Lamia B. Ilalh 4 Experimant OL: Serial Dioton Tibroduction ie Serial civhors are perbrmed! on bacterial somples fo rece a very Wigh contenbaton of. Lacheria f a manageable amownt for bose A counting and analysis. Once grown in colfure, te backrial, colonies an be combed and pe diltin pacer Can be used 40 calevlafe be number of be Cel tia fle onginal sample. fen-fold Silvbon 1s mash ofhen carried wr, from [0''ybo OF Kesults ie L gam of soil was mired tat sober solfon ps crafe Be shotk solution R- fe resulls offer serial dofjon are shown Selw. Fque 1: Serial dilion & soil Me see Hal te samph gradvally Lecames or and mont Abed as fle concenbrabjon of soil LecreovesLama Liffah 2 Discussion : Ae frst pigpared fe stock solution 6 ohhh ue tnLolls— 10", 107 10° 10" andl lashy 10° J ml A shock soln R tos “ptaken by a ruceqpipelke, andl etched thfy Fuld saline water jn a febe. The pube 10s closed offer Jeobne. hover fre. fabelled (0 anel pot onfo a vorfer mixer fo polly ineoporafe. len again Lml. of He 10" solvhon wes added to a tube confaming, Ql saline walker Jo make a (0 solutitn, all steps are repeater! Th Mis wamer, fe process is carrie! on unfl (07. Geral rule 4 aseohe pechnigre mut be followed i order 4 aver conhirrnabn. Sbrile hp 8 ved every firme. . Conclus ion. The twer He chitin pachor, feser fe amoonf A batheria is possent, Me solufions 10°, 103 107 10 £10" ax row ready to be used fr ging bacteria va colkure B cod dbus be Cay gta More accvirafe cnreration trol it makes backrial solation casienLamia Lilleh 3 vernal 02: Poor Plate Thtroduction : oor-plabag Js ont fle processes Sy whith we Can /solate and count viable microbes fom diferent. samples | [his whol feces fhe Glonth a Soth acrobic and | anacpobic mittoorganisms , which grow on Pe surface on f He agar ancl inderveath /f pepechrely. The process is cone by simply wiring. Ahk sample with fgg hu foen fp agar tn a pefer lish and hehing a tacobate » ¢ Lesu/fs , | Figure : Poor plate ooLama Byileh 4 As observed aber 24 hours, He dah with 0 ohlebon fos He bighes! number A Lackriol cobnies. The amount gradvally decrecaes fom 10" fo (07 considering 10” 25 anomalous. Tle enumeotion of. Sackria & splayed ta foe Fable Lelow Ibu] CFU [Nomber a baclerin | Lo") 813 ome) | 13 x10 Crum 10" 796 (rm) | 796 x 10" CFUAnl | fea areca — or 260 | 260 x10" crim 17) 196 1a x10 Crum | Tb ll coun) = CFU x Pili alr *| Discussion: Lach dha Le rerbas. in oor Group worleel on ove chloton Pyctor for the experivent. tal dt be dhbhis | were adlded fo agar solition by a merapipele.and swirled clockwise - counterclockwise f obssohe. Te rot olay cer inclthation, fe 10" dish had abrorrally fos CFU, Possibly dhe fo micropicebe misanoling #é. fess solhen | feted. The counts may hare been patonsislen/ due fy striel oblton errors. 2 few dishes had a ssiinf polern trom inadeguate Pang. Conchsion ! LP done properly, the poor plate mefgod allows easier counting , Collechion and analysis fr olikeren Apes of 2trbic and anatrohic microbesLamia Billah Ezperinnd 03: Sorted Hole Tpbroclvetion : Spree plobna is anopler phibng wtfhods by whch microorganisms Can be ologed are! conked. Unlike fo pour ple mathod , spread plate only favilfates fhe grouth atrobes, vhich grow on the siface fa sohdied agar plate. Hore a serially dilubedl somphe is sproded frely over He agar plate using A spread, anc! febf fo procreafe for 24 hours. G Kesults : Tle vert doy, wr experivenf yielded fle following rests. Figure 2 Spread plate 4s seen in He image, backericl colonies are Armed on the ogar surface, A cohur with mostly vrlar shape pete yelou Lereammia Lillah ° Te enomeration fo backria is prstnied on fhe fable low: Dillion | CFU | Number of backeria in sample 10° | TNTC TNTC 10 | [02 |roaxl™ = 102 X 10 crUAnL 10° 4O — |4ox®F =40 X [O%cem 10" 10 |rox BE =10 x 107A [107 | 29 jaax Sr = 29 x10‘crumt Amount d sobtion used = 100uL + ou fe . - Diltion Factor Tobol viabe cell comb = CFU X Gnoonl wt soln) Discussion: The experiment is stocked! by ooing. serial dltion ofa Soil samppe , similar fo experiment 02. Sohobfed agar plates were popared beforehand Lack of fle fae Group members selected! a chlution factor to work with, and prepared G spread plates in foto: 10QuL of solifion tos ree onto oe as “sing A ratere- - pipette. (bing a sherile spreackr (ohpped K bens) and shied tig Oe Hle solifon 15 es ae ee / agar. Gene yP-clun mobo ts vsed while rotating Phe plate simulfereouly. Tf fe sprachr is too hol, microbes ul/ de and the count will be matturale as seen jin [7 Alben. Pecouhors mst be fakery whife working with ang *| Conelusiont Spread plate Sa sinphe and relatively as/ er Hhocl polafing bacleria. HF is moch easier fo oon ee fom compared - pour Plate. bith Proper Pechargue be Gan get rehable reso lb.a —Leverinen 04: Sheak Hae. Steak plalng is a method by which pure culrs A vcroorganisms Can be soleted from avr inihially Ata Sampfe. his petro alows separapion f. SPCIES for a wore deboiled aralysis. Tf 1s elore by scraping A a y ynixeel sample and ronning long sheaks long & soliljfed ager surface, using on tnoclating trop. Tifrooliction : +| Lull : Figure : Sheok plate 1-2 Abler 24 hors, flere were no observable colonies on He fist atempt. Howeyer, on He second attenpl a very nse incubation ct off-whife wolhni'es ean be Seen Here He Lf quackant 1s gyhpcly core wi backrin. Te secon and! Hird quackonts show Ines fo Chnies along where streaking vos done. The Sfh guadran f 5 abo chase with cobnie pith lhe b ro serarcha,Lamia Blah £ " Discussion! TL sherk plabing webhod took fle hast atnount ft tine fo carry out. Tt 15 abo He only melted of plating Wt practiced frat oes not peguire Serial, Mebion. Fhe olihon is on Pack done on fhe agar plate Melk, Firs, an inoculahng (oop is Kealed unl glowing rect Ys Serifice if Then, some of the bacterial colonies 1 scraped off Foma mixed plate. fle scrapings are sheabed on a posh i) ou plake. fla ff pradhan! oe one lpetng dose speaks. Then, fle foop 1 heated and cok an £o ainiliabe eek Loorio. The secon! and Ate! guactad consist ch 3-9 separated Streaks. Ay secn th fhe mage, ny Pecnger wos Copsisbent bith fhe rule. Loshly He Joop iss hated 5 dle Sth geadranf. and a Wavy sek S made. foweler, I mm off foo much bacteria on my Ln Ke ee why the colonies are not oshinguishable. » Which 1s th te offer Jan 0 colonies were seen on my Arsh afenpt. This Covll be de to He loop not coofng property Lehre He mocvlation. resulting in all bacteria pe olie- Conchsion: Sheak plating Can seve 23 a guich method! for jsolaling. Inicroorganismns. Homerer, excyohonal Reabniger is KiuINd 40 obtain perfect, Confable colnjes at tee end:| Lamia Lihah 7 Lxverinenf. 8: Gram Shining . Lphoolction; Gram sfairmg is o process fr difrentokng Layeen phe 2 prnciok grops bf backria ~ Gram Posifie anol ram negate. [his ts ohne on fe boss of Heir Alf walf compasion. Named offer Dr. Cristian Gram, fois Aopular staining method savohes ¢ reagenls over 4 SHPS_ 1) Crystal violet ston 2) Grams jodie sfain (mordant) ) 96L ety| aleobol decolorizer 4) Safranin Courker-shaiy | Bosal : Figure : Gran Shain ing Upon inspection. we observe fyck. perp =shained rod “shaped Sackeria iin clysfers, 05 well cs smaller, pink-staved. pocdt-shaped Lackria scafere! Hroughout. A magnikeotion th 00x 5 reguirtd +o wer this properh,Love bailleh 16 I. jon: - ib cary owt His everett, cach sholut fook a shell oss shed and pachad o snear on tt. A dup A saline solution wos put on te stde using a sterile hnoculabing hoop. thing te bop. a sample was falen bam Gg mixed culure, arc sneared tia a Cirlar motion on Phe gloss she. lath. fhe shite twos feat fired. Cr tushector casi shed us tn aching fle reagents. Arsh was costal violet, fle Primacy Shain foal sbawed all cals wolet offer Tmin- Then, Grams Tockue wos cpofed br anofrer. mule offer breshing ol the ceyshal violef This ach hke & mordant. forming an rasolubse compoend with He prinard stan, olled crystel viokt raclne compks CC). whith bind perranently fo mognesiom —riLowckic acd, only present ya Gram pos'hve coll walls. Lue fo Abs. only Gran- -paifie backrio are able fo refain Pe violet cobur. an after decolerising uith ofhyf aleohol. Lastly, He shde js wesked end sofpanin 1s opphed fr $Sveconck fo visvellze He Gram regabre bacterin. The purple rod shaped bacteria are fitely Bouiles or lactobaceles, mbyle swaler pink rods coll be Ecol. "|Z onclusion : Through fle Gram Shaining process, he Can Porefove Ashaguish ano! vdnb ly varius species of Gram - park aac! Gran- ragafre bacteria, and ofo visvablzo Heir relate shape and size. Xx