7 6 3 IW A P u b lis h in g 2 0 1 4 W a t e r S c ie n c e & T e c h n o lo g y | 7 0 .

5 | 2 0 1 4

T h e u s e o f a n t ib a c t e r ia l a c t iv it y o f Z n O n a n o p a r t ic le s in
t h e t r e a t m e n t o f m u n ic ip a l w a s t e w a t e r
F a t e m e h E lm i, H e s h m a t o lla h A lin e z h a d , Z a h r a M o u la n a , F a t e m e h S a le h ia n ,
S a h a r M o h s e n i T a v a k k o li, F a r ib a A s g h a r p o u r , H o r r ie h F a lla h a n d M a r y a m
M it r a E lm i

A B S T R A C T
F a t e m e h E lm i
N a n o t e c h n o lo g y h o ld s g r e a t p o t e n t ia l in a d v a n c e d w a t e r a n d w a s t e w a t e r t r e a t m e n t t o im p r o v e
H e s h m a t o lla h A lin e z h a d
t r e a t m e n t e f c i e n c y . Z i n c o x i d e n a n o p a r t i c l e s ( Z n O N P s ) h a v e r e c e i v e d c o n s i d e r a b l e a t t e n t i o n d u e t o F a t e m e h S a le h ia n
S a h a r M o h s e n i T a v a k k o li
t h e ir u n iq u e a n t ib a c t e r ia l a c t iv it ie s t o w a r d v a r io u s m ic r o o r g a n is m s t h a t a r e c o m m o n ly f o u n d in t h e D e p a r t m e n t o f C h e m is t r y ,
U n iv e r s it y o f M a z a n d a r a n ,
e n v ir o n m e n t . In t h e p r e s e n t s t u d y , Z n O N P s w e r e s y n t h e s iz e d t h r o u g h b o t h m e c h a n o - c h e m ic a l a n d P O B o x : 4 7 4 1 6 9 5 4 4 7 , B a b o l s a r ,
Ir a n
s o l g e l m e t h o d s . T h e s y n t h e s i z e d Z n O N P s w e r e c h a r a c t e r i z e d t h r o u g h X - r a y d i f f r a c t i o n a n d
Z a h r a M o u la n a
t r a n s m is s io n e le c t r o n m ic r o s c o p y t e c h n iq u e s . T h e n , t h e ir a n t ib a c t e r ia l a c t iv it ie s a g a in s t s e p a r a t e d
In f e c t io u s D is e a s e s a n d T r o p ic a l M e d ic in e
w a s t e w a t e r b a c t e r ia w e r e e v a lu a t e d b y d e t e r m in in g t h e z o n e in h ib it o r , t h e m in im u m in h ib it o r y R e s e a rc h C e n te r,
B a b o l U n iv e r s it y o f M e d ic a l S c ie n c e s ,
c o n c e n t r a t io n , a n d t h e m in im u m b a c t e r ic id a l c o n c e n t r a t io n . T h e r e s u lt s w e r e c o m p a r e d w it h t h o s e G a n g A f r o o z S t r e e t , B a b o l,
Ir a n
o b t a in e d f r o m w a s t e w a t e r a ft e r c h lo r in e d is in f e c t io n a n d u lt r a v io le t (U V ) d is in f e c t io n . T h e s e s t u d ie s
F a r ib a A s g h a r p o u r
d e m o n s t r a t e d t h a t t h e a n t ib a c t e r ia l a c t iv it y o f Z n O N P s d e p e n d s o n t h e t y p e a n d t h e s t r a in o f H o r r ie h F a lla h
P a r a m e d ic a l F a c u lty ,
b a c t e r ia . T h e y h a v e a ls o d e m o n s t r a t e d t h a t t h e a c t iv it y in c r e a s e s a s t h e c o n c e n t r a t io n o f Z n O N P s
B a b o l U n iv e r s it y o f M e d ic a l S c ie n c e s ,
in c r e a s e s . O v e r a ll, t h e e x p e r im e n t a l r e s u lt s s u g g e s t t h a t Z n O N P s c a n p o t e n t ia lly b e a n a n t ib a c t e r ia l G a n g A f r o o z S t r e e t , B a b o l,
Ir a n
r e a g e n t t o t r e a t w a s t e w a t e r . T h e y c a n p a r t ic u la r ly b e a p p lie d a s a c o m p le m e n t a r y m e t h o d w it h U V
M a r y a m M it r a E lm i (c o r r e s p o n d in g a u t h o r )
d is in f e c t io n . T h u s , t h e y c a n b e d e v e lo p e d a s a n t ib a c t e r ia l a g e n t s t o im p r o v e w a s t e w a t e r q u a lit y . C e llu la r a n d M o le c u la r B io lo g y R e s e a r c h C e n t e r ,

K e y w o r d s | a n t ib a c t e r ia l, c h lo r in e , d is in f e c t io n , U V , w a s t e w a t e r , Z n O n a n o p a r t ic le B a b o l U n iv e r s it y o f M e d ic a l S c ie n c e s ,
G a n g A f r o o z S t r e e t , B a b o l,
Ir a n
E - m a il: m .e lm i@ m u b a b o l.a c .ir

IN T R O D U C T IO N

T h e g r o w th o f p o p u la tio n a n d h u m a n c iv iliz a tio n fo r c e s th e o n t h e t o t a l s u s p e n d e d s o l i d s i n e f u e n t , w h i c h w i l l r e d u c e

w o r l d s s c i e n t i s t s t o l o o k f o r e f c i e n t w a y s t o t r e a t w a t e r a n d
w a s te w a te r . S o m e p h y s ic a l, c h e m ic a l, a n d b io lo g ic a l
m e th o d s c a n b e a p p lie d fo r th e d is in fe c tio n o f w a te r a n d
w a s te w a te r . S in c e th e la te 1 9 4 0 s , c h lo r in a tio n h a s b e e n c o n -
s id e r e d to h a v e a m a jo r r o le in in a c tiv a tin g w a te r b o r n e
p a th o g e n s in m a n y c o u n tr ie s . H o w e v e r , m a n y h a lo g e n a te d
d is in fe c t io n b y -p r o d u c t s ( D B P s ) h a v e b e e n c h a r a c t e r iz e d
d u r in g c h lo r in a tio n (Z h a n g e t a l. a , b ), w h ic h
h a v e b e e n s h o w n to b e m u ta g e n ic , a n d c a n in c r e a s e th e
r is k o f c a n c e r (M o n a r c a e t a l. ) . A ls o , c h lo r in e r e s id u a ls
a r e to x ic to m a r in e life (W a r d & D e G r a e v e ). A n e ffe c -
t iv e a lte r n a tiv e to c h lo r in a tio n is th e d is in fe c tio n o f w a te r
a n d w a s te w a te r th r o u g h u ltr a v io le t ( U V ) ir r a d ia tio n to in a c -
t iv a te a v a r ie ty o f m ic r o o r g a n is m s (C h a n g e t a l. ; D a s
; B e r g m a n n e t a l. ; H ijn e n e t a l. ). H o w e v e r ,
t h e g e r m i c i d a l e f c i e n c y o f U V i r r a d i a t i o n s t r o n g l y d e p e n d s
g e r m i c i d a l e f c i e n c y o r U V e f f e c t ( C h a n g e t a l . ) ,
a lth o u g h lo w -p r e s s u r e U V p r o d u c e s u n d e s ir a b le D B P s
( B e r g m a n n e t a l. ; H ijn e n e t a l. ) . D e s p it e t h e f a c t
th a t m u n ic ip a l w a s te w a te r c o n ta in s a v a r ie ty o f p a th o g e n ic
o r g a n is m s o f h u m a n o r ig in , it is u s e d fo r th e ir r ig a tio n o f
p la n ts . T h e r e fo r e , it is h ig h ly n e c e s s a r y to c o m e u p w ith
a n a d v a n c e d m e th o d o f d is in fe c tio n to r e u s e th e m u n ic ip a l
w a s te w a te r in a g r ic u ltu r e ( M u je r ie g o & A s a n o ).
T o o v e r c o m e th e lim ita t io n s o f tr a d itio n a l d is in fe c t io n
m e th o d s , re s e a rc h h a s fo c u s e d o n d e v e lo p in g a lte r n a tiv e
o n e s (Q u e t a l. ). M a n y n a n o m a te r ia ls , s u c h a s n a n o -
A g , n a n o -Z n O , n a n o -T iO 2 , a n d c a r b o n n a n o tu b e s h a v e
th e p o te n tia l to b e u s e d in th e d is in fe c tio n o f w a te r a n d
w a s te w a te r (L i e t a l. ; V a h a b i e t a l. ; B a r u a h
e t a l. ). T h e y d is p la y a n tim ic ro b ia l p ro p e r tie s w ith o u t
s t r o n g o x id a tio n . T h e r e fo r e , th e y h a v e a lo w e r te n d e n c y

d o i: 1 0 .2 1 6 6 / w s t .2 0 1 4 .2 3 2
 7 6 4 F . E lm i e t a l. | A n t ib a c t e r ia l a c t iv it y o f Z n O n a n o p a r t ic le s in w a t e r t r e a t m e n t W a t e r S c ie n c e & T e c h n o lo g y | 7 0 .5 | 2 0 1 4

to fo r m D B P s (Q u e t a l. ). A m o n g th e m , Z n O n a n o p a r - a n d th e tra n s m is s io n e le c tr o n m ic r o s c o p y (T E M ) m ic ro -
tic le s (N P s ) a re o f p a r tic u la r in te re s t d u e to th e ir e x te n s iv e g ra p h o f th e Z n O N P s a re s h o w n in th e stu d y b y
a p p lic a tio n a s a n a n tim ic r o b ia l a g e n t o n a w id e r a n g e o f A lin e z h a d e t a l. ( ). T h e a v e r a g e c r y s ta llite s iz e o f th e
m ic ro o r g a n is m s . P re v io u s s tu d ie s re v e a le d th a t Z n O N P s Z n O N P s w a s c a lc u la t e d b y X R D lin e b r o a d e n in g te c h n iq u e
h a v e b a c te r ic id a l a c tiv itie s o n b o th G r a m -p o s itiv e a n d u s in g th e S c h e r re r e q u a tio n (C u llity ).
G r a m -n e g a tiv e b a c te r ia (A d a m s e t a l. b ; Jo n e s e t a l.
; L i e t a l. ; L iu e t a l. ; Z h a n g e t a l. ). It S o l g e l m e t h o d
h a s b e e n s u g g e s te d th a t th e a n tib a c te r ia l m e c h a n is m o f
Z n O N P s is d u e to th e e le c tr o s ta tic in te r a c tio n b e tw e e n Z i n c a c e t a t e ( 0 . 1 M ) i n e t h a n o l i c s o l u t i o n w a s r e u x e d f o r
th e n a n o p a r tic le s a n d th e b a c te r ia l m e m b r a n e (Z h a n g 1 8 0 m in a t 8 0 C . T h e n , 0 .2 L o f t h e o b t a in e d r e a c t io n p r o -
W

e t a l. ). T h e in te ra c tio n b e tw e e n H 2 O 2 a n d m e m b r a n e d u c t ( p r e c u r s o r ) w a s d ilu te d to 0 .5 L w ith a b s o lu te

p r o te in s is a ls o r e g a r d e d a s a n o th e r p r o p o s e d m e c h a n is m
fo r th e b a c te r ic id a l p ro p e r tie s o f Z n O (Z h a n g e t a l. , c u r s o r t o g i v e a n a l l i t h i u m c o n c e n t r a t i o n o f 0 . 1 4 M . T h e
). T h e r e la tio n s h ip b e tw e e n th e s iz e o f Z n O N P s a n d
th e a n t ib a c t e r ia l a c tiv it y is a m b ig u o u s . S o m e r e p o r t s in d i-
c a te d th a t d is in fe c tio n b e h a v io r in c re a s e s th ro u g h
re d u c in g th e s iz e o f Z n O N P s ( Jo n e s e t a l. ; N a ir
e t a l. ). F u r th e r m o re , it w a s fo u n d th a t a h ig h e r c o n -
c e n tr a tio n o f s m a lle r p a r tic le s w ith a h ig h e r s u r fa c e a r e a
g iv e s b e tte r a n tib a c te r ia l a c tiv ity (M a k h lu f e t a l. ; tr o n m ic r o s c o p y ( S E M ) m ic r o g r a p h o f th e Z n O N P s w a s
Z h a n g e t a l. ). H o w e v e r , F ra n k lin e t a l. d id n o t o b s e r v e
a n y r e la tio n s h ip b e tw e e n th e s iz e o f Z n O N P s a n d th e a n ti-
b a c te r ia l a c tiv ity (F r a n k lin e t a l. ).
I n th is s tu d y , th e s y n th e s iz e d Z n O N P s w e r e p r o d u c e d
t h r o u g h m e c h a n o - c h e m i c a l a n d s o l g e l m e t h o d s . T h e n ,
th e y w e r e c h a r a c te r iz e d a n d te s te d fo r th e ir a n tib a c te r ia l
a c tiv ity a g a in s t K le b s ie lla p n e u m o n ia e , E s c h e ric h ia c o li,
P ro te u s , a n d S ta p h y lo c o c c u s e p id e rm id is b a c te r ia in m u n ic i-
p a l w a s te w a te r c o lle c te d fro m tw o a re a s in M a z a n d a ra n ,
I r a n . T h e b a c te r ic id a l a c tiv ity o f Z n O N P s w a s a ls o c o m -
p a r e d w ith t h e c o n v e n t io n a l d is in fe c t io n m e t h o d s , i.e .,
c h lo r in e a n d U V d is in fe c tio n , w h ic h a r e c o m m o n ly u s e d
fo r w a s te w a te r tr e a tm e n t in th e s e tw o a r e a s . I n a d d itio n ,
th e r e la tio n s h ip b e tw e e n th e c o n c e n tr a tio n o f Z n O N P s
a n d th e a n tib a c te r ia l a c tiv ity w a s in v e s tig a te d in th is s tu d y .
M A T E R IA L S A N D M E T H O D S
S y n t h e s is o f Z n O N P s
e th a n o l. L ith iu m h y d r o x id e p o w d e r w a s a d d e d to th e p r e -
m ix tu r e w a s th e n h y d r o ly z e d in a n u ltr a s o n ic b a th . T h e
h y d r o ly s is r e a c tio n w a s c o n tin u e d a t r o o m te m p e r a tu r e fo r
a b o u t 1 h . T h e Z n O c o l l o i d a l s u s p e n s i o n w a s t h e n l t e r e d
t h r o u g h a 0 . 1 - p m g l a s s b e r l t e r t o r e m o v e d u s t a n d a n y
u n d is s o lv e d lith iu m h y d r o x id e . T h e a v e r a g e s iz e o f th e
Z n O N P s w a s e s tim a te d b y X R D p a tte r n . T h e s c a n n in g e le c -
r e c o r d e d .
M u n ic ip a l w a s t e w a t e r c o lle c t io n
T h e w a s te w a te r s a m p le s w e r e c o lle c te d fr o m tw o d iffe r e n t
w a s te w a te r c e n te r s in M a z a n d a r a n (G h a e m s h a h r a n d
B a b o ls a r ), I r a n . T h e c h lo r in e d is in fe c tio n is a p p lie d in
G h a e m s h a h r , s ta tio n I , a n d th e U V d is in fe c tio n is u s e d in
B a b o ls a r , s ta tio n I I . I n b o th s ta tio n s , la r g e p a r tic le s a r e
r e m o v e d t h r o u g h s e t t l i n g , o c c u l a t i o n , a n d s e d i m e n t a t i o n .
I n s ta tio n I , c h lo r in e is a d d e d to m u n ic ip a l w a s te w a te r a t
1 k g d 1 . T h e r e m a i n i n g c h l o r i n e i n t h e e f u e n t i s a b o u t
0 . 4 0 . 6 p p m . I n s t a t i o n I I , 3 2 l o w - p r e s s u r e l a m p s e m i t e s s e n -
tia lly m o n o c h r o m a tic lig h t a t a w a v e le n g th o f 2 5 4 n m a n d
1 2 W s c m 2 d o s e s o f U V . T h e e f u e n t i s r e u s e d i n t r e a t -
m e n t p la n ts . T h e w a s te w a te r s a m p le s w e r e ta k e n in 1 L
g la s s b o ttle s ju s t b e fo r e a n d a fte r b e in g d is in fe c te d b y c h lo r -
in e o r U V , a n d w e r e im m e d ia te ly tr a n s fe r r e d to th e
la b o r a to r y . M a in c h e m ic a l c h a r a c te r is tic s o f th e w a s te w a te r
fro m b o th s ta tio n s a r e s h o w n in T a b le 1 .

M e c h a n o -c h e m ic a l m e th o d B a c t e r ia l s e p a r a t io n

Z in c a c e ta te w a s d r ie d a t 1 2 0 C in a ir fo r 2 4 h p r io r to its T h e w a s te w a te r s a m p le s (b e fo re a n d a ft e r b e in g d is in fe c te d
W

u s e . Z in c a c e ta te a n d o x a lic a c id w e r e m ix e d in a n a g a te b y c h lo r in e o r U V ) w e re c u ltu r e d o n n u tr ie n t a g a r
m o rta r w ith m o la r r a tio o f 2 :3 a n d w e r e p o w d e r e d fo r m e d iu m . A ft e r b e in g in c u b a t e d a t 3 7 C f o r 2 4 4 8 h , o n e
W

4 5 m in a t r o o m te m p e ra tu re . T h e w h ite p r e c u rs o r w a s c a l- lo o p fro m e a c h c o lo n y w a s tra n s fe r re d e v e n ly o n E M B

c in e d a t 4 5 0 C in a ir in a p o r c e la in c r u c ib l e fo r 3 0 m in to (e o s i n m e t h y le n e b lu e ) a n d b lo o d a g a r m e d iu m th r o u g h a
W

p re p a re th e Z n O N P s . T h e X -r a y d if fr a c t io n (X R D ) p a tte r n s te r ile g la s s ro d . A fte r b e in g in c u b a te d a t 3 7 C fo r 2 4 h ,

W
 7 6 5 F . E lm i e t a l. | A n t ib a c t e r ia l a c t iv it y o f Z n O n a n o p a r t ic le s in w a t e r t r e a t m e n t W a t e r S c ie n c e & T e c h n o lo g y | 7 0 .5 | 2 0 1 4

T a b le 1 | M a in c h e m ic a l c h a r a c t e r is t ic s o f t h e w a s t e w a t e r M in im u m b a c te r ic id a l c o n c e n tr a tio n

S t a t io n I S t a t io n II
T h e m in im u m b a c te r ic id a l c o n c e n tr a tio n ( M B C ) is th e l o w e st
M a in c h e m ic a l c h a r a c t e r is t ic s In p u t O u t p u t In p u t O u t p u t c o n c e n tr a tio n a t w h ic h th e b a c te r ia d o n o t g r o w w h e n th e y a re
o f t h e w a s t e w a t e r ( p p m ) ( p p m ) ( p p m ) ( p p m )
tr a n s fe r r e d to fr e s h a g a r p la te a fte r th e in c u b a tio n p e r io d
C h e m ic a l o x y g e n d e m a n d 3 2 6 3 2 1 5 0 2 0 ( 9 9 .9 % k ille d ) . T e n m ic r o litr e s o f t h e c o n te n t w ith o u t t u r b id ity
B io c h e m ic a l o x y g e n 2 3 3 1 4 7 5 1 0 w e l l s w a s c u l t i v a t e d o n t h e M e l l e r H i n t o n p l a t e . T h e n u m b e r
d e m a n d o f c o l o n i e s w a s c o u n t e d a f t e r 2 4 h a t 3 7 C . T h e r s t
W
w e ll,
T o ta l d is s o lv e d s o lid s 1 ,0 0 0 w h ic h h a d th r e e o r fe w e r c o lo n ie s , w a s r e g a r d e d a s M B C .
T o ta l s u s p e n d e d s o lid s 1 3 3 0 4 0

( ) T h e d a t a a r e n o t a v a i l a b l e .
R E S U L T S

G r a m -n e g a t iv e b a c t e r ia w e r e c h a r a c t e r iz e d b y a n a p i-2 0 A
k it a n d G r a m -p o s itiv e b a c te r ia w e r e r e c o g n iz e d b y a b io -
c h e m ic a l te s t.
A n t ib a c t e r ia l a c t iv it y o f Z n O N P s
D is k d iffu s io n a s s a y
Z n O N P s ( p r e p a r e d fr o m tw o m e th o d s ) w e r e s te r iliz e d
th r o u g h U V lig h t. T h e n , th e y w e r e s u s p e n d e d in s te r ile
n o r m a l s a lin e a n d s t ir r e d u n t il a u n ifo r m c o llo id a l s u s p e n s io n
w a s f o r m e d , a f t e r w h i c h 1 . 5 1 0 8 C F U m L 1 o f e a c h b a c t e r -
iu m (K . p n e u m o n ia e , E . c o li, P ro te u s , a n d S . e p id e rm id is
s e p a r a te d fr o m e a c h s ta tio n ) w e r e s p r a y e d o n M e lle r
H in to n m e d iu m . A fte r le ttin g th e b a c te r ia d r y ( w ith in
5 1 0 m i n ) , 2 0 L o f n a n o - Z n O s u s p e n s i o n s , w h o s e c o n c e n -
t r a t io n s v a r ie d fr o m 0 .0 1 to 5 0 m M , w e r e d r o p p e d in t o a
d is k o f 5 m m in d ia m e te r . T h e p u r e s o lv e n t w a s a ls o u s e d
fo r c o n tr o l. T h e z o n e o f in h ib itio n w a s m e a s u r e d a fte r th e
in c u b a tio n p ro c e s s a t 3 7 C fo r 2 4 h . T h is e x p e r im e n t w a s
W
C h a r a c t e r iz a t io n o f Z n O N P s
S e v e r a l m e th o d s a r e u s e d to s y n th e s iz e Z n O N P s . S o m e o f
t h e s e m e t h o d s a r e s o l g e l ( R i s t i c e t a l . ) , t w o s t e p
m e c h a n o -c h e m ic a l th e r m a l s y n t h e s is ( R a je s h e t a l. ) , p r e -
c ip ita tio n m e th o d ( K u m a r e t a l. ), a n d m ic r o w a v e ( K h o z a
e t a l. ). H o w e v e r , in m o s t o f th e s e te c h n iq u e s , th e h ig h
s u r fa c e e n e r g y o f n a n o p a r tic le s c a u s e s th e m to a g g lo m e r a te .
T h e s iz e o f Z n O N P s d e p e n d s o n th e ty p e o f p r e c u r s o r , th e
s o lv e n t, th e p H , a n d th e te m p e ra tu re o f th e re a c tin g s o lu tio n
( P a d m a v a th y & V ija y a r a g h a v a n ) . I n th is s tu d y , Z n O
N P s w e r e p r e p a r e d b y t w o m e t h o d s ( m e c h a n o -c h e m ic a l
a n d s o l g e l m e t h o d s ) . A s w a s r e p o r t e d e a r l i e r , t h e Z n O
N P s , s y n t h e s iz e d b y m e c h a n o -c h e m ic a l m e t h o d s ( Z n O -1 ) ,
w e r e c h a r a c te r iz e d b y X R D a n d T E M . T h e d iffr a c tio n a n g le
a n d th e in te n s ity o f th e c h a r a c te r is tic p e a k s o f th e ty p ic a l
Z n O N P s c o r r e s p o n d to JC P D S c a r d N o . 3 6 -1 4 5 1 . A ll p e a k
lo c a tio n s a n d r e la tiv e p e a k in te n s itie s o f th e Z n O N P s
a g r e e w ith th o s e o f th e s ta n d a r d X R D p a tte r n . I n a d d itio n ,

r e p e a te d th r e e tim e s a n d th e a v e ra g e n u m b e r w a s c a lc u la te d .
M in im u m in h ib ito r y c o n c e n tr a tio n
T h e m in im u m in h ib ito r y c o n c e n tr a tio n (M I C ) is th e lo w e s t
c o n c e n tr a tio n w h ic h c a n c a u s e c o m p le te g r o w th in h ib itio n .
T h e b r o th m ic r o d ilu tio n m e th o d w a s a p p lie d to e v a lu a te th e
M I C (W ie g a n d e t a l. ) . O n e h u n d r e d m ic r o litr e s o f
M e l l e r H i n t o n b r o t h , c o n t a i n i n g 1 0 5 C F U m L 1 o f e a c h
b a c te r iu m , w a s a d d e d to e a c h w e ll. O n e h u n d r e d m ic r o litr e s
o f Z n O s u s p e n s io n s ( c o n c e n t r a tio n s v a r ie d f r o m 0 .0 1 to
5 0 m M ) w a s a ls o p o u r e d in to 9 6 s te r iliz e d w e ll m ic r o p la te s
a n d t h e n i n c u b a t e d a t 3 7 C f o r 2 4 h . T h e r s t w e l l , w h i c h
W
th e r e a r e n o p e a k s o f im p u r itie s , in d ic a tin g th a t th e y a r e o f
h ig h p u r ity . T h e a v e r a g e c r y s ta llin e s iz e o f th e Z n O N P s p r o -
d u c t , c a l c u l a t e d t h r o u g h u s i n g S c h e r r e r s e q u a t i o n ( C u l l i t y
) , w a s fo u n d to b e 1 5 .8 n m . T E M im a g e s w e r e a ls o u s e d
to d e te r m in e th e s iz e s a n d s tr u c tu r a l m o r p h o lo g ie s o f th e s y n -
th e s iz e d Z n O N P s . T E M im a g e s s h o w e d th a t Z n O N P s a re
m a i n l y s p h e r i c a l i n s h a p e a n d 1 0 2 0 n m i n s i z e , w h i c h
m a k e s th is r e s u lt c o n s is t e n t w ith X R D d a ta .
T h e Z n O N P s ( Z n O - 2 ) f r o m t h e s o l g e l m e t h o d w e r e e s t i -
m a te d b y X R D d a ta (F ig u r e 1 ). A c c o r d in g to th e r e s u lts , th e
X R D p a tte rn o f th e ty p ic a l Z n O N P s a lso m a tc h e d JC P D S
c a r d N o . 3 6 -1 4 5 1 . I n a d d it io n , th e r e w e r e n o d iffr a c tio n
p e a k s fr o m o th e r ty p e s d e t e c te d , w h ic h illu s tr a te s h ig h p u r ity .
T h e a v e r a g e s iz e o f th e Z n O N P s , a s e s tim a te d th r o u g h S c h e r -

w a s c o m p le te ly tr a n s p a r e n t a n d h a d n o b a c te r ia l g r o w th , r e r s e q u a t i o n , w a s f o u n d t o b e a b o u t 5 7 n m . T h e S E M
w a s c o n s id e re d fo r M I C . E a c h c o n c e n tra tio n w a s te s te d m ic r o g r a p h o f th e Z n O -2 N P s is s h o w n in F ig u r e 2 . T h e m o r -
th r e e tim e s a n d th e r e s u lts w e r e a v e r a g e d . p h o lo g ic a l o b s e r v a tio n a ls o d e m o n s tr a te d th e n a n o p a r tic le s
 7 6 6 F . E lm i e t a l. | A n t ib a c t e r ia l a c t iv it y o f Z n O n a n o p a r t ic le s in w a t e r t r e a t m e n t W a t e r S c ie n c e & T e c h n o lo g y | 7 0 .5 | 2 0 1 4

F ig u r e 1 | X R D p a t t e r n o f Z n O N P s y n t h e s i z e d v i a t h e s o l g e l m e t h o d .

T a b le 2 | C u lt u r e d b a c t e r ia f r o m m u n ic ip a l w a s t e w a t e r s f r o m t w o s t a t io n s

B a c t e r ia

W a s t e w a t e r B e f o r e d is in f e c t io n A f t e r d is in fe c t io n

S ta tio n I (c h lo r in e E s c h e ric h ia c o li K le b s ie lla

d is in fe c tio n ) p n e u m o n ia e
P ro te u s
K le b s ie lla
p n e u m o n ia e
S ta tio n I I (U V K le b s ie lla S ta p h y lo c o c c u s
d is in fe c tio n ) p n e u m o n ia e e p id e rm id is
E s c h e ric h ia c o li
S ta p h y lo c o c c u s
F ig u r e 2 | S E M m i c r o g r a p h o f Z n O N P s y n t h e s i z e d v i a t h e s o l g e l m e t h o d .
e p id e rm id is

h e x a g o n a l w u rtz ite s tr u c tu re . F u r th e r m o r e , th e im a g e s h o w s
th e a g g lo m e r a tio n o f p a r tic le s , w h ic h w a s m u c h h ig h e r in th is G r a m -n e g a tiv e b a c te r ia ( K . p n e u m o n ia e , E . c o li, a n d P ro -
m e t h o d o f p r e p a r a t io n t h a n in t h e m e c h a n o -c h e m ic a l m e t h o d . te u s ) w e re fo u n d in s ta tio n I s a m p le s , a n d K . p n e u m o n ia e ,
E . c o li a n d G r a m -p o s it iv e b a c te ria (S . e p id e rm id is ) w e re s e p -
S e p a r a t io n o f b a c t e r ia f r o m w a s t e w a t e r a ra te d fro m s ta tio n II s a m p le s . A fte r th e c h lo r in e
d is in fe c tio n p r o c e s s , K . p n e u m o n ia e w a s th e o n ly b a c te r iu m
T h e m u n ic ip a l w a s te w a te r s a m p le s w e r e c o lle c te d fro m tw o th a t r e m a in e d , a n d a fte r U V d is in fe c tio n S . e p id e r m id is w a s
d iffe r e n t w a s te w a te r s ta tio n s b e fo r e a n d a fte r th e d is in fe c - th e o n ly b a c te r iu m d e te c te d .
tio n p r o c e s s . A s m e n tio n e d a b o v e , c h lo r in e is a p p lie d in
s ta tio n I a n d U V is u s e d in s ta tio n I I fo r d is in fe c tio n . T h e A n t ib a c t e r ia l a c t iv it y o f Z n O N P s
s a m p le s w e r e d ir e c tly tr a n s fe r r e d to t h e la b o r a to r y a t 4 C .
W

T h e n , th e s a m p le s w e r e c u ltu r e d e v e n ly o n E M B a n d I n th is s tu d y , th e a n tib a c te r ia l a c tiv ity o f v a r io u s c o n c e n -

b lo o d a g a r m e d iu m . A s T a b le 2 s h o w s , b e fo re d is in fe c tio n , t r a t i o n s ( 0 . 0 1 5 0 m M ) o f Z n O N P s ( Z n O - 1 a n d Z n O - 2 )
 7 6 7 F . E lm i e t a l. | A n t ib a c t e r ia l a c t iv it y o f Z n O n a n o p a r t ic le s in w a t e r t r e a t m e n t W a t e r S c ie n c e & T e c h n o lo g y | 7 0 .5 | 2 0 1 4

to w a r d s th e w a s te w a te r s e p a r a te d b a c te r ia w a s in v e s tig a te d T a b le 4 | Z o n e o f in h ib it io n ( m m ) a g a in s t K . p n e u m o n ia e , E . c o li, a n d S . e p id e r m id is
(s e p a r a t e d f r o m s t a t io n II) b y Z n O - 1 a n d Z n O - 2 N P s
b y s u c h b a c te r io lo g ic a l te s ts a s d is k d iffu s io n a s s a y , M I C ,
a n d M B C . D is k d iffu s io n a s s a y o f Z n O N P s a g a in s t K . p n e u -
m o n ia e , E . c o li, a n d P ro te u s ( s e p a r a te d fr o m s ta tio n I ) is Z o n e o f in h ib it io n (m m )

s h o w n o n th e b a s is o f th e in h ib itio n z o n e (m m ) s iz e in
C o n c e n t r a t io n o f Z n O -1 (m M ) K. p n eu m on iae E. co li S. ep i d er m id is
T a b le s 3 (a ) a n d 3 (b ). A n tib a c te r ia l a c tiv ity o f Z n O -1 N P s p r o -
Z n O -1
d u c e s z o n e s o f in h ib itio n o f 8 , 1 0 , a n d 1 2 m m o n E . c o li a t 1 5 ,
0 .0 1
2 5 , a n d 5 0 m M c o n c e n tr a tio n s a n d 1 0 a n d 1 1 m m o n P ro te u s
a t 2 5 a n d 5 0 m M , r e s p e c t iv e ly . F o r t h e Z n O -2 N P s , it p r o - 1

d u c e s a z o n e o f in h ib itio n o f 1 0 m m o n b o th E . c o li a n d 2 .5 1 0
P ro te u s a t 5 0 m M c o n c e n tr a tio n . I n th e c a s e o f K . p n e u m o - 5 1 2
n ia e , b o th Z n O N P s d o n o t s h o w a n y a c tiv ity . 1 0 1 0 1 2
A s s h o w n in T a b le s 4 (a ) a n d 4 (b ), a n tib a c te r ia l a c tiv ity o f 1 5 1 0 1 5
b o t h Z n O N P s c a n p r o v id e m a x im u m z o n e o f in h ib itio n 2 5 1 2 1 6
a g a in s t E . c o li a t 5 0 m M c o n c e n t r a tio n . A n tib a c t e r ia l a c tiv ity 5 0 1 4 1 8
o f Z n O N P s p r o d u c e s th e b e s t z o n e o f in h ib itio n a g a in s t S . Z o n e o f in h ib it io n (m m )
e p id e rm id is (1 8 m m ) a n d (2 1 m m ) a t 5 0 m M c o n c e n tra tio n
C o n c e n t r a t io n o f Z n O -2 (m M ) K. p n eu m on iae E. co li S. ep i d er m id is
o f Z n O -1 a n d Z n O -2 , r e s p e c tiv e ly . N o in h ib ito r y e ff e c t w a s
f o u n d a g a in s t K . p n e u m o n ia e a t c o n c e n t r a t io n s le s s t h a n Z n O -2 N P s
0 .0 0 1

T a b le 3 | Z o n e o f in h ib it io n (m m ) a g a in s t K . p n e u m o n ia e , E . c o li, a n d P r o t e u s ( s e p a r a t e d
1
fr o m s t a t io n I) b y Z n O - 1 a n d Z n O - 2 N P s 2 .5
5 8
Z o n e o f in h ib it io n (m m )
1 0 1 2
C o n c e n t r a t io n o f Z n O -1 (m M ) K. p n eu m o n i ae E. co li Prot eu s
1 5 1 0 1 5
Z n O -1 2 5 1 2 1 8
0 .0 1 5 0 9 1 3 2 1
1
( ) N o a c t i v i t y i n t h e s e c o n c e n t r a t i o n s .
2 .5
5
5 0 m M , w h e r e a s , a t 5 0 m M c o n c e n tr a tio n o f Z n O -2 N P s ,
1 0
th e z o n e o f in h ib itio n p ro d u c e s 9 m m o n K . p n e u m o n ia e .
1 5 8
A c c o r d in g to th e r e s u lts , Z n O N P s g iv e th e b e s t a c tiv ity
2 5 1 0 1 0
a g a in s t G r a m -p o s it iv e b a c t e r ia , S . e p id e r m id is .
5 0 1 2 1 1
I n a d d itio n , th e M I C a n d M B C o f Z n O N P s fo r a ll b a c -
Z o n e o f in h ib it io n (m m )
te r ia a r e s u m m a r iz e d in T a b le 5 . A s T a b le 5 s h o w s ,
C o n c e n t r a t io n o f Z n O -2 (m M ) K. p n eu m o n i ae E. co li Prot eu s a n tib a c te r ia l a c tiv ity o f Z n O N P s a g a in s t E . c o li w a s h ig h e s t
a m o n g G r a m -n e g a tiv e b a c te r ia . T h e M I C a n d M B C fo r
Z n O -2 N P s 1
b o t h Z n O N P s w a s c a l c u l a t e d t o b e 2 0 3 g m L a n d
0 .0 1 > 1 , 0 1 7 g m L 1 f o r S . e p i d e r m i d i s , r e s p e c t i v e l y .
1
2 .5
5
D IS C U S S IO N
1 0
1 5
T h e la c k o f w a te r a n d th e e n v ir o n m e n ta l q u a lity o f w a te r a re
2 5
re g a rd e d a s tw o im p o r ta n t p ro b le m s fo r th e n e a r fu tu re .
5 0 1 0 1 0
A d v a n c e s i n t h e e l d o f n a n o te c h n o lo g y m a y s h o w n e w w a y s
( ) N o a c t i v i t y i n t h e s e c o n c e n t r a t i o n s . fo r tre a tm e n t o f w a te r a n d w a ste w a te r . T h is re p o rt e v a lu a te d
 7 6 8 F . E lm i e t a l. | A n t ib a c t e r ia l a c t iv it y o f Z n O n a n o p a r t ic le s in w a t e r t r e a t m e n t W a t e r S c ie n c e & T e c h n o lo g y | 7 0 .5 | 2 0 1 4

T a b le 5 | D e t e r m in a t io n o f M IC a n d M B C o f Z n O - 1 a n d Z n O - 2 N P s a g a in s t K . p n e u m o n ia e , E . c o li, S . e p id e r m id is , a n d P r o t e u s b a c t e r ia

S t a t io n I S t a t io n II

B a c t e r ia M I C ( g m L 1
) M B C ( g m L 1
) M I C ( g m L 1
) M B C ( g m L 1
)

Z n O -1
K . p n e u m o n ia e
E . c o li 1 ,0 1 7 > 1 ,0 1 7 2 ,0 3 5 > 2 ,0 3 5
S . e p id e rm id is 2 0 3 >1 , 0 1 7
P ro te u s
S t a t io n I S t a t io n II

B a c t e r ia M I C ( g m L 1
) M B C ( g m L 1
) M I C ( g m L 1
) M B C ( g m L 1
)

Z n O -2 N P s
K . p n e u m o n ia e
E . c o li 1 ,0 1 7 > 1 ,0 1 7 1 ,0 1 7 > 2 ,0 3 5
S . e p id e rm id is 2 0 3 1 ,0 1 7
P ro te u s

T h e b la c k s q u a r e s s h o w t h e b a c t e r iu m w a s n o t p r e s e n t in t h a t s t a t io n .
( ) N o a c t i v i t y i n t h e s e c o n c e n t r a t i o n s .

th e r e la tiv e a n tib a c t e r ia l a c tiv ity o f Z n O N P s w ith a v e r a g e s iz e s t h a t Z n O N P s w e r e m o r e to x ic t o w a r d G r a m -p o s itiv e b a c -

o f 1 5 . 8 n m a n d 5 7 n m , w h i c h w e r e s y n t h e s i z e d b y m e c h a n o - te r ia (S a w a i ; T a m e t a l. ; P r e m a n a th a n e t a l.
c h e m i c a l a n d s o l g e l , r e s p e c t i v e l y , o n b a c t e r i a s e p a r a t e d f r o m ). H o w e v e r, th e y a re c o n tra d ic to ry to th e o n e s th a t
w a s te w a te r . I n o u r s tu d y , th e s iz e o f th e s y n th e s iz e d Z n O u s e d P E G y la te d Z n O N P , w h ic h h a d le s s e ffe c t o n G ra m -
N P s fr o m th e s e tw o m e th o d s w a s c o m p a tib le w it h th o s e o f p o s itiv e b a c te ria (N a ir e t a l. ).
o th e r s (S p a n h e l & A n d e r s o n ; M e u le n k a m p ; A o F u r th e r m o r e , th e r e s u lts s h o w e d th a t th e a n tib a c te r ia l
e t a l. ; S h e n e t a l. ; L u e t a l. ). A ls o , in o u r a c tiv ity in c r e a s e d a s th e c o n c e n tr a tio n o f Z n O N P s
s t u d y , t h e a g g l o m e r a t i o n s o f s o l g e l w e r e m u c h h i g h e r t h a n in c re a s e d . I t s e e m s th a t Z n O N P s c o u ld d a m a g e th e m e m -
th e m e c h a n o -c h e m ic a l m e t h o d . T h e a g g r e g a t io n o b s e r v e d in b r a n e , le a d to th e le a k a g e o f c y to s o lic c o m p o n e n ts a n d
o u r s tu d y w a s a ls o c ite d in o th e r r e p o r ts fo r Z n O a n d , th e re fo re , d e s tr o y th e b a c te r ia l c e lls (W a n g e t a l. ) . T h e r e s u lts o f
is c o n s id e r e d to b e a n in n a te p r o p e r ty o f u n c o a te d N P s (A d a m s o u r s tu d y a ls o d e m o n s tr a te d th a t Z n O N P s e x h ib ite d n o a n ti-
e t a l. a ; K e lle r e t a l. ; T s o e t a l. ). b a c te r ia l a c tiv ity in a c o n c e n t r a tio n lo w e r th a n 1 m M . S a w a i
A c c o r d in g to th e r e s u lts , b o th Z n O N P s p r o v e d to h a v e m a in ta in s th a t th is m a y b e d u e to lo w e r c o n c e n tr a tio n s o f
M B C a n d M I C a c tiv itie s a g a in s t E . c o li a n d S . e p id e rm id is Z n 2 io n s , w h ic h m ig h t a c t a s a n u tr ie n t (S a w a i ).
b u t th e y h a d n o e ffe c t o n K . p n e u m o n ia e a n d P ro te u s b a c - M a n y re s e a rc h e rs h a v e re p o rte d th e e ffe c t o f s iz e o n th e
te r ia . O n ly d is k d iffu s io n a s s a y s h o w e d a m o d e r a te e ffe c t to x ic ity (N a ir e t a l. ; N a p ie r s k a e t a l. ). O u r r e s u lt
o n K . p n e u m o n ia e in h ig h e r c o n c e n tr a tio n . T h is r e s u lt is d o e s n o t s h o w a n o b v io u s r e la tio n s h ip b e tw e e n to x ic ity
c o n s is te n t w ith th e r e s u lts o b ta in e d b y Jo n e s e t a l. ( ), a n d s i z e o f Z n O N P s . S m a l l e r Z n O N P s ( f r o m t h e s o l g e l
P a d m a v a th y & V ija y a r a g h a v a n ( ) , G o r d o n e t a l. ( ), m e th o d ) w e r e a little b it m o r e e ffe c tiv e . I t s e e m s th a t o u r
M e r u v u e t a l. ( ), W a n g e t a l. ( ) a n d Y o u s e f & r e s u lts m a y b e r e la te d to fo r m e d a g g r e g a tio n s a n d , th u s ,
D a n ia l ( ) , b u t it d o e s n o t e m p ir ic a lly s u p p o r t th e r e s u lts th e a g g r e g a te d s iz e c o u ld a ls o b e r e g a r d e d a s a fa c to r a ffe c t-
o f s tu d ie s in w h ic h Z n O N P h a s m o d e r a te M B C a n d M I C in g th e p a r tic le s to x ic ity (H s ia o & H u a n g ).
e ffe c ts (Y o u s e f & D a n ia l ). T h e s e a u th o r s p r o p o s e d T h e a n tib a c te r ia l m e c h a n is m o f Z n O N P s is s till u n d e r
t h a t t h e a n t i b a c t e r i a l a c t i v i t y o f Z n O N P s c a n b e i n u e n c e d in v e s tig a t io n . H o w e v e r , s e v e r a l m e c h a n is m s h a v e b e e n p r o -
b y th e p r e p a r a tio n m e th o d in a d d itio n to th e e ffe c t o f p a r - p o s e d to in te r p r e t th e d is in fe c tio n a c tiv ity o f Z n O N P s
tic le s iz e a n d th e ty p e o f b a c te r ia l s tr a in s . A ls o , th e s e (Z h a n g e t a l. , , ) . C o n s id e r in g th e s m a ll s iz e o f
n d i n g s a r e c o n s i s t e n t w i t h t h e r e s u l t s t h a t d e m o n s t r a t e Z n O N P s , th e a n tib a c te r ia l a c tiv ity m ig h t r e la te to th e a b ility
 7 6 9 F . E lm i e t a l. | A n t ib a c t e r ia l a c t iv it y o f Z n O n a n o p a r t ic le s in w a t e r t r e a t m e n t W a t e r S c ie n c e & T e c h n o lo g y | 7 0 .5 | 2 0 1 4

t o a d h e r e to t h e c e ll w a ll e a s ily , c a u s in g t h e d e s t r u c tio n o f its A lin e z h a d , H ., S a le h ia n , F . & B ip a r v a , P . S y n th e s is o f

b a c te r ia l m e m b r a n e a n d le a d in g to th e d e a th o f th e c e ll b e n z im id a z o le d e r iv a tiv e s u s in g h e te r o g e n e o u s Z n O
n a n o p a r t i c l e s . S y n t h . C o m m u n . 4 2 , 1 0 2 1 0 8 .
(Z h a n g e t a l. ) . I n a d d itio n , th e p r o d u c tio n o f d iffe r e n t
A o , W ., L i, J ., Y a n g , H ., Z e n g , X . & M a , X . M e c h a n o c h e m ic a l
a m o u n ts o f H 2 O 2 a n d r a d ic a l o x y g e n s p e c ie s (R O S ) is a n o th e r
s y n th e s is o f z in c o x id e n a n o c r y s ta llin e . P o w d e r T e c h n o l. 1 6 8 ,
p r o p o s e d m e c h a n i s m o f N P s a c t i v i t y ( S a w a i ; Z h a n g e t a l . 1 4 8 1 5 1 .
, , ; R a g h u p a th i e t a l. ). G ra m -p o s itiv e b a c te r ia A tm a c a , S ., G u e l, K . & C ic e k , R . T h e e f fe c t o f z in c o n
w e r e fo u n d to b e m o r e s u s c e p tib le to Z n O N P s th a n w e r e m i c r o b i a l g r o w t h . T u r k i s h J . M e d . S c i . 2 8 , 5 9 5 5 9 8 .
G r a m -n e g a tiv e b a c te r ia , p r o b a b ly b e c a u s e th e G r a m -p o s itiv e B a r u a h , S . K ., P a l, S . & D u tt a , J . N a n o s t r u c t u r e d z in c o x id e
f o r w a t e r t r e a t m e n t . N a n o s c i . N a n o t e c h n o l . - A s i a 2 , 9 0 1 0 2 .
b a c te r ia c o n ta in a w e a k e r a n tio x id a n t c e llu la r c o n te n t,
B e r g m a n n , H ., I o u r tc h o u k , T ., S c h o p s , K . & B o u z e k , K . N e w
m a k in g th e m le s s r e s is ta n t to R O S . H o w e v e r , it h a s b e e n U V ir r a d ia tio n a n d d ir e c t e le c tr o ly s is p r o m is in g m e th o d s
s u g g e s te d th a t Z n 2 io n , r e le a s e d fr o m d is s o lu tio n o f Z n O f o r w a t e r d i s i n f e c t i o n . C h e m . E n g . J . 8 5 , 1 1 1 1 1 7 .
N P s , c a n b in d to th e b a c te r ia l m e m b r a n e a n d p ro lo n g th e la g C h a n g , J . C ., O s s o ff, S . F ., L o b e , D . C ., D o r fm a n , M . H ., D u m a is ,
p h a s e o f th e g r o w th c y c le o f b a c te r ia (A tm a c a e t a l. ) . C . M ., Q u a lls , R . G . & J o h n s o n , J . D . U V in a c t iv a t io n o f
p a th o g e n ic a n d in d ic a to r m ic r o o r g a n is m s . A p p l. E n v iro n .
M i c r o b i o l . 4 9 , 1 3 6 1 1 3 6 5 .
C u llit y , B . E le m e n t s o f X - r a y D iffr a c t io n . A d d is o n -W e s le y ,
C O N C L U S IO N L o n d o n , p . 1 0 2 .
D a s , T . K . U ltr a v io le t d is in fe c tio n a p p lic a tio n to a w a s te w a te r
O u r e x p e r im e n t s o u g h t to e x p lo r e th e a n tib a c te r ia l p r o p e r - t r e a t m e n t p l a n t . C l e a n P r o d . P r o c e s s . 3 , 6 9 8 0 .
F r a n k lin , N . M ., R o g e r s , N . J ., A p te , S . C ., B a tle y , G . E ., G a d d ,
tie s o f b o th Z n O N P s to w a r d s im p o r ta n t w a te r b o r n e
G . E . & C a s e y , P . S . C o m p a r a tiv e to x ic ity o f
p a th o g e n s , E . c o li, P ro te u s , S . e p id e rm id is a n d K . p n e u m o - n a n o p a r tic u la te Z n O , b u lk Z n O , a n d Z n C l2 to a fr e s h w a te r
n ia e . T h e r e s u lts in d ic a te d th a t th e in h ib ito r y e ffe c ts m ic r o a lg a (P s e u d o k irc h n e rie lla s u b c a p ita ta ): th e im p o r ta n c e
in c re a s e d a s th e c o n c e n tra tio n s o f Z n O N P s in c re a s e d . o f p a r t i c l e s o l u b i l i t y . E n v i r o n . S c i . T e c h n o l . 4 1 , 8 4 8 4 8 4 9 0 .
O u r s tu d y d o e s n o t, h o w e v e r , s h o w th e im p o r ta n t r e la tio n - G o r d o n , T ., P e r ls te in , B ., H o u b a r a , O ., F e ln e r , I ., B a n in , E . &
M a r g e l, S . S y n th e s is a n d c h a r a c te r iz a tio n o f z in c / ir o n
s h ip b e tw e e n th e s iz e o f Z n O N P s a n d a n tib a c te ria l
o x id e c o m p o s ite n a n o p a r tic le s a n d th e ir a n tib a c te r ia l
p ro p e rtie s . T h e s e r e s u lts s h o w th a t Z n O N P s c a n b e p o te n - p r o p e r t i e s . C o l l o i d . S u r f . A 3 7 4 , 1 8 .
tia lly r e g a r d e d a s a n e ffe c tiv e a n tib a c te r ia l a g e n t fo r H ijn e n , W ., B e e r e n d o n k , E . & M e d e m a , G . J . I n a c tiv a tio n
w a te r b o r n e p a th o g e n s a n d c a n b e c o n s id e r e d a s a c o m p - c r e d it o f U V r a d ia tio n fo r v ir u s e s , b a c te r ia a n d p r o to z o a n
le m e n ta ry m e th o d fo r w a te r a n d w a s te w a te r tre a tm e n t. ( o o ) c y s t s i n w a t e r : a r e v i e w . W a t e r R e s . 4 0 , 3 2 2 .
H s ia o , I .-L . & H u a n g , Y .-J . E ffe c t s o f v a r io u s p h y s ic o c h e m ic a l
c h a r a c te r is tic s o n th e to x ic itie s o f Z n O a n d T iO 2
n a n o p a r tic le s to w a r d h u m a n lu n g e p ith e lia l c e lls . S c i. T o ta l
A C K N O W L E D G E M E N T S E n v i r o n . 4 0 9 , 1 2 1 9 1 2 2 8 .
J o n e s , N ., R a y , B ., R a n jit , K . T . & M a n n a , A . C . A n tib a c t e r ia l
T h e w o r k w a s n a n c e d b y th e R e s e a rc h C o u n c il o f B a b o l a c tiv ity o f Z n O n a n o p a r tic le s u s p e n s io n s o n a b r o a d
s p e c tr u m o f m ic r o o r g a n is m s . F E M S M ic ro b io l. L e tt. 2 7 9 ,
U n iv e r s ity o f M e d ic a l S c ie n c e s . T h is in v e s tig a tio n w a s a c o l-
7 1 7 6 .
la b o ra tiv e w o r k o f th e C e llu la r a n d M o le c u la r B io lo g y K e lle r , A . A ., W a n g , H ., Z h o u , D ., L e n ih a n , H . S ., C h e r r , G .,
R e s e a rc h C e n te r , B a b o l U n iv e rs ity o f M e d ic a l S c ie n c e s , C a r d in a le , B . J ., M ille r , R . & J i, Z . S t a b ility a n d
a n d th e N a n o te c h n o lo g y B io te c h n o lo g y R e s e a rc h G r o u p , a g g r e g a tio n o f m e ta l o x id e n a n o p a r tic le s in n a tu r a l a q u e o u s
M a z a n d a r a n U n iv e r s ity , B a b o ls a r . T h e a u th o rs h a v e n o c o n - m a t r i c e s . E n v i r o n . S c i . T e c h n o l . 4 4 , 1 9 6 2 1 9 6 7 .
K h o z a , P . B ., M o lo t o , M . J . & S ik h w iv h ilu , L . M . T h e e ffe c t o f
i c t o f i n t e r e s t t o d e c l a r e .
s o lv e n ts , a c e to n e , w a te r , a n d e th a n o l, o n th e m o r p h o lo g ic a l
a n d o p tic a l p r o p e r tie s o f Z n O n a n o p a r tic le s p r e p a r e d b y
m ic ro w a v e . J. N a n o te c h n o l. 2 0 1 2 , a rt. ID 1 9 5 1 0 6 . h ttp :// d x .
R E F E R E N C E S d o i.o r g / 1 0 .1 1 5 5 / 2 0 1 2 / 1 9 5 1 0 6
K u m a r , S . S ., V e n k a t e s w a r lu , P ., R a o , V . R . & R a o , G . N .
A d a m s , L ., L y o n , D ., M c I n to s h , A . & A lv a r e z , P . a S y n th e s is , c h a r a c te r iz a tio n a n d o p tic a l p r o p e r tie s o f z in c
C o m p a r a tiv e to x ic ity o f n a n o -s c a le T iO 2 , S iO 2 a n d Z n O o x i d e n a n o p a r t i c l e s . I n t . N a n o . L e t t . 3 , 1 6 .
w a te r s u s p e n s i o n s . W a t e r S c i . T e c h n o l . 5 4 ( 1 1 1 2 ) , 3 2 7 3 3 4 . L i, Q ., M a h e n d r a , S ., L y o n , D . Y ., B r u n e t, L ., L ig a , M . V ., L i, D . &
A d a m s , L . K ., L y o n , D . Y . & A la v a r e z , P . J. b C o m p a ra tiv e A lv a r e z , P . J. A n tim ic r o b ia l n a n o m a te r ia ls fo r w a te r
e c o -t o x ic ity o f n a n o s c a le T iO 2 , S iO 2 , a n d Z n O w a te r d is in fe c tio n a n d m ic r o b ia l c o n tr o l: P o te n tia l a p p lic a tio n s
s u s p e n s io n s . W a t e r R e s . 4 0 , 3 5 2 7 3 5 3 2 . a n d i m p l i c a t i o n s . W a t e r R e s . 4 2 , 4 5 9 1 4 6 0 2 .
 7 7 0 F . E lm i e t a l. | A n t ib a c t e r ia l a c t iv it y o f Z n O n a n o p a r t ic le s in w a t e r t r e a t m e n t
L iu , Y ., H e , L ., M u s t a p h a , A ., L i , H ., H u , Z . & L i n , M .
A n tib a c te r ia l a c tiv itie s o f z in c o x id e n a n o p a r tic le s a g a in s t
E s c h e r i c h i a c o l i O 1 5 7 : H 7 . J . A p p l . M i c r o b i o l . 1 0 7 , 1 1 9 3 1 2 0 1 .
L u , J . , N g , K . M . & Y a n g , S . E f c i e n t , o n e - s t e p
m e c h a n o c h e m ic a l p ro c e s s fo r th e s y n th e s is o f Z n O
n a n o p a r t i c l e s . I n d . E n g . C h e m . R e s . 4 7 , 1 0 9 5 1 1 0 1 .
M a k h lu f, S ., D r o r , R ., N it z a n , Y ., A b r a m o v ic h , Y ., J e lin e k , R . &
G e d a n k e n , A . M ic r o w a v e -a s s is te d s y n th e s is o f
n a n o c r y s ta llin e M g O a n d its u s e a s a b a c te r io c id e . A d v .
F u n c t . M a t e r . 1 5 , 1 7 0 8 1 7 1 5 .
M e r u v u , H ., V a n g a la p a t i, M ., C h ip p a d a , S . C . & B a m m id i, S . R .
S y n th e s is a n d c h a r a c te r iz a tio n o f z in c o x id e
n a n o p a r tic le s a n d its a n tim ic r o b ia l a c tiv ity a g a in s t B a c illu s
s u b t i l i s a n d E s c h e r i c h i a c o l i . C h e m i s t r y 4 , 2 1 7 2 2 2 .
M e u le n k a m p , E . A . S y n th e s is a n d g r o w th o f Z n O
n a n o p a r t i c l e s . J . P h y s . C h e m . B 1 0 2 , 5 5 6 6 5 5 7 2 .
M o n a r c a , S ., Z a n i, C ., R ic h a r d s o n , S . D ., T h r u s to n J r ., A . D .,
M o r e t ti, M ., F e r e t ti, D . & V illa r in i, M . A n e w a p p r o a c h
to e v a lu a tin g th e to x ic ity a n d g e n o to x ic ity o f d is in fe c te d
d r i n k i n g w a t e r . W a t e r R e s . 3 8 , 3 8 0 9 3 8 1 9 .
M u je r ie g o , R . & A s a n o , T . T h e r o le o f a d v a n c e d tr e a tm e n t in
w a s te w a te r r e c la m a tio n a n d r e u s e . W a te r S c i. T e c h n o l. 4 0
( 4 5 ) , 1 9 .
N a ir , S ., S a s id h a r a n , A ., R a n i, V . D ., M e n o n , D ., N a ir , S .,
M a n z o o r , K . & R a in a , A . S . R o le o f s iz e s c a le o f Z n O
n a n o p a r tic le s a n d m ic r o p a r tic le s o n to x ic ity to w a r d b a c te r ia
a n d o s te o b la s t c a n c e r c e lls . J. M a te r. S c i. M a te r. M e d . 2 0 ,
2 3 5 2 4 1 .
N a p ie r s k a , D ., T h o m a s s e n , L . C ., R a b o lli, V ., L is o n , D ., G o n z a le z ,
L ., K ir s c h -V o ld e r s , M ., M a r te n s , J . A . & H o e t, P . H .
S iz e -d e p e n d e n t c y t o t o x ic ity o f m o n o d is p e r s e s ilic a
n a n o p a r t i c l e s i n h u m a n e n d o t h e l i a l c e l l s . S m a l l 5 , 8 4 6 8 5 3 .
P a d m a v a t h y , N . & V ija y a r a g h a v a n , R . E n h a n c e d b io a c t iv it y
o f Z n O n a n o p a r tic le s a n a n tim ic r o b ia l s tu d y . S c i. T e c h n o l.
A d v . M a te r. 9 , 0 3 5 0 0 4 .
P r e m a n a th a n , M ., K a r t h ik e y a n , K ., J e y a s u b r a m a n ia n , K . &
M a n iv a n n a n , G . S e le c tiv e to x ic ity o f Z n O n a n o p a r tic le s
to w a r d G r a m -p o s it iv e b a c t e r ia a n d c a n c e r c e lls b y a p o p to s is
th r o u g h lip id p e r o x id a tio n . N a n o m e d . N a n o te c h n o l. B io l.
M e d . 7 , 1 8 4 1 9 2 .
Q u , X ., A lv a r e z , P . J . & L i, Q . A p p lic a tio n s o f n a n o t e c h n o lo g y
i n w a t e r a n d w a s t e w a t e r t r e a t m e n t . W a t e r R e s . 4 7 , 3 9 3 1 3 9 4 6 .
R a g h u p a t h i, K . R ., K o o d a li, R . T . & M a n n a , A . C . S iz e -
d e p e n d e n t b a c te r ia l g r o w th in h ib itio n a n d m e c h a n is m o f
a n tib a c te r ia l a c tiv ity o f z in c o x id e n a n o p a r tic le s . L a n g m u ir
2 7 ( 7 ) , 4 0 2 0 4 0 2 8 .
R a je s h , D ., V a r a L a k s h m i, B . & S u n a n d a n a , C . T w o -s te p
s y n th e s is a n d c h a r a c te r iz a tio n o f Z n O n a n o p a r tic le s . P h y s ic a
B : C o n d e n s . M a t t e r 2 3 , 4 5 3 7 4 5 3 9 .
R i s t i c , M . , M u s i c , S . , I v a n d a , M . & P o p o v i c , S . S o l g e l
s y n th e s is a n d c h a r a c te r iz a tio n o f n a n o c r y s ta llin e Z n O
p o w d e r s . J . A l l o y C o m p d 3 9 7 , L 1 L 4 .
S a w a i, J. Q u a n tita tiv e e v a lu a tio n o f a n tib a c te r ia l a c tiv itie s o f
m e ta llic o x id e p o w d e r s (Z n O , M g O a n d C a O ) b y
c o n d u c t i m e t r i c a s s a y . J . M i c r o b i o l . M e t h . 5 4 , 1 7 7 1 8 2 .
F ir s t r e c e iv e d 1 0 F e b r u a r y 2 0 1 4 ; a c c e p te d in r e v is e d fo r m 7 M a y 2 0 1 4 . A v a ila b le o n lin e 1 7 Ju n e 2 0 1 4
W a t e r S c ie n c e & T e c h n o lo g y | 7 0 .5 | 2 0 1 4
S h e n , L ., B a o , N ., Y a n a g is a w a , K ., D o m e n , K ., G u p t a , A . & G r im e s ,
C . A . D ir e c t s y n th e s is o f Z n O n a n o p a r tic le s b y a s o lu tio n -
f r e e m e c h a n o c h e m i c a l r e a c t i o n . N a n o t e c h n o l o g y 1 7 , 5 1 1 7 5 1 2 3 .
S p a n h e l, L . & A n d e r s o n , M . A . S e m ic o n d u c to r c lu s te r s in th e
s o l g e l p r o c e s s : q u a n t i z e d a g g r e g a t i o n , g e l a t i o n , a n d c r y s t a l
g r o w th in c o n c e n tr a te d z in c o x id e c o llo id s . J. A m . C h e m . S o c .
1 1 3 , 2 8 2 6 2 8 3 3 .
T a m , K ., D ju r is ic , A ., C h a n , C ., X i , Y ., T s e , C ., L e u n g , Y ., C h a n , W .,
L e u n g , F . & A u , D . A n tib a c te r ia l a c tiv ity o f Z n O
n a n o ro d s p r e p a re d b y a h y d ro th e rm a l m e th o d . T h in S o lid
F i l m s 5 1 6 , 6 1 6 7 6 1 7 4 .
T s o , C .-P ., Z h u n g , C .-M ., S h ih , Y .-H ., T s e n g , Y .-M ., W u , S .-C . &
D o o n g , R .-A . S t a b ility o f m e ta l o x id e n a n o p a r t ic le s in
a q u e o u s s o l u t i o n s . W a t e r S c i . T e c h n o l . 6 1 ( 1 ) , 1 2 7 1 3 3 .
V a h a b i, K ., M a n s o o r i, G . A . & K a r im i, S . B io s y n t h e s is o f
s ilv e r n a n o p a r tic le s b y fu n g u s T ric h o d e rm a re e s e i (a r o u te fo r
l a r g e - s c a l e p r o d u c t i o n o f A g N P s ) . I n s c i e n c e s J . 1 , 6 5 7 9 .
W a n g , C ., L iu , L .-L ., Z h a n g , A .-T ., X ie , P ., L u , J .-J . & Z o u , X .-T .
A n tib a c te r ia l e ffe c ts o f z in c o x id e n a n o p a r tic le s o n
E s c h e r i c h i a c o l i K 8 8 . A f r . J . B i o t e c h n o l . 1 1 , 1 0 2 4 8 1 0 2 5 4 .
W a r d , R . & D e g r a e v e , G . R e s id u a l to x ic ity o f s e v e r a l
d is in fe c ta n ts in d o m e s tic w a s te w a te r . J. W a te r P o llu t. C o n tro l
F e d . 5 0 , 4 6 6 0 .
W ie g a n d , I ., H ilp e r t, K . & H a n c o c k , R . E . A g a r a n d b r o th
d ilu tio n m e th o d s to d e te r m in e th e m in im a l in h ib ito r y
c o n c e n tr a tio n (M I C ) o f a n tim ic r o b ia l s u b s ta n c e s . N a tu re
P r o t o c o l . 3 , 1 6 3 1 7 5 .
Y o u s e f, J . M . & D a n ia l, E . N . I n v itro a n tib a c te r ia l a c tiv ity a n d
m in im u m in h ib ito r y c o n c e n tr a tio n o f z in c o x id e a n d n a n o -
p a r tic le z in c o x id e a g a in s t p a th o g e n ic s tr a in s . J. H e a lth S c i. 2 ,
3 8 4 2 .
Z h a n g , L ., J ia n g , Y ., D in g , Y ., P o v e y , M . & Y o r k , D . I n v e s t ig a t io n
in to th e a n tib a c te r ia l b e h a v io u r o f s u s p e n s io n s o f Z n O
n a n o p a r t i c l e s ( Z n O n a n o u i d s ) . J . N a n o p a r t i c l e R e s . 9 , 4 7 9 4 8 9 .
Z h a n g , L . , D i n g , Y . , P o v e y , M . & Y o r k , D . Z n O n a n o u i d s
A p o t e n t i a l a n t i b a c t e r i a l a g e n t . P r o g . N a t . S c i . 1 8 , 9 3 9 9 4 4 .
Z h a n g , H ., Q u , J ., L iu , H . & W e i, D . a C h a r a c te r iz a t io n o f
d is s o lv e d o r g a n ic m a tte r fr a c tio n s a n d its r e la tio n s h ip
w ith t h e d is in fe c tio n b y -p r o d u c t fo r m a tio n . J . E n v iro n . S c i .
2 1 , 5 4 6 1 .
Z h a n g , H ., Q u , J ., L iu , H . & Z h a o , X . b C h a r a c t e r iz a tio n o f
is o la te d fr a c tio n s o f d is s o lv e d o r g a n ic m a tte r fr o m s e w a g e
tr e a tm e n t p la n t a n d t h e r e la t e d d is in fe c tio n b y -p r o d u c ts
f o r m a t i o n p o t e n t i a l . J . H a z a r d . M a t e r . 1 6 4 , 1 4 3 3 1 4 3 8 .
Z h a n g , L ., J ia n g , Y ., D i n g , Y ., D a s k a l a k is , N ., J e u k e n , L ., P o v e y , M .,
O N e i l l , A . J . & Y o r k , D . W . M e c h a n i s t i c i n v e s t i g a t i o n i n t o
a n tib a c te r ia l b e h a v io u r o f s u s p e n s io n s o f Z n O n a n o p a r tic le s
a g a i n s t E . c o l i . J . N a n o p a r t i c l e R e s . 1 2 , 1 6 2 5 1 6 3 6 .
Z h a n g , L . L ., C h e n , B ., X ie , L . L . & L i, Z . F . S t u d y o n th e
a n tim ic r o b ia l p r o p e r tie s o f Z n O s u s p e n s io n a g a in s t G r a m -
p o s itiv e a n d G r a m -n e g a tiv e b a c te r ia s tr a in s . A d v . M a te r . R e s .
3 9 3 , 1 4 8 8 1 4 9 1 .
Z h a n g , L ., L i, Y ., L iu , X ., Z h a o , L ., D in g , Y ., P o v e y , M . & C a n g , D .
T h e p r o p e r t i e s o f Z n O n a n o u i d s a n d t h e r o l e o f H 2 O 2 i n t h e
d i s i n f e c t i o n a c t i v i t y a g a i n s t E . c o l i . W a t e r R e s . 4 7 , 4 0 1 3 4 0 2 1 .
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