GGeeoorrggiiaa SSttaattee UUnniivveerrssiittyy SScchhoollaarrWWoorrkkss @@ GGeeoorrggiiaa SSttaattee UUnniivveerrssiittyy Public Health Theses School of Public Health Spring 5-15-2015 TThhee EEffifficcaaccyy ooff UUttiilliizziinngg CChhiittoossaann aass aann AAnnttiivviirraall AAggeenntt iinn WWaatteerr TTrreeaattmmeenntt Jessica Wilson Follow this and additional works at: https://scholarworks.gsu.edu/iph_theses RReeccoommmmeennddeedd CCiittaattiioonn Wilson, Jessica, "The Efficacy of Utilizing Chitosan as an Antiviral Agent in Water Treatment." Thesis, Georgia State University, 2015. doi: https://doi.org/10.57709/7055668 This Thesis is brought to you for free and open access by the School of Public Health at ScholarWorks @ Georgia State University. It has been accepted for inclusion in Public Health Theses by an authorized administrator of ScholarWorks @ Georgia State University. For more information, please contact [email protected]. ! ! ! ! ! ! ! ! ! ! ! The$Efficacy$of$Utilizing$Chitosan$as$an$Antiviral$Agent$in$Water$Treatment! ! ! ! ! ! ! ! ! ! By! ! Jessica!Wilson! ! B.S.!in!Anthropology! EMORY!UNIVERSITY!! ! ! A!Thesis!Submitted!to!the!Graduate!Faculty! of!Georgia!State!University!in!Partial!Fulfillment! of!the! Requirements!for!the!Degree! ! MASTER!OF!PUBLIC!HEALTH! ! ! ATLANTA,!GEORGIA! 30303! ! ! ! ! ! ! ! ! TABLE!OF!CONTENTS! ! ACKNOWLEDGMENTS!................................................................................................3! LIST!OF!TABLES……………………………………………………………………….4! LIST!OF!FIGURES……………………………………………………………………….5! ABSTRACT………………………………………………………………….…………….7! INTRODUCTION............................................................................................................10! !!!!!1.1!Current!Options!for!Water!treatment………………………………………..11! !!!!!1.2!Chitosan!Properties………………………………………………..12! !!!!!1.3!Why!Chitosan?……………………………………………………………….13! ! METHODS!AND!PROCEDURES…………………................................................................!15! !!!!!2.1!Preparation!of!Chitosan!Samples………………………………………………………….15! !!!!!2.2!MS2!Titer………………………………………………………………..15! !!!!!2.3!Chitosan!Antimicrobial!Assay………….......................................................16! !!!!!2.4!Spot!Plates..................................................................................................17! ! !RESULTS.........................................................................................................................18! !!!!!3.1!Chitosan!effectiveness!in!viral!removal!……………………………………………...18! !!!!!! 3.1.1!Modified!Chitosan!effects!of!viral!removal…………..18! ! 3.1.2!Chitosan)DD)effects)of)viral)removal……….)19! ! 3.1.3.!!Molecular!weight!effects!on!viral!removal…..19! !!!!!3.2!Efficacy!of!chitosan!viral!removal!across!sample!types...........20! ! !DISCUSSION.........................................................................................................................21!!!!!!!!!!!!!!!!!!!!!! !!!!!4.1!Effectiveness!of!Modified!Chitosan!on!Viral!Removal............................21! !!!!!4.2!Effectiveness!of!Degrees!of!Deacetylation!of!Chitosan!on!Viral!Removal...........22! !!!!!4.3!Effectiveness!of!Molecular!Weight!on!Viral!Removal………………………………23! !!!!!4.4!Justification!for!Methodology!…………………………….24! !!!!!4.5!A!Comparison!of!Chitosan!Effectiveness!By!Comparing!Antimicrobial!Data!and!!!!!!!!!!!!!!!!!!!! Coagulation!Data!…………………………………….25! ! 4.5.1!Chitosan!Effectiveness!of!Bacterial!Reduction!in!Coagulation……….25! 4.5.2!Chitosan!Effectiveness!of!Viral!Reduction!in!Coagulation…………..28! ! CONCLUSION............................................................................................................................31! REFERENCES..............................................................................................................................32! ! ! 2! ! ! ! Acknowledgements! ! Dr.!Casanova,!I!cannot!articulate!enough!how!I!appreciate!your!constant!support!and!guidance! throughout!this!process.!Your!contributions!to!this!project!and!my!growth!as!a!researcher!have! been!invaluable.!! ! I!would!like!to!thank!Dr.!Stauber!for!feedback!and!support!on!this!project!as!my!committee! member.! ! To!Ampai,!your!patience!and!assistance!helped!make!this!project!possible,!thank!you.!! ! 3! ! ! ! List!of!Tables! Table!1:!Chitosan!Samples……………………………………………………………………………………………15! Table!2:!Modified!Chitosan!Samples…………………………………………………………………………….18! Table!3:!Chitosan!DD………………………………………………………………………………………………......19! Table!4:!!Molecular!Weight!Chitosan……………………………………………………………………………19! ! ! ! ! ! ! 4! ! ! ! List!of!Figures!! Figure!1:!Structure!of!Chitin!and!Chitosan……………………………………………………………………12! Figure!2:!A!Comparison!of!MIC!Across!Chitosan!Samples…………………………………………….20! Figure!3:!!E.)Coli!inhibition!by!chitosan………………………………………………………………………..26! Figure!4:!MS2!inhibition!by!chitosan……………………………………………………………………………28! ! ! ! 5! ! ! ! ! APPROVAL!PAGE! ! The#Efficacy#of#Utilizing%Chitosan%as%an%Antiviral%Agent%in%Water%Treatment! ! ! ! By! ! Jessica!Wilson!! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! Approved:!! _________Approved_________________________!!! Committee!Chair!! _____Dr.!Lisa!Casanova!_______________________!! Committee!Member!! ___Dr.!Christine!Stauber______________________! Date:!April!23,!2015! ! ! ! ! 6! ! ! ! Abstract! ! Chitosan!is!naturally!occurring!polysaccharide!that!is!related!to!the!molecule!chitin!and!has! been!shown!to!act!as!an!antifungal!and!antibacterial!agent,!as!well!as!a!coagulant!for!microbial! removal!from!water.!However,!little!research!has!explored!the!antiviral!properties!of!chitosan,! particularly!as!an!as!an!antimicrobial!agent!to!treat!drinking!water.!!In!this!study!we! investigated!the!efficacy!of!utilizing!chitosan!as!an!antiviral!agent!for!the!purposes!of!water! treatment!by!measuring!the!minimum!inhibitory!concentrations!(MIC)!of!seven!chitosan! samplese!chitosan!lactate,!chitosan!acetate,!chitosan!HCl,!70DD,!85DD,!95DD,!and!100K.!An! antimicrobial!assay!was!conducted!utilizing!seven!chitosan!samples!with!varying!degrees!of! deacetylation,!different!molecular!weights,!and!different!added!functional!groups.!We! determined!that!amongst!all!chitosan!samples!only!three!samplese!85DD,!95DD,!and!100K!Da! had!the!lowest!MIC,!or!the!concentration!at!which!there!were!no!detectable!viral!components! at!a!concentration!of!5,000!µg/ml.!The!results!attained!in!this!experiment!add!to!the!body!of! knowledge!about!the!potential!benefits!of!utilizing!chitosan!as!an!antimicrobial!agent!in!areas! with!limited!water!and!sanitation!infrastructure.! ! ! ! ! ! ! ! ! ! ! ! ! 7! ! ! ! Author’s!Statement!Page! ! In!presenting!this!thesis!as!a!partial!fulfillment!of!the!requirements!for!an!advanced!degree! from!Georgia!State!University,!I!agree!that!the!Library!of!the!University!shall!make!it!available! for!inspection!and!circulation!in!accordance!with!its!regulations!governing!materials!of!this! type.!I!agree!that!permission!to!quote!from,!to!copy!from,!or!to!publish!this!thesis!may!be! granted!by!the!author!or,!in!his/her!absence,!by!the!professor!under!whose!direction!it!was! written,!or!in!his/her!absence,!by!the!Associate!Dean,!College!of!Health!and!Human!Sciences.! Such!quoting,!copying,!or!publishing!must!be!solely!for!scholarly!purposes!and!will!not!involve! potential!financial!gain.!It!is!understood!that!any!copying!from!or!publication!of!this! dissertation!which!involves!potential!financial!gain!will!not!be!allowed!without!written! permission!of!the!author.!! ______________________________!! Signature!of!Author!! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! ! 8! ! ! ! ! Notice!to!Borrowers!Page! ! All!theses!deposited!in!the!Georgia!State!University!Library!must!be!used!in!accordance!with! the!stipulations!prescribed!by!the!author!in!the!preceding!statement.!! ! The!author!of!this!thesis!is:!! Student’s!Name:!Jessica!Wilson! Street!Address:!!!6011!Tate!Drive! City,!State,!and!Zip!Code:!!Austell,!GA!30106! The!Chair!of!the!committee!for!this!thesis!is:!!Dr.!Lisa!Casanova!! Department:!School!of!Public!Health! College:!Health!and!Human!Sciences! Georgia!State!University!! P.O.!Box!3995!! Atlanta,!Georgia!30302[3995!! Users!of!this!thesis!who!not!regularly!enrolled!as!students!at!Georgia!State!University!are! required!to!attest!acceptance!of!the!preceding!stipulation!by!signing!below.!Libraries! borrowing!this!thesis!for!the!use!of!their!patrons!are!required!to!see!that!each!user!records! here!the!information!requested.! ! NAME%OF%USER% ADDRESS% DATE% TYPE%OF%USE% (EXAMINATION%ONLY%OR% COPY)% % % % % % % % % % % % % !! ! ! ! 9!