dissol%e$ e)tract and potato de)trose agar mi)ed and homogenized.

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6. the media poured into a test tube and then erlemenyer or ready for

sterilization. C. Tin)auan Pu%ta!a (he working principle and function of laboratory equipment medical student must be known in order to a%oid errors when the use of laboratory equipment . In addition to the safety of laboratory equipment must be taken to ensure that quality is maintained . 1aboratory equipment is di%ided into two ma>or groups $ namely light tools and hea%y equipment . 1ightweight tools are usually made of wood $ glass $ plastic $ rubber 5Anonima !"#!6. Autocla%es ha%e the form as panic hit $ but has a temperature and pressure gauges . ,owe%er$ this tool has a weakness $ namely its capacity a bit and the price is %ery e)pensi%e 50arisno !"#"6. Aseptic technique is needed in microbiology work that requires precision and accuracy in addition to sterility to be maintained to be free of contaminants that can pollute . ne of the basic techniques in microbiological analysis is aseptic transfer technique 5 a method or technique in the culture memindahlan or transferring bacteria from one place to another aseptically to a%oid contamination by other microbes into the culture . Besides use of aseptic techniques during sampling in order no pollution should use sterile tools . aseptic transfer techniques include techniques such as inoculating 5 inoculation 6 with needle loop $ transferring with a pipette and alcohol &lamming 5 transfer with forceps were burned with alcohol 6 5Ahyari !""<6. 8ulti%ate a seed or microorganisms can be done by creating a culture medium alone . making should be done in aseptic media with nenggunakan tekik microbiology . Equipment is required and one must be in a sterile condition free from unwanted contaminants 50idya !""<6. (he purpose of doing work aseptic basically bred to pre%ent microorganisms from contaminants or pre%ent the entry of unwanted contaminants . Basic use of aseptic technique is a lot of dust particles that contain microorganisms 5 bacteria or spores 6 that may be entered into the cup $ mouth erlenmeyer $ work or settle in the area . 2nwanted microbial growth which may affect or interfere with the results of an e)periment . (he use

: of aseptic techniques to minimize the material used to contaminants agent . ,owe%er$ the technical work aseptically not absolutely perfect to eliminate microorganism contaminants 5?iffiani !"#"6. 'terilization in microbiology means freeing any ob>ect or substance of all life in any form . &or microbiological purposes in an attempt to obtain sterile conditions $ microorganisms can be turned off by the local thermal 5 heat 6 $ gases such as formaldehyde $ etilenoksida or betapriolakton by a %ariety of chemical solutions @ by ultra %iolet rays or gamma rays . Microorganisms can also be mechanically remo%ed by high speed centrifugation or by filtration . In principle$ sterilization can be done in . ways* mechanical $ physical and chemical 5Budhiyanto !"#!6. Media and all equipment must be sterilized before use . If not sterile then the whole media will be contaminated by bacteria and fungi . (here are se%eral kinds of sterilization $ which should be used depending on the tool and the material . 'terilization by dry heat $ for e)ample$ this method is only used for tools and equipment glass made of metal or other material that is not damaged in high temperature . (his sterilization method used usually by using a drying o%en . Baking o%en can also be used . (he temperature is appro)imately #:""8 for 7 hours . (he tools will be sterilized carefully wrapped using aluminum foil before inserting it into the o%en . In addition to the dry heating $ sterilization can also be done with a wet heating . (his method of using a tool called an autocla%e $ which works with the %apor pressure 5Auliarti !"#"6. 'terilization is a way to free things such as tools $ groceries $ ingredients 3 chemicals from pathogenic microorganisms and non- pathogenic . 'ome of the ways in which such sterilization sterilization tau wet dry $ tyndalisasi $ pasteurization $ direct flame 5 Bunsen lamp 6 and steaming hot water and pressure . 'terilization is needed in the manufacture of medium and the other aimed at freeing a de%ice $ or substance of microorganisms . 'terilization can be performed such as by using a Bunsen lamp or autocla%e $ media or tools that are free of pathogenic microorganisms or not . Based on the obser%ation of the way in a de%ice or substance mensterilisasikan one of them is by using an autocla%e which helps sterilize tools and media that will be used 5?idha !"#!6. 'eaweed is an abundant natural resource in Indonesia but has not been optimally utilized by the community . Marine algae contains many ma>or nutrients such as

; carbohydrates $ proteins and fats . 8arbohydrates are found in many marine algae contain cellulose and hemicellulose $ which is part of a nutrient that can not be completely digested by enzymes in the body 5 gi%e a little energy 6 . 4ow with the ad%ancement of science and technology $ the use of seaweed has e)panded into %arious fields . In agriculture seaweed useful as organic fertilizer and the manufacture of one of the growing medium 5(riastinurmiatiningsih !""<6. -otato 9e)trose Agar 5 -9A 6 is a %ery common medium used to breed and culti%ate fungi and yeasts . -otato 9e)trose Agar composition consists of potato powder $ de)trose $ and also in order . -otato de)trose powder and also a food source for fungi and yeasts . Because functions that can breed mold $ now also widely used by -9A mushrooms like oyster mushroom culti%ation . (o ma)imize the growth of mushroom seed $ farmers usually set low p, conditions 5 around ../ 6 and also add acid or antibiotics to inhibit the growth of bacteria 5Anonimb !"#!6. (he most common way used to calculate the number of bacteria is counting the number of li%e bacteria . -enegnceran glow of samples containing microorganisms grown on appropriate growth media . 'uspension was spread on the surface of agar plates or mi)ed with a liquid in order $ which is then poured into petri dishes and allowed to solidify . Agar plates were then incubated under conditions that allow microorganisms to reproduce and form colonies %isible without the aid of a microscope . It is assumed that each bacterial colony will emerge from one bacterial cell . (herefore $ by counting the number of colonies and the dilution factor into account $ the number of bacteria in the original sample can be determined 5,armita !"":6.