1(2)58-66 !!!!!!!!!!!!!!!!!! CONTACT&:&EVI&LAUW&&&&&&&evi.lauw09@gmail.com! ©!International!Journal!of!Applied!Biology! 58! Abstract& Lactic!acid!bacteria!are!known!to!have!potential!in!producing!antimicrobial! compounds,! including! antifungal! agents.! This! study! aims! to! determine! whether!the!lactic!acid!bacteria!from!human!oral!mucosa!have!the!potential! to! produce! the! secondary! metabolite! to! inhibit! the! candidiasis,! which! caused! by! pathogen! fungi,! Candida& albicans.! Lactic! acid! bacteria! were! isolated!from!three!healthy!respondents!based!on!age!difference!that!is!an! infant,! child,! and! adult.! Lactic! acid! bacteria! isolation! using! the! deMann! Rogosa!Sharpe!Broth!(MRSB)Jliquid!medium,!followed!by!purification!stage! with!the!quadrant!scratch!method!using!the!deMann!Rogosa!Sharpe!Agar! (MRSA)Jsolid! medium.! The! purified! isolates! obtained! were! characterized! and! identified! by! Gram! staining,! biochemical! tests,! and! inhibition! zone! diameter! measurement! toward! pathogenic! fungi! Candida& albicans.! The! results!were!obtained!from!each!of!the!two!lactic!acid!bacteria!isolates!from! each!respondent!with!characteristics!including!gramJpositive!bacteria!with! short!stem!cell!shape,!possibly!as!Lactobacillus&sp.!It!is!conclude!that!the! lactic! acid! bacteria! of! oral! mucosa! from! adult! had! greater! anticandidal! activity!than!infant!and!child.! & ISSN!:!2580J2410! eISSN!:!2580J2119! ! ! ! Study&of&Lactic&Acid&Bacteria&Activities&from&Human&Oral& Mucosa&for&Candida&albicans&Inhibition& ! Evi&Lauw1,&Khusnul&Hatimah&Ilham1,&Febliani&Taruk&Palinggi1,&Desy&Setiady2&and&Sartini1& ! 1!Faculty!of!Pharmacy,!Hasanuddin!University,!Makassar,!Indonesia! 2!Faculty!of!Dental!Medicine,!Hasanuddin!University,!Makassar,!Indonesia! ! ! & & & & & & & & & & & & & & & Introduction& During! the! last! one! decade,! the& prevalence! of! infectious! diseases! caused! by! pathogenic!fungi,!which!is!candidiasis,!has!increased!sharply!(Fortún&et&al.,!2012).!Candidiasis! is! a! mucosal! fungal! infection! on! humans! of! all! ages! that! affect! the! skin! and! mucous! membranes!of!oropharynx,!oesophagus,!and!vagina!(Fortún&et&al.,!2012;!Gajer!et&al.,!2012).! There!are!at!least!15!distinct!Candida!species!that!cause!human!disease,!but!more!than!90%! of!invasive!disease!is!caused!by!the!five!most!common!pathogens,!C.&albicans,&C.&glabrata,&C.& tropicalis,&C.¶psilosis,&and&C.&krusei!(SilvaJDias!et&al.,!2014).!Candida&albicans&continues!to! be!the!most!prevalent!and!problematic!of!all!Candida!species!(Perera!et&al.,!2015;!Paul!et&al.,! 2014).!The!pathogenicity!of!Candida!species!is!attributed!to!certain!virulence!factors,!such!as! the!ability!to!evade!host!defences,!adhesion!and!biofilm!formation!(on!host!tissues!and!or!on! medical! devices),! and! the! production! of! tissueJdamaging! hydrolytic! enzymes,! such! as! proteases,!phospholipases!and!haemolysins!(Paul!et&al.,!2014).!Moreover,!the!morbidity!and! &&&&&&&&OPEN&ACCESS& & & &&&&&&&&&&International&Journal&of&Applied&Biology& Keyword! Lactic&Acid&Bacteria&& Human&Oral&Mucosa& Antifungal&Activity& Candida&albicans! Article&History! Received!23!May!2017! Accepted!10!November!2017!! International Journal of Applied Biology is licensed under a Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ! International Journal of Applied Biology, p-ISSN : 2580-2410 e-ISSN : 2580-2119. Journal homepage : http://journal.unhas.ac.id/index.php/ijoab International&Journal&of&Applied&Biology,&1(2),&2017& ! 59! mortality!associated!with!candidiasis!are!still!very!high,!even!using!the!actual!antifungal!drugs.! Annually,! 50%! of! adults! and! up! to! 30%! of! children! die! of! candidiasis! (Paul! et& al.,! 2016;! Martinez!et&al.,&2015).! The!major!classes!of!antifungal!drugs!used!for!treatment!of!Candida!species!infections! are! azoles,! polyenes,! and! echinocandins! (Martinez! et& al.,! 2015).! According! to! previous! experimental!results,!irrational!use!may!lead!to!colonization!of!Candida&sp.!that!are!resistant! to!these!drugs.!It!has!been!reported!that!conventional!treatment!for!treating!candidiasis!with! fluconazole!has!a!case!of!resistance!(Martinez!et&al.,!2015;!Costa!et&al.,!2015).!The!occurrence! of! Candida! species! infections! becoming! more! difficult! to! treat! due! to! the! growth! of! immunogenic! diseases,! endocrine! disorders,! malnutrition,! the! disproportionate! use! of! immunosuppressive!drugs,!and!broad!spectrum!antibiotics!(Costa!et&al.,!2015).!It!is!believed! that!the!increase!of!C.&albicans&infections!is!due!to!its!intrinsically!low!susceptibility!to!azoles,! including! the! imidazole! and! the! oralJparenteral! triazoles! (e.g.,! fluconazole,! voriconazole)! (Gayford! JJ! &! Harskell! R,! 2010).! Additionally,! it! is! known! that! the! acquired! resistance! is! resulted!of!rare!mutations!that!are!selected!by!drug!pressure!(Samaranayake,!2012).! The!recent!efforts!of!various!excellent!researches!tremendously!broadened!on!the! complex!mechanisms!of!normal!flora!(Scatassa!et&al.,!2015).!They!were!shown!that!living! creatures!without!normal!flora!are!more!susceptible!to!pathogenic!microbes!(Kanmani!et&al.,! 2013;! Amadou! et& al.,! 2013;! Amenu,! 2013).! In! this! sense,! the! authors! highlighted! the! importance!of!normal!flora!and!that!the!addition!of!secondary!metabolite!to!improve!the! efficacy!of!candidasis.!The!result!of!a!study!showed!that!there!was!an!increase!in!oral!and! dental!health,!as!well!as!a!decrease!in!the!number!of!Candida&albicans&fungi!in!school!children! who!consumed!milk!or!cheese!containing!Lactobacillus&rhamnosus&strain&GG.3!which!belongs! to!the!lactic!acid!bacteria!group!(Canabarro!et&al.,!2012).!According!to!the!authors,!some! strains! of! lactic! acid! bacteria! have! a! positive! activity! in! reducing! the! amount! of! Candida& albicans!in!the!saliva!of!the!human!oral!cavity!(Amenu,!2013;!Canabarro!et&al.,!2012).! Recent! studies! revealed! the! involvement! lactic! acid! bacteria,! especially! the! genus! Lactobacillus,! can! be! isolated! from! animal! and! human! digestive! tracts,! including! upper! gastrointestinal! organs! such! as! the! oral! cavity! (Corona! et& al.,! 2016).! It! has! obtained! Lactobacillus&gasseri&isolates!identified!using!16SJrRNA!sequencing!from!oral!mucosa!of!a!4J monthJold!infant!(Settanni!et&al.,!2013).!However,!this!article!showed!antiJfungal!activity!of! lactic!acid!bacteria!isolates!from!three!most!potent!sources!as!producers!of!anti!candidiasis! metabolite!compounds!that!are!an!infant,!child,!and!adult.!The!significance!of!this!study!is! due!to!its!contribution!to!knowledge!on!the!determination!of!the!ability!of!lactic!acid!bacteria! isolates!from!the!oral!cavity!of!humans!as!anti!candidiasis!agents.! & Materials&and&Methods& Preparation&of&Tools&and&Materials! The!tools!used!in!this!research!were!Autoclave!(All!American),!Incubator!(Memmert),! Laminary!Air!Flow,!Microscope!(Olympus),!Oven,!Caliper,!Analytical!Scales!(Ohauss),!Sterile! Swab! (OneMed),! petri! disc,! Chemical! Glass! (Pyrex),! Bunsen,! Cotton,! Measuring! cylinder! (Pyrex),!handscoon,!electric!stove,!cloth!mask,!round!loop,!straight!loop,!spoon,!test!tube,!vial! tube,!etc.! The!materials!used!in!this!research!were!70%!alcohol,!aqua!distillate,!pure!Candida& albicans!culture!(obtained!from!Microbiology!Laboratory,!Faculty!of!Pharmacy,!Hasanuddin! International&Journal&of&Applied&Biology,&1(2),&2017& ! 60! University),!ketoconazole,!deMann!Rogosa!Sharpe!Agar!and!Broth!(MRSA!and!MRSB)!media,! and!Potato!Dextrose!Agar!(PDA)!medium.! ! Experimental&Methods& Isolation&of&Lactic&Acid&Bacteria& Oral!mucous!samples!were!taken!from!each!of!the!three!samples!individually,!infant,! child,!and!adult.!Sampling!is!done!by!rubbing!into!the!inside!cheeks!in!samples!using!a!sterile! swab!sticker.!The!swab!result!is!then!inserted!into!a!vial!containing!the!MRSB!medium!as!a! transport!medium,!then!immediately!taken!to!the!laboratory.! Samples!of!the!oral!mucosa!were!diluted!by!1!mL!pelleted!and!fed!into!9!mL!of!sterile! distilled!water,!then!diluting!the!dilutions!from!10J1!to!10J5.!A!total!of!1!mL!of!liquid!from!each! dilution!was!introduced!into!Petri!dishes!and!10!mL!of!MRSA!and!CaCO3!1%!were!added!and! homogenized,!then!incubated!at!37!°C!for!1x24!hours.!When!a!clear!zone!is!formed!around! the!colony,!then!the!colony!is!suspected!as!lactic!acid!bacteria.! ! Purification&of&Lactic&Acid&Bacteria&Isolates! Lactate!Acid!Bacteria!isolates!were!then!inoculated!by!scraping!them!onto!the!medium! in!a!petri!dish!then!incubated!for!1x24!hours!at!37!°C!at!the!aerobic!incubator.!Then!purified! by!inoculating!the!MRSA!medium!inclined!in!the!test!tube,!after!which!incubated!for!1!×!24! hours!at!37!°C!in!the!aerobic!incubator.! ! Characterizations&and&Identifications&of&Lactic&Acid&Bacteria! Macroscopic&Test! Pure!lactic!acid!bacteria!isolates!were!inoculated!on!MRSA!medium!in!petri!dishes,! after!which!incubated!for!1x24!hours!at!37!°C!in!aerobic!incubators.!Then,!observed!colonies.! & Microscopic&Test& Gram!staining!method!was!used!by!isolate!growing!on!MRSB!medium!placed!on!the! preparation.!Then,!fixed!and!added!2J3!drops!of!gram!A!(crystal!violet),!left!for!1!minute.! Wash!with!running!water,!dried!with!filter!paper.!Add!gram!B!(mordant).!Left!1!minute.!Spilled! with!gram!C!(95%!ethanol)!for!10J20!seconds.!Wash!with!running!water,!dried!with!filter! paper.!Add!gram!D!(safranin)!for!1!minute.!Wash!with!running!water,!dried!with!filter!paper.! It!was!observed!under!a!microscope!with!100x!widening!by!using!immersion!oil.!Gram!positive! bacteria!will!be!purple!and!gram!negative!will!be!red!(Ventimiglia!et&al.,!2015).! & Biochemical&Test! Biochemical!tests!in!this!study!consist!of!the!several!tests!as!following;! & Carbohydrate&Fermentation&Test! Cultures!of!lactic!acid!bacteria!inoculated!into!Lactose!Broth,!Sucrose!Broth,!Glucose! Broth,!and!Mannitol!Broth!media!were!incubated!for!1!x!24!hours!at!37!°C.!Thus,!observed! changes!in!medium!color!and!gas!formation.! & Motility&Test! The!culture!of!lactic!acid!bacteria!was!inoculated!puncture!on!an!erect!SIM!medium! and!incubated!for!1!x!24!hours.!If!isolate!growth!spreads!far!from!puncture!indicates!motile! isolate,!whereas!if!growth!only!occurs!puncture!discharge,!it!indicates!no!motile!isolate.! International&Journal&of&Applied&Biology,&1(2),&2017& ! 61! Catalase&Test! Suspension!isolates!drops!as!much!as!two!drops!on!glass!object,!and!then!spilled!3%! H2S2!solution.!The!positive!catalase!reaction!is!shown!by!the!presence!of!foam!or!foam!that! occurs!after!the!addition!of!the!solution!for!1!min.! & Test&of&Optimum&Growth&Temperature& A! total! of! 1! mL! of! pure! culture! suspension! were! each! inoculated! into! 4! tubes! containing! 9! mL! MRSB! media! on! which! the! purple! bromJkresol! indicator! was! split.! The! medium!is!incubated!at!a!temperature!of!10!°C,!15!°C,!37!°C!and!45!°C!for!1J5!x!24!hours!under! suitable!conditions.!The!media!color!change!from!purple!to!yellow!signifies!the!growth!of! culture.! & Test&of&Optimum&Growth&pH! A!total!of!1!loop!of!pure!isolates!was!each!inoculated!into!4!tubes!containing!MRSB! media!on!which!the!purple!bromJkresol!indicator!was!split.!The!medium!was!incubated!at!pH! 2,!4,!6!and!8!for!1J!5!x!24!hours!at!37!°C.!The!media!color!change!from!purple!to!yellow!signifies! the!growth!of!culture.! & Test&of&Isolate&Activity&against&Candida&albicans! The!isolated!lactic!acid!bacteria!were!grown!in!MRS!Broth!media!for!24J48!hours.!The! result! of! the! fermentation! will! be! precipitated! its! cell! mass.! Then! the! supernatant! fluid! containing! the! bioactive! component! was! tested! for! its! anticandidal! activity! of! Candida& albicans!by!the!paper!discJdiffusion!method.!The!isolates!were!prepared!for!the!test!with!two! replications!of!each!sample.& & Results&and&discussion&& Lactic&Acid&Bacteria&Isolation&& This! research! began! with! sterilization! as! the! most! important! stage! in! the! implementation!of!research!related!to!microorganisms.!Furthermore,!the!manufacture!of!a! special!medium!for!the!growth!of!lactic!acid!bacteria!is!MRSB!(deMann!Rogosa!Sharpe!Broth)! as! the! transport! medium.! The! next! stage! is! the! oral! mucosal! sampling! of! three! people! samples;! infant,! child,! and! adult! from! three! volunteers! with! healthy! dental! and! mouth! conditions.!Sampling!is!done!by!using!sterile!swab!swabs!and!immediately!inserted!into!the! transport!medium!immediately!after!taking!to!minimize!the!occurrence!of!contamination.! The!purpose!of!this!early!stage!is!to!obtain!lactic!acid!bacteria!from!the!oral!mucosa!and! increase!the!number!of!cells!(cultivation)!before!the!next!stage.& This!process!used!MRSA!(deMann!Rogosa!Sharpe!Agar)!medium!in!a!petri!dish!and! calcium!carbonate!(CaCO3!1%)!added!to!the!medium!in!order!to!identify!differences!of!lactic! acid!bacteria!compared!to!the!other!types!of!bacteria.!The!validation!of!the!lactic!acid!bacteria! was!performed!by!the!muddiness!of!medium!colour!(Vestman!et&al.,!2013).!If!it!contained! lactic!acid!bacteria,!the!calcium!from!CaCO3!in!MRSA!can!react!with!lactic!acid!produced!by! lactic!acid!bacteria!to!form!soluble!calcium!lactate,!which!makes!the!colour!of!the!medium! becomes!transparent!(Vestman!et&al.,!2013).! ! & & International&Journal&of&Applied&Biology,&1(2),&2017& ! 62! Lactic&Acid&Bacteria&Purification! The!next!stage!was!the!purification!of!isolates.!This!isolation!was!carried!out!each!into! 2!petri!dishes!for!each!sample.!The!observation!of!colony!growth!will!be!done!on!days!1,!3,!5,! and!7.!On!the!5!the!day,!a!transparent!zone!has!been!formed!on!the!MRSA!medium!that!is! thought!to!be!a!colony!of!lactic!acid!bacteria.!The!purification!results!are!repeated!twice!with! the!aim!of!obtaining!more!pure!isolates!and!increasingly!free!from!mixtures!of!other!types!of! bacterial!colonies.! Subsequently,! the! isolates! were! moved! into! the! test! tube! containing! the! inclined! MRSA!in!the!test!tube!for!the!further!stages,!i.e.!identification!of!isolates.!Isolates!were!named! by!code,!i.e.!infant’s!mouth!isolates!(MB,!MB2),!child’s!mouth!isolates!(MA,!MA),!and!adult’s! mouth!isolates!(MD,!MD2).! & Lactic&Acid&Bacteria&Characterizations&and&Identifications& In!this!study,!the!identification!of!lactic!acid!bacteria!microscopically!was!done!by!the! gram!staining!method.!The!staining!results!for!the!three!types!of! isolates!(MB,!MA,!MD)! showed!purple!bacterial!cells,!so!the!isolates!in!this!study!included!gramJpositive!bacteria.! This!supports!the!theory!that!lactic!acid!bacteria,!one!of!which!is!from!the!genus!Lactobacillus,! is!a!type!of!gramJpositive!bacteria!that!in!gram!staining!will!have!a!purple!appearance!under! an!optical!microscope!(Ventimiglia!et&al.,!2015;!Norder!et&al.,!2013).! In!addition,!besides!the!microscopic!tests!and!gene!identification!above,!the!isolates! were!tested!for!their!antimicrobial!activity!by!the!diffusion!method!in!order!to!use!paper!discs.! It!was!measured!from!horizontal,!vertical,!and!diagonal,!then!the!calculated!for!the!average! diameter.!Based!on!antimicrobial!activity!test!from!oral!mucosa!isolate!to!Candida&albicans! fungi!causing!infectious!disease!of!candidiasis,!and!obtained!the!result!of!the!zone!diameter! as!follows.! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! Figure&1.&Inhibition&zone&diameter&of&lactic&acid&bacteria&isolates&from&human&oral&mucosa& & Kontrol!1! MA!1! MA!2!Kontrol!2!MB!1! MB!2! MD!2! MD!1!2! International&Journal&of&Applied&Biology,&1(2),&2017& ! 63! Table&1.&Inhibition&zone&diameter&measurement&of&lactic&acid&bacteria&isolates&from&human& oral&mucosa& Types&of&Isolate& Replication& Diameter&±&Standard&Deviation&(mm)& I& II& Ketoconazole&(Control&+)& 19.1! 20.4! & &18.7! 19.3! 19.4! 18.3! Average& 19.07& 19.30& 19.19&±&0.16& Infant&(MB)& 8.1! 7.3! & &6.5! 7.5! 7.1! 6.8! Average& 7.23& 7.20& 7.22&±&0.02& Child&(MA)& 9.3! 10.3! & &9.5! 9.9! 9.1! 10.2! Average& 9.30& 10.13& 9.72&±&0.58& Adult&(MD)& 16.4! 15.8! & &16.2! 16.1! 15.9! 16.2! Average& 16.17& 16.03& 16.10&±&0.09& ! From!the!table!above,!it!can!be!seen!that!the!average!inhibitory!zone!diameter!of!the! oral!mucosa!of!infants!(MB!and!MB2)!is!7.23!mm!and!7.20!mm!(average!7.22!mm!±!0.02!mm),! lactic!acid!bacteria!isolate!from!the!oral!mucosa!of!children!(MA)!is!9.30!mm!and!10.13!mm! (average!9.72!mm!±!0.58!mm),!and!lactic!acid!bacteria!isolate!from!adult!mouth!mucosa!(MD)! were!16.17!mm!and!16.03!mm!(average!16.10!mm!±!0.09!mm).!While!the!results!of!inhibitory! power!using!ketoconazole!antifungal!drugs!as!a!comparison!were!19.07!mm!and!19.30!mm! (average!19.19!mm!±!0.16!mm).!It!is!showed!that!lactic!acid!bacteria!isolates!from!adult!mouth! mucosa!(MD)!had!the!best!inhibitory!power!seen!from!their!larger!inhibitory!diameter!zones! compared!to!both!infant!isolate!(MB)!and!child!isolate!(MA).! ! ! ! & & & & & & & & & & & & & & & Figure&2.&The&collection&of&potent&lactic&acid&bacteria&isolates&as&anti&candidiasis& International&Journal&of&Applied&Biology,&1(2),&2017& ! 64! For!additional!data,!biochemical!tests!consisting!of!carbohydrate!fermentation!test,! motility!test,!catalase!test,!optimum!pH!test,!and!optimum!temperature!test!were!conducted.! The!aim!of!these!five!biochemical!tests!is!to!confirm!that!the!isolates!obtained!from!the!oral! mucosa!of!humans!in!this!study!can!satisfy!the!characteristics!of!lactic!acid!bacteria!with! specific!physiological!and!metabolic!activities!specifically.! The!results!of!carbohydrate!fermentation!test!on!the!three!isolates!(MB,!MA,!MD)!with! medium! LB,! GB,! and! SB! showed! the! three! results! of! each! positive! able! to! ferment! carbohydrate.!The!positive!result!is!marked!by!the!colour!change!of!the!medium!to!yellow! and!the!formation!of!gas!in!the!Durham&tube.!This!is!consistent!with!the!reference!that!lactic! acid!bacteria!include!heterofermentative!bacteria,!which!are!capable!of!fermenting!sugars! (monosaccharaides!and!disaccharides)!to!produce!acidic!and!CO2!gases.!The!acidic!conditions! in!the!medium!result!in!a!change!of!medium!colour!and!bubble!formation!(Minervini!et&al.,! 2012;!Pepe!et&al.,!2013).! The!result!of!the!motility!test!of!all! three! isolates!showed!the!growth!of!colonies! perpendicularly!in!the!direction!of!inoculation!using!a!loop,!and!without!spreading!around!the! inoculants.! If!the!bacterial!cell! is!motile,! its!colonies!will!spread!throughout!the!medium.! Therefore,!the!three!isolates!(MB,!MA,!MD)!are!nonJmotile!bacteria!(Minervini!et&al.,!2012;! Pepe&et&al.,!2013).! The!result!of!catalase!test!of!lactic!acid!bacteria!isolated!from!oral!mucosa!of!the!three! samples!(MB,!MA,!MD)!was!negative!catalase.!This!is!in!accordance!with!the!reference!to!the! fact!that!lactic!acid!bacteria!do!not!produce!catalase!enzymes!so!that!the!test!results!are! negative!as!a!result!of!the!absence!of!H2O2!degradation!by!catalase!enzymes!(Minervini!et&al.,! 2012;!Pepe!et&al.,!2013).! The!optimum!pH!test!results!in!this!study!were!pH!4.!At!pH!4,!the!most!turbid!visible! medium!was!thought!to!be!the!highest!growth!rate!of!lactic!acid!bacteria!in!this!study.!The! previous! study! stated! that! lactic! acid! compounds! capable! of! inhibiting! the! growth! of! pathogenic! microbes! by! lowering! the! pH! of! the! environment! to! 3! to! 4.5! so! that! other! microbial!growth!will!be!hampered!(Minervini!et&al.,!2012;!Pepe!et&al.,!2013).! !The!optimum!temperature!test!results!are!at!45!°C,!which!is!the!highest!temperature! of!this!test.!The!highest!growth!of!colonies!in!the!form!of!the!cloudyJlooking!medium!occurred! at!45!°C!compared!to!25!°C!and!37!°C.!Based!on!the!reference,!lactic!acid!bacteria!can!grow! at!a!temperature!of!20J50!°C!(Minervini!et&al.,!2012;!Pepe!et&al.,!2013).! Based!on!the!five!biochemical!tests,!the!three!types!of!isolates!are!catalase!negative,! nonmotile,!and!include!Gram!positive!(+)!bacteria!with!basil!(stem)!morphology.!In!this!study,! it!can!be!said!that!the!isolates!obtained!from!the!mucosa!of!the!human!mouth!(MB,!MA,!MD)! belong!to!the!lactic!acid!bacteria!group,!were!from!the!genus!Lactobacillus!based!on!the! biochemical!test!results.! ! Conclusions! The!anti!candidiasis!activity!of!lactic!acid!bacteria!isolates!from!the!adult!oral!mucosa! (MD)! most! potentially! inhibited! the! growth! of! Candida& albicans! that! is! 16.10! mm,! when! compared!to!lactic!acid!bacteria!isolates!from!the!infant!(MB)!and!the!child!(MA)! & Acknowledgment&& The!authors!are!grateful!to!Ministry!of!Research,!Technology,!and!Higher!Education! Indonesia!which!has!funded!this!research!through!Student!Creativity!ProgramJExact!Research! 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