Antibacterial electrospun chitosan-based nanofibers: A bacterial membrane perforator

Titre:

Title

:

Antibacterial electrospun chitosan-based nanofibers: A bacterial

membrane perforator

Auteurs:

Authors

:

Mounia Arkoun, France Daigle, Marie-Claude Heuzey et Abdellah

Ajji

Date: 2017

Type:

Article de revue / Journal article

Référence:

Citation

:

Arkoun, M., Daigle, F., Heuzey, M.-C. & Ajji, A. (2017). Antibacterial electrospun

chitosan-based nanofibers: A bacterial membrane perforator. Food Science &

Nutrition, 5(4), p. 865-874. doi:

10.1002/fsn3.468

Document en libre accès dans PolyPublie

Open Access document in PolyPublie

URL de PolyPublie:

PolyPublie URL:

https://publications.polymtl.ca/4753/

Version: Version officielle de l'éditeur / Published version

_{Révisé par les pairs / Refereed}

Conditions d’utilisation:

Terms of Use

: CC BY

Document publié chez l’éditeur officiel

Document issued by the official publisher

Titre de la revue:

Journal Title:

Food Science & Nutrition (vol. 5, no 4)

Maison d’édition:

Publisher:

Wiley

URL officiel:

Official URL:

https://doi.org/10.1002/fsn3.468

Mention légale:

Legal notice:

Ce fichier a été téléchargé à partir de PolyPublie, le dépôt institutionnel de Polytechnique Montréal

This file has been downloaded from PolyPublie, the institutional repository of Polytechnique Montréal

Food Sci Nutr. 2017;5:865–874. www.foodscience-nutrition.com  

|

  865 Received: 1 December 2016 

|

  Revised: 15 January 2017 

|

  Accepted: 16 January 2017

DOI: 10.1002/fsn3.468

O R I G I N A L R E S E A R C H

Antibacterial electrospun chitosan- based nanofibers: A

bacterial membrane perforator

Mounia Arkoun

1

_{| France Daigle}

2

_{| Marie-Claude Heuzey}

1

_{| Abdellah Ajji}

1

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

© 2017 The Authors. Food Science & Nutrition published by Wiley Periodicals, Inc. 1_{CREPEC Department of Chemical} Engineering, École Polytechnique de Montréal, Montréal, QC, Canada 2_{Department of microbiology, infectiology} and immunology, Pavillon Roger-Gaudry, Université de Montréal, Montréal, QC, Canada Correspondence Marie Claude Heuzey and Abdellah Ajji, CREPEC Department of Chemical Engineering, École Polytechnique de Montréal, Montréal, QC, Canada. Emails: [email protected]; [email protected]

Abstract

This study investigates the antibacterial action of chitosan- based nanofibers (CNFs) obtained by the electrospinning process on the permeability of bacterial membranes. The bactericidal efficiency of CNFs was first determined against Gram- negative

Escherichia coli and Salmonella Typhimurium, and Gram- positive Staphylococcus aureus

and Listeria innocua bacteria as a baseline. The results strongly suggest that CNFs interact with the negatively charged bacterial cell wall causing membrane rupture and inducing leakage of intracellular components among which are proteins and DNA. Results clearly indicate that the release of such components after contact with CNFs is an indication of membrane permeabilization and perforation, as pore formation was observed in transmission electron microscopy (TEM). This work suggests a plausible antibacterial mechanism of action of CNFs and also provides clear evidence in favor of chitosan as a bacterial membrane disruptor and perforator. As a result, CNFs can find promising applications as bioactive food packaging materials capable to extend shelf life of food products while inhibiting the spread of alteration flora and foodborne pathogens. K E Y W O R D S antibacterial, electrospun chitosan-based nanofibers, membrane perforation, membrane permeability

1 | INTRODUCTION

Electrospinning of chitosan, with the aim of producing nanofibers with diameters ranging from few tens of nanometers to micrometers, has been the subject of several recent studies (Desai, Kit, Li, & Zivanovic, 2008; Doğan, Özyıldız, Başal, & Uzel, 2013; Elsabee, Naguib, & Morsi, 2012; Geng, Kwon, & Jang, 2005; Homayoni, Ravandi, & Valizadeh, 2009; Kriegel, Kit, McClements, & Weiss, 2009; Pakravan, Heuzey, & Ajji, 2011; Rieger, Birch, & Schiffman, 2016; Ziani et al., 2011). The resulting chitosan nanofiber (CNF) mats exhibit a remarkably high porosity (in the range of 80%–90%) and surface area per unit mass (between 10 and 500 m2

/g) and display good biocompatibility and bio-functionality (Ardila et al., 2016; Greiner & Wendorff, 2007). Therefore,

CNFs may have promising applications in biomedical (cell culture, wound healing, tissue engineering) (Ignatova, Manolova, Markova, & Rashkov, 2009), pharmaceutics (controlled drug release, gene ther-apy) (Jayakumar, Prabaharan, Nair, & Tamura, 2010), water filtration (chelation of metal ions) (Haider & Park, 2009), and food packaging (Martínez- Camacho et al., 2011), among others. However, achieving high yield and quality fiber formation from neat chitosan solutions is a challenging task. This is mainly due to the very rigid structure of chi-tosan chains, which does not promote entanglements that are required for the formation of the Taylor cone, which in turn generates nano-fibers. For example, some authors reported the preparation of neat CNFs using trifluoroacetic acid (TFA) as a solvent or its mixtures with dichloromethane (DCM) (Gu et al., 2013; Lee et al., 2014). However,

ѶѵѵՊ

|

ՊՊՍ ARKOUN ETAL.

$ bv_b]_Ѵ‹1‹|o|oŠb1ķ1ouuovbˆ;ķ-m7;mˆbuoml;m|-ѴѴ‹_-ul=†Ѵķl-hbm] |_;†v;o=v†1_l-|;ub-Ѵvbm1olr-|b0Ѵ;‰b|_-rrѴb1-|bomv-v7;Ѵb1-|;-v =oo7r-1h-]bm]ĺou;oˆ;uķ;Ѵ;1|uovrbmmbm]bv-l†Ѵ|b=-1|oub-Ѵruo1;vv |_-|bmˆoѴˆ;vv;ˆ;u-Ѵr-u-l;|;uv-lom]‰_b1_ruo1;vvbm]1om7b|bomv v†1_ -v =Ѵo‰ u-|;ķ ;Ѵ;1|ub1 =b;Ѵ7ķ 1oѴѴ;1|bm] 7bv|-m1;ķ |;lr;u-|†u; -m7 _†lb7b|‹ķ-v‰;ѴѴ-vbm|ubmvb1voѴ†|bomr-u-l;|;uvbm1Ѵ†7bm]1om7†1|bˆ-b|‹ķv†u=-1;|;mvbomķ-m7ˆbv1o;Ѵ-v|b1b|‹ĺ$_†vķbmou7;u|oblruoˆ;|_; ;Ѵ;1|uovrbmm-0bѴb|‹o=1_b|ov-mķ-1ovrbmmbm]-];m|-|lo7;u-|;1om|;m| bvo=|;mm;;7;7-m7†v;7-v-1-uub;uroѴ‹l;u|o|ub]];u=b0;u=oul-|bom Őo-‹;ubşffbķƑƏƐƔĸ!b;];u;|-ѴĺķƑƏƐѵőĺ

"|†7b;v _-ˆ; 7;lomv|u-|;7 |_-| 1_b|ov-mķ bm |_; =oul o= voѴ†|bom -m7=bѴlvķ;Š_b0b|v;==b1b;m|-m|blb1uo0b-Ѵ-1|bˆb|‹Ő†ŒŒ-u;ѴѴb;|-ѴĺķƐƖѶѶĸ -rbm;-†ķ ooˆ;uķ mouuķ ş -uh-vķ ƐƖƖƐĸ "_-_b7bķ u-1_1_bķ ş ;omķ ƐƖƖƖĸ"†7-uv_-mķooˆ;uķşmouuķƐƖƖƑĸ+o†m]ķक़_Ѵ;ķş-†vvķƐƖѶƑőĺ o‰;ˆ;uķ =;‰ _-ˆ; ;Š-lbm;7 |_; -m|b0-1|;ub-Ѵ ruor;u|b;v o=  vĺ m - u;ˆb;‰ -u|b1Ѵ;ķ -u|झm;ŒŊ-l-1_o ;|-Ѵĺ ŐƑƏƐƐő robm| o†| |_-| lov| u;rou|vom|_;-m|blb1uo0b-Ѵ-1|bˆb|‹o= v_-ˆ;†v;71_b|ov-mvoѴ†|bomv bmv|;-7ĺmlov|1-v;vķ|_;ruorov;7l;1_-mbvl=ou v‰-vbm7bu;1|Ѵ‹ u;Ѵ-|;7|o|_;ru;v;m1;-m7u;Ѵ;-v;o=ruo|om-|;7-lbmo]uo†rv=uol  vl-|vķ‰_b1_‰;u;moѴom];um-mo=b0;uvĺ$_;-†|_ouv_b]_Ѵb]_|;7 |_-|=†u|_;ubmˆ;v|b]-|bom‰o†Ѵ70;†v;=†Ѵbmou7;u|o7;|;ulbm;‰_;|_;u  v=oѴѴo‰|_;v-l;ru;v†l;7l;1_-mbvlķvbm1;b|lb]_|0;-==;1|;7 0‹ |_; v|u†1|†u-Ѵ 1om=oul-|bom |_;v; m-mol-|;ub-Ѵv 1-m -7or| Őom] ;|-Ѵĺķ ƑƏƏѶőĺ$_; l;1_-mbvl o= -1|bom 0‹ ‰_b1_ 1_b|ov-mķ bm voѴ†|bom v|-|;ķbv-0Ѵ;|obm_b0b|ouhbѴѴ0-1|;ub-bv-1olrѴ;Šr_;mol;mom|_-|_-v mo|0;;m=†ѴѴ‹;ŠrѴ-bm;7;b|_;uŐ-ll;u;|-ѴĺķƑƏƐƏĸom]ķ_;mķ*bm]ķ ş-uhķƑƏƐƏĸ!--=-|ķ(om-u];mķ--vķş"-_ѴķƑƏƏѶőĺou;oˆ;uķmo bm=oul-|bom bv -ˆ-bѴ-0Ѵ; u;]-u7bm] |_; l;1_-mbvl †m7;uѴ‹bm] |_; -m|b-lb1uo0b-Ѵ-1|bˆb|‹o= vĺ$oo†uhmo‰Ѵ;7];ķmov|†7‹_-vu;rou|;7|_; ;==;1|o= vom0-1|;ub-Ѵ1;ѴѴl;l0u-m;bm|;]ub|‹ķmou|_;bulo7;o= -1|bomĺ1‹|oѴo]b1-Ѵv|†7‹o=|_;;==;1|o= vom|_;0-1|;ub-Ѵl;l-0u-m; r;ul;-0bѴb|‹ bv m;1;vv-u‹ |o †m7;uv|-m7 |_;bu ;Š-1| l;1_-mbvl o=-1|bom-m7|o-ˆob7|_;o†|0u;-ho=ro|;m|b-Ѵu;vbv|-m1;r_;mol;m-ĺ m |_bv v|†7‹ķ‰; bmˆ;v|b]-|; |_; -m|b0-1|;ub-Ѵ l;1_-mbvl o= -1|bom o=  v-]-bmv|=o†u1ollom-Ѵ|;u-|bom=Ѵou--m7=oo70oum;r-|_o];mvķ lov|=u;t†;m|Ѵ‹bm1ublbm-|;7bm=oo7vrobѴ-];-m7=oo7robvombm]ķu;-vr;1|bˆ;Ѵ‹ĺѴѴ|;v|v‰;u;r;u=oul;7†m7;uv|-m7-u7bŒ;7-m71om|uoѴѴ;7 ;Šr;ubl;m|-Ѵ1om7b|bomv|o=-1bѴb|-|;u;ruo7†1b0bѴb|‹-m7-ѴѴo‰1olr-u--|bˆ;v|†7b;vĺrѴ-†vb0Ѵ;lo7;o=-1|bombm‰_b1_ v-1|-vl;l0u-m; r;ul;-0bѴb|‹7bvu†r|ou-m7;ˆ;mr;u=ou-|oubvrov|†Ѵ-|;7ĺm|_bv1om|;Š|ķ  vu;ru;v;m|b7;-Ѵ0bol-|;ub-Ѵv|_-|1-m0;†v;7-vv†b|-0Ѵ;0-1|;-ub1b7-Ѵ 0-uub;uv |o ru;ˆ;m| 0-1|;ub-Ѵ bm=;1|bomv bm v;ˆ;u-Ѵ -u;-vķ bm1Ѵ†7-bm]=oo7r-1h-]bm]-m70bol;7b1-Ѵ-rrѴb1-|bomvĺvr-u|o=-1|bˆ;=oo7 r-1h-]bm]ķ v1-m0;-rrѴb;7|o;Š|;m7|_;v_;Ѵ=Ѵb=;o==oo7ruo7†1|v -m7ru;ˆ;m|vrobѴ-];-m7=oo70oum;7bv;-v;v1-†v;70‹Escherichia coli, ListeriaķStaphylococcus,-m7Salmonella.

ƑՊ|Պ$!" $ "

ƑĺƐՊ|Պ_;lb1-Ѵv-m7roѴ‹l;uv

)-|;uŊvoѴ†0Ѵ; 1_b|ov-m Ő"őķ - (;mŒ‹lŤ ]u-7; o0|-bm;7 via ;mŒ‹-l-|b1|u;-|l;m|o=1_b|bm7;ubˆ;7=uolv_ublrv_;ѴѴv‰-v];m;uo†vѴ‹

7om-|;70‹ˆ;mv-Őm|-uboķ-m-7-őĺ$_;‰-|;uŊvoѴ†0bѴb|‹o=|_bv" ]u-7;bv7†;|o|_;ru;v;m1;o=-Ѵo‰-lo†m|o=u;vb7†-Ѵ-1;|b1-1b7 Ő1őķ -v 1om=bul;7 0‹ |_; v†rrѴb;uĺ $_; 1ouu;vrom7bm] 7;]u;; o= 7;-1;|‹Ѵ-|bomŐ ő-m7m†l0;u-ˆ;u-];loѴ;1†Ѵ-u‰;b]_|Őnő-u; ƖƔѷ-m7ƔƏh -ķu;vr;1|bˆ;Ѵ‹ķ‰b|_-m-uuo‰loѴ;1†Ѵ-u‰;b]_|7bv|ub-0†|bomĺ oѴ‹Ő;|_‹Ѵ;m; oŠb7;ő Őőķ - 1ovrbmmbm] -];m| =ou 1_b|ov-mķ ‰b|_ - loѴ;1†Ѵ-u ‰;b]_| o= ѵƏƏh -ķ -m7 -1;|b1 -1b7 Ő1ķ ]Ѵ-1b-Ѵķ ƖƖĺƕѷő ‰;u; r†u1_-v;7 =uol bv_;u "1b;m|b=b1 Ő"-bm|Ŋ-†u;m|ķ ķ -m-7-őĺѴѴl-|;ub-Ѵv‰;u;o=-m-Ѵ‹|b1-Ѵ]u-7;-m7†v;7-vu;1;bˆ;7ĺ

ƑĺƑՊ|Պb1uoou]-mbvlvķ1†Ѵ|†u;l;7b--m71om7b|bomv

ƑĺƑĺƐՊ|Պ-1|;ub-Ѵv|u-bmv

Escherichia coli Ő Ɣαőķ Salmonella $‹r_bl†ub†l Ő"ƐƒƓƓőķ Staphylococcus aureus ŐƔƓŊƕƒőķ -m7 Listeria innocua Ő" ƒƑѶƓő ‰;u; v†rrѴb;70‹|_;-0ou-|ou‹o=b1uo0boѴo]‹ķ&mbˆ;uvb|࣐7;om|u࣐-Ѵ Ő †࣐0;1ķ-m-7-őĺ†Ѵ|†u;v‰;u;l-bm|-bm;7-|ƓŦrubou|o†v;ķ|_;m |u-mv=;uu;7 bm|o - 1†Ѵ|†u; l;7b†l -m7 =bm-ѴѴ‹ bm1†0-|;7 -| ƒƕŦ =ou ƑƓ_ubm-mou0b|-Ѵv_-h;uŐ;‰u†mv‰b1hő|o-1_b;ˆ;-mbmb|b-Ѵ1om-1;m|u-|bomo=ƐƏ91oѴom‹=oulbm]†mb|r;ulbѴѴbѴb|;uŐ &ņlѴőĺ

ƑĺƑĺƑՊ|Պ†Ѵ|†u;l;7b-†ub-ŋ;u|-mb Őő 0uo|_ -m7 0u-bm _;-u| bm=†vbom Őő ‰;u; †v;7 -v ]uo‰|_ l;7b- |o v|-u| |_; 0-1|;ub-Ѵ 1†Ѵ|†u;vĺ $o 1_ |_; u;-t†bu;7 =bm-Ѵ 1om1;m|u-|bomķ 1†Ѵ|†u;v ‰;u; 7bѴ†|;7 †vbm] r_ovr_-|; 0†==;u v-Ѵbm; Ő"ķ r ƔĺѶķ -7f†v|;7 ‰b|_ ƐloѴņ Ѵőĺ  -]-u -m7  v†rrѴ;l;m|;7 ‰b|_ -]-u ŐƐƔ]ņő ‰;u; †v;7 -v voѴb7 l;7b- =ou 1o†m|bm]|_;v†uˆbˆbm]0-1|;ub-ĺ

ƑĺƒՊ|Պu;r-u-|bomo=1_b|ov-m-m7

stock solutions

_b|ov-m Ő"ő -m7  v|o1h voѴ†|bomv Őƕѷ ‰ņˆ -m7 ƒѷ ‰ņˆķ u;vr;1|bˆ;Ѵ‹ő ‰;u; bm7bˆb7†-ѴѴ‹ ru;r-u;7 0‹ 7bvvoѴˆbm] roѴ‹l;u ro‰7;uv bm ƔƏѷ Ոņˆő 1 †m7;u oˆ;umb]_| l-]m;|b1 v|buubm]ĺ $_; "ņ0Ѵ;m7v‰;u;o0|-bm;70‹l-]m;|b1v|buubm]o=|_;|‰oroѴ‹l;u voѴ†|bomvbm-ruorou|bomo=ѶƏņƑƏՉņ‰őu-|bo=ouƓ_u-]b|-|bomĺ$_; -7ˆ-m|-];o=†vbm]-t†;o†v-1;|b1-1b7voѴ†|bomvbv|_;bumom|oŠb1-m7 ;1o=ub;m7Ѵ‹1_-u-1|;uĺ

ƑĺƓՊ|Պu;r-u-|bomo=1_b|ov-mŊ0-v;7m-mo=b0;uvvia

;Ѵ;1|uovrbmmbm]

"ņm-mo=b0;uv‰;u;ru;r-u;7-11ou7bm]|o-hu-ˆ-m;|-ѴĺŐƑƏƐƐő †vbm]|_;;Ѵ;1|uovrbmmbm]ruo1;vvĺѴ;1|uovrbmmbm]o=|_;0Ѵ;m7voѴ†-|bom ‰-v r;u=oul;7 †vbm] - _oubŒom|-Ѵ _ol;l-7; v;|†r 1om|-bmbm] ŐƐő-_b]_ˆoѴ|-];ro‰;uv†rrѴ‹Ő-ll-b]_(oѴ|-];!;v;-u1_ķ ķ &"őķ ŐƑő - ruo]u-ll-0Ѵ; r†lr Ő-uˆ-u7 rr-u-|†vķ   ƑƏƏƏő |o 7;Ѵbˆ;u |_; roѴ‹l;u voѴ†|bom -| |_; u;t†bu;7 =Ѵo‰ u-|;ķ -m7 Őƒő - l;|-ѴѴb1uo|-|bm]7u†l‰u-rr;7‰b|_-m-Ѵ†lbm†l=obѴ|o1oѴѴ;1||_; m-mo=b0;uvĺ  v1_;l-|b1 u;ru;v;m|-|bom o= |_; v;| †r bv v_o‰m bm

ՊՍՊ

|

ՊѶѵƕ

ARKOUN ETAL.

b]†u;Ɛĺ$_;;Ѵ;1|uovrbmmbm]0Ѵ;m7voѴ†|bom‰-vro†u;7bm|o-ƐƏlѴ v‹ubm]; ‰b|_ †;uŋo1h 1omm;1|bom |o -m ƐѶŊ]-†]; 0Ѵ†m| |br m;;7Ѵ; Ő-7;m1;"1b;m1;ķ&"őĺ$_;v‹ubm];‰-vlo†m|;7om|_;r†lr‰b|_ -]ubr-m7]uo†m7;70‹†v;o=-m-ѴѴb]-|ou1Ѵbrĺ$_;or|bl-Ѵruo1;vv r-u-l;|;uv‰;u;=Ѵo‰u-|;o=ƏĺƔlѴņ_uķˆoѴ|-];o=ƑƏh(ķ-m7m;;7Ѵ; |brŊ|oŊ1oѴѴ;1|ou 7bv|-m1; o= ƐƔ1lĺ ѴѴ ;Šr;ubl;m|v ‰;u; 1om7†1|;7 -|uool|;lr;u-|†u;ŐƑƑƼƐŦőķu;Ѵ-|bˆ;_†lb7b|‹o=ƑƏѷķ-m7†m7;u -|lovr_;ub1ru;vv†u;ĺ$_;1oѴѴ;1|;7m-mo=b0;uv‰;u;7ub;7oˆ;umb]_| †m7;u-_oo7|o;mv†u;1olrѴ;|;;ˆ-rou-|bomo=|_;voѴˆ;m|ĺ

ƑĺƔՊ|Պ"1-mmbm];Ѵ;1|uomlb1uov1or‹Ő"ő

$_;lour_oѴo]‹o=|_;;Ѵ;1|uovr†m1_b|ov-mŊ0-v;7m-mo=b0;uvŐ vő ‰-v ;Š-lbm;7 -11ou7bm] |o - vѴb]_| lo7b=b;7 l;|_o7 o= o-‹;ub -m7 ffb ŐƑƏƐƔőķ †vbm] - =b;Ѵ7 ;lbvvbom v1-mmbm] ;Ѵ;1|uom lb1uov1or; Ő "  "ŊƕѵƏƏ$ őķ or;u-|;7 -| ƐĺƔh(ĺ "-lrѴ;v ‰;u; o0v;uˆ;7-v1oѴѴ;1|;7om-m-Ѵ†lbm†l=obѴ-=|;uƑ_u;Ѵ;1|uovrbmmbm]ĺ "u;v†Ѵ|vu;ˆ;-Ѵ;7|_-|†mb=oul-m70;-7Ѵ;vv=b0;uv‰;u;o0|-bm;7 bm |_; ru;v;m1; o= |_; 1ovrbmmbm] -];m|ķ  bm |_bv vr;1b=b1 1-v;ĺ $_; -ˆ;u-]; =b0;u 7b-l;|;u ‰-v ;ˆ-Ѵ†-|;7 †vbm] l-];Ŋuo Ѵ†v® vo=|‰-u;ĺrruoŠbl-|;Ѵ‹ѵƏƏm-mo=b0;uvu-m7olѴ‹1_ov;m=uol|_u;; bm7;r;m7;m| ;Ѵ;1|uovr†m l-|v ŐƑƏƏ =b0;uv =uol ;-1_ v-lrѴ;ő ‰;u; †v;7=ou|_;t†-m|b=b1-|bomo==b0;ulour_oѴo]‹r-u-l;|;uvĺ

ƑĺѵՊ|Պm|b0-1|;ub-Ѵ;==b1b;m1‹o= v

$_;-m|b0-1|;ub-Ѵ-1|bˆb|‹o=;Ѵ;1|uovr†m v‰-v;ˆ-Ѵ†-|;7bmˆb|uo =oѴѴo‰bm] |_; l;ub1-m v|-m7-u7 |;v| l;|_o7 Ő"$ ƑƐƓƖƴƐƒ-ķ ƑƏƐƒőĺ ollomѴ‹ =o†m7 0-1|;ub-ķ E. coliķ S. aureusķ L. innocuaķ -m7 S. $‹r_bl†ub†lķbm=oo71om|-lbm-|bom-m7vhbmbm=;1|bomv‰;u;v;-Ѵ;1|;7 =ou |_bv r†urov;ĺ "-lrѴ;v o= Ɛ1l2 -m7 ƑĺƔ1l2 v‰-|1_;v o=  v ‰;u; ru;r-u;7 bm -v;r|b1 1om7b|bomvĺ -1|;ub-Ѵ v†vr;mvbomv ŐƐƏ6 &ņlѴķƔlѴ"ķrƔĺѶő‰;u;r†|bm1om|-1|‰b|_ vĺ|bv mo|;‰ou|_‹ |_-| ;ˆ;m |_o†]_ |_; " ]u-7; †v;7 bm |_bv v|†7‹ ‰-v ‰-|;uŊvoѴ†0Ѵ;ķ |_; u;v†Ѵ|bm] m-mo=b0;uv ‰;u; ˆbv†-ѴѴ‹ bmvoѴ†0Ѵ; bm -t†;o†vl;7b-rov|Ŋ;Ѵ;1|uovrbmmbm]7†;|ovoѴˆ;m|;ˆ-rou-|bom7†u- bm]ruo1;vvbm]ĺ;]-|bˆ;1om|uoѴvo=0-1|;ub-v†vr;m7;7bm"‰b|_-o†| v‰;u;-Ѵvoru;r-u;7ĺѴѴ|†0;v‰;u;rѴ-1;7-|ƒƕŦķor|bl-Ѵ |;lr;u-|†u; =ou 0-1|;ub-Ѵ ]uo‰|_ķ =ou Ɠ_u bm1†0-|bom bm -m ou0b|-Ѵ v_-h;uĺ ";ub-Ѵ 7bѴ†|bomv ‰;u; r;u=oul;7 -m7 vru;-7 om -]-u rѴ-|;v

bm1†0-|;7oˆ;umb]_|-|ƒƕŦ=ou=†u|_;u1o†m|bm]o=v†uˆbˆouvĺѴѴ|;v|v ‰;u;1om7†1|;7bm|ubrѴb1-|;ĺ bm-ѴѴ‹ķ|_;-m|b0-1|;ub-Ѵ;==b1b;m1‹‰-v ;Šru;vv;7-v-=†m1|bomo=|_;u;7†1|bomu-|;ŐRőo=|_;|o|-Ѵm†l0;uo= |;v|0-1|;ub-ĺR‰-v1-Ѵ1†Ѵ-|;7-11ou7bm]|o;Ѵ-Ѵb-ķu;Ѵb;uķ;m-bvv-ķ -m7ol-ŐƑƏƏѶő†vbm]|_;=oѴѴo‰bm];t†-|bomĹ ‰_;u;ķA-m7B-u;|_;m†l0;uvo=v†uˆbˆbm]0-1|;ub-bm|_;1om|uoѴv -m7|;v|v-lrѴ;vķu;vr;1|bˆ;Ѵ‹ĺ

ƑĺƕՊ|Պ==;1|o=1_b|ov-mŊ0-v;7m-mo=b0;uvom

l;l0u-m;r;ul;-0bѴb|‹

ƑĺƕĺƐՊ|Պ"o7b†l7o7;1‹Ѵv†Ѵ=-|;ŊroѴ‹-1u‹Ѵ-lb7;];Ѵ

;Ѵ;1|uor_ou;vbvŐ" "Ŋő

$_; u;Ѵ;-v; o= bm|u-1;ѴѴ†Ѵ-u ruo|;bmv =uol  Ŋ|u;-|;7 0-1|;ub- ‰-v bmˆ;v|b]-|;7 0‹ " "Ŋĺ m |_bv v;1|bomķ E. coli Őu-lŊm;]-|bˆ;ő -m7S. aureusŐu-lŊrovb|bˆ;ő‰;u;v;Ѵ;1|;7bmou7;u|o-rru-bv;|_; ;==;1|o=u-lŊ|‹r;om|_;v|u-bmvĽv†v1;r|b0bѴb|‹ņu;vbv|-m1;|o vĺ ˆ;umb]_| 1†Ѵ|†u;v o= E. coli -m7 S. aureus ‰;u; u;v†vr;m7;7 bm " ŐŜƐƏ8 &ņlѴő-m7bm1†0-|;7-|ƒƕŦbm|_;ru;v;m1;o= vĺ=|;u Əķ Ɛķ Ƒķ ƒķ -m7 Ɠ_u 1om|-1| |bl;ķ ƔlѴ -Ѵbt†o|v ‰;u; ‰b|_7u-‰m -m7 1;m|ub=†];7 -| ƒķƏƏƏgņƐƏlbm -| ƓŦĺ $_; v†r;um-|-m|v ‰;u; |_;m lbŠ;7 ‰b|_ |ub1_Ѵouo-1;|b1 -1b7 Ő$ ƐƏĹƐő -m7 Ѵ;=| =ou ru;1brb|-|bom -|ƓŦoˆ;umb]_|ĺ=|;u-v;ub;vo=‰-v_ķv-lrѴ;v‰;u;u;v†vr;m7;7 bm " "ŊѴo-7bm] 0†==;u -m7 v†0f;1|;7 |o " "Ŋ -11ou7bm] |o |_; l;|_o7o=-;llѴbŐƐƖƕƏőĺovb|bˆ;1om|uoѴvŐ|uѴ+ őo=;Š|u-1|;7ruo-teins from E. coli-m7S. aureus‰;u;-Ѵvoru;r-u;70‹1_;lb1-ѴѴ‹vbv o=0o|_0-1|;ub-†vbm]-Ѵ‹vbvvoѴ†|bom1om|-bmbm]ƔƏμѴo=1_Ѵouo=oul -m7ƑƔ μѴo=" "ŐƏĺƔѷˆņˆőĺ oulou;v;mvb|bˆb|‹ķu;ˆ;Ѵ-|bom‰-vr;u-=oul;7†vbm]vbѴˆ;umb|u-|;v|-bmbm]o=ruo|;bmvĺ

ƑĺƕĺƑՊ|Պ]-uov;];Ѵ;Ѵ;1|uor_ou;vbvo=u;Ѵ;-v;7 

;1-†v; o= b|v blrou|-m1; bm =†m7-l;m|-Ѵ u;v;-u1_ķ b|v †v; bm |_; bm7†v|ub-Ѵ=b;Ѵ7-m7b|vbmˆoѴˆ;l;m|bm|_;-]ubŊ=oo7v;1|ouķ|_;E. coli Ѵ-0ou-|ou‹v|u-bm_-v0;;m=†ѴѴ‹v;t†;m1;7-m7b|v];mol;bv1†uu;m|Ѵ‹ ƐƏƏѷhmo‰mĺm|_;=oѴѴo‰bm]v;1|bomķE. coliŐ Ɣαő0-1|;ub†l‰-v 1_ov;m |o v|†7‹ |_; ;==;1| o=  v om l;l0u-m; r;ul;-0bѴb|‹ -m7 v†0v;t†;m| Ѵ;-h-];ĺ

$_; Ѵ;-h-]; o=  =uol  Ŋ|u;-|;7 E. coli ‰-v bmˆ;v|b]-|;7 0‹ -]-uov; ];Ѵ ;Ѵ;1|uor_ou;vbv -v -m bm7b1-|bom o= l;l0u-m; 7-l--];ĺ ‰-v;Š|u-1|;7=uol Ŋ|u;-|;7E. coli1†Ѵ|†u;v-11ou7bm] |o|_;ruo|o1oѴo=u;;m-m7"-l0uoohŐƑƏƐƑőĺub;=Ѵ‹ķƔlѴ-Ѵbt†o|v ‰;u;v†0f;1|;7|o1;m|ub=†]-|bomŐѵķƏƏƏurlķƐƏlbm-|ƓŦőķ=bѴ|u-|bom ŐƏĺƑƑμl rou; vbŒ;ő -m7 oˆ;umb]_| ru;1brb|-|bom -| ƴƑƏŦ bm vo7b†l -1;|-|;Ő-1ƒloѴņrƔĺƑő-m7;|_-moѴŐ|ƐƏƏѷķƴƑƏŦķƑĺƔ ƵˆoѴ†l;őĺ"-lrѴ;v‰;u;1;m|ub=†];7ŐƖķƏƏƏgķƐƔlbmķƓŦő-m7|_; u;v†Ѵ|bm]r;ѴѴ;|v‰;u;v†vr;m7;7bm;|_-moѴŐƕƏѷķƴƑƏŦőķ1;m|ub=†];7 -]-bmķ7ub;7†m7;u|_;_oo7-m7u;v†vr;m7;7bmlbѴѴb ‰-|;uĺovb|bˆ; 1om|uoѴv o= 0-1|;ub-Ѵ  ;Š|u-1|;7 =uol E. coli -=|;u 1_;lb1-Ѵ -m7 ŐƐő R(%) = A−B

A ×100

  & !   Ɛ Պ "1_;l-|b1u;ru;v;m|-|bomo=|_;_ol;l-7; ;Ѵ;1|uovrbmmbm]v;|†r

ѶѵѶՊ

|

ՊՊՍ ARKOUN ETAL.

_;-||u;-|l;m|Ő|uѴ+-m7|uѴ+ķu;vr;1|bˆ;Ѵ‹ő‰;u;-Ѵvoru;r-u;7ĺm -77b|bom-Ѵv|;ro=r-7f†v|l;m|ŐrƕĺƏő‰b|_ƐloѴņ-bmou7;u |o7;ruo|om-|;|_; v-m70u;-h†r"Ŋ bm|;u-1|bomv‰-vm;1-;vv-u‹ĺroѴ‹l;u-v;1_-bmu;-1|bomŐ!ő=ou|_;rrnB];m;Ɛѵ"! ‰-vr;u=oul;7bmou7;u|o-lrѴb=‹|_;u;Ѵ;-v;7 =u-]l;m|v=uol 1_b|ov-mŊ|u;-|;7 1†Ѵ|†u;vĺ bm-ѴѴ‹ķ  ;Š|u-1|;7 v;t†;m1;v ‰;u; Ѵo-7;7 om - Ƒѷ Չņˆő -]-uov; ];Ѵ -m7 lb]u-|;7 =ou ƑƏlbm -| ƖƏ(ĺ  t†-m|b=b1-|bom ‰-v -Ѵvo r;u=oul;7 †vbm] - -mo uor vr;1|uo-r_o|ol;|;uŐ ŊƐƏƏƏķ$_;ulo"1b;m|b=b1őĺ

ƑĺƕĺƒՊ|Պ

βŊ-Ѵ-1|ovb7-v;-vv-‹

m |_bv v;1|bomķ E. coli Ɣ _Š| ƔƔѵƒƑŋLac Z+ķ - v|u-bm |_-| oˆ;u;Š-ru;vv;v|_;];m;;m1o7bm]|_;βŊ]-Ѵ-1|ovb7-v;ŐβŊ]-Ѵő-1|bˆb|‹Չb|_o†| -77b|bomo=Ѵ-1|ov;|o|_;l;7b†lő‰-vv;Ѵ;1|;7|o-vv;vv|_;;==;1|o=  voml;l0u-m;r;ul;-0bѴbŒ-|bomĺ$o|_bv;m7ķ|_;u;Ѵ;-v;o=bm|u--1;ѴѴ†Ѵ-uβŊ]-Ѵ‰-v;ˆ-Ѵ†-|;70‹;mŒ‹l-|b1|b|u-|bom-11ou7bm]|obѴѴ;u ŐƐƖƖƑőĺmoˆ;umb]_|1†Ѵ|†u;‰-v7bѴ†|;7bm-m70uo†]_||o-mor|b-1-Ѵ 7;mvb|‹ Ő ₆₀₀ő o= Əĺѵķ †vbm] - vr;1|uor_o|ol;|;u Ő"r;1|uo|omb1 ƑƏƏĸ$_;ulo bv1_;uőĺ$_;v†vr;mvbom‰-v|_;mbm1†0-|;7-|ƒƕŦķbm |_;ru;v;m1;Ő|u;-|;7v-lrѴ;vő-m7-0v;m1;Őm;]-|bˆ;1om|uoѴķ|uѴƴő o= v-|7b==;u;m|1om|-1||bl;vĺrovb|bˆ;1om|uoѴŐ|uѴ+őo=Ѵ‹v;7 1;ѴѴv ‰-v ru;r-u;7 0‹ -77bm] ƔƏμѴo=1_Ѵouo=oul -m7ƑƔμѴo= " " ŐƏĺƐѷ ˆņˆő |o |_; 1†Ѵ|†u;ĺ  ˆoѴ†l; Ővő o= ƔƏμѴo=;-1_v-lrѴ;‰-v 7bѴ†|;7bmƖƔƏμѴo=m;†|u-Ѵ0†==;uŐ,0†==;uķrƕĺƏőoˆ;u-mb1;0-|_ĺ "-lrѴ;v‰;u;rѴ-1;7=ouƔlbm-|ƑѶŦbm-‰-|;u0-|_0;=ou;v|-u|bm] |_; u;-1|bomĺ $o ;-1_ v-lrѴ;ķ ƑƏƏμѴ o= oŊmb|uor_;m‹ѴŊβŊ]-Ѵ-1|ovb7; ŐķƓl]ņlѴő‰-v-77;7-m7|_;u;-1|bom‰-v|bl;7ĺ)_;mv-l-rѴ;v|†um;7‹;ѴѴo‰bv_ķ|_;u;-1|bom‰-vv|orr;70‹-77bm]ƔƏƏμѴo= ƐloѴņ -23 -m7 |_; |bl; u;1ou7;7 Őtőĺ $†0;v ‰;u; |_;m 1;m|ub- =†];7Ƒlbm-|ƐƒķƏƏƏg|ou;loˆ;1;ѴѴu;vb7†;v-m7|_;or|b1-Ѵ7;m-vb|‹o=|_;v†r;um-|-m|‰-vl;-v†u;7-|ƓƑƏml-m7ƔƔƏmlŐ 420 -m7  ₅₅₀őĺ bm-ѴѴ‹ķ |_;βŊ]-Ѵ-1|ovb7-v; -1|bˆb|‹ķ ;Šru;vv;7 bm βŊ]-Ѵ †mb|voubѴѴ;u†mb|v‰-v1-Ѵ1†Ѵ-|;7†vbm]|_;=oѴѴo‰bm];t†-|bomĹ

ƑĺѶՊ|Պ$u-mvlbvvbom;Ѵ;1|uomlb1uov1or‹-m-Ѵ‹vbvo=

0-1|;ub-Ѵl;l0u-m;bm|;]ub|‹

$u-mvlbvvbom ;Ѵ;1|uom lb1uov1or‹ Ő$ő ‰-v r;u=oul;7 |o bmˆ;v|b-]-|;|_;;==;1|o= vom1;ѴѴlour_oѴo]‹-m7l;l0u-m;bm|;]ub|‹ĺ "-lrѴ; ru;r-u-|bom ‰-v r;u=oul;7 =oѴѴo‰bm] |_; ]†b7;Ѵbm;v o= $-oķ b-mķ -m7 *b; ŐƑƏƐƐő -m7 *bm] ;|-Ѵĺ ŐƑƏƏƖ-ő ‰b|_ - vѴb]_| lo7b=b1--|bomĺ ˆ;umb]_| 1†Ѵ|†u;v ŐƐƏ6 &ņlѴő o= |_; v;Ѵ;1|;7 0-1|;ub- ‰;u; ;Šrov;7 |o  v =ou ƐƏķ ƑƏķ -m7 ƒƏlbmĺ †Ѵ|†u;v ‰;u; |_;m 1;m-|ub=†];7 ŐѵķƏƏƏ gņƒlbmő -m7 |_; u;v†Ѵ|bm] r;ѴѴ;|v ‰;u; u;v†vr;m7;7 bm - Ƒѷ Ոņˆő ]Ѵ†|-u-Ѵ7;_‹7; voѴ†|bom 1om|-bm;7 bm " Őr ƕĺƓő =ou oˆ;umb]_|=bŠ-|bomo=|_;1;ѴѴv-|ƓŦĺt†-m|b|‹o=ƐƏμѴo=;-1_v-lrѴ; ‰-v7;rovb|;7om oulˆ-u1-u0omŊ1o-|;7]ub7v1om|-bmbm]om;7uor o= Ɛѷ Ѵ1b-m Ѵ†;ĺ ;ѴѴv ‰;u; |_;m v†0f;1|;7 |o Ɣlbm rov|Ŋ=bŠ-|bom ‰b|_Ƒѷr-u-=oul-Ѵ7;_‹7;bm"ķ-m7]ub7v‰;u;v|-bm;7†vbm]-7uor o==bѴ|;u;7Ƒѷr_ovr_o|†m]v|b1-1b7Ő$ķrƕĺƏő=ouƒƏvĺv;ub;vo=

=bѴ|u-|bom -m7ņou ‰-v_bm] |u;-|l;m| ‰-v r;u=oul;7 -=|;u ;-1_ v|;r |o u;loˆ; ;Š1;vv Ѵbt†b7ķ =bŠ-|bˆ;ķ -m7 v|-bmbm]ĺ &m|u;-|;7 0-1|;ub- v-lrѴ;v‰;u;-Ѵvoru;r-u;70‹|_;v-l;l;|_o7ĺ bm-ѴѴ‹ķ$o0v;u-ˆ-|bom ‰-v r;u=oul;7 †vbm] - _bѴbrv ƐƏƏ |u-mvlbvvbom ;Ѵ;1|uom lb1uov1or;Ő_bѴbrvѴ;1|uomr|b1vķbm7_oˆ;mķ$_;;|_;uѴ-m7vő-m7 7b]b|-Ѵ lb1uo]u-r_v ‰;u; 1-r|†u;7 †vbm] -m $ *!ѶƏ   7b]b|-Ѵ 1-l;u-Ő7ˆ-m1;7b1uov1or‹$;1_mbt†;vķ)o0†umķ&"őĺ

ƒՊ|Պ!"&$"  "&""

ƒĺƐՊ|Պour_oѴo]‹o=;Ѵ;1|uovr†m v

b]†u;Ƒv_o‰v"bl-];vo=;Ѵ;1|uovr†m v=uolƕѷՉņˆő" voѴ†|bombmƔƏѷՈņˆő1ķ-m7ѶƏņƑƏ‰|u-|bo"ņ0Ѵ;m7bmƔƏѷ Ոņˆő 1ĺ v v_o‰m bm b]†u;Ƒ-ķ ;Ѵ;1|uovrbmmbm] m;-| " ‰-v - 7b==b1†Ѵ||-vh-m7lov|Ѵ‹]-ˆ;ubv;|or-u|b1Ѵ;vo=m-mol;|;u-m7lb-1uol;|;u vbŒ; Ő;Ѵ;1|uovru-‹bm]őĺ ;m1;ķ |_; -77b|bom o= - 1ovrbmmbm] -];m|ķv†1_-vķ|o=-1bѴb|-|;|_;;Ѵ;1|uovrbmm-0bѴb|‹o="‰-v†m--ˆob7-0Ѵ;ĺ)_;m-77;7|o"voѴ†|bombm-lo7;u-|;ruorou|bomŐ"ņ ‰|u-|boĹѶƏņƑƏőķ1o†Ѵ7-1|-v-1-uub;u-m7blruoˆ;|_;ˆbv-1o;Ѵ-v|b1ruor;u|b;vo="voѴ†|bom-v‰;ѴѴ-v1_-bm;m|-m]Ѵ;l;m|-m7 =Ѵ;Šb0bѴb|‹Ő-hu-ˆ-m;|-ѴĺķƑƏƐƐőķ|‰osine qua non conditions for fiber =oul-|bomĺomv;t†;m|Ѵ‹ķ_olo];m;o†v-m70;-7Ѵ;vv1_b|ov-mŊ0-v;7 m-mo=b0;uv‰b|_-ˆ;u-];=b0;u7b-l;|;uo=ƕѶmlƼƑƑ‰;u;v†11;vv-=†ѴѴ‹o0|-bm;7ĺ

ƒĺƑՊ|Պm|b0-1|;ub-Ѵ;==b1b;m1‹o= v

$-0Ѵ;Ɛ -m7 b]†u;ƒ 7bvrѴ-‹ķ u;vr;1|bˆ;Ѵ‹ķ |_; 0-1|;ub-Ѵ u;7†1|bom u-|; ŐRő -m7 |_; bm ˆb|uo -m|b0-1|;ub-Ѵ -1|bˆb|‹ o=  vķ t†-m|b|--|bˆ;Ѵ‹ -vv;vv;7 0‹ |_;  & l;|_o7 -]-bmv| E. coliķ S. $‹r_bl†ub†lķ L. innocuaķ-m7S. aureusĺ=|;uƓ_u1om|-1|-|ƒƕŦbm"ŐrƔĺѶőķ  v ŐƐ1l2ő v_o‰;7 vb]mb=b1-m| u;7†1|bom u-|; ŐR > ƖƖѷő o= 0-1|;-ub-Ѵ]uo‰|_o=E. coliķL. innocuaķ-m7S. aureusŐ$-0Ѵ;Ɛőķˆ;uv†vƖѵĺƖƐѷ for S. $‹r_bl†ub†lĺ)_;m"1om|;m|‰-vbm1u;-v;7ŐƑĺƔ1l2

bmv|;-7 of 1 cm2

őķ  v ‰;u; -0Ѵ; |o 1olrѴ;|;Ѵ‹ v|or |_; ]uo‰|_ o= E. coli -m7L. innocuaķŐƐƏƏѷRķ$-0Ѵ;Ɛőķ-vv_o‰m0‹|_;-uuo‰vŐ b]†u;ƒőĺ o‰;ˆ;uķ S. aureus -m7 S. $‹r_bl†ub†l v_o‰;7 Ѵo‰;u v†v1;r|b0bѴb|‹ |o |_; -1|bom o=  vĺ ;ˆ;u|_;Ѵ;vvķ - vb]mb=b1-m| 7ov;Ŋ7;r;m7;m| 7;1u;-v;o=0-1|;ub-Ѵror†Ѵ-|bomŐƔѴo]v-m7ƓѴo]vķu;vr;1|bˆ;Ѵ‹ő‰-v v|bѴѴ o0v;uˆ;7 Ő b]†u;ƒőĺ †u|_;ulou;ķ bm ou7;u |o bm1u;-v; |_; -m|bŊ v-Ѵlom;ѴѴ- ou -m|bŊv|-r_‹Ѵo1o11-Ѵ -1|bˆb|‹ o=  vķ b| bv rovvb0Ѵ; |o 1ol0bm; 1_b|ov-m ‰b|_ o|_;u -m|blb1uo0b-Ѵ -];m|v v†1_ -v ;|_‹Ѵ;m-;7b-lbm;|;|u--1;|b1-1b7Ő $ķƏĺƑѷőŐѴ-bl-|şoѴѴ;‹ķƑƏƐƔő-m7 ;vv;m|b-ѴobѴvŐ"_-_0-Œbş"_-ˆbvbķƑƏƐѵőķ=ou-v‹m;u]bv|b1;==;1|ĺ

ƒĺƒՊ|Պuo|;bmvѴ;-h-];

$_; u;Ѵ;-v; o= bm|u-1;ѴѴ†Ѵ-u ruo|;bmv bv -m bm7b1-|bom o= l;l0u-m; 7;|;ubou-|bomĺ b]†u;Ɠ v_o‰v " "Ŋ r-||;umv o= u;Ѵ;-v;7 1‹|o-rѴ-vlb1voѴ†0Ѵ;ruo|;bmv=uol1_b|ov-mŊ|u;-|;7E. coli-m7S. aureus. In |_;1-v;o=E. coliķ|_;ruo|;bm1om|;m|bm|_;1;ѴѴŊ=u;;v†r;um-|-m|‰-v vblbѴ-u |o |_-| o= |_; rovb|bˆ; 1om|uoѴ Ő|uѴ+ő |_-| u;=;uv |o 0-1|;ub-Ѵ ŐƑő

β−galactosidase units =1000×(OD420−1.75×OD550)

ՊՍՊ

|

ՊѶѵƖ

ARKOUN ETAL.

v†vr;mvbom -=|;u 1;ѴѴ Ѵ‹vbv |u;-|l;m|ĺ $_bv u;v†Ѵ| bm7b1-|;v |_-| |_; ;==;1|o= v‰-vbmv|-m|-m;o†vŐbm|_;=buv|_o†uo=|u;-|l;m|ő-m7 -Ѵlov| =†ѴѴ‹ 1olrѴ;|;7 vbm1; -ѴѴ bm|u-1;ѴѴ†Ѵ-u ruo|;bmv ‰;u; u;Ѵ;-v;7 |o |_; ;Š|u-1;ѴѴ†Ѵ-u l;7b†lķ -v f†7];7 0‹ |_; 1olr-ubvom 0;|‰;;m

|_; Ŋ|u;-|;7-m7|_;1_;lb1-ѴѴ‹Ѵ‹v;7v-lrѴ;vĺ ouS. aureus0-1-|;ub†lķ |_; ;==;1| ‰-v ]u-7†-Ѵĺ $_; ;Ѵ;1|uor_ou;vbv r-||;um v_o‰;7 |_-| |_; bm|;mvb|‹ o= |_; 0-m7v bm1u;-v;7 ‰b|_ |bl;ķ bm7b1-|bm] |_-| ruo|;bmѴ;-h-];‰-vѴom];u-m7ruo]u;vvbˆ;ĺ)_;m1olr-u;7|o|_; rovb|bˆ;1om|uoѴŐ|uѴ+őķ|_;;==;1| o= v‰-vbm1olrѴ;|; -m7v;ˆ-;u-Ѵ0-m7v7b7mo|-rr;-u;ˆ;m-=|;uƓ_u;Šrov†u;ĺ$_bvbm7b1-|;v|_-| l-m‹ruo|;bmvu;l-bm;7bm|_;1‹|orѴ-vlo=Ѵbˆbm]1;ѴѴvĺ$_;v;u;v†Ѵ|v v†]];v||_-|1_b|ov-mrѴ-‹v-m-1|bˆ;uoѴ;bml;l0u-m;r;ul;-0bѴbŒ--|bomĺ o‰;ˆ;uķ |_; o0v;uˆ;7 -m|b0-1|;ub-Ѵ ;==;1| o=  v om l;l-0u-m;7-l-];ķ-vu;rou|;70‹ruo|;bmu;Ѵ;-v;‰-vlou;ruomo†m1;7 bm|_;1-v;o=E. coli|_-mS. aureusķv†]];v|bm]-_b]_;uv†v1;r|b0bѴb|‹ of E. coliķ-vu;rou|;7bm-mo|_;uv|†7‹Őuho†mķ -b]Ѵ;ķ;†Œ;‹ķşffbķ ƑƏƐƕőĺ

ƒĺƓՊ|Պ Ѵ;-h-];

$_;u;Ѵ;-v;o=0-1|;ub-Ѵ];molb1 bm|_;v†r;um-|-m|‰-v7;|;1|;7 0‹!-lrѴb=b1-|bomo=|_;rrnB];m;ŐƐѵ"ő=ouE. coliŐ b]†u;Ɣőĺ$_; -77b|bom-Ѵ v|;r o= r -7f†v|l;m| |o m;†|u-Ѵb|‹ l;m|bom;7 bm |_; l;|_o7oѴo]‹ ‰-v m;1;vv-u‹ |o _bm7;u 1olrѴ;Š-|bom o=  ‰b|_  vĺ|_;u‰bv;ķmo|u-1;o=|_;=oul;u1o†Ѵ70;7;|;1|;7ĺ ;|;1|bom o= bm|_;;Š|u-1;ѴѴ†Ѵ-ul;7b†lŐv†r;um-|-m|ő‰-v-1omv;t†;m1;   & !   Ƒ Պ "lb1uo]u-r_vo=Ő-őĹ ;Ѵ;1|uovru-‹;7ƕѷՉņˆő"bmƔƏѷ Ոņˆő1-m7Ő0őĹ;Ѵ;1|uovr†mƕѷ Չņˆő"ņŐѶƏņƑƏőbmƔƏѷՈņˆő 1-|ƑƐŦķƕѷu;Ѵ-|bˆ;_†lb7b|‹ķ Ő1őĹ b0;u7b-l;|;u7bv|ub0†|bomo=0ĺ uo1;vvr-u-l;|;uvĹ|brŊ|oŊ1oѴѴ;1|ou 7bv|-m1;ƷƐƔ1lķ=Ѵo‰u-|;ƷƏĺƔlѴņ_uķ ˆoѴ|-];ƷƑƏh(ĺ"1-Ѵ;0-uvu;ru;v;m|Ɛμm 7b-l;|;u-m7l-]mb=b1-|bomƵѵ-m7ƵƐƏ=ou v-lrѴ;vƑ--m7Ƒ0ķu;vr;1|bˆ;Ѵ‹ (a) (b) (c) !;7†1|bomu-|;Őѷő

-mo=b0;u‰;0v E. coli S. $‹r_bl†ub†l S. aureus L. innocua

1 cm2

 v ƖƖĺƖƒƼƏĺƔ ƖѵĺѶƐƼƑĺƒ ƖƖĺƐƓƼƐĺѶ ƖƖĺƖƏƼƏĺƏƑ

2.5 cm2

 v ƐƏƏƼƏ ƖѶĺƖƕƼƐķƑ ƖƖĺƖѶƼƏķƔ ƐƏƏƼƏ

$      Ɛ Պ -1|;ub-Ѵu;7†1|bomu-|;Ő!őo=  v-]-bmv|E. coliķS. $‹r_bl†ub†lķ L. innocuaķ-m7S. aureusķ-vt†-m|b|-|bˆ;Ѵ‹ -vv;vv;70‹|_; &l;|_o7ķ-=|;uƓ_u bm1†0-|bom-|ƒƕŦbm"ŐrƔĺѶő

  & !   ƒ Պ m|b0-1|;ub-Ѵ-1|bˆb|‹o= v-]-bmv|E. coliķL. innocuaķ S. aureusķ-m7S. $‹r_bl†ub†lķ-v;ˆ-Ѵ†-|;70‹|_;7‹m-lb1 & l;|_o7-=|;uƓ_ubm1†0-|bom-|ƒƕŦbm"ŐrƔĺѶőĺ$_;-uuo‰v robm|-||_;1olrѴ;|;bm_b0b|bomo=0-1|;ub-Ѵ]uo‰|_ŐRƷƐƏƏѷő

ѶƕƏՊ

|

ՊՊՍ ARKOUN ETAL.

o=|_;7bvu†r|bomo=l;l0u-m;r;ul;-0bѴb|‹1-†v;70‹ vŐ b]†u;Ɣ -m7őĺm1om|u-v|ķmo ‰-v7;|;1|;7bm|_;;Š|u-1;ѴѴ†Ѵ-ul;7b†l o= †m|u;-|;7 v-lrѴ; Ő|uѴƴķ b]†u;Ɣ őķ ‰_b1_ ‰-v v‹mom‹lo†v ‰b|_ l;l0u-m;bm|;]ub|‹ĺ$_;o0v;uˆ;70ub]_|m;vv-||_;Ѵo-7bm]vro|vo= |_;|u;-|;7v-lrѴ;v‰-vruo0-0Ѵ‹7†;|o-7;rovb|bomo=vl-ѴѴ1-|bomb1 1_-bmvo="b|v;Ѵ=ķ‰_b1_7b7mo|lb]u-|;|o‰-u7v|_;1-|_o7;ĺ$_bv 1-m0;-Ѵvo-||ub0†|;7|o-7;1;Ѵ;u-|bomo=|_;;Ѵ;1|uor_ou;|b1lo0bѴb|‹ o=];molb1 1-†v;70‹|_;1_;Ѵ-|bom;==;1|o=1_b|ov-mķ-vv†]-];v|;70‹*bm]ķ_;mķb†ķ_-ķ-m7-uhŐƑƏƏƖ0őĺ;]-|bˆ;Ѵ‹1_-u];7 r_ovr_-|; ]uo†rv ru;v;m| bm m†1Ѵ;b1 -1b7vķ v†1_ -v  -m7 !ķ lb]_|0;-mbm|u-1;ѴѴ†Ѵ-u|-u];|=ou"-m71om|ub0†|;|ob|vbm|;u-1|bom ‰b|_0-1|;ub-Ѵ1;ѴѴvĺ$_bv1omf;1|†u;‰-vˆ;ub=b;7‰_;m"‰-v7;ruo-|om-|;7 Ő-| m;†|u-Ѵ rő bm ou7;u |o ru;ˆ;m| "Ŋ  1olrѴ;Š-|bomĺ v-1omv;t†;m1;ķ];molb1 ‰-v7;|;1|;70o|_t†-Ѵb|-|bˆ;Ѵ‹-m7 t†-m|b|-|bˆ;Ѵ‹ĺ $_;v; u;v†Ѵ|v robm| o†| |_-| |_; Ѵ;-h-]; o= 0-1|;ub-Ѵ  ‰o†Ѵ7 mo| o11†u ‰b|_o†| l;l0u-m; r;u=ou-|bom -m7 v|uom]Ѵ‹ v†]];v| - l;l0u-moѴ‹|b1 ;==;1| bm  vĽ l;1_-mbvl o= -1|bomĺ $_; 1om1;m|u-|bomvo=u;Ѵ;-v;7 -=|;u;Šrov†u;|o vķ-vl;-v†u;7 †vbm]--mo uorvr;1|uor_o|ol;|;uŐ ŊƐƏƏƏķ$_;ulo"1b;m|b=b1őķ -=|;u!‰;u;ƐѶĺƑķƐƖĺƔķƑƏĺƖķѵƏĺƑķ-m7ƐƕƑĺƒm]ņμѴķ-=|;uƏķƐķƑķƒķ -m7Ɠ_u;Šrov†u;|bl;vķu;vr;1|bˆ;Ѵ‹ĺ †-m|b=b1-|bomo=u;Ѵ;-v;7  =uol Ŋ|u;-|;7E. coli1Ѵ;-uѴ‹bm7b1-|;v|_-|];molb1 1o†Ѵ70; 7;|;1|;7bm|_;;Š|u-1;ѴѴ†Ѵ-ul;7b†l-m7b|v1om1;m|u-|bom‰-vruo-rou|bom-Ѵ|o|_;1om|-1||bl;0;|‰;;mE. coli-m7 vĺ

ƒĺƔՊ|Պ!;Ѵ;-v;o=bm|u-1;ѴѴ†Ѵ-u

βŊ]-Ѵ-1|ovb7-v;;mŒ‹l;

$_;u;Ѵ;-v;o=1‹|orѴ-vlb1βŊ]-Ѵ-1|ovb7-v;ŐβŊ]-Ѵő‰-v-Ѵvo-m;ˆb7;m1; o=l;l0u-m;r;ul;-0bѴbŒ-|bomĺ b]†u;ѵv_o‰v|_;u;Ѵ;-v;o=βŊ]-Ѵ;m-Œ‹l;=uolE. coli-=|;u7b==;u;m|1om|-1||bl;v‰b|_ vĺ$_;u;v†Ѵ|v u;ˆ;-Ѵ;7|_-|m;]-|bˆ;1om|uoѴvo=†m|u;-|;70-1|;ub-Ő0Ѵ-1hvt†-u;vő v_o‰;7mo;mŒ‹l-|b1-1|bˆb|‹ķ‰_b1_‰-v-mbm7b1-|bomo=l;l0u-m; bm|;]ub|‹ĺ )_;m  v ‰;u; -77;7 |o |_; 0-1|;ub-Ѵ v†vr;mvbomķ - ruo]u;vvbˆ; |bl;Ŋ7;r;m7;m| ;mŒ‹l-|b1 -1|bˆb|‹ ‰-v o0v;uˆ;7 Őu;7 1bu1Ѵ;vőķ-1omv;t†;m1;o=l;l0u-m;Ѵ;vbomĺo‰;ˆ;uķu;v†Ѵ|v7;lom-v|u-|;7|_-|b|‰-vmo|rovvb0Ѵ;|ou;-1_|_;l-Šbl†l;Šr;1|;7Ѵ;ˆ;Ѵ o=u;Ѵ;-v;7βŊ]-Ѵ=uol1_;lb1-ѴѴ‹Ѵ‹v;71;ѴѴvŐŜƓƑƏβŊ]-Ѵ†mb|vőĺ$_bv v†]];v|v|_-||_;-m|b0-1|;ub-Ѵ;==;1|o= v‰-vmo|1olrѴ;|;7-m7 |_; u;Ѵ;-v; o= |_; ;mŒ‹l; bv - Ѵom];u ruo1;vv o11†uubm] -=|;u 7;-|_ -m7Ѵ‹vbvo=|_;1;ѴѴĺ$_;v;u;v†Ѵ|vu;-vom-0Ѵ‹7;lomv|u-|;|_;-0bѴb|‹

  & !   Ɠ Պ " "Ŋr-||;umvo= u;Ѵ;-v;7bm|u-1;ѴѴ†Ѵ-uruo|;bmv=uol Ŋ |u;-|;7E. coli-m7S. aureusķ-=|;uƏķƐķƑķƒķ -m7Ɠ_u1om|-1||bl;-|ƒƕŦbm"ĺ|uѴ+ u;=;uv|o|o|-Ѵruo|;bmv1_;lb1-ѴѴ‹;Š|u-1|;7 -=|;u|u;-|l;m|o=1;ѴѴv‰b|_-Ѵ‹vbvvoѴ†|bom 1om|-bmbm]ƔƏμѴ1_Ѵouo=oul-m7ƑƔμѴ" " ŐƏĺƔѷˆņˆő   & !   Ɣ Պ ]-uov;];Ѵ;Ѵ;1|uor_ou;vbvo=u;Ѵ;-v;7];molb1 =uol Ŋ|u;-|;7E. coli-=|;uĹƓ_u-m7ĹƑƓ_u1om|-1| |bl;ĺ"-lrѴ;v-m7u;=;u|o|uѴ+-m7|uѴ+rovb|bˆ;1om|uoѴv of E. coli -=|;u1_;lb1-Ѵ-m7_;-||u;-|l;m|o=0-1|;ub-Ѵ1;ѴѴvķ u;vr;1|bˆ;Ѵ‹ĺ"-lrѴ; u;=;uv|om;]-|bˆ;1om|uoѴo=];molb1 ;Š|u-1|;7=uol†m|u;-|;70-1|;ub-Ѵ1;ѴѴvĺu;=;uv|oѴ-77;uĽv =u-]l;m|v‰_ov;loѴ;1†Ѵ-u‰;b]_|v-u;]bˆ;mbm0-v;r-buŐ0rő   & !   ѵ Պ !;Ѵ;-v;o=1‹|orѴ-vlb1βŊ]-Ѵ-1|ovb7-v;ŐβŊ]-Ѵő;mŒ‹l; from E. coli Ɣ_Š|ƔƔѵƒƑŋLac Z+ ķ-=|;u7b==;u;m|;Šrov†u;|bl; |o vĺ|uѴƴŐm;]-|bˆ;1om|uoѴőu;=;uv|o|_;Ѵ;ˆ;Ѵo=u;Ѵ;-v;7βŊ]-Ѵ bm|_;-0v;m1;o= vĺ|uѴ+Őrovb|bˆ;1om|uoѴőu;=;uv|o|_;Ѵ;ˆ;Ѵo= βŊ]-Ѵu;Ѵ;-v;70‹1_;lb1-ѴѴ‹Ѵ‹v;71;ѴѴvŐru;r-u;70‹-77bm]ƔƏμѴo= 1_Ѵouo=oul-m7ƑƔμѴo=" "|o|_;1†Ѵ|†u;ő

ՊՍՊ

|

ՊѶƕƐ ARKOUN ETAL. o= v|or;ul;-|;0-1|;ub-Ѵl;l0u-m;-m71obm1b7;‰b|_|_;=bm7-bm]vo=$-o;|-ѴĺŐƑƏƐƐőķ‰_ou;rou|;7vblbѴ-uu;v†Ѵ|v=ou"voѴ†|bomvĺ

ƒĺѵՊ|Պ$u-mvlbvvbom;Ѵ;1|uomlb1uov1or‹-m-Ѵ‹vbvo=

l;l0u-m;r;ul;-0bѴbŒ-|bom;==;1|o= v

$_; ;==;1| o=  v om l;l0u-m; lour_oѴo]‹ -m7 bm|;]ub|‹ ‰-v bmˆ;v|b]-|;7 0‹ $ Ő b]†u;ƕőĺ &m|u;-|;7 1;ѴѴv o= E. coli Őu-lŊ m;]-|bˆ;ő -m7 S. aureus Őu-lŊrovb|bˆ;ő ‰;u; bm|-1| -m7 7b7 mo|

v_o‰ -m‹ l;l0u-m; Ѵ;vbom ou -mol-Ѵ‹ Ő b]†u;ƕ- -m7 ѵ;őĺ =|;u ;Šrov†u; |o  vķ - u;l-uh-0Ѵ; -Ѵ|;u-|bom o= l;l0u-m; bm|;]-ub|‹ ‰-v o0v;uˆ;7ĺ $ bl-];v o= ;Šrov;7 1;ѴѴv |o  v u;ˆ;-Ѵ;7 |_-| -=|;u ƐƏlbm 1om|-1|ķ 0o|_ E. coli -m7 S. aureus v|u-bmv v_o‰;7 l;l0u-m; r;ul;-0bѴbŒ-|bom 0‹ r;u=ou-|bom Ő b]†u;ƕ0 -m7 =őĺ =|;u ƑƏlbm ;Šrov†u;ķ 0o|_ 0-1|;ub- ‰;u; Ѵ;-hbm] 1‹|ovoѴb1 1olrom;m|v Ő b]†u;ƕ1-m7]őĺo‰;ˆ;uķl;l0u-m;7;|-1_l;m|o11†uu;7omѴ‹bm E. coliŐ b]†u;ƕ7őĺu-7†-Ѵl;l0u-m;7;|-1_l;m|=uol|_;1;ѴѴ‰-ѴѴ of E. coliķ-m7v_ubmh-];o=|_;1‹|orѴ-vl‰-vo0v;uˆ;7-=|;uƒƏlbm   & !   ƕ Պ $u-mvlbvvbom;Ѵ;1|uom lb1uov1or‹Ő$őlb1uo]u-r_vo=-ķ0ķ1ķ d: t₀ķƐƏķƑƏķ-m7ƒƏlbm;Šrov†u;o=E. coli 1;ѴѴv|o vķ-m7;ķ=ķ]-m7_Ĺ|0ķƐƏķƑƏķ -m7ƒƏlbm;Šrov†u;o=S. aureus1;ѴѴv |o vķu;vr;1|bˆ;Ѵ‹ĺ$_;‹;ѴѴo‰ķ]u;;m -m70Ѵ†;-uuo‰vķu;vr;1|bˆ;Ѵ‹ķrobm|-| l;l0u-m;r;u=ou-|bomķѴ;-h-];o=1‹|ovoѴķ -m71;ѴѴѴ‹vbv (a) (b) (c) (d) (e) (f) (g) (h)

872 

|

     ARKOUN et Al.

contact time with CNFs, as pointed by the blue arrows (Figure 7d). This detachment of the plasma membrane was due to desorption of the cytosol, subsequent to leakage of intracellular compounds (Figure 7c), making cells look transparent and empty (Figure 7d). After 30 min contact time, the cytoplasmic membrane of E. coli col-lapsed (Figure 7d) and S. aureus cells were completely disintegrated (Figure 7h). Adsorption of molecules to bacterial cell walls, of both E. coli and S. aureus was also observed and was proportional to con-tact time (Figure 7b, 7c and 7d). This might be due to (1) the release of intracellular components that can attach to the surface of bac-teria, reflecting local cell rupture, or (2) to small soluble CS chains surrounding the bacterial cells via electrostatic and hydrophobic interactions, or (3) both possibilities. A simple visual inspection of the CS/PEO nanofiber mats, before and after the antibacterial tests indicated that the fibers were stable after 48 hr at 37°C, pH 5.8 in PBS. This suggests that resolubilization was only partial, as the mats remained intact. However, as nanofibers contain PEO, which is solu-ble in water, a certain solubility of PEO is expected. In addition, due to the pH of the medium (5.8), chitosan may solubilize partially, as verified by Ardila, daigle, Heuzey, and Ajji (2017). Consequently, both the released chitosan in the medium and the one remaining in the na-nofiber mats may contribute to the antibacterial effect of the CNFs. The second conjuncture coincides with the findings of other au-thors (Chung et al., 2004; Helander, Nurmiaho- Lassila, Ahvenainen, Rhoades, & Roller, 2001), who studied the adsorptive characteristics of bacterial cells to chitosan solutions. This suggests that the mecha-nism of action of CNFs may be also due to partial resolubilization of CS in the media, even though visually the mats looked intact after 48 hr in PBS or LB. Short CS chains might, thereby, penetrate the cell wall and perforate the plasma membrane, while longer chains could enclose bacteria and prevent cell exchange with the extracellular me-dium. Accordingly, Figure 7b, c, d, and f, show that CS formed an impermeable envelope surrounding the bacteria which might block the absorption of essential elements into the cells (Choi et al., 2001; Eaton, Fernandes, Pereira, Pintado, & Malcata, 2008). Ultimately, it can be inferred that the bactericidal effect of CNFs may be the result of membrane perforation. Our results are in agreement with those of other authors (Tao et al., 2011; Xing et al., 2009a), who observed membrane perforation of E. coli caused by CS solutions and particles, respectively. However, our experiments conducted on CNF- treated E. coli (Gram- negative) and S. aureus (Gram- positive), revealed vari-ous surface characteristics and cell stages in response to treatment with CNFs. This suggests that the mechanism of action of CNFs is a complex combination of different bactericidal effects that can occur at different stages: (1) CNFs inhibit bacterial growth through mem-brane pervasion and perforation, (2) partly resolubilized CS chains can kill bacteria by causing membrane rupture and/or suppressing cell exchange and nutrient uptake, (3) CS nanofibers and/or solutions can cause osmotic stress by chelating trace elements such as metallic ions, essential to bacterial growth. However, the common mecha-nism behind these different modes of action is undeniably due to the protonated functional groups of CS. The results clearly demonstrate that CNFs’ bactericidal effect involves permeabilization of bacterial membrane with pore formation, contrary to what has been reported so far. However, no evidence of penetration of the membrane can be inferred, even though pore formation assuredly occurred. The next challenge should aim at clarifying the molecular mechanisms behind the bactericidal activity of CNFs and identifying the membrane ele- ments and metabolic pathways involved in the internalization of chi-tosan into the bacterial cell wall. These further studies will not only be critical for the application of such materials in food packaging, but also for the prevention of outbreak of resistance phenomena toward chitosan.

4 | CONCLUSIONS

The results of this study show that the antibacterial activity of chi-tosan nanofibers (CNFs) can be attributed to membrane disruption and perforation. Consequently, this resulted in the leakage of intra- cellular components such as proteins and nucleotides. The bioavail-ability of NH₃+ _{functional groups on CNFs favored and maximized}

cell adhesion and attachment to the surface of the mats. The model established here, regarding CNFs’ mode of action suggests that bac-teria migrate to the surface of the nanofibers and not the reverse. Since bacteria use adhesion and attachment surfaces to better grow and multiply, CNFs showed the ability to efficiently attract and trap bacteria through electrostatic interactions, on account of their large surface- to- mass ratio and high porosity. Our results also suggest that adsorption of CS to the bacterial surface is the first step in CNFs’ mechanism of action, followed by membrane perforation, leak-age of cytosolic compounds, and finally cell lysis and disintegration. Nevertheless, it is not excluded that part of the antibacterial activity might be due to partial dissolution of the nanofibers, making chitosan available in solution. As promising practical application, CNFs can be used as part of active food packaging in order to extend the shelf life of food products along with preventing spoilage by bacteria such as E. coli, and foodborne diseases caused by Listeria, Staphylococcus and Salmonella.

ACKNOWLEDGMENTS

This work was supported by the Fonds de Recherche du Québec - Nature et Technologies (FRQNT) and the Natural Sciences and Engineering Research Council of Canada (NSERC). The authors acknowledge Dr. Philippe Tremblay from Ovensa Inc. for providing the chitosan grade used in this work. We are grateful to Dr. Ève- Lyne Quévillon of UdeM for her precious help in performing PCR and NanoDrop analysis. Finally, the authors also acknowledge the review-ers of Food Science & Nutrition for their valuable comments to improve this manuscript.

CONFLICT OF INTEREST

ՊՍՊ

|

ՊѶƕƒ ARKOUN ETAL. !   !     " u7bѴ-ķĺķ7-b]Ѵ;ķ ĺķ;†Œ;‹ķĺŋĺķşffbķĺŐƑƏƐƕőĺm|b0-1|;ub-Ѵ-1|bˆb|‹ o=m;-|1_b|ov-mro‰7;u-m7=Ѵ-h;vĺMoleculesķ22ŐƐƏƏőķƐŋƐƖĺ u7bѴ-ķ ĺķ ;7bm-ķ ĺķ uho†mķ ĺķ ;†Œ;‹ķ ĺŊĺķ ffbķ ĺķ ş -m1_-Ѵķ ĺ ĺŐƑƏƐѵőĺ_b|ov-mŋ0-1|;ub-Ѵm-mo1;ѴѴ†Ѵov;m-mo=b0uo†vv|u†1|†u;v=ou ro|;m|b-Ѵ‰o†m77u;vvbm]-rrѴb1-|bomvĺCelluloseķ23ŐƔőķƒƏѶƖŋƒƐƏƓĺ uho†mķĺķ -b]Ѵ;ķ ĺķ;†Œ;‹ķĺĺķşffbķĺŐƑƏƐƕőĺ;1_-mbvlo=-1|bom

o= ;Ѵ;1|uovr†m 1_b|ov-mŊ0-v;7 m-mo=b0;uv -]-bmv| l;-| vrobѴ-]; -m7 r-|_o];mb10-1|;ub-ĺMoleculesķv†0lb||;7ĺ

"$ƑƐƓƖƴƐƒ-ĺŐƑƏƐƒőĺStandard test method for determining the

antimi-crobial activity of antimiantimi-crobial agents under dynamic contact conditions.

);v|omv_o_o1h;mķĹl;ub1-m"o1b;|‹=ou$;v|bm]-m7-|;ub-Ѵvĺ

;Ѵ-Ѵb-ķ!ĺķu;Ѵb;uķ"ĺķ;m-bvv-ķĺķşol-ķ(ĺŐƑƏƏѶőĺ;‰0bo-1|bˆ;0bo-l-|;ub-Ѵv0-v;7omt†-|;umbŒ;71_b|ov-mĺJournal of Agriculture and Food

Chemistryķ56ŐƔőķƐƔѶƑŋƐƔѶѶĺ

_obķĺŊĺķblķĺŊ+ĺķ+ooķ+ĺŊĺķ_ķ"ĺŊĺķ_obķĺŊĺķşblķĺŊ+ĺŐƑƏƏƐőĺ mˆb|uo-m|blb1uo0b-Ѵ-1|bˆb|‹o=-1_b|ooѴb]ov-11_-ub7;lbŠ|†u;-]-bmv| 1|bmo0-1bѴѴ†v -1|bmol‹1;|;l1olb|-mv -m7 "|u;r|o1o11†v l†|-mvĺ

International Journal of Antimicrobial Agentsķ18ŐѵőķƔƔƒŋƔƔƕĺ

_†m]ķ+ĺŊĺķ"†ķ+ĺŊĺķ_;mķĺŊĺķb-ķĺķ)-m]ķĺŋѴĺķ)†ķĺĺķşbmķ ĺŊĺ ŐƑƏƏƓőĺ !;Ѵ-|bomv_br 0;|‰;;m -m|b0-1|;ub-Ѵ -1|bˆb|‹ o= 1_b|ov-m -m7v†u=-1;1_-u-1|;ubv|b1vo=1;ѴѴ‰-ѴѴĺActa Pharmacologica Sinicaķ25ķ 932–936.

;v-bķĺķb|ķĺķbķĺķş,bˆ-moˆb1ķ"ĺŐƑƏƏѶőĺour_oѴo]b1-Ѵ-m7v†u=-1; ruor;u|b;vo=;Ѵ;1|uovr†m1_b|ov-mm-mo=b0;uvĺBiomacromoleculesķ9Őƒőķ 1000–1006.

oऀ-mķ ĺķ ࠑŒ‹dѴ7dŒķ ĺķ -w-Ѵķ ĺķ ş &Œ;Ѵķ ĺ ŐƑƏƐƒőĺ -0ub1-|bom o= ;Ѵ;1-|uovr†m 1_b|ov-m -m7 1_b|ov-mņroѴ‹ Ő;|_‹Ѵ;m; oŠb7;ő m-mo=b0;u ‰;0v -m7 -vv;vvl;m| o= |_;bu -m|blb1uo0b-Ѵ -1|bˆb|‹ĺ International Polymer

Processingķ28ŐƑőķƐƓƒŋƐƔƏĺ

-|omķĺķ ;um-m7;vķĺĺķ;u;bu-ķĺķbm|-7oķĺĺķş-Ѵ1-|-ķ ĺ*ĺŐƑƏƏѶőĺ |olb1 =ou1; lb1uov1or‹ v|†7‹ o= |_; -m|b0-1|;ub-Ѵ ;==;1|v o= 1_b-|ov-mvomv1_;ub1_b-1oѴb-m7"|-r_‹Ѵo1o11†v-†u;†vĺUltramicroscopyķ

108ŐƐƏőķƐƐƑѶŋƐƐƒƓĺ

Ѵv-0;;ķĺ,ĺķ-]†b0ķĺ ĺķşouvbķ!ĺĺŐƑƏƐƑőĺ_b|ov-m0-v;7m-mo-=b0;uvķu;ˆb;‰ĺMaterials Science and Engineering: Cķ32ŐƕőķƐƕƐƐŋƐƕƑѵĺ

;m]ķ*ĺķ‰omķĺŊĺķş-m]ķĺŐƑƏƏƔőĺѴ;1|uovrbmmbm]o=1_b|ov-m7bv-voѴˆ;7 bm 1om1;m|u-|;7 -1;|b1 -1b7 voѴ†|bomĺ Biomaterialsķ 26ŐƑƕőķ 5427–5432.

u;;mķ ĺķ ş "-l0uoohķ ĺ ŐƑƏƐƑőĺ Molecular cloning: A laboratory manual

(Vol. 4th edition)ĺ;‰+ouhĹoѴ7"rubm]-u0ou-0ou-|ou‹u;vvŐ_-r

Ɛķruo|o1oѴƓőĺ

u;bm;uķĺķş);m7ou==ķĺĺŐƑƏƏƕőĺѴ;1|uovrbmmbm]Ĺ=-v1bm-|bm]l;|_o7 =ou|_;ru;r-u-|bomo=†Ѵ|u-|_bm=b0;uvĺAngewandte Chemie International

Editionķ46ŐƒƏőķƔѵƕƏŋƔƕƏƒĺ

†ķĺĺķ-uhķ"ĺĺķblķĺ"ĺķ-m]ķĺĺķblķĺŊĺķşblķĺŊĺŐƑƏƐƒőĺ -0ub1-|bomo=vomb1-|;71_b|ov-mm-mo=b0;ul-|‰b|_;mѴ-u];7rouovb|‹ =ou†v;-v_;lov|-|b1l-|;ub-ѴvĺCarbohydrate Polymersķ97ŐƐőķѵƔŋƕƒĺ -b7;uķ "ĺķ ş -uhķ "ĺŊ+ĺ ŐƑƏƏƖőĺ u;r-u-|bom o= |_; ;Ѵ;1|uovr†m 1_b|ov-m

m-mo=b0;uv -m7 |_;bu -rrѴb1-|bomv |o |_; -7vour|bom o= † Őő -m7 0 Őőbomv=uol-m-t†;o†vvoѴ†|bomĺJournal of Membrane Scienceķ328ŐƐőķ 90–96.

-ll;uķ ĺ &ĺķ u-†v;uķ ĺķ Ѵ-hķ ĺķ u;Œ;vbmvhbķ ĺķ oѴ7l-mmķ $ĺķ †|vl-mmķ $ĺķ ş m7u࢜ķ ĺ ŐƑƏƐƏőĺ broroѴ‹v-11_-ub7; bm|;u-1|bom bv 7;1bvbˆ; =ou |_; -1|bˆb|‹ o= |_; -m|blb1uo0b-Ѵ r;r|b7; ŊƑ -]-bmv| v1_;ub1_b- 1oѴb -m7 uo|;†v lbu-0bѴbvĺ Biochemical Journalķ 427Őƒőķ 477–488.

;Ѵ-m7;uķĺķ†ulb-_oŊ-vvbѴ-ķĺŊĺķ_ˆ;m-bm;mķ!ĺķ!_o-7;vķĺķş!oѴѴ;uķ"ĺ ŐƑƏƏƐőĺ_b|ov-m7bvu†r|v|_;0-uub;uruor;u|b;vo=|_;o†|;ul;l0u-m; o= u-lŊm;]-|bˆ; 0-1|;ub-ĺ International Journal of Food Microbiologyķ

71ŐƑőķƑƒƔŋƑƓƓĺ

ol-‹ombķĺķ!-ˆ-m7bķ"ĺĺĺķş(-ѴbŒ-7;_ķĺŐƑƏƏƖőĺѴ;1|uovrbmmbm]o= 1_b|ov-m m-mo=b0;uvĹ uo1;vvbm] or|blbŒ-|bomĺ Carbohydrate Polymersķ

77ŐƒőķѵƔѵŋѵѵƐĺ

]m-|oˆ-ķĺķ-moѴoˆ-ķĺķ-uhoˆ-ķĺķş!-v_hoˆķĺŐƑƏƏƖőĺѴ;1|uovr†m omŊ)oˆ;m -mo=b0uo†v ‹0ub7 -|v -v;7 om _b|ov-m -m7  =ou )o†m7Ŋ u;vvbm] rrѴb1-|bomvĺ Macromolecular Bioscienceķ 9ŐƐőķ 102–111.

-‹-h†l-uķ !ĺķ u-0-_-u-mķ ĺķ -buķ "ĺķ ş $-l†u-ķ ĺ ŐƑƏƐƏőĺ oˆ;Ѵ 1_b-|bm -m7 1_b|ov-m m-mo=b0;uv bm 0bol;7b1-Ѵ -rrѴb1-|bomvĺ Biotechnology

Advancesķ28ŐƐőķƐƓƑŋƐƔƏĺ

om]ķĺķ_;mķ*ĺĺķb†ķĺ"ĺķb†ķĺĺķ;m]ķ*ĺĺķş+†ķĺĺŐƑƏƏѶőĺ m|b0-1|;ub-Ѵ l;1_-mbvl o= 1_b|ov-m lb1uovr_;u;v bm - voѴb7 7bvr;uv-bm]v‹v|;l-]-bmv|ĺ1oѴbĺColloids and Surfaces B: Biointerfacesķ65ŐƑőķ 197–202.

om]ķĺķ_;mķ*ĺĺķ*bm]ķĺķş-uhķĺĺŐƑƏƐƏőĺm|blb1uo0b-Ѵruor;u|b;v o=1_b|ov-m-m7lo7;o=-1|bomĹv|-|;o=|_;-u|u;ˆb;‰ĺInternational

Journal of Food Microbiologyķ144ŐƐőķƔƐŋѵƒĺ

ub;];Ѵķĺķb|ķĺķ1Ѵ;l;m|vķ ĺĺķş);bvvķĺŐƑƏƏƖőĺѴ;1|uovrbmmbm] o=1_b|ov-mŋroѴ‹Ő;|_‹Ѵ;m;oŠb7;ő0Ѵ;m7m-mo=b0;uvbm|_;ru;v;m1;o= lb1;ѴѴ-uv†u=-1|-m|voѴ†|bomvĺPolymerķ50ŐƐőķƐѶƖŋƑƏƏĺ -;llѴbķ&ĺĺŐƐƖƕƏőĺѴ;-ˆ-];o=v|u†1|†u-Ѵruo|;bmv7†ubm]|_;-vv;l0Ѵ‹ o=|_;_;-7o=0-1|;ubor_-];$ƓĺNatureķ227ķѵѶƏŋѵѶƔĺ ;;ķ"ĺĺķ;oķ ĺĺķoomķĺŊĺķoķ)ĺŊĺķ;;ķĺĺķ-;ķĺ"ĺķĻblķ ĺŊĺŐƑƏƐƓőĺѴ;1|uovr†m1_b|ov-mm-mo=b0;uv‰b|_1om|uoѴѴ;7Ѵ;ˆ;Ѵvo= vbѴˆ;um-mor-u|b1Ѵ;vĺu;r-u-|bomķ1_-u-1|;ubŒ-|bom-m7-m|b0-1|;ub-Ѵ-1-|bˆb|‹ĺCarbohydrate Polymersķ111ķƔƒƏŋƔƒƕĺ

-u|झm;ŒŊ-l-1_oķ ĺ ĺķ ou|;ŒŊ!o1_-ķ ĺ ĺķ -v|bѴѴoŊu|;]-ķ ĺ ĺķ †u]ovŊ;um࢙m7;Œķĺķ Œt†;uu-Ŋu-†;uķĺ ĺķ ş Ѵ-v1;m1b-Ŋ-|ol;-ķ ĺ ŐƑƏƐƐőĺ m|blb1uo0b-Ѵ -1|bˆb|‹ o= 1_b|ov-m m-mo=b0;uv o0|-bm;7 0‹ ;Ѵ;1|uovrbmmbm]ĺPolymer Internationalķ60ŐƐƑőķƐѵѵƒŋƐѵѵƖĺ

bѴѴ;uķĺĺŐƐƖƖƑőĺA short course in bacterial genetics: A laboratory manual

and handbook for Escherichia coli and related bacteriaĺ Ѵ-bmˆb;‰ķ ĺ+ĺĹ

oѴ7"rubm]-u0ou-0ou-|ou‹u;vvĺ

o-‹;ubķ ĺķ ş ffbķ ĺ ŐƑƏƐƔőĺ -0ub1-|bom o= roѴ‹-mbѴbm;ņroѴ‹ Ő;|_‹Ѵ;m; oŠb7;őņmomŊ1oˆ-Ѵ;m|Ѵ‹=†m1|bom-ѴbŒ;7]u-r_;m;m-mo=b0;uvˆb-;Ѵ;1|uo-vrbmmbm]ĺSynthetic Metalsķ200ķƕŋƐƔĺ

†ŒŒ-u;ѴѴbķ!ĺķ-Ѵ7-vv-uu;ķ(ĺķom|bķ ĺķ ;uu-u-ķĺķb-]bmbķĺķ-ŒŒ-m;ѴѴbķĺķ ş(-vbķ(ĺŐƐƖѶѶőĺboѴo]b1-Ѵ-1|bˆb|‹o=1_b|ov-mĹ&Ѵ|u-v|u†1|†u-Ѵv|†7‹ĺ

Biomaterialsķ9ŐƒőķƑƓƕŋƑƔƑĺ

Ѵ-bl-|ķ ĺ ĺķ ş oѴѴ;‹ķ !ĺ ĺ ŐƑƏƐƔőĺ om|uoѴ o= "-Ѵlom;ѴѴ- om =u;v_ 1_b1h;m0u;-v|v0‹κŊ1-uu-];;m-mņ1_b|ov-mŊ0-v;71o-|bm]v1om|-bmbm] -ѴѴ‹Ѵbvo|_bo1‹-m-|;ou7;o7oubŒ;7ub;m|-Ѵl†v|-u7;Š|u-1|rѴ†v $ĺ Food Microbiologyķ48ķѶƒŋѶѶĺ -hu-ˆ-mķĺķ;†Œ;‹ķĺŊĺķşffbķĺŐƑƏƐƐőĺ=†m7-l;m|-Ѵv|†7‹o=1_b-|ov-mņ;Ѵ;1|uovrbmmbm]ĺPolymerķ52ŐƑƐőķƓѶƐƒŋƓѶƑƓĺ -rbm;-†ķĺĺķooˆ;uķ ĺĺķmouuķ ĺķş -uh-vķ ĺ ĺŐƐƖƖƐőĺm|blb1uo0b-Ѵ ;==;1|o=‰-|;uŊvoѴ†0Ѵ;1_b|ov-mv‰b|__b]__‹7uov|-|b1ru;vv†u;ĺFood Biotechnologyķ5ŐƐőķƓƔŋƔƕĺ !--=-|ķ ĺķ(om-u];mķĺķ--vķĺķş"-_ѴķĺŊĺŐƑƏƏѶőĺmvb]_|vbm|o|_; lo7;o=-1|bomo=1_b|ov-m-v-m-m|b0-1|;ub-Ѵ1olro†m7ĺApplied and Environment Microbiologyķ74ŐƐƑőķƒƕѵƓŋƒƕƕƒĺ

!b;];uķ ĺĺķ bu1_ķ ĺ ĺķ ş "1_b==l-mķĺ ĺ ŐƑƏƐѵőĺ Ѵ;1|uovrbmmbm] 1_b-|ov-mņroѴ‹ Ő;|_‹Ѵ;m; oŠb7;ő voѴ†|bomv ‰b|_ ;vv;m|b-Ѵ obѴvĹ ouu;Ѵ-|bm] voѴ†|bomu_;oѴo]‹|om-mo=b0;u=oul-|bomĺCarbohydrate Polymersķ139ķ 131–138.

"_-_0-Œbķ +ĺķ ş "_-ˆbvbķ ĺ ŐƑƏƐѵőĺ m|;u-1|bomv o= ,bŒbr_ou- 1Ѵbmoro7b-ob7;v -m7 ;m|_- vrb1-|- ;vv;m|b-Ѵ obѴv ‰b|_ 1_b|ov-m -m7 1bruo=ѴoŠ--1bm -]-bmv| 1ollom =oo7Ŋu;Ѵ-|;7 r-|_o];mvĺ LWT- Food Science and

Technologyķ71ķƒѵƓŋƒѵƖĺ

"_-_b7bķ ĺķu-1_1_bķĺĺ(ĺķş;omķ+ĺŊĺŐƐƖƖƖőĺ oo7-rrѴb1-|bomvo=1_b|bm -m71_b|ov-mvĺTrends in Food Science & Technologyķ10ŐƑőķƒƕŋƔƐĺ

"†7-uv_-mķĺķooˆ;uķ ĺķşmouuķ ĺŐƐƖƖƑőĺm|b0-1|;ub-Ѵ-1|bomo=1_b-|ov-mĺFood Biotechnologyķ6ŐƒőķƑƔƕŋƑƕƑĺ

$-oķ +ĺķ b-mķ ĺŊĺķ ş *b;ķ ĺ ŐƑƏƐƐőĺ ==;1| o= 1_b|ov-m om l;l0u-m; r;ul;-0bѴb|‹ -m7 1;ѴѴ lour_oѴo]‹ o= v;†7olom-v -;u†]bmov- -m7 "|-r_‹Ѵo11o1†v-†u;†vĺCarbohydrate Polymersķ86ŐƑőķƖѵƖŋƖƕƓĺ *bm]ķĺķ_;mķ*ĺĺķom]ķĺķb†ķĺ"ĺķ_-ķ ĺ"ĺķş-uhķĺĺŐƑƏƏƖ-őĺ

874 

|

     ARKOUN et Al.

dispersion system on viability, membrane permeability and cell mor-phology of Escherichia coli and Staphylococcus aureus. Carbohydrate Polymers, 76(1), 17–22.

Xing, K., Chen, X. G., Liu, C. S., Cha, D. S., & Park, H. J. (2009b). Oleoyl- chitosan nanoparticles inhibits Escherichia coli and Staphylococcus aureus by damaging the cell membrane and putative binding to extracellular or intracellular targets. International Journal of Food Microbiology, 132(2), 127–133.

Young, D. H., Köhle, H., & Kauss, H. (1982). Effect of chitosan on membrane permeability of suspension- cultured Glycine max and Phaseolus vul-garis cells. Plant Physiology, 70(5), 1449–1454.

Ziani, K., Henrist, C., Jérôme, C., Aqil, A., Maté, J. I., & Cloots, R. (2011). Effect of nonionic surfactant and acidity on chitosan nanofibers with different molecular weights. Carbohydrate Polymers, 83(2), 470–476.

How to cite this article: Arkoun M, Daigle F, Heuzey M-C,

Ajji A. Antibacterial electrospun chitosan- based nanofibers: A bacterial membrane perforator. Food Sci Nutr. 2017; 5:865–874. https://doi.org/10.1002/fsn3.468