Prosiding Seminar Nasional Biologi FMIPA UNM. ISBN: 978-602-52965-8 ‘Inovasi Peneltian Biologi dan Pembelajarannya di Era Merdeka Belajar Makassar, 8 Agustus 2020 IDENTIFIKASI MORFOLOGI MIKROBA PADA RUANGAN WATER CLOSET JURUSAN BIOLOGI UNIVERSITAS NEGERI MAKASSAR Identification of Microbial Morphology in The Water Closet Room Department of Biology Universitas Negeri Makassar Deny Romadhon Badaring"), M. Fiqriansyah W®, Arsad Bahri? » Jurusan Biologi,Fakultas Matematika dan Iimu Pengetahuan Alam, Universitas Negeri Makassar, Makassar, 2 Jurusan Biologi, Fakultas Matematika dan llmu Pengetahuan Alam, Universitas Negeri Makassar, Makassar. » Jurusan Biolog, Fakultas Matematika dan Iimu Pengetahuan Alam. Universitas Negeri Makassar, Makassar. Email korespondensi: deny 181299@gmail.com 2\wahabfigriansyah(a'gmail.com arsad.bahri@unm.ac.id ABSTRAK Water closet merupakan tempat umum yang memiliki potensi untuk pertumbuhan berbagai jenis mikroba. Tujuan dari penelitian ini adalah untuk mengisolasi. dan mengidentifikasi morflogi koloni mikroba terdapat disalah satu ruangan water closet Jurusan Biologi. Metode yang digunakan adalah isolasi mikroba menggunakan media TEA (Tauge Ekstrak Agar) dengan komposisi untuk setiap bahan di konversi menjadi 100. Selanjutnya diletakkan pada ruangan water closet selama 15 menit dan di inkubasi dengan suhu 37°C selama 2 hari. Pengamatan dan identifikasi morfologi koloni mikroba pada media dilakukan dengan bantuan flash. Hasil yang didapatkan adalah pertumbuhan 2 koloni mikroba MI dan M2. MI memiliki ciri-ciri morfologi warna putih, size moderate, form rhizoid, margin filamentous, elevation convex. M2 memiliki ciri-ciri morfologi warna putih, size small, form circular, margin entire, elevation convex. Penggunaan media TEA yang telah dikonversi 100 dapat menumbuhkan dua jenis koloni mikroba. Kata kunci: Identifikasi Morfologi, Isolasi Mikroba, TEA (Tauge Ekstrak Agar) ABSTRACT A Water closet is a public place that has the potential for the growth of various types of microbes. The purpose of this study was to isolate and identify the morphology of microbial colonies in one of the water closet rooms of the Biology Department. The method used was microbial isolation using TEA (Tauge Ekstrak Agar) media with the composition for each material being converted to 100. Then placed in a water closet for 15 minutes and incubated at 37°C for 2 days. Observation and identification of microbial colony morphology on the media were carried out with the aid of light. The results obtained were the growth of 2 microbial colonies M1 and M2. MI has morphological characteristics of white color, moderate size, rhizoid form, filamentous margins, elevation convex. M2 has the characteristics of white morphology, small size, circular form, margin entire, elevation 161Prosiding Seminar Nasional Biologi FMIPA UNM ISBN: 978-602-52965-8 ‘novasi Penelitian Biologi dan Pembelajarannya di Era Merdeka Belajar ‘Makassar, 8 Agustus 2020 convex. The use of TEA media that has been converted to 100 can grow two types of microbial colonies. Keywords: Morphological Identification, Microbial Isolation, TEA (Tauge Ekstrak Agar) PENDAHULUAN Mikroba merupakan salah satu organisme yang memiliki jumlah yang melimpah dan ‘memiliki ukuran renik. Mikroba hidup bebas di lingkungan, menyebar di udara, tanah, air, makanan, bahkan mikroba yang hidup dalam tubuh manusia, Water closet merupakan tempat yang sering bersentuhan dengan manusia. Water closet diketahui memiliki jumlah mikroba yang melimpah dikarenakan faktor lingkungan yang memungkinkan berbagai mikroba dapat tumbuh dan berkembang biak dengan baik. Mikroba ada yang bermanfaat dan ada yang merugikan yang bersifat patogen. Mikroba yang bermanfaat dan mikroba yang merugikan untuk membedakannya tentu sulit, ‘mengingat mikroba tersebut dalam bentuk populasi campuran, Hal ini dapat diatasi dengan proses identifikasi antara mikroba bermanfaat dan mikroba yang merugikan dapat melalui pemisahan populasi campuran dari lingkungannya, Pemisahan ini lebih dikenal dengan nama isolasi mikroba, Pengamatan mikroba hanya dapat dilakukan jika mikroba yang diamati di isolasi di tempat-tempat tertentu schingga mereka mudah diamati. Pengamatan terhadap mikroba tertentu hanya dapat dilakukan jika mikroba dipisahkan dari lingkungan dan mikroba lainnya, Ini bisa dilakukan dengan teknik isolasi. [Lestari dan Hartati 2017). Pentingnya mengisolasi mikroba dari lingkungan, seperti makanan (substrat padat), ‘minuman (substrat cair), dan diri Anda sendiri karena banyaknya mikroba yang sulit diamati atau dibedakan secara langsung menggunakan panca indera, Schingga isolasi akan membuat lebih mudah untuk melihat dan mengamati bentuk-bentuk pertumbuhan mikroba dalam beberapa media dan dapat melihat morfologi mikroba, yaitu inokulasi yang merupakan 162Bites eae ile | sieve) I) MICROBIOLOGICAL APPLICATIONS Laboratory Manual in General Microbiology ie ie a SL)Mc Eire [BENSON'S MICROBIOLOGICAL APPLICATIONS; LABORATORY MANUAL IN GENERAL MICROBIOLOGY, SHORT VERSION, THIR- ‘TENTH EDITION Pblished by McGraw-Hill Education, 2 Peon Pza, New York, NY 10121, Copytigh © 2015 by McGraw-Hill Education AI sighs reserved. 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Theobald [Executive Marketing Manager: Purch E.R Disectr, Content Production: Terri Schies! [Content Project Manager: Laura. Bies Bayer: Mchole Birkenbole Designer Morgane Reynolds (Cover nage: Seence Pro Libary RFZGety Images Senior Content Licensing Specialist: Lavi Hancock (Composite Lachine Pablihing Servicer ‘Typeface: 11/2 Times LT Sud Primer Quad'Graphics All rei apeatngon page ora the en ofthe ook te considered o ean extension ofthe copyright pags (Crt: Banner image (etre: CDCnice Haney Care Some ofthe laboratory experiments include in his text may be hazardous if materi re handled improper if pocedies ae conducted incor ret Safety precautions are necessary when you are working With chemicals, lastest tubes, hot water ut, shar instruments, ad the ik, o or ‘ny procedures that generally regi cauton. Your school may hve se regulations gain safety procedures thet your instructor wil explain You ‘Shosld you have any problems with materials or procedures, please ask Your suc or help. “The erm arses tinted in he text wee accurate athe time of publication. The incon of « website dos not nae an emdonement by the authors MGraw- Hl Eden, and McGraw-Hill Education Joes not guarantce the accuracy of the information resented at hese sites. wwe com2 ¥ () A small amount of Vaseline is placed near each comer of| the cover glass with a toothpick =, (8) Depression slide is pressed against Vaseline on cover glass and quickly invented 2 Motility Determination EXERCISE 17 ‘Two loopfuls of organisms are placed in the cover glass. Vase Sa Organisms (4) The competed preparation can be examined under il Figure 17.3. The hanging drop slide. ‘molecules bombarding bacterial cells. Ifthe only ‘movement you see is vibrational and not direc- tional, the organism is nonmotile. + If you see only a few cells exhibiting motility, consider the organism to be motile. Characteristi- cally, only a few of the cells will be motile at a given moment, + Don’t confuse water current movements with true motility. Water currents are due to capillary action caused by temperature changes and dry- ing out, All objects move in a straight line in one + And, finally, always examine a wet mount imme- once it has been prepared, because motil- ity decreases with time alter preparation, Hanging Drop Slides By referring to figure 17.3, pre- ppare hanging drop slides of each organism. Be sure to use clean cover glasses and label each slide with a china marking pen. When placing loopfuls of organisms on the cover glass, be sure 10 flame the loop between applications. Once the slide is placed on the microscope stage, do as follows: 1. Examine the slide first with the low-power objec- tive. If your microscope is equipped with an auto- ‘matic stop, avoid using the stop: instead, use the course adjustment knob for bringing the image into focus. The greater thickness of the depression slide prevents one from being able to focus at the stop point. . Once the image is visible under low power, swing the high-dry objective into position and readjust the lighting. Since most bacteria are drawn to the cedge of the drop by surface tension, focus near the edge of the drop. If your microscope has phase-contrast optic switch to high-dry phase. Although a hanging drop does not provide the shallow field desired for phase contrast, you may find that it works fairly well - Ifyou wish to use oil immersion, simply rotate the high-dry objective out of position, add immersion oil to the cover glass, and swing the oil immersion lens into position, . Avoid delay in using this setup. Water conden- sation may develop to decrease clarity, and the ‘organisms become less motile with time Review all the characteristics of bacterial motility that are stated on page 122 under wet mounts. 123EXERCISE 17. Motility Determination (1) Wire with organisms is broweht into tube without touching walls of tube. Figure 17.4. Stab technique for motility test. Plate Method Mark the bottom of a plate of soft agar le with two 1/2" circles about 1" apart, Label one ci ML and the other PV. These circles will be targets for 124 2) Wiee penetrates medium oo.) thirds of is depth is withdrawn from medium and tbe, Neck of tube i lamed and plugged. ‘Assemble the following materials that were inocu- lated during the last period and incubated. + culture tubes of motility medium that have been incubated + inoculated petri plate that has been incubated Compare the two tubes that were inoculated with M, luteus and P. vulgaris. Look for cloudiness as evi- dence of motility. Proteus should exhibit motility. Does it? Record your results on the Laboratory Repon. ‘Compare the appearance of the two stabs in the soft agar, Describe the differences that exist in the two stabs. Does the plate method provide any better differ~ entiation of results than the tube method? Complete Laboratory Report 17 for this exerciseAcquisitions Editor: Kelsey Churchman Director of Development: Barbara Yien Senior Project Development Editor: Marie Beaugureau Editorial Assistant: Ashley Williams Managing Editor: Deborah Cogan Production Project Manager: Megan Power Production Management and Composition: Integra Interior Designer: Jerilyn Bockorick Cover Designer: Yvo Riezebos Image Management: Travis Amos Manufacturing Buyer: Stacey Weinberger Executive Marketing Manager: Neena Bali ‘Text Printer: Weberafters, Inc. 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Where those designations appear in this book, and the publisher was aware of a trademark claim, the designations have been printed in initial caps or all caps. Library of Congress Cappuccino, James G. Microbiology : laboratory manual/James G. Cappuceino, Natalie Sherman. —10th ed, psem. Includes bibliographical references and index. ISBN-13: 978-0921-81022.6 ISBN-10- 0-321-84022-4 L Sherman, Natalie. IL. Tie. [DNLM: 1. Microbiology—Laboratory Manuals. QW 25) LC Classification not assigned 579.078 —de23 2012097782 ‘Student edition ISBN 10: 0921-84024 ISBN 13: 978-0.321-84022.6 Instructor's Review Copy ISBN 10: 0921-84515 ISBN 13: 978-0.921-88451.0 PEARS MO cies ttt ssc orn© Place the tubes in the paim of your hand, secure wath your thumb, and separate to form a V. © Flame the necks of the tubes by rapidly passing them through the flame once. Broth-to-slant transfer: Obtain 3 loopful (of broth and place at base of slant. Withdraw the loop in a zigzag mation. @ Fiame the necks of the tubes by rapidly passing them through the flame once.This International Student Edition is for use outside of the U.S. Microbiolo n Marjorie Kelly ee nel a abe Fide 4Published by McGraw Hill LLC, 1525 Avene ofthe Americas, New York, NY 10019. Copysight ©2028 by “MeGaw Hil LLC. Aleit eerved. Print in the United Sates of America. No part ths puiation may te reprediced or distibted i any ferwor by at eeu, osord in adaabas oe Fetieva yes, without the price writen consent of McGraw Hill LLC, icing, bat ot ined, nay network or ter electronic Sorageor transmission, or broads for distance learning. Some anciaries,ncaing electronic and print components may ot be aaah customers outside the ned Stats. “This ook i pete on aid ree pape 1254567 8OLWE2627 2625 2425, ISBN OTE L266.177330 MID 1266-17798 Cover Image: Viarume/CorbisGey Images All eredits appearing on page or atthe endothe book are considered to be an extension ofthe copyright page. “The ftemet addreses stein the eat were acct the ine of publication. The incasion ofa website des rot incite sn endorement by the naar oe Mera Hill LLC. an MeGraw Hill LLC des no guarantee the ccuracy ofthe information presented a these ses ‘bedcaton convhighered54 Chapter3. Tools ofthe Laboratory 1 Whats the intended message of the article? 1 What s your critical reading of the summary ofthe article provided above? Remember hatin this context, “critical reading" does not necessarily mean What rcism do you have? but asks you to apply your knowledge to interpret whether the article is factual and wether the facts support the intended message. 1 How would you interpret the news tem for your nonmicrobiologis fiend? 1 What s your overall grade fr the news itemtaking into account ts accuracy and the accuracy ofits intended effect? Outline and Learning Outcomes 34 Methods of Culturing Microorganisms: The Five I's 4. Explain what the Five 's mean and what each step ental 2. Discuss three physical states of media and when each is useful [2 Compare and contrat selective and differential media, and give an example of each. 4, Provide bret defintions for dined and complex media 3.2 The Microscope: Window on an Invisible Realm '5. Convert among liferent lengths within the metric system 6. Describe the earliest microscopes. 7, Ust and descrize the three elements of good microscopy. 8. Differentiate between the principles of ght and electron microscopy. ‘9. Compare and contrast the three main categories of stains, and provide examples of each. should be noted that inthe last decade, advances in molecu lar techniques mean thatthe Five I's are no longer necessary in all eases. For example, it has become possible to identify micro: forganisms without growing them, But there are many things that 3.1 Methods of Culturing Microorganisms: The Five I's “Microbiologists are confronted by some unique problems. First, in the natural setting, whether in a living host or inthe environment, different species of microbes are mixed together in complex associations, This can make it difficult to study the characteristics fof just one of them. Second, because unseen microbes are everywhere—in the air, on every surface—it is very easy 10 introduce unwanted microbes when working with your intended microbe, ‘The “Five I's" represent five asc techniques to manipulate, stow, examine, and characterize microorganisms inthe laboratory. They are inoculation, incubation, isolation, inspection, and Identification (the Five's: figure 3.1), These procedures ae time: tested procedures to handle and maintain microorganisms. It comes to specimen callection, timing is everthing In der to accurately idently the microorganism, and to use the appropriate antibiotics, proper specimen collection s crucial aly, cultures should be collected price to veating a patient ith antbotics. Antibiotic reatment may eliminate microbes inthe ample affecting the accuracy of the patient’ diagnoss. Astrea continues, a repeated fever or continuing symptoms may ciclo the need to change the antbiotc, and ational cures ‘need tobe collected before administering new antimicrobial, gs clinical microbiologists and researchers do with microbes that, requite the techniques described here. Incubation ‘Once container of medium hasbeen inoculated with specimen itis incubated, which means its placed ina temperature-controlled cham ber (incubator) fo encourage growth, Although there are microbes that grow optimally at temperatures ranging from lreezing to boil- ing, the usual temperatures used in nborstory growth fall between56 Chapter’3_Tools othe Laboratory ‘Types of Artificial Media Media can be classified aecording to three properties table 3.2): 1. physical stat, 2. chemical composition, and 3. functional type (purpose) Table 3 Incubation Conditions of Various Bacterial Pathogens Listeria monocytogenes 25-30 Preudomonas fluorescens 25-30 12 Srepococcus pyogenes ” 12 Mycobacterim nbereutoss u 28 Table 3.2 Three Categories of Media Classification Physical States of Media Liquid media are water-based solutions that do not solidity at temperatures above freezing and that tend to flow freely when the container i tilted (figure 32a). These media, termed broths, ‘milks, or infusions, are made by dissolving various solutes in dis- tiled water. A common laboratory medium, nutrient broth, con: tins beef extract and peptone dissolved in water. Methylene blue mill and litmus milk are opaque liquids containing whole milk and dyes, Fluid thioglycollate isa slightly viscous broth used for determining the oxygen requirement of different microbes At ordinary room temperature, semisolid media exhibit 2 lotike consistency (figure 3.26) because they contain an amount of solidifying agent (agar or gelatin) that thickens them but does ‘ot produce a firm surface. Semisolid media are used to determine ‘the motility of bacteria and to localize a reaction at a specific site. Solid media provide a firm surface on which cells ean form discrete colonies (igure 3.2e) and are useful for isolating and iui 1. Chemically defined (sytetie) Genera pupose 3. Reducing 2. Semisotia 2 Complex: ntchemialy defined 2. riche 6. Specimen tanspor 3. Solid (eam be iii) 3. Soecive 1. Asay 4 Sold (annot be guetid) 4. ital 8. Eoumeation Seni at .264 Chapter 9 Microbial Nutrition and Growth Figure 9.6 Red snow. (a) An early summer snowbnk proveles ‘perfect habitat fr payctroptic photosyhetcorgansams ie CChiamydomones avis.) Mitescoptevew ofthis snow alga actualy Classiled a a “green” alga attrough 2rd pigment dominates at ths stage of ecco generally cannot grow above 20°C. Laboratory work with true psychrophiles can be a real challenge. Manipulations have to be done in a cold room because room temperature can be lethal to the organisms. Unlike most laboratory cultures, storage in the feftigerator incubates, eather than inhibits, them, As one may predict, the habitats of psychrophilic bacteria, fungi and algae are lakes and rivers, snowfields (figure 9.6), polar ice, and the ‘deep ocean, Rarely if ever, ate they pathogenic, A less extreme group of cold-loving bacteria, called psychrotolerant, grow slowly in cold but have an optimum temperature between 15°C and 30°C, ‘The majority of medically significant microorganisms are :mesophiles (mez -ob-f)2), organisms that grow at intermediate tem- peratures, Although an individual species ean grow atthe extremes 10f 10°C or S0°C, the optimum growth temperatures (optima) of mast rmesophiles fall into the range of 20°C wo 40°C. Organisms in this group inhabit animals and plants as well as soil and water in tem- erate, subtropical, and topical regions. Most human pathogens have optima somewhere between 30°C and 40°C (human body temperature is 37°C). Some mesophilic bacteria, such as Staphy~ ococeus aureus, gow optimally at body temperature but are also psyehrovolerant, meaning they can survive and multiply slowly at refrigerator temperatures, causing concem for food storage. Lis- feria monoeyiogenes is a human pathogen that is psychrotolerant and often grows in ice eream and refrigerated meat. Thermoduric microbes, which can survive short exposure to high temperatures but are normally mesophiles, are common contaminants of heated ‘or pasteurized foods, Examples include protozoa that produce Deat-resistanteysts such as Giardia, or sporeformers such as Bac lus and Clostridium. ‘A thermophile (hur-mob-yl) is a microbe that grows opt rally at temperatures greater than 45°C. Such heat-loving microbes live in soil and water associated with volcanic activity in compost piles, and in habitats directly exposed tothe sun, Thermophils vary imheatrequirements, with a general range of growth of 45°C 80°C. Most eukaryotic microbes cannot survive above 60°C, bat a few thermophilic bacteria, called extreme thermophiles, grow between 80°C and 121°C (currently thought to be the temperature limit because of enzymes and cell structures). Extreme thermophiles are ‘so heat tolerant that autocaves can be used to isolate them in culture, Gases ‘The atmospheric gases that most influence microbial growth are Op and CO. OF these, oxygen gas has the greatest impact on microbial growth, Not only isi an important respiratory as, but itis also a powerful oxidizing agent that can exist in many toxic forms. In general, microbes fll into one of three categories: + those that use oxygen and can detoxily it, ‘+ those that can neither use oxygen nor detoxify it, and + those that do not use oxygen but can detox it How Microbes Process Oxygen [As oxygen enters into cellular reactions, it is transformed into several toxie products. Singlet oxygen (written either as (O) or as ©.) is an extremely reactive molecule produced by both living and nonliving processes. Notably, itis one ofthe substances pro- ‘duced by phagocytes (cells of our immune system) to kill inva ing bacteria, The buildup of singlet oxygen and the oxidation of membrane lipids and other molecules can damage and destroy cell. The highly reactive superoxide ion (;"), hydrogen per ‘oxide (H.0,), and hydroxyl radicals (OH) are other destructive by-products of oxygen. To protect themselves against damage, most cells have developed enzymes whose job itis to scavenge ‘and neutralize these chemicals. The complete conversion of superoxide ion into harmless oxygen requires two-step process and at least two enzymes: Step 1. 205 +21 “1.0, cnydrogen prone) + 0; Step2. 2,0, "21,0 +0,Table 3.3A Chemically Defined Medium for Growth ‘and Maintenance of Pathogenic ‘Stophylococcus aureus Cystine Arginine Asai acid Histidine Glycine (Glue acd Levcine Irolencine Phenylalanine Lysine Proline Methionine ‘Typtophan Serine ‘Tyrosine Threonine Valine (0.005 moe nicotinamide (0.005 moe thine 0.005 mee pyridoxine 0.5 — microgram biotin 1.28. prams magnesium sulfate 1.25. grams dipoasium hydrogen phorphte 1.25. prams sodium chloride Vitamins sas (0.125 pram ion chloride Ingredients dissed in 1,000 milter of died water nd afer 0 final pH of 7.0. Chemical Content of Media Media whose exact chemical compositions are known are termed defined (also known as synheric). Such media contain pure organic and inorganic compounds that vary litle from one source to another and have a molecular content specified by means of an exact formula. Defined media may contain nothing more than a few essential compounds such as salts and amino acids dissolved in water, or may be composed of a variety of defined organic and inorganic chemicals (table 3.3). Such standardized and reproduc- ible media are most useful in research when the exaet nutritional needs of the test organisms are known If even one component ofa given medium is not chemically definable, the medium belongs in the complex category. Complex media contain extracts of animals, plants, or yeasts, including such materials as ground-up cells, tissues, and secretions. Examples are blood, serum, and meat extracts or infusions. Other ronsynthetic ingredients are milk, yeas extract, soybean digests tnd peptone. Nutrient broth, blood agar, and! MacConkey agar, 231 Methods of Cuturng Miroorganisms:The Five's $7 Table 3.38 Brain-Heart Infusion Broth: A Complex, Nonsynthetic Medium for Growth ‘and Maintenance of Pathogenic ‘Stophylococcus aureus 27'S rams brain, heart extract, peptone extract 2 grams lucose 5 grams sodium chlorite 2.5 grams disodium hydrogen phosphate Ingredients solved in 1,000 milters of dsl wae and ater 10 final pH of 70. though different in function and appearance, are all complex nonsynthetic media, ‘Table 3.3 provides an illustration of chemically defined and complex media forthe growth of Staphylococcus species Figure 3.3 Examples of enriched media. (a) 81000 aga pate ‘growing bacteria frm te human trout. Enzymes fom te bacterl Colonies break davn te red blood cells nthe agar, leaving a clear hale” This calles hemolysis (0) Culture of Neissei spon chocolate ‘9908 Chocolate ag gests brownish color rom cooked biood (not chocolate) and doesnot produce hemlysiz.80 Chapter 4 Bacteria and Archaea hhave a cell wall and some form of surface coating or glycocalyx. Specific structures that are found in some, but not all, bacteria ate flagella, pli fimbriae, an $ layer, a cytoskeleton, inclusions, microcompariments, endospores, nanowires, and intracellular ‘membranes The Structure of a Generalized Bacterial Cell Bacterial cells look very simple and two-dimensional when viewed with an ordinary microscope. But the electron microscope, as well as 150 years of laboratory research, has shown us how intricate and functionally complex they actually are. Figure 4.1 presents a three-dimensional anatomical view of a generalized, rod-shaped, bacterial cel. Bacterial Arrangements and Sizes In most bacterial species, each individual bacterial coli fully capable of carrying out all necessary life activities, such as Feproduction, metabolism, and nutrient processing, unlike the ‘more specialized cells of a multicellular organism. On the other hand, sometimes bacteria can act as a group. When bacteria ane close to one another in colonies or in biofilms, they con municate with each other through chemicals that cause them to behave differently than if they were living singly. More surprisingly, some bacteria have structures called nanowires, ‘which are appendages that can be many micrometers long and ae used for transferring clecrons or other substances outside the cell onto metas. The wires also intertwine wih the wires Of neighboring bacteria and can be used for exchanging nur tents, This is not the same as being a multicellular organism, bot it seems that bacteria can have it both ways: or cooperating with other microorg Bacteria come in many different shapes, sizes, and arrange- tment. Interms of size, bacterial cellshave an average sizeof about 1 micron um). As with everything in nature, hough, there isa great deal of vatiaton in microbial size. Uni the summer of 2022, the Jargestnon-eukaryote(akaryote) ye iscovered was Thiomargarita rnamibienss, which can be as large as 750jm (igure 42), In 2022 mother Thiomargarita was discovered that was more than 10 times larger than T: namibienss. Thiomargarita magrifica was found in underwater mangrove forests and is nearly 1 cm in Length. On the ater end ofthe size spectrum are the so-called ulr-small, bacteria. They are between 200 and 30) nanometers. One hundred fifty of them coud iin an (average sized) E.coli cel. They ate ‘ery common in nature. Scientists believe they are approaching the Tower imit possible for ving organisms because the important things like cles acids and lbosomes barely fi inside ch col en though bacteria come in many differen shapes, there a three most common shapes (figure 4.3). First, there i the occus (kokus), in which the cell is spherical or ball-shaped. Cocei thoks-eye) can be perfect spheres, but in some species the spheres are more oval, bean-shaped, or even pointed. The second type of shape is @ rod, of bacillus (bah-sil-us). (Note: ‘There is also a genus named Bacillus.) Rods can also show dif- ferent variations in shape. Depending on the species, they canbe blocky, spindle-shaped, round-ended, ong and threalike (called filamentous), or even club-shaped or drumstick-shaped. When a rod is short and plump, itis called acoccobacillus, whieh, you can tel, is a combination ofthe words coccus and bacillus, The thitd general shape for bacterial cells is eurved. If itis gently curved, itis avibrio(vib'-ree-oh). A bacterium having a curled or spiral- shaped cylinder is called a spirillum (spy-til-em), a tigi helix, twisted twice or more along its axis (like a corkscrew). Another spiral cell isthe spirochete, more flexible form that resembles asp It is somewhat common for cells of a single species to vary in shape and size. This phenomenon, called pleomorphism, is due ‘o individual variations in cell wal structure caused by nutritional or slight genetic differences. For example, although the cells of Cormebacterium diphuheriae ate generally considered rod- shaped, in culture they display variations such as club-shaped, swollen, curved, filamentous, and coccoid. The mycoplasmas are bacteria that entirely lack cell walls, and for that reason they dis- play extreme variations in shape (figure 4.4. Bacterial cells can also be categorized according to how they are arranged (see figure 4.3). The main factors influencing the arrangement of a particular cell type ae itspaltern of division and how the cells remain attached afterward. The greatest variety in arrangement accurs in coeei, which can be single, in pairs (diplo- cocci, in tetrad (groups of fou), in irregular clusters (as in staph- ylococe! and micrococei, or in chains of a few to hundreds of cells (as in streptococci). An even more complex grouping is a cubical packet of 8,16, or more cells called a sareina sar-sh-nah). These Aiferent coceal roupings depend on whether the cells divide along single plane, in two perpendicular planes, or in several intesect- ing planes. Because bacteria usually reproduce by spliting in two (overand over again) ifthe divided cells remain attached, they lead to these diverse arrangements. Rods are less varied in arrangement because they divide only in the transverse plane (perpendicular to the axis). They ‘occur either as single cells, as a pair of cells with their ends attached (diplobacilli), or as a chain of several cells (streptoba- ‘A palisades arrangement is formed when cells ofa chain remain partially attached at the ends. This hinge area can fold back, creating a side-by-side row of cells. Spirilla are occasion- ally found in short chains, bt spirochetes rarely remain atached alter division, Biofilms. Bacteria often do not exist in a planktonie oF single-cell form but rather live in cooperative associations that can include other organisms of the same species as well as other species of bacteria, archaea, fungi, and algae, These biofilms are microbial habitats with access to food, water, atmosphere, and other environmental factors that are beneficial to each type of organism living there Often, the biofilm is stratified, with the aerobic microbes near the surface where the oxygen levels are high and the anaerobic microbes near the botiom where oxygen levels are low. Each mem ber ofthe biofilm community finds its niche Biofilms can form on numerous inert substances, usually \when the surice is moist and has developed thin ayer of organicneeded to process toxic oxygen products, An organism that must have oxygen is an obligate aerobe. Most fungi and protozoa, a5 ‘well as many bacteria (and humans!) ae obligate aerobes. ‘A facultative anaerobe is a microbe that does not require ‘oxygen for its metabolism and is eapable of growth in the absence of it. This type of organism metabolizes by aerobic respiration when oxygen is present. In its absence, it can tog- gle to an anaerobic mode of metabolism such as fermentation, Facultative anaerobes usually have both catalase and superoxide dismutase. A large number of bacterial pathogens fall into this group (eg.. gram-negative intestinal bacteria and staphylo- cocci). A microaerophile (myk"-roh-air-oh-fyl) does not grow At normal atmospheric concentrations of oxygen but requires 4 small amount of it in metabolism, Most organisms in this category live ina habitat (soil, water, or the human body) that provides small amounts of oxygen but is not directly exposed to the atmosphere. ‘An anaerobe (anaerobic microorganism) lacks the meta- bolic enzyme systems for using oxygen in respiration. Because obligate anaerobes (also Known as strict anaerobes) lack the ‘enzymes for processing toxic oxygen, they cannot tolerate any free ‘oxygen in the immediate environment and wil die if exposed to it ‘Obligate anaerobes live in habitats suchas deep mul, lakes, oceans, and sol, Even though human cells use oxygen and oxygen is foundin the blood and tissues, some body sites provide anaerobic pockets oF microhabitats where colonization or infection ean occur: One region that i an important site for anaerobic infections is the oral cavity, ‘The pockets between the gums and the teeth can be anaerobic atthe ‘tom of them, and this fosters the growth of microbes that cause ‘gingivitis and periodontal disease, Another common site for anaero- bic infections isthe large intestine, a relatively oxygen-free habitat that harbors a rich assortment of strictly anaerobic bacteria. Grow: ing anaerobic bacteria usually requires special media, methods of incubation, and handling chambers that exclude oxygen (figure 9.7). { Disease Connection “Tetanus, also known as lockjaw, is caused by an anaerobic bac terium, Clostridium teton:. You are aware that you may be at risk {or this infection through puncture wounds such as stepping on '@ nail. This reflects the oxygen requirements of the bacterium Deep puncture wounds contain Imited oxygen atthe “nottom” of the wound and often are not perfused with alot of blood (ancther “source of oxygen}. n that situation, Clostridum endospores can return toa growing stage and mutiply Aerotolerant anaerobes do not utilize oxygen but ean sure vive and grow toa limited extent in its presence. These anaerobes are not harmed by oxygen, mainly because they possess alternate ‘mechanisms for breaking down peroxides and superoxide, Certain Iactobacili and streptococci use manganese ions or peroxidases 10 perform this task, Determining the oxygen requirements of a microbe from a biochemical standpoint can be a very time-consuming process, One 9.2 Environmental Factor That Intuence Microbes 235, Catalyst chamber contains palladium pene, which scavenge excess oxygen outers Inner i Rubber gasket Gas geertr elope, Water Soria San hss protic 0 ich Anaerobic inceatr stip ‘als ofthe chamber ‘sence of, o Figure 9.7 Culturing techniques for anaerobes. (8) special ‘anaerobic envranmetal chamber makes & posible to hance sc specton in a completely O-ee system. (B)A simpler anaerobic, oF CO, incubator syter.T igh. The gas react wth avalable oxygen to procice wate Carbon dioxide can aso be added to the system fr growth of organisms cing high concenvations oft way to do it i to grow the microbe using reducing media (those that contain an oxygen-absorbing chemical), Figure 9.8 depicts the growth of different microbes in tubes of fluid thioglycollate ‘The location of growth indicates the oxygen requirements of the microbes, Insight 9.1 tells the tale of how microbes that were able to produce oxygen via photosynthesis were responsible for turning ‘our planet from an anaerobic one to an aerobic one ‘Now let's talk COs, Although all microbes require some ‘carbon dioxide in their metabolism, cupnophiles grow best at aUji Bioaktivitas Minyak Cengkeh (Syzygium aromaticum) Terhadap Pertumbuhan Bakteri Streptococcus mutans Penyebab Karier Gigi Antibacterial Activity Of Clove Oil (Syzygium Aromaticum) In Inhibiting The Growth Of Streptococcus mutans causing Dental Disease A.R.Pratiwi Hasanuddin"’,dan Subakir Salnus! ‘Prodi Dill Teknik Laboratorium Medik, STIKES Panrita Husada Bulukumba “Corresponding Author: a.r.pratiwihasanuddin@gmail.com Abstrak Kesehatan gigi dan mulut merupakan faktor yang dapat menimbulkan masalah kesehatan jika diabaikan. Salah satu masalah yang ditimbulkan adalah gigi berlubang (karier gigi), Jenis bakteri yang dapat menyebabkan terbentuknya plak, yaitu bakteri dari genus Streptococcus. Tanaman cengkeh (Syzygium aromaticum) merupakan tanaman tropis. Bunga cengkeh mengandung minyak atsiri (clove essential ofl) dengan kandungan utama jalah Eugenol yang dianggap mampu membunuh bakteri Streptococcus mutans penyebab karier gigi. Penelitian ini bertujuan untuk mengetahui kemampuan ekstrak minyak cengkeh (Syzygium aromaticum) dalam menghambat pertumbuhan bakteri Streptococcus mutans.Hasil penelitian menunjukkan terdapat perbedaan. signifikan antara Kelompok konsentrasi tertinggi dengan kelompok konsentrasi rendah dan kontrol (P<0.05). Sedangkan, kelompok perlakuan ciprofioxacin jika dibandingkan dengan kelompok konsentrasi 100% tidak terdapat perbedaan signifikan (p>0.05).Ekstrak minyak cengkeh dapat menghambat pertumbuhan bakteri Streptococcus mutans. Semakin tinggi konsentrasi ekstrak minyak cengkeh yang digunakan maka semakin besar diameter zona hambat (zona bening) yang terbentuk. Kata Kunci:Aktivitas Antibakteri, Ekstrak Minyak Cengkeh, Bakteri Streptococcus mutans. Abstract Oral health is a factor that can cause health problems. One of the problems caused cavities (dental disease). The type of bacteria caused plaque is a bacteria from the genus Streptococcus. Cloves (Syzygium aromaticum) is a tropical plant. Clove flowers contain. clove essential oil, with Eugenol has antibacterial activity. This study aimed to determine the effect of the clove oil (Syzygium aromaticum) in inhibiting the growth of Streptococcus ‘mutans. The results showed that there was a significant difference between the group with the higher concentration than lower concentration and control (p<0.05). The clove oil effective in inhibitng the growth of Streptococcus mutans. The higher concentration of the love oil can inhibit with larger diameter of the inhibition zone (clear zone) is formed. Keywords: intibacterial Activity, The Clove Oil, Streptococcus mutans. http://journal.unhas.ac.id/index php/bioma 241Bioma volume 5 (2) : 241-250, Juli - Desember 2020 antara 80-85% (Hadi, 2012). Dari setiap bagian-bagian cengkeh yaitu bunga cengkeh, tangkai cengkeh dan daun cengkeh, kandungan minyak atsiri dan kadar eugenolnya yang paling banyak terdapat pada bunga cengkeh. Eugenol merupakan salah satu ‘senyawa golongan fenol yang diketahui memiliki efek toksik terhadap bakteri. Senyawa fenol dapat menembus membran sel bakteri kemudian berinteraksi dengan enzim dan protein pada membrane tersebut maka dapat menroton yang berlawanan sehingga dapat, merusak sel bakteri (Xing ef al, 2012).Mekanisme eugenol sebagai antibakteri dengan cara menembus bagian membrane sitoplasma kemudian mengganggu atau merusak kemampuan permeabiltas dinding sel bakteri. Selain itu, sitat hydrophobic (tidak larut dalam air)yang dimitiki eugenol lebih memudahkannya menembus lipopolisakarida dari membrane sel bakteri. dan mengubah struktur dinding sel, struktur dinding sel yang berubah kemudian menyebabkan kebocoran pada bagian intrasel sehingga menghambat pertumbuhan bakter, kemampuan inilah akhimya ekstrak minyak cengkeh dapat dijadikan sebagai antibakteri (Posangi, 2016). Pada penelitian ini diketahui bahwa ekstrak minyak cengkeh menghambat pertumbuhan bakteri Streptococcus mutans. Streptococcus mutans adalah salah satu bakteri gram positif, yang memiliki peptidoglikan lebih tebal. Penelitian ini serupa dengan Penelitian yang dilakukan oleh Andries ef el. (2014) yang menyebutkan bahwa ‘Streptococcus mutanssebagai bakteri gram positif dihambat pertumbuhannya secara in vitro oleh ekstrak bunga cengkeh. Eugenolyang terkandung di dalam cengkeh merupakan ‘senyawa antibakteri yang dapat menghambat pertumbuhan bakteri pathogen yaitu bakteri gram negatif dan bakteri gram positif. salah satu bakteri yang termasuk bakteri (Pertiwi et al., 2017). Selain terhadap beberapa bakteri, minyak atsiri cengkeh juga telah diujikan terhadap jamur Candida Albicans. Penelitian yang dilakukan oleh Pratiwi et al. (2015) menunjukkan adanya daya hambat minyak atsiri cengkeh terhadap Candida albicans. Penelitian tersebut menggunakan minyak atsiri dari ekstrak daun cengkeh yang juga memilki komponen utama eugendl. Zona hambat yang terbentuk dapat klasifkasikan seberapa besar respon hambatannya dengan mencocokkan pada tabel Klasifikasi respon zona hambat menurut Alfath ef al. 2013). Peneliian ini respon kuat terdapat pada konsentrasi 100%, 80%, dan. 60% dalam menghambat bakteri Streptococcus mutans karena memiliki rerata diameter zona hambat lebih dari 20 mm (Alfath et al, 2013). Ciprofloxacin yang digunakan sebagai pembanding memiiki rerata diameter zona hambat lebih dari 20 mm sehingga diklasifikasikan dengan respon kuat.Pemilinan ciprofloxacin sebagai kontrol positif karena ciprofioxacin merupakan golongan obat flouroquinolon yang memiliki berfungsi untuk menghambat sintesis DNA bakteri sehingga menghambat resistensi mikroba dan merupakan antimikroba berspektrum luas. Kesimpulan Ekstrak minyak cengkeh dapat menghambat pertumbuhan bakteri Streptococcus mutans. Semakin tinggi konsentrasi ekstrak minyak cengkeh yang digunakan maka semakin besar diameter zona hambat (zona bening) yang terbentuk. http://jpurnal.unhas.ac.id/index php/bioma 248Cakradonya Dent J 2022; 14(2): 95-99 ORAL THRUSH PADA BAYI: GAMBARAN KLINIS DAN TATALAKSANA. (LAPORAN KASUS) ORAL THRUSH IN BABIES: CLINICAL APPEARANCE AND CASE MANAGEMENT (CASE REPORT) ‘Taufiqi Hidayatullah!+ Laxmi Nurul Suci2 'Bagian lmu Kedokteran Gigi Anak, Fakultas Kedokteran Gigi Universitas Syiah Kuala Banda Acch Staf Bagian Instalasi Gawat Darurat Rumah Sakit Pertamedika Ummi Rosnati Banda Acch Correspondence email to: taupq.drg@unsyiah.ac.id ABSTRAK Oral thrush merupakan infeksi jamur didalam mulut yang umum terjadi pada bayi. Etiologi utama oral trush disebabkan kebersihan yang tidak adekuat. Faktor tersebut dapat berasal dari rongga mulut bayi, kulit puting ibu, kebersihan tangan ibu dan bayi serta peralatan makan dan minum bayi Diagnosis kerja umumnya diperoleh melalui anamnesis dan pemeriksaan klinis. Biopsi dapat dilakukan untuk konfirmasi diagnosis apabila oral thrush menyerupai kondisi oral lain. Gambaran Klinis dapat berupa bercak putih menyerupai krim yang dapat diseka pada mukosa mulut serta pada beberapa kondisi dapat meninggalkan dasar hiperemis setelah diseka. Oral higiene yang adekuat mampu menghilangkan infeksi jamur jika belum terlalu parah. Apabila sudah meninggalkan permukaan hiperemis saat discka dan lokasi jamur makin meluas dapat dibantu dengan obat anti jamur seperti miconazole cream. Selain itu, diperlukan pula mengurangi faktor predisposisi yang terlibat, sepert sanitasi bayi dan ibu guna menunjang efektivitas obat mengeliminir jamur. Cara pemakaian obat yang tidak tepat akan memyebabkan perawatan tidak berhasil. Oleh Karena itu, dokter gigi harus memberi edukasi secara baik dan benar mengenai tata cara penggunaan obat dan pembersihan mulut bayi agar terapi yang diberi sesuai harapan. Kata kunei: Oral thrush, kebersihan mulut, infeksi jamur ABSTRACT Oral thrush is common fungal infection in the mouth of infants. The primary etiology of oral thrush is, caused by inadequate oral hygiene, These factors can come from the baby’s oral cavity, the skin of the mother’s nipples, the cleanliness of the mother’s and baby’s hands, as well as from the baby’s eating and drinking utensils. The working diagnosis is generally obtained through history taking and clinical examination, A biopsy may be performed to confirm the diagnosis if oral thrush resembles other oral conditions. The clinical picture can be white patches resembling cream that can be wiped on the oral ‘mucosa and, in some conditions, can leave a hyperemic base after wiping. Adequate oral hygiene can climinate fungal infections if they are not too severe. If it has left a hyperemic surface when wiped and the location of the fungus is expanding, it can be helped with antifungal drugs such as miconazole cream, In addition, it is also necessary to reduce the predisposing factors involved, such as the sanitation of infants and mothers, to support the effectiveness of drugs to eliminate fungi. Improper use of the drug will result in unsuccessful treatment. Therefore, dentist must educate properly and correctly about the procedures for using drugs and cleaning the baby’s mouth so that the given therapy is success Keywords: Oral thrush, oral hygiene, fungal infection 95 | Cakradonya Dental Journal p-ISSN: 2085-546X; e-ISSN; 2622-4720. Available at http://www jural.unsyiah.ac.id/CDJPENDAHULUAN i akan dibahas suatu Dalam laporan kasus oral thrush yang tidak kunjung sembuh, Lesi-lesi_putih bertambah banyak walaupun orangtua telah diresepkan obat oleh dokter. Edukasi mengenai tatalaksana kasus yang tidak tepat akan -menyebabkan perawatan tidak maksimal, Setelah diberikan edukasi yang tepat, —infeksi—_jamur menghilang dalam 3 hari berangsur-angsur LAPORAN K ASUS Pada tanggal 10 Juni 2021 seorang bayi berusia 11 bulan datang bersama orangtwanya ke IGD rumah sakit dengan keluhan bayi rewel tidak mau menyusui dan demam. Orangtua menginformasikan ada gumpalan putih yang banyak didalam mulut bayi. Gumpalan tersebut bertambah banyak dalam 1 minggu terakhir, Temuan bercak tersebut pertama kali dijumpai sekitar 10 hari yang lalu, Orangtua sudah dibekali nistatin gel oleh dokter tetapi tidak kunjung sembuh Gumpalan tersebut sempat berkurang setelah 2 hari pemakaian obat tetapi berangsur-angsur memenuhi mulut bayi sampai ke bagian langit- langit, Area pertama yang terkena adalah lidah, Cakradonya Dent J 2022; 14(2): 95-99 Pemeriksaan menunjukkan ada lesi putih ‘menyerupai krim di palatum mole, Saat diseka menggunakan kasa, lesi_tethapus dan meninggalkan eritema. Lesi putin lainnya dijumpai pada mukosa bibir dan pipi. Lesi tersebut dapat dihapus tanpa meninggalkan eritema, Bayi tampak rewel dan diketahui malas menyusui sejak 2 hari terakhir. Kasus ini Kemudian dirujuk ke dokter gigi spesialis anak untuk ditindaklanjuti Orangtua —-menjelaskan_——_sudah. memberikan nistatin gel pada bayi. Orangtua ‘mengaplikasikan nistatin tersebut 2 kali. sehari dengan cara meneteskannya di lokasi_lesi. Tidak ada upaya menyeka mulut anak pasca menyusui dan makan karena khawatir anak akan rewel. Akibat kondisi pandemi, dokter ‘memberikan instruksi khusus kepada orangtua Setelah menganalisis hasil anamnesis dan pemeriksaan klinis, maka ditegakkan diagnosis yaitu pseudomebran kandidiasis akut atau oral thrush, Orangtua khususnya Ibu diajarkan dan diinstruksikan untuk menjaga _kebersihan tangan, kulit puting saat menyusui, peralatan ‘makan minum bayi dan kebersihan mulut bayi tersebut, Setiap kali memberikan makan minum kepada bayi, Ibu ditekankan untuk mencuci tanga dengan sabun dan membersihkan putting dengan air matang sebelum menyusui. Selain itu, juga menyeka mulut anak kasa yang, dibasahi air matang sebelum dan sesudah ‘menyusi atau makan. Pemberian obat anti jamur _diganti dengan miconazole oral gel sebanyak 1,25 ml yang dibagi 4 kali schari. Mempertimbangkan faktor bayi yang belum koperatif saat diberikan bat, maka orangtua disarankan meneteskan gel pada kasa dan menggulungkan kasa tersebut pada jari telunjuk yang bersih—lalu mengoleskannya tepat pada lesi-lesi putih Kepalabayi dikontrol pergerakannya agar pengaplikasian obat optimal, Bayi tidak diberikan makan ataupun minum selama 30 menit, Agar memudahkan orangtua, dokter ‘menginstruksiakn memberikan obat_tersebut pada pukul 8 pagi, pukul 12 siang, pukul 4 sore dan pukul 8 malam. Setelah 1 jam pemberian terakhir, orangtua disarankan membersihkan mulut bayi menggunakan kasa lembab yang dibasahi cairan khlorheksidine glukonat 0,2 % dan diseka lembut pada seluruh permukaan mulut bayi, termasuk bagian luar bibir. Tiga hari 96 | Cakradonya Dental Journal p-ISSN: 2085-S46X; eISSN: 2622-4720. Available at http:/www.jumal.unsyiah.ac.id/CD)