Removal and selective recovery of heavy metals from electroplating effluents

! ! " " # " ! $ " % " ! & " " # # $ " ! $ " ' ( )* # + ,- . * / & 0 * 1 * 12343516776 '%18 19:3551677;# !) # ) + . 8 < = / ) $ > <? @) * '# ) " ' 8 ! # ! ! ? 8 9768" " " A >- # * ! ! " ! #

) " )C " D " 0 " ) " ' " ' " " EF " ### * ! # ! " " # * " ) " " G " H" ! " # * " C " ! # " ! " " " #

$ " , L ,J # @ M @ M # 0 ,-,- ,- = C # 0 & M L " K " ,- @ " # ,- < & D N :9;2" " ,-,- ,- C M @ # & " . ,- " , G 6O" 6O" D6O" P 6O 9OH ,- = # " = < ,- @ J " @ = < 25 Q # " & ,-" @ ,J # < ,- " ,- # ) " = ,- @ = ,J $ ,- " I L . # F G " " " " H" " . ,- #

< M GD H # < < = ,- D " C " " . ,- ,- " # ,- 4RS @ # " M G " D P H G " " D P H @ ,- @ % ;#7 C G < H# C " ,J $ T ,J U 4R" R:" R6 R2S D " " P " # D " < B ,- ;O 9O_{" & % ;"7} < # " ;O % 6"9 GR4SH % 6"9T M & % ;"7 < # " = L T " =" @ @ # 0 " L " C # 0 @ = " C$ L " ,-= = # 0 C " ,- " C $ ,-9O ;O _{,J # ,- =} C # - " M ,-,- C " " @ = #

# $ " # $ # * M # * M " " " # " D N :9;2" " M # " " G 6O" 6O" D6O" P 6O 9O_{H !} = # " ! " ! = M! 25 Q # " $ # " # ? " = $ " # G " " " " H" " # $ 9O #

D " # #M " # * ! 4RS# " G " D P H G " " D P H " % ;#7 M G M ! H# " T V 4R" R:" R6 R2S D " " P " # " B ;O 9O" % & ;#7 # " ;O GR4SH % 6#9T " % ;#7 # " T " " # " " # D ! = " $ ! " # " " $ 9O ;O _! # # " " " ! = #

< $ # . < " < < $ < < $ " W. < < W # ? M $ < < # ? X < $ <. W < < < < < < < $ W # ?W & X < W < " K W< " W " < < < < $ W # " W X D N :9;2" " < $ < M " <# & " W " < < < $ G 6O_" 6O_{" D}6O_{" P} 6O 9O_{H <} < " < < <# < " < < W< < < " ! = K 25Q # " < < < < & W< < < W # ? < < < < W < $ < < M W # < " < < < < X K $ < K < " < < < $ < < # < < W G " " " <" H" . <

< < M<< GD H < # ? < < < < D K < < < " X X W W < " # K 4RS ! # " $ < << G " D P H G " " D P H < W W % ;#7 < < G H# X X " < $ X K < $ < < " U 4R" R:" R6 R2S D " " P " # " $ < <<B < ;O _K 9O_{" & % K ;"7} < # " ;O , < <K % 6"9 GR4SH K % 6"9" W& % K ;"7 < # " < $ << << W X < M " < $ < < <<" <" < # ? < <<" < " < K Y<# ? ! = < <<" $ < < < . < . <# ? " <" X $ 9O_K ;O _{# ? <} . < # " < < < < < < " " ! = W #

" # " " = # " # * ! " M # * = :7 " T W # D $ " " " # 6" " " $ T " ! # G " ! = H 9#

" = " " $ 2# 5" # ; 3" D N :9;2 " " # " ! = " ! G ;H# " ! $ G 3H# * 4T " " " 4 G " ! = H R G " ! H# " ! :7 !#

!( * ) ) * * * + ** % ( ** * !(( *** $ $, - $ $,$, . ' $ $,", / $,",$, / $,",$,$, 8 / $,",$,$,$, 0 $,",$,$,", 1 $,",$,", 8 2 $,",$,",$, 2 $,",/, 3 $,",0, 4 $,",1, $# $,",2, $0 $,",3, $1 $,",3,$, 8 = $1 $,",3,", $2 $,",5, _$3 $,",4, $5 $,",$#! "#

" ", ( ' 6 7 // !( // ",$, - /0 ",", /3 ",",$, " /3 ",",", # /3 ",",/, /5 ",",0, $ /4 ",",1, /4 ",",2, ! 0# ",/, 0$ ",/,$, 0$ ",/,", 0" ",/,/, 00 ",/,0, % 01 ",/,1, ! 04 ",0, 1# 1" / /, ( ' 6 7 13 !( 13 /,$, - 15 /,", 2#

/,",/, & 2# /,",0, 2$ /,",0,$, Z 2$ /,",0,", 2" /,",0,/, 2" /,",1, '( _2" /,",2, $ 2/ /,",3, 2/ /,",5, 20 /,",4, ) 20 /,",$#, * 21 /,/, 21 /,/,$, + 21 /,/,", , 24 /,/,/, $ 3$ /,/,0, ) '(_"-'( _.'( ₃₁ /,0, 32 5" 0 0, ( ' 8 7 53 !( 53 0,$, - 55 0,", 4# 0,",$, " 4# 0,",", # 4$ 0,",/, & 4$ 0,",0, ) / 4$

0,",3, ! 40 0,",5, * 40 0,/, 41 0,/,$, # 0 / 41 0,/,", 0 / 43 0,/,/, ! $#1 0,0, $#5 $$# 1 1, -( ' 6 $$1 !( $$1 1,$, - $$1 1,", $$3 1,",$, " $$3 1,",", # $$3 1,",/, , $$5 1,",0, $$5 1,",1, ! $$5 1,",2, * $$4 1,/, $$4 1,0, $"1 $"2 2 2, ( ' 8 7 $"4

2,", $/" 2,",$, " $/" 2,",", # $/" 2,",/, & $/" 2,",0, / $/" 2,",1, $/0 2,",2, $/0 2,",3, * $/1 2,/, $/1 2,/,$, / $/1 2,/,", 1 $/3 2,0, $0" 2,1, $05 $04 3 3, & ( ' 8 $1/ !( $1/ 3,$, - $1/ 3,", $10 3,",$, " $10 3,",", # $11 3,",/, , $11 3,",0, $11 3,",1, ! 0 $12 3,/, $12 3,0, $2$ $2"

( $21 !( $21 5,$, - $21 5,", $23 5,",$, ) $23 5,",", $25 5,",/, $25 5,",0, $24 5,",1, % / / 2 34 $24 5,/, $3# 5,/,$, ) $3$ 5,/,", $ $3" 5,/,/, " % / $30 5,/,/,$, $30 5,/,",", ! = $33 5,/,0, " % / $5/ 5,0, $51 $52 4 4, & 9 : $54 !( $54 4,$, - $4# 4,", $4$ 4,",$, ) $4$ 4,",", $4" 4,",/, $4"

4,/, $40 4,0, "## "#$ $# $#, ' & _"#1 $#,$, "#1 $#,", + ' & "#5 !; ; < ! * ! !:$7 D N :9;2 !:"7 D N :9;2 !:/7 25 Q D N :9;2 ! * = =:$7 ! * :$7 & % ;

+ ",$, D N :9;2 G H# M G8H# /1 + ",", ;2;M:8 G H" D N ::R7 G8H" D N ::R5 G H D N :9;2 G H# N G5 1 H B G[H :7 ) ) " % ;#7" 2 ) 6O G HT G\H 6#9 ) 6O_{" :#5 ) D}6O _{7#4 ) P} 6O_{" &} % ;#7 G]H & % ;#7 2 ) 6O_{# E} $ T G ^;H# 0" + ",/, * D N :9;2 # 5 1 ) % G:7 )" % ;#7H" 7#6 ) B G\H 6O_{O D}6O_{T G[H} 6O_{O P} 6O_T G_H 6O _O 6O_{T G]H} 6O _O 9O<sub>T G`H</sub> 6O <sub>G H# E</sub> $ T G ^;H# 0/ + ",0, D N :9;2# G\H 7#65" G]H 7#5" G[H :#7" G`H 6#7" G_H 5#7 GaH :7#7 1 " :7 ) ) " % ;#7" 2 ) 6O_{# E $ T} G ^;H# 01 + ",1, ! # N G5 1 H D N :9;2 :7 ) ) " % ;#7" 2 ) 6O_{B G]H G H} G_H 25 Q " GaH 55 Q G\H 47 Q # E $ T G ^;H# 02 + ",2, M ! D N :9;2# b T 8" b 25" 55 47 Q " # 03 + ",3, ! D N :9;2# E $ # $ " 977 G 25" 55 47 Q H # 03 + ",5, D N :9;2" 47 Q # A # M G H" G8H M G H # 05 + ",4, S ! D N :9;2 # :6 1 1#

# E $ T G ^2H# + /,$, 6O_{G H" D}6O_{G8H P} 6O_{G H G]H G\H} D N :9;2# :7 ) ) % ;#7 2 1 T 5 1 # $ T G V 2H# 23 + /,", D6O_{G H P} 6O _{G8H G]H G\H} D N :9;2# :7 ) ) % ;#7 2 1 T :77 1 # $ T G V 2H# 25 + /,/, 6O_{G H" D}6O_{G8H P} 6O_{G H G]H G\H} D N :9;2# :7 ) ) % ;#7 2 1 # * : # $ T G V 2H# 3# + /,0, D N :9;2 M! 25 Q G H# M G8H# 3" + /,1, D N :9;2 M! 25 Q )6' G HT B # M G8H# 3" + /,2, D N :9;2# ? G H M! 25 Q G8H# B # 3/ + /,3, ' D N :9;2# 25 Q G H G HT G8H# 30 + /,5, ' D N :9;2 $ # G H G HT P 6O _{G8H" D}6O_{G H} 6O $ G H# :7 ) ) % ;#7 6 1 # * B 66#6 6O _{1 " 96#9 D}6O _{1 6;#6 P} 6O _{1 #} ;7 65 Q # 31 + 0,$, M # ) B $##

# + 0,", ! :#5 1 D N :9;2# ! G HT ! G HT ! G ! H# * ! G) D (?OHT # $#" + 0,/, ! # M :#5 1 T 8 b :6 1 # D ! G HT ! D N :9;2 G HT ! G ! H# ! 2#6# ! B 6#7 1 G92µΜH# $#0 + 0,0, ! D N :9;2 $ # 8 B T B : G HT = B 6 G HT ! B # * 2#6# $ :6 1 * 2#9# * ! ! :57 " 65 Q # 97 " " # E $ G ^;H, $#3 + 1,$, S M D N :9;2 # ) B 6O G HT D6O_{G = HT} 9O_{G ! H# 8} :4 1 " % ;#7# * ! ! :57 " 65Q # 97 " " # E $ T G ^ 2H# $"/ + 1,", 9O_" 6O _D6O_# G!H G=H # G HT G HT G ! H# * G) D (?OHT 5#6#2# $"0

G HT G ! HT G H# G) D (?OH# + 2,", $ D N :9;2 $ # b T 8 b 8# * B T $ B ! # * " ;#6#5# $ :4 1 # E $ G ^ ;H# $0" + 3,$, M D N :9;2 # ) B G HT ! G ! HT G HT GE H G = H# 8 :4 1 " % 6#9 G H % ;#7 G H# E $ T G ^ 2H# $14 + 3,", 9#5 $ :7M2 ₁ ;O _% # * ;O G) D (?OHB 026M G]H" 6036M G\H" % 02M G[H" %6 02GaH# $2# + 5,$, # :5 W " " 557 Q # # $3" + 5,", # * % & 6 M7#; E1c 1 GZ 9)Hd" 95 Q G H 27 Q G8H# $31 + 5,/, ! = % # 6#4 1 ! G H 2#9 1 = G8H # * G) D (?OH B " )6O_{G H G`H" )G0%H} 6G H G_H )G0%H9MG H G\H )G0%H26MG HG]H# $34 + 5,0, ! $5#

+ 5,1, = $ % :7# G H" G H" ! $ G = H $ G ! H" = $ # $5" + 5,2, ! " ! = # $50 + 4,$, !# * " " ! = # $41 + 4,", 6#4 1 D G H" 7#6 1 P G8H 9#4 1 9O_{G H} ;O_{G H" % # *} G) D (?OH # D P B " )6O_{G H} G`H" )G0%H6G H G_H" )G0%H9MG H G\H )G0%H26MG HG]H# 9O B 9OG H G`H" G0%H6O_{G H G\H" G0%H9G H G_H G0%H2}M_{G H G]H#} ;O _{B 02}6M_{G H G`H"} 6036MG HG_H % 02MG HG\H# $45

% ( /,$, ? 6O_{" D}6O _P 6O _{# 3$} % ( /,", ! # 5# % ( 0,$, M = # * G M :R42H G M 69;1R4H # 42 % ( 0,", " Z" " ! = " % ;#7" % 5#7e# 44 % ( 0,/, 8 D N :9;2" # * G M :R42H G M 69;1R4H # $#2 % ( 1,$, D N :9;2 # $"$ % ( 1,", # $"" % ( 2,$, ! G % & ;#7H# * G M " :R42H G M 69;1R4H # $/2 % ( 2,", 8 D N :9;2" # * G M :R42H G M 69;1R4H # $0$ % ( 3,$, 8 D N :9;2" # $15 % ( 5,$, ' ! $ # $5# % ( 5,", ' = $ # $5$ % ( 5,/, ! $ # $5/ % ( 5,0, = $ # $5/

?

S !! !! :+! =

(

? G H %! :"6 b $ @ ) % ! DEAE ;! $ ! %! ' $ +% -- M

A

G H

B

B 1# ! 57S $

B

) M $ 6MGDM H ; C D ;%! D =

-@

) !

@

* ) $ ! '

,

% * %. * : ! M E $ C N $ $

-$,$, . ' ! " T # 8 $ ! G " % H" Gf 677;H" " M # % G !H " = # * B H G " P " " " " " ) " D H GZ !677;HT H G " " " " D " " " " H G 6772HT H G " H G8 6777HT H G " " ) " P H GE ! 677:HT H G " H G) ! 677RHT H G " " " P " D " H Gf 677:H# $ " " M # " B

G 1 = H G $ " H# * G 1 = H G $ H Gf 677;H# * M GD & E ! 67:7H# $ M # * G " " H Gf 677RH# " " $ " 6O_" 6O_" 6O_{" %} 6O_" ;O_" 9O_" 6O_" 6O_{" D}6O_" 6O _P 6O_{G M & :RRRT} 6776T A 6779T g= g= 6779T E 6779T = 6772T % 677;T ) 6773T 6774T & 677RT f 67:7H" # " & & T # $ G :R4:T f :RR;T = 6772HT "

" # # % # D " B G H " G H " G H = G H # * # $,", :#6#:# ) B M G H " " M G H# :#6#:#:# 8 * " ! " G H ! G8 ! :RR5H# % G 5M97 Q HT "

! G8 ! :RR5H# G M H " " " ! # % " " " # * $ M 9# :#6#:#:#:# * " :5M65S " $ G47M R7SH cαM G H βM d" G5M:7SH G:M6SH GZ 6776H# " " = = M G :RR2H# = # * βM # βM βG:h 9H βG:h ;HM ! T βG:h 2HM ! DM GZ 6776H# G 677:H# * αM G H" 97M57 S G 677:T ? 8 677;HT ! $ M " ! #

:#6#:#:#6# * " " " " G8 :RR2H# * ! " $ $ 6O_" 6O _{G8 :RR2 H# f G6776H %} G677;H $ ! ! # " ? G6775H $ $ 9O _{# $ "} G 6773 H# " " $ " " " " ! # 8 ! $ " " $ T " " GA :RR7T E ! :RR7H# G6774H $ G H T " f # G H $ G 6773 H# D6O ;O Gg= g= 6779H#

N $ GZO_{" D} O_{" )} 6O_" 6O_{H # f G6773H} = " $ " = # 0 = " G8 :RR2 T > 6774T 677RH# ) T " # Z G6779H * $ # :#6#:#6# 8 8 " " G8 ! :RR5H# 8 GD Z :R33T 8 M :R4:H# " GD Z :R33T * :RR6H# :#6#:#6#:# " #

" " %O_{M *} G8 M :R4:T A :RR9H# ) GD Z :R33T * :RR6T 8 ! :RR5HT " " " T Gf A :R43H# :#6#9# ) %" " # % G ' $ :RR6H# %" $ " G8 ! :RR5H# $ " % G D 6773H # * % 02 M _{% G 6H 0} 2 6M _{% G} 3H# 0 " % G % i 9H" # " %O _{G D 6773H"} ;O Gg= g= 6779H# ) T " :#6#2#

* G* * :RR2T 8 ! :RR5H# = * $ G:RRRH $ " " %M # # A " " T = G H" $ GA 677RH# A # G6779H G ;O 6O_{H "} " " " " ! T ! # * T G :RR;H# " # ! ! " " " GA :RR7T * :RR6T ? f ! :RR;T ! = :RR3T 8 6772T Aj! 6775H # $ GA :RR7H# * # # 8 Gf 677;H#

* # 8 G:RR2 H 5M27 Q # :#6#2# ) G H $ G* 6777H# * " " # %" $ G H" 1 " G% :RR:H# % " $ # % " # % $ " GA :RR9T 8 ! :RR5H# ' $ T $ " $ ;O 9O # * G # # " " " H $ G% :RR:T * :RR9T A :RR9H# $ " $ G :R46H# 0 # G:R46H $ # 8 "

" G :#6#9H# ) = # $ G # # H G " " %H" # " & $ " 2# :#6#5# " # % " ! ? " # 0 " ! 8 M M* G8 *H" & ! # " $ " # " T " ! # " # " ? #

8 * " " " ? " GE ! 6779 H# * ? B , 9 , :;3 ( ,< c:M:d , ! G 1 H" , $ ! G 1 H" G H G 1 H " # * = c:M:d B 3:, 9 3:, ( =3:;, <> ;3: ,< c:M6d * ! ;3:,< ;3: _,< , c:M6d# * c:M9d B , 9 % 3: _c:M9d = " c:M2d" B , 9 %( ;3: < , c:M2d

? " , , ! ;%< ;3: < T , , ? # ) $ ! = # , G H G H ? # 8 * ? c:M5dB , 9 ,, : =; 0 ,< =3 ( ; 03< ,: >> c:M5d G 1 HT ," " , , ? # * = c:M5d B ,:=; 0 ,< ,> 9 3 :; , < ( =; 03< : , > ; ,: < c:M;d * , : =; 0 ,< ,> ,: , c:M;d# ' = = M # "

$ :B: " " c:M3dB ( c:M3d " M $# " + B + 9 = >:= >= > c:M4d = >" = > = > M $" G H# $ " ' = " c:MRd c:M:7d " # ' = B = >:= > 93:;+ = >< ( ;3:= ><= > c:MRd * = >:= > = > " = > + # B = >:= > 9 + = > 0 + = > c:M:7d * = >:= > = > " + = > #

* " +"G H = # Z T # # " ! # 9 G 9#2" * 9#6H G % H ! # :#6#;# * # $ " " $ % GZ E :RR5H# " " G8 A 6775H# ? T " G :RR4H# % " " " G' :R33T :RR;T 8 ! * :RRRH#

= " G H# * " G* = :RR7H# " ! 1 M & # * # $ M " G H Gf 677;H# * " " # :#6#3# * " = " T " " " G* = :RR7H# :#6#3#:# 8 = * = # * " = # f G:RR5H " = " " ! = ! M # P G:RR3H " = M !

# k = $ ! G* 6777H# ) " = M G 67:7H# = $ = # D " = B " " %" G :RR;T f 677RH# :#6#3#6# G H # * " T GE 6779H# * " $ " ! " # $ B G " H" G " %" $ H GE 6779H# " G H" = G H G) ! :R46H# * #

* " " " M # G * :RR6T = * $ :RRRT ) :RRRT ) 6777T = 6772H# #G6776H ! M T M " # " k # " ! T # :#6#4# 8 # * ! " " G ! 67:7H# " ! " M ! " = " " M M G* = :RR7T E ! :RR7T * :RR2T A 6772T 6774H#

" " G ! H M G " " " H" # ) " G H ! GE ! :RR7T * :RR2H# " B 1 T " " GE ! 6779 H# ! M # % " " ! G ! 67:7H# 8 " M ! " ! " G ! 67:7H# $ GE ! 677:H# :#6#R# 8 " " M

G $ " ! H# % " " # G:RR;H G 6O_" ;O_" 6O_{" D}6O P 6O_H # = M " " " D P M # " ! G :RR3H# 8 ;O G$ " $ # 7 H # ;O GZ 6774H# ) 6O_" 6O_" 6O_" 6O _" M G' 67:7H# 0 = # G6773 H # " ! " % " = ! # * % G 6773 H# 8 = M G D 6773H#

* ! ! " " " # $ $ # :#6#:7# ! 0 # * " # " # " " G H # ' M # * G $ H" G 1 H # G% " %D09 %6 02H" GD 0%" D 6 09H G7#: )H $

G * H G8 6779T = 6772T 6774T 67:7H# D " M # $,/, % M # % " # G H G* ! 6779T ! 6772T * ! 6774T Z 67:7T 67:7H# * " ! = ! G6772H * ! G6774H # ' " G67:7H " ! = # % RR#5 4:#2S = " " ! # Z G67:7H " ! " = $ # " R;#; R:#5S G % :#5H P G % 2#5H

" # $ " ;2S ! $ # % " $ # " " $ G% 677;H# D #

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f # # %# G:R4:H M* N # ! ! $ 21#G6M9H" 44M:63#

8 " )#" 8 & " ># " '# # G6777H M 1' 1R61:67# * 0 $ D 0 = # 8 " # " ># '# G:RR2 H 8 # / $2" ;99M;94# 8 " # " ># '# G:RR2 H 8 M # $ ! 0$G:H" :2RM:52# 8 " # " >#'# G:RR2 H M # ! $2G5H" 529M524# 8 " #" " # " ># '# G:RR2H = $ # ! ! 00" 6R3M976# " #" ' " #" " )#" " ?# 8# E " # G677:H * # ?! "32" :R;3RM:R;46# " # > " f # G6774H = )M l l # $ ! 34" 6R9M6RR# " )# 8#" ? " %# * " ># *# G:RR;H = B # ?) $2" 3RM:7:# " # f " ># ?# G6773H P 6O # ! "#G;H" 234M246# " # f " ># ?# G67:7H ' # ? 0$ $/2G:H" R5M:76# " # %# Z " # N# G6779H = 6O * # ! ! "1G5H" 63:M633# & " #" " )#" " #" " ># E " E# G677RH 8 G H # $0 # $ //1G:M9H" :4:M:44#

" ?# P#" f " A# # > " *# # G67:7H ! k # ? 55G:H" :7RM::5# " # >#" f " # P#" P " # (#" > " # N#" f " N# ># ? " f # A# G6774H ' # - $5G2H" :774M:7:9# " D#" E " '# Z ! " # G6774H 8 M # ) ?! 3G6H" :5RM:;R# " ?# )#" " >#" 8 " l#" N " D# ?#" " # %# P " N# G677RH # ;))< # '88@ ) ) $ "A )"# # # 3:6M3:5# =" # #" * " *# * $ " ># # G6772H G H # ?$#1" ::M67# =" # # * $ " ># # G:RRRH * k G H G H # ! "$G5H" 29:M293# " *# )# G:R46H $ " " = # $ $ $0$" 39MR2# " # ' $" ># # G:RR6H % M M M %# $ ! /3G9H" 9RRM 279# ! " *#" " *#" " Z#" Z & " N#" ) " %# " # G6772H " P D # + + * /#G6H" 663M696# A " A# )# G:RR7H # * # " %# ?# 8 " # ?# # 62RM635# D N !B ) A M% # A " A# )# G:RR9H $ # - # $"0G:H" 65M;7#

A " A# )# G677RH 8 B " # ? ! 50G:H" :9M64# A " )# G6772H ' # 0 D/E"6:RM696# Aj! " N#" m " # An ," # G6775H 8 ! k # ! 42" :79M :7R# A " D#" > " # # 8 & " # # G6779H # $ * 3G6H" 9::M9:R# % " '# #" ? " %# Z#" ? " N# %#" P " ># %#" l " %# ># " ># G677;H 8 # ?2 / $/3G9H" :5;RM:53;# % " )# D# " '# Z# G:RR:H ) M M G H # ?B $/3" 365M392# % " ># ?#" " A# =" # G677;H 8 " # 2 5/G:M2H" :7;M::9# Z " # E " *# G:RR5H M B # ! 1/G9H" :R5M67;# Z " # )#" ) " #" % " Z# 8 " # G6776H * "2G9H" 69RM65;# Z " #" ! " *#" Z " )# ) " %# G67:7H ' " P " D $ # ) ? "G6H" ;6M;;# Z " '#" 8 " D# '#" A 8 " Z# G6774H 8 GE H # ?$/1G9H" 676M674#

Z !" D# G677;H # C '886"# 0 # # 6;3M63;# ! E B ) o ) # ? " A# 8 " %# G677;H # ! * 3#9:3M929# ? " P# N#" f " >#)#" l " '# N " N# G6775H = 9O _{# $ #} $0 ! 2$G2H" 3;:M3;5# ? " N# f ! " # G:RR;H % M = # ?2 / 04" :;5M:3R# ) ! " )#" D " ># % " %# G677RH ' k ? # ) ? 2G6H" :49M:R7# ) " #" " %# )# ' " )# # G6773H G H $ M # & "$0G:M9H" 929M95:# ) " # #" " %# )#" * $ " ># # ' " )# # G:RRRH ' 6O_" 6O 6O _{B %} # & $"0" :93M:22# ) " # # # #" ' " )# # " %# )# G6777H % 6O_" 6O 6O # ! "/" :95M:2:# ) !" 8# ?# #" " D# %#" > " # # " E# ?# G:R46H D3 # ?! $1#" 434M44R# D &" A# )# E ! " 8# G67:7H 0 1 # 2 % 2 # f " ?# Z#" " ># #" % " N# *# " D# Z# # 623M6R:# 8 ' " ?B * o ' #

D " # '# Z " # # G:R33H # ?B 44" 9:3M962# g= " # g= " # G6779H G H" D G H" GE H B # ?2 / ! $##" 6:RM66R# " E# G6774H 8 ! G H ! k B Z " # ! 44G4H" 9:77M9:7R# !" #" N " N# # !" ># )# G67:7H * " " # ! ! $1G:H" 4;M:76# " Z# D " '# G6773H 8 M # 0"G9H" ;65M;94# " Z#" D " '# D ! " '# G6773 H ? M " # ?! $#G:H" R6M:75# " Z#" D " '# D ! " '# G6773 H % # ? ) * 22G4H" ;35M;3R# " (# (#" ? " N# A#" P " A# )#" l " f # %#" N " # # P " >#>#G67:7H 8 G H = M # ? 2 / $33G:M9H" ;3;M;46# ' " # # G:R33H ! $ # ! 24" 33M4:# ' " ?#" " #" ) " )#" " # " # # G67:7H ' p! ! =p # $ ! 51G9H" 3;9M33:# " )# G6772H # + %0D # # 347M344# > f o " #

" # " # G:RR;H # $ ! 01" 634M645# " # E#" !" A#" " # " %# G6776H 6O _B # ! "0G4H" ;;9M;;;# " Z#" ) " D#" Z " #" Z " )#" ) " %# ! " *# G67:7H " = ! # ) ? "G6H" R9MR3# " # " ># '# G:RR;H " # ! $5" :67RM:6:6# " # " ># '# G:RR3H M # # 43G9H" 623M65:# " A# f #" " # # >" ># '# G:R4:H ) B # # $ 0$" 693M625# " )# G:RR2H ! M M # $#G:9H" :32:M:356# " ># %#" N " ># f # Z " # # G:RR4H 6O $ # ! "#G9H" 623M65:# * !" %# %#" " '#" 8 ! " >#" Z " # Z# A " '# G6779H B :# ) # ! $0G;H" 269M29;# * " # )#" " #" " '#" " ?# A#" ) " %#" E ? " %# # ? !" '# G6777H A # " " G ' 6777H# # $ 3"G4H" :259M:237#

* " N# # " A# G6777H = $ # ? ! 31" 52:M52;# * " N# # * " f # Z# G:RR2H ! = M M M # ! 1#G6H" ::9M ::3# * " ># )#" f " # A " A# )# G:RR2H ) M # ?) $/G6H" :6;M:97# * ! " %#" Z " #" ) " D#" Z " )#" ) " %# ! " *# G6774H ! " P " D %6 M M # 3/GRH" :224M :256# * = " )# G:RR7H # # " %# ?# 8 " # ?# # 965M99R# B ) A M% # E " #" " E# " # # G6779H Z 8 ! k # ! 54G9H" 64:M643# E " Z# >#" !$" A#" E " %# $" # '# G6779H N B ! # $ ! 2$" :R3M675# E ! " 8# G:RR7H ! ! # E ! " 8# # 9M5# 8 ' " ?B ' # E ! " 8# G677:H $ M B $ # 2 14" 679M6:;# E ! " 8# G6779 H ! # # E ! " 8# # :3RM:R7# ) M # ? " ( " B 8E $" # E ! " 8# G6779 H , # # E ! " 8# # :79M:67# ) M # ? " ( " B 8E $" # f " %# ?# " # G677;H 8 B # ! $ "0G5H" 263M25:# f " ># ?# G6776H 8 G H # ! /3G4H" 423M457#

f " ># ?# " # G677RH 8 # ! $ "3G6H" :R5M66;# f " # #" ! " f # ># E A " # >#G677:H # # * # ? "5G9H" 92RM952# f " # A " A# )# G:R43H * ! M = # ?B $//" 363M393# f " 8# # " ># '# G:RR5H ) # ! $3GRH" :773M:7:6# f " 8# # " ># '# G:RR;H ' M # ! $5G5H" 59:M59;# P " )# " ># '# G:RR3H M ! # ! $4G:7H" R59MR55#

( ' 6 7 $ !( !" W # * M # * 6O_{" D}6O_{" P} 6O_" 6O 9O_" # 7#5 1 " R5S 5 T M # 25 Q " 47 Q G27SH # * M " " D6O G6 1 HT R:S# D6O _GR;SH# * " ! M # 1 ₈ * RR G6774H 6:73M6::5

",$, -? # * " M # * G " " $ H $ T ! M GE ! :RR7T E ! 677:H# * GE ! :RR7T E E ! 6777T E ! 677:H# $ # N $ G H" $ G8 ! :RR5H# * " # $ G* = :RR7H# M # $ " " ! " M = " ! " ! # = " G H"

G ! H G H GD k :RR2T * 6777T 8 L ! 677:T A & 6772H# 8 = " B " %" = " ! = G* = :RR7T :RR;H# " ! GP ) :RR3T ? = 6772H# * # % G H # # G :RR6H# * " " " M G 6#:H#

B

A

_B

_B

A

A

+ ",$, D N :9;2 G!H# M G=H#

M ! " G H" " G H T G) ! :R46 T :RR6H# N # GE 6779H# ' " 6O _! G $ ! D3H M G 6776H# ! 6O " M M G 6776H# A " " $ ! T " # " GE ! :RR9T E ! ) M :RR5T :RR;T 8 ! * :RRRT ? 677;H" G * :RR6T = * $ :RRRT ) :RRRT 6777T = 6772H" $ ! # ! " " ! # " $ #

",", 6#6#:# " !# * M D N GD N H ::R7" D N ::R5T D N :9;2 ;2;M:8 G) * 1α" 2D:2D" D3: D3" 3: 3H# * D N " Z" $ ;2;M:8 ) # * 2 Q N G HB $ " :7 T " 67 T " 67 T " 67 # M 27 N :77 !# 65 Q ! A ! 0 277 Gf $" ZH" :57 " 62 # N 57 1 : " 6 !" 2S G 1 H M # M 24 # 6#6#6# # " G6777 " 5 H" 97 ) G * H # " " :7 ) c6MGDM H d ) % " % ;#7 ) % # ) % ! $ " " " = G :RRR T :RRR H# "

24 G 6777H# 25" 55 47 Q " # 6#6#9# * " G ) :RR3H# 8 " 65 " 5 1 G H" ) % G:7 )" % ;#7H" 2 ) 6OG H & % ;#7 2 ) 6O# _" ) % G:7 )" % ;#7H 7#6 ) 6O" D6O" P 6O" 6O 9O ! GD09H6" D 6" P 6" 6 GD09H9# G:4 H # " " ! $ G 67 H 97 ) * # G H ;77 # G H # " 97 ) * " 65 # * G:4 H# ! 97 ) * " ;77 G H# * S GE H B

# " " # " ! # * $ # 6#6#2# $ " 2577 " : " G 8 H G57 )" % 3#2H" 5$:73 _{1 #} G H G " " A H 9µ 1 # " !" 67 # $ ? ?)8 G? ) " f = A %" A H %80M :77 9 G $ 8 257M2R7" 5:7 ? 5:5H ? # ? 977 G? ) " % " = H D $ :77 & T ? ) 57M # 6#6#5# 25" 55 47 Q ? 977 # " 67 ! " D $ :77 & # = 977

# * ? ) 57M # * # B A 9 !'_{G:6 c6M6d} ! " # 6#6#;# ! " 25 Q " $ # :6 1 D6O" 6O 9O ! 577 ! :57 " ! " 65 Q # 97 " " # $ " T $ ! G 9SH !# % = G M H ! 277 GD !" *" H" #

",/, 6#9#:# # % G9MRH G :R4RT 6777H# * % % # B G H & % ;#7" G H 2 ) 6O _{G H ) % G % ;#7"} :7 )H 2 ) 6O_T G 6O H# 6#6 G R5S 67 H" 6O_# " B 6#9 ) 6O" :#5 ) D6O 7#4 ) P 6O# % " D N ::R7 D N :9;2 G 6#6M8 H# " " 6O" " R5S 5 G 68 6 H# D N :9;2 6O_{G7#75 )H " D N} ::R7 G7#: )H G 6777H" ! T " D N :9;2 #

0 20 40 60 80 100 0 5 10 15 20 0 20 40 60 80 100 0 5 10 15 20 Se tt le d ce lls (% ) 0 20 40 60 80 100 0 5 10 15 20

Settling time (min)

A

B

D

0 20 40 60 80 100 0 5 10 15 20

Settling time (min)

Se tt le d ce lls ( % )

C

0 20 40 60 80 100 0 5 10 15 20 0 20 40 60 80 100 0 5 10 15 20 Se tt le d ce lls (% ) 0 20 40 60 80 100 0 5 10 15 20

Settling time (min)

A

B

D

0 20 40 60 80 100 0 5 10 15 20

Settling time (min)

Se tt le d ce lls ( % )

C

+ ",", ;2;M:8 G H" D N ::R7 G8H" D N ::R5 G H D N :9;2 G H# N G5 1 H B G[H :7 ) ) " % ;#7" 2 ) 6O_{G HT G\H 6#9} ) 6O_{" :#5 ) D}6O _{7#4 ) P} 6O_{" & % ;#7 G]H & %} ;#7 2 ) 6O_{# E} $ T G ^;H# 6#9#6# * " " B 6O" D6O" P 6O" 6O 9O_{# " 7#6 ) # 6#9} D N :9;2 G

$ M: H# * 6#9#5" " 2" 2#9#: 8 ;" ;#9#: G $ M6H# 0 20 40 60 80 100 0 5 10 15 20

Settling time (min)

Se tt le d ce lls (% ) + ",/, * D N :9;2 # 5 1 ) % G:7 )" % ;#7H" 7#6 ) B G\H 6O_{O D}6O_{T G[H} 6O_{O P} 6O_{T G_H} 6O _O 6O_{T G]H} 6O_O 9O_{T G`H} 6O_{G H# E} $ T G ^;H# * P 6O" 6O 6O D6O _{G 6776H# "} 6O T " 6O _GA 6772H#

* " # * # $ ! " $ # " # " # * # * " = M # 6#9#9# = " # 6 $ :7; 1 G 7#: 1 H G HT 7#65 1 " " 7#5 1 " G 6#2H# * T + / ) ! # G) ! :R46 T :R46 H# " 1 = # !" 7#5 1 M #

0 20 40 60 80 100 0 5 10 15 20

Settling time (min)

Se tt le d ce lls (% ) + ",0, D N :9;2# G\H 7#65" G]H 7#5" G[H :#7" G`H 6#7" G_H 5#7 GaH :7#7 1 " :7 ) ) " % ;#7" 2 ) 6O_{# E} $ T G ^;H# 6#9#2# % N " " G :RR6H# 0 " G* = :RR7H# * " ! # N ! 47 Q 27S G 6#5HT " 25 55 Q G 6#5H# 0 " + / 57M;7 Q " G) :R;2T * 0 :R35H# * !

47Q ! ! # 0 20 40 60 80 100 0 5 10 15 20

Settling time (min)

Se tt le d ce lls (% ) + "#5# ! # N G5 1 H D N :9;2 :7 ) ) " % ;#7" 2 ) 6O_{B G]H G H G_H 25 Q " GaH 55 Q} G\H 47 Q # E $ T G ^;H# N 25 Q 97 ) * G H 6#9#5" 2" 2#9#: 8 ;" ;#9#: G $ M9H# * # G 6776H" 6O _" #

* ! " 47Q G 6#;H" ::5 O 4µ 9 _{59 O 9}µ 9 _{! 47 Q G 6#3H#}

D

A

B

C

D

D

A

B

B

C

C

+ ",2, M ! D N :9;2# b T 8" b 25" 55 47 Q " # 0 20 40 60 80 100 120 140 Live 45ºC 55ºC 80ºC C el l v ol um e ( µµµµ m 3 ) + ",3, ! D N :9;2# E $ # $ " 977 G 25" 55 47 Q H #

& G M H " M G% 6775H# ! 25 Q G 9" 9#9#9" 9#2H# ! 55 Q 47 Q G 6#4H#

A

B

C

A

A

B

C

C

+ ",5, D N :9;2" 47 Q # A # M G H" G8H M G H #

* ! " # #" " G 6#4H# * " " 55 Q 47 Q # * ! 25 Q # * # 6#9#5# ! * 25 Q " " D6O_" 6O 9O_{# *} ! $ ! G* 6#:H# % ( ",$, 8 # & ' B . ! $ ( ! " ( ! / ( ; 7#76 6#7 66 O 6 2#4 O 7#: :#R7 O 7#7R 7#R4 O 7#7R 7#776 :#7 7#63 O 7#76 D D D 7#75 6#7 i 7#; M M M D >EF * ! G " R4149 H G 69;1R4H# F_{) $} ; 94<# D B T # " D6O _{34S G 6#RHT} " D6O

:#R 1 G* 6#:H" G6 1 H" G M 69;1R4H# " D6O G* 6#:H R;S D6O G 6#RH# * " # 0 20 40 60 80 100 1 2 3 Number of batch M et al r em ov al (% ) + ",4, S ! D N :9;2 # :6 1 66 1 D6O_{" 577 ! ! :57 "} 65Q # 97 " " # E $ T G ^2H# ",0, " " D " P " # * R5S 5 # * ! T D N :9;2" !"

V 7#5 1 M # " # 25 Q = # # * " " T R:S D6O_# R;S D6O _# ) " W " ! # * " # " #

" # E# * " ># )# G:RR6H ) ! # $ 15" 9449M 944R# 8 ! " Z# >#" " # * " >#)# G:RR5H ) ! B # $ ! 0/" 53RM542# 8 ! " Z# ># * " ># )# G:RRRH # ?) ! "/" 672M674# 8 L !" *#" E " # #" ) M =" ># )# * $ " ># # G677:H b = # ! "/" :739M:734# " )# 8#" ? " %# * " ># *# G:RR;H = B # ?) $2" 3RM:7:# M 69;1R4 G:RR4H $ lE # E -L # L ' I " < M " :3;1R4# # 9::3M9::4# M " R41491 9 D :RR4 " 0 > ? 997" 751:61:RR4# =" # #" * " *# * $ " ># # G6772H G H # ?$#1" ::M67# =" # # * $ " ># # G:RRRH * k G H G H # ! "$G5H" 29:M293# A & " *#" ) " # A ! " Z# G6772H $ M 6O _{M = #} C ? ! "#" 639M63R# A " # " # E# G6772H 6O # ?) ! $$#" :2:M:25#

% " '# # G6775H $ " # 2 %F $ "38 # " )# *# P# # ;RRM33;# B # ? = " D# Z#" ! " # D#" Z " *# # ) " Z# # G6772H # 2 30" :2RM:5;# ? " N# %#" % " # # >#" " f # # " Z# # G677;H = # ? $ # 44" 394M329# ) " # #" " %# )#" * $ " ># # ' " )# # G:RRRH ' 6O " 6O 6O B % # & $"0" :93M:22# ) " # # # #" ' " )# # " %# )# G6777H % 6O_" 6O 6O # ! "/" :95M:2:# ) !" 8# ?# #" " D# %#" > " # # " E# ?# G:R46 H D3 # ?! $1#" 434M44R# ) !" 8# ?# #" " D# %# " E# ?# G:R46 H ' # ?! $1#" 4R7M4RR# ) " # ># G:R;2H * # ?B /1" ;:M;4# D " # k " *# G:RR2H = B # / $2" 9;5M935# " # " # G:RR;H # $ ! 01" 634M645# " # E#" !" A#" " # " %# )# E# )# G6776H 6O _B # ! "0" ;;9M;;;#

" # E#" " # # '# # " %# )# E# )# G6777H 6MGDM H % # ! $"" 5RM;5# " # E# ) " )# G:RR3H ( # ?) ! $#/" R9MR4# " # E# " ># G6777H k % # ! """ :463M:496# " %# )# E# )#" " # # # ?#" " # # D# E " )# *# # # G:RRR H DM % # $ $ /40" 965M995# " %# )# E# )#" " # # 8 " )# A# '# *# )# G:RRR H DM % $ = B # $$" :9:6M:9:3# " )# G:R4RH N B # 1" 243M2R;# " )# G:RR6H N B # $ # //" :M36# * " D# f # 0 " f # ?# G:R35H # ?) ! 5$" 59M53# * " N# # " A# G6777H = $ # ? ! 31" 52:M52;# * = " )# G:RR7H # # " %# ?# 8 " # ?# # 965M99R# B ) A M% # E " Z# >#" !$" A#" E " %# $" # '# G6779H N B ! # $ ! 2$" :R3M675# E " '# %# # # E ! " 8# G6777H 8 B q) /" :3M62#

E ! " 8# G:RR7H ! ! # E ! " 8# # 9M5# 8 ' " ?B ' # E ! " 8# G677:H $ M B $ # 2 14" 679M6:;# E ! " 8# ) M " %# # G:RR5H 8 # $ ! 0"" 3R3M47;# E ! " 8#" ) " %# % " P# '# G:RR9H # ! ! 0$" 46;M46R# P " # # ) " Z# # G:RR3H ' # * "3" :R5M695#

( ' 6 7 ( " !( * M! 25 Q 6O_{" D}6O _P 6O _{# Z} $ D6O _P 6O _:7 " 6O" 97 ;7 " # P 6O _D6O _# 6O_" # " ! # * ! ! $ " $ M # % M! T ! 25 Q !# $ M # 6_{> ) :7; G677RH :3R6M:472}

/,$, -% = # " " " " $ # * B H $ ! G " % H" T H T " " # ) M M # " " M " $ # " G H 1 # " " $ # * " = GE ! 6779H# 8 # " " GE ! % :RR5T 8 !! :RR4T E E ! 6777T f 677;H# * " = 1 $ GE ! 677:H# ! "

G = * $ :RRRT Z 6775T f 677;H# * # ? " " GD Z :R33T * :RR6T E ! :RR9H# 0 " B " $ GA :RR7T E ! :RR7T A :RR9H# ! " ! " ! " " " " GA :RR7T * :RR6T ? f ! :RR;T ! = :RR3T 8 6772T Aj! 6775H# % " ! 6O _{" G H G8} :RR2 HT " 6O_G * :RR6H 6O _{! G Z 6777H #} !" G ! ! H G! 25 Q H ! " = # * ! " ! " " ! # " $ # " M! #

/,", 9#6#: " !" D N GD N H :9;2 # " M G 6" 6#6#:H# 9#6#6# # 6 G 6#6#6#H# 25 Q # 9#6#9# & G " % rH ;77 * G97 )H # G H #

9#6#2# 9#6#2#:# Z Z 577 ! :77 G2 1 H ) % G:7 ) % ;#7H# ) G " D P H 5 :77 1 6777 1 c GD09H6 P 6d :777 1 cD 6d ! D 6 G) !H# :57 " ! G8 " ) " A H" 65 Q # " G:7 H ! 7#25 µ M = # $ $ T G :SH ! # ! " " ! M " # % M ! 277 GD !" *" H" # * ! " , Gµ 1 H" B , ^ A $ G b H $ :7771G $ H c9M:d " A $ G H" G 1 H " T G H #

9#6#2#6# :77 ! 97 G2 1 H ) % G:7 ) % ;#7H G 5 677 1 ! 5 57 1 = H# ! :57 65 Q # : " 7#25 µ M = # # 9#6#2#9# * ? " : G :#6#5H" ! 25 Q B ,9, ,: ;3 ( ,< c9M6d " , ! Gµ 1 H" , Gµ 1 H $ ! " ? G 1µ HT , Gµ 1 H# 9#6#5# '( 25 Q :$:74 _{1 # :77}µ 65 Q !#

6O_{" :7 ) )} % G % ;#7H 2 1 T # #" ! # 8 5 1 " G :77 µ H ! " N G :77M677 µ H# 9M2 65 Q " T !# 9#6#;# $ T 6 G 6#6#2H# 9#6#3# ? " :$:73 _{1 "} # )6' G " " A H 65 µ)# ! 97 # * " $ %80M:77 G $ 8 927M947" 277 ? 265H ? # 6 G 6#6#2H#

9#6#4# $ 9S G 1 H G ! H 6 :77 # * " G577 " 5 H" 7#: ) G % ;#4H G H $ G6SH G ! H# : :77 " 7#: ) G % ;#4H" # 7#:S G 1 H M?M G H# $ c:7" 67" 97" 57" 37 :77S G 1 Hd :7 # D $ " $ 8 = k # * " ) G 577H $ > 0? >)M95 :5 !E $ :7777 # 9#6#R# ) G 25 Q H" $ " ! = " # ? G6 1 H # " ) % G:7 ) % ;#7H 66#6 1 " 96#9 1 D 6;#6 1 P # :57 : " 65 Q " ! 65 Q G * 'H G8 )8M :52 H# ! :77

Z8 9S G 1 H $ # 2777 ;57 M: f M8 9#72 # 9#6#:7# * $ # ! " # /,/, 9#9#:# + * ! G 6O" D6O P 6OH " % ;#7" B 5 1 :77 1 # * G5 1 H G:77 1 H # ! % ; G% :R46H# Z " ! # 5 1 " :5 " 94 22S # * $ 3#7 4#9 µ 1 " G 9#:M H# 97 " B 2: 57S "

$ 3#; R#; 6O µ ₁ " # " 97 ;7 $ R#: µ 1 " 2RS 6O _{G 9#:M H# $} G H 62 " ! # " ! = ! # ! " $ ! ! :7 G 9#:M8 9#6M H# 5 1 ! " ! 9#3 ;#2 D6O µ ₁ " G 9#:M8HT :77 1 ! " M! 62 42 D6O µ _{1 "} G 9#6M H# = G5 1 H" B M! 9#4 ::µ 1 " " :7 G 9#:M H# = G:77 1 H" c22 24S" $ :26 :;7 P 6O µ 1 " G 9#6M8Hd :7 c27S" $ :64 P 6O µ M: G 9#6M8Hd#

0 2 4 6 8 10 12 14 0 30 60 90 120 150 180 C u 2+ a cc um ul at ed (µµµµ m ol /g d ry w ei gh t b io m as s) . 0 2 4 6 8 10 12 14 0 30 60 90 120 150 180 N i 2+ a cc um ul at ed (µµµµ m ol /g d ry w ei gh t b io m as s) . 0 2 4 6 8 10 12 14 0 30 60 90 120 150 180 Time (min) Z n 2+ a cc um ul at ed (µµµµ m ol /g d ry w ei gh t b io m as s) . A B C 0 2 4 6 8 10 12 14 0 30 60 90 120 150 180 C u 2+ a cc um ul at ed (µµµµ m ol /g d ry w ei gh t b io m as s) . 0 2 4 6 8 10 12 14 0 30 60 90 120 150 180 N i 2+ a cc um ul at ed (µµµµ m ol /g d ry w ei gh t b io m as s) . 0 2 4 6 8 10 12 14 0 30 60 90 120 150 180 Time (min) Z n 2+ a cc um ul at ed (µµµµ m ol /g d ry w ei gh t b io m as s) . A B C + /,$, 6O_{G H" D}6O_{G8H P} 6O_{G H G]H G\H} D N :9;2# :7 ) ) % ;#7 2 1 T 5 1 # $ T G V 2H#

" $ 62 ! = # 0 40 80 120 160 200 0 30 60 90 120 150 180 N i 2+ a cc um ul at ed (µµµµ m ol /g d ry w ei gh t b io m as s) . 0 40 80 120 160 200 0 30 60 90 120 150 180 Time (min) Z n 2+ a cc um ul at ed (µµµµ m ol / g d ry w ei gh t b io m as s) .

A

B

0 40 80 120 160 200 0 30 60 90 120 150 180 N i 2+ a cc um ul at ed (µµµµ m ol /g d ry w ei gh t b io m as s) . 0 40 80 120 160 200 0 30 60 90 120 150 180 Time (min) Z n 2+ a cc um ul at ed (µµµµ m ol / g d ry w ei gh t b io m as s) .

A

B

+ /,", D6O _{G H P} 6O _{G8H G]H G\H} D N :9;2# :7 ) ) % ;#7 2 1 T :77 1 # $ T G V 2H#

9#9#6# , & G 6O" D6O P 6OH ! M D N :9;2" $ $ % G;#7H G65 Q H# ! G 9#: 9#6H" ;7 # ) ! G,H G ,H G H G 9#9H# " D6O _P 6O _! G 9#9M8 9#9M H# % M D6O _P 6O G 9#9M8 9#9M H# 6O_" T " G 9#9M H# = ! " ? $ # P 6O_{" G ^ 7#452H" ?} $ G* 9#:H# * , G $ H D6O _P 6O _{G :3 :2 " H} G* 9#:H#

0 20 40 60 80 100 120 0 100 200 300 400 500 600 700 Ceq (µµµµM Zn2+₎ q (µµµµ m ol Z n 2+ /g d ry w ei gh t b io m as s) . 0 20 40 60 80 100 120 0 100 200 300 400 500 Ceq (µµµµM Cu2+) q (µµµµ m ol C u 2+ /g d ry w ei gh t b io m as s) . 0 20 40 60 80 100 120 0 1000 2000 3000 4000 Ceq (µµµµM Ni2+) q (µµµµ m ol N i 2+ /g d ry w ei gh t b io m as s) . A B C 0 20 40 60 80 100 120 0 100 200 300 400 500 600 700 Ceq (µµµµM Zn2+₎ q (µµµµ m ol Z n 2+ /g d ry w ei gh t b io m as s) . 0 20 40 60 80 100 120 0 100 200 300 400 500 Ceq (µµµµM Cu2+) q (µµµµ m ol C u 2+ /g d ry w ei gh t b io m as s) . 0 20 40 60 80 100 120 0 1000 2000 3000 4000 Ceq (µµµµM Ni2+) q (µµµµ m ol N i 2+ /g d ry w ei gh t b io m as s) . A B C + /,/, 6O_{G H" D}6O_{G8H P} 6O_{G H G]H G\H} D N :9;2# :7 ) ) % ;#7 2 1 # * : #

" B :66 µ 6O_{1 :5;} µ 6O_{1 "} G* 9#:H# " ! B 6O _{VV P} 6O _D6O_{T "} # % ( /,$, ? 6O_{" D}6O _P 6O _# B B B "G :66 :5; 6#: :#9 7#RR6 7#R25 ; "G 4 :92 :: :#3 7#R53 7#RRR H "G _{:6 :;9 5 6#4 7#452 7#RR7} @ * ( Dµµµµ > ' ( E D >µµµµ E 9#9#9# $ $ ! " M # ) G H# $ # T D $ M G% 6775H# 9#2" M 25 Q # * ! B 25 Q ! 65 Q N T " #

A

B

A

A

B

B

+ /,0, D N :9;2 M! 25 Q G H# M G8H# # G :RR:H# G 677:H# " " $ M G 9#5H# * #

A

_B

A

_B

_B

+ /,1, D N :9;2 M! 25 Q )6' G HT B # M G8H#

) ) G )H# D T M G 9#;H# 1 µµµµm 1 µµµµm

A

B

1 µµµµm 1 µµµµm 1 µµµµm 1 µµµµm 1 µµµµm 1 µµµµm

A

B

+ /,2,Scanning electron microscopy of S. cerevisiae NCYC 1364. Live (A) or

heat-killed cells at 45 ºC (B). Arrow: bud car.

) M * ' # * " ! B 3R7 ::47 M: T :677 b :6R7 T :277 :377 M: T 6577 b 9477 M: _{M0% MD%} T " ! 6R77 M: _G8 677:T A 677:T 6779H# *

! G 9#3H# 600 1600 2600 3600 λ λ λ λ (cm-1₎ A bs or ba nc e 34 16 15 41 14 58 14 04 12 40 10 74 29 28 17 42 16 34 28 68 34 21 15 43 14 04 12 42 1082 29 28 ₁₇45 16 34 28 72 14 58

A

B

600 1600 2600 3600 λ λ λ λ (cm-1₎ A bs or ba nc e 34 16 15 41 14 58 14 04 12 40 10 74 29 28 17 42 16 34 28 68 34 21 15 43 14 04 12 42 1082 29 28 ₁₇45 16 34 28 72 14 58 600 1600 2600 3600 λ λ λ λ (cm-1₎ A bs or ba nc e 34 16 15 41 14 58 14 04 12 40 10 74 29 28 17 42 16 34 28 68 34 21 15 43 14 04 12 42 1082 29 28 ₁₇45 16 34 28 72 14 58

A

B

+ /,3,IR spectra of live and dead cells of S. cerevisiae NCYC 1364. Cells heated at

45 ºC (dead cells) (A); live cells (B).

) " ! $ :732 M: G H" βG:h 9H " & G97M25S H# " ! B ! :52: M: G MD DM% H ! :;92 M: _{G ^0 DM} % H# * ! $ 92:; M:" " $ D% G 6779H#

9#9#2# ) '("-'( . '( * 6O" D6O P 6O * ' # 9#4" !# 600 1600 2600 3600 λ λ λ λ (nm) A bs or ba nc e 29 26 16 51 15 41 32 85 ₂₉26 16 49 15 39 12 40 10 80 33 64 32 94 15 41 16 34 29 26 12 42 1074 29 28 34 16 12 40 10 74 16 49 15 39 12 40 1074

A

B

D

C

600 1600 2600 3600 λ λ λ λ (nm) A bs or ba nc e 29 26 16 51 15 41 32 85 ₂₉26 16 49 15 39 12 40 10 80 33 64 32 94 15 41 16 34 29 26 12 42 1074 29 28 34 16 12 40 10 74 16 49 15 39 12 40 1074 600 1600 2600 3600 λ λ λ λ (nm) A bs or ba nc e 29 26 16 51 15 41 32 85 ₂₉26 16 49 15 39 12 40 10 80 33 64 32 94 15 41 16 34 29 26 12 42 1074 29 28 34 16 12 40 10 74 16 49 15 39 12 40 1074

A

B

D

C

+ /,5, ' D N :9;2 $ # G H G HT P 6O_{G8H" D}6O_{G H} 6O _{$ G H#} :7 ) ) % ;#7 6 1 # * B 66#6 6O _{1 " 96#9 D}6O ₁ 6;#6 P 6O _{1 # ;7 65 Q #} " ! 6577M9477 M:G 9#4H $ ! 92:; M:_{" " 99;2" 96R2 9645} M: _" 6O " P 6O D6O" # * bD% b0% #

! 6R64 M: G 9#4H# * " " # " ! " :;92 :52: M:_{G H"} $ :;2R M: G D6O P 6O_{H" :;5:} M: _G 6O_{H :59R} M: _G P 6O_{H # " !} " :732 M:G H" $ :747 M: _G 6O_{H # *} G 9#4H# /,0, * ! # Z # N ! B G H G H" T G H G H ! G8 ! :RR5H# D ! = D N :9;2 :7 T G ' :R;4H" G 9#:H $ G5 1 H G) A

:R49T 6779H# * $ " ! # ) ! " ! ! = # " D6O _P 6O _! M G H G ' :R;4T :RR;H# !" ! G25 Q H G) 6774H# " " " ! M ! G) 6774H# !" G 25 Q H ! # * ! $ ! G( $#H 65 Q % ;#7" D6O P 6O G:3 :2 " H G* 9#:HT 6O_{" ! # *} 25 Q # " " " # * M # * T $ = # * βM # βM βG:h 9H βG:h ;HM ! T βG:h 2HM ! DM # G:#5 M ;SH

# * αM G H" 97M57S G 677:T ? 8 677;HT G8 :RR2 H# G :RR:H# " B G 9#5H# * )" G 9#;H# " ' B ! G 9#3H# M ! " G 9#2H" ! = ! $ M # " 6O _# = GA :RR7H % %0 % 6G :R4:H" " # " # * $ $ " ! G 6779H# !" 5 1 6O _{57S 67} 37S ;7 G $ 8M:H# * " G 9#:H# $ ! G 9#4H# *

$ " " $ G ! " H ! # * # G6779H" D6O# 8 G:RR2H ! " $ $ 6O_{# *} 6O # * ! D N :9;2 B M " " % # * 9#6 # * $ ! D N :9;2 ! * 9#6# " * 9#6 T ! D N :9;2 ! # ! = T $ # * " " G ! H# N ,

5 2 1 80 % ( /,", ! # & % , ( _Dµµµµ > ' ( E % I D E ? ! W 677 2 :; GE ! ) M :RR5H ? W :;7 2 :; GE ! ) M :RR5H W 34 5#5 : G8 !! :RR4H ! R7 ee 6 G8 ) % :RR4H ? ! W :77 ; 62 G M & :RRRH ; W 65 5#5 : G8 !! :RR4H ? ! W :9; ; 62 G M & :RRRH 34R 5 62 Gg= g= 6779H ! W :R2 ;#35 62 G 6779H H W 59 ; : G8 !! :RR4H ! 654 ee 6 G8 ) % :RR4H ? ! W 6R7 5 62 G M & :RRRH I_{* #} II_{D #}

" G 25 Q H ! # * ! 25 Q # * ! ! $ T ! $ M # Z ;7 # " ' " ! # $ " " $ ! #

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n M) " f #" X " ># ' " )# G677:H = # # 0"" 95;RM9547#

8 " )# ) % " # # G:RR4H 8 M # ! $4" 95:M959# " #" ' " #" " )#" " ?# 8# E " # G677:H * # ?! "32" :R;3RM:R;46# =" # # * $ " ># # G:RRRH * k G H G H # ! "$G5H" 29:M293# " A#M # ' " # G:R;4H * P 6O" 6O D6O # ! ! $ $2/" 965M997# A " A# )# G:RR7H # * # " %# ?# 8 " # ?# # 62RM635# D N !B ) A M% # A " A# )# G:RR9H $ # - # $"0G:H" 65M;7# A " #" ! " A# #" 8 " # ) " )# G677:H * ' B $ !M # $43" :3RM:4;# Aj! " N#" m " # An ," # G6775H 8 ! k # ! 42" :79M :7R# % " '# # G6775H $ " # 2 %F $ "38 # " )# *# P# # ;RRM33;# B # % " # G:R46H 0 ) 0# $1) # # # 2R# 0$ # Z " *# N#" !" # Z#" " # N#" Z " %# 8#" Z " N#" Z " # # Z " # ># G6775H # + ? "" R:MR4# ? " A# 8 " %# G677;H # ! * 3#9:3M929#

? " N# f ! " # G:RR;H % M = # ?2 / 04" :;5M:3R# ) " )# #" " )# # #" A " #" " %# )# )# " # E# G6774H ' k B * # ! 44G3H" 6:73M6::5# ) " ># ?# A " A# )# G:R49H P ! $ # ?B $"4" 926:M9265# D " # '# Z " # # G:R33H # ?B 44" 9:3M962# g= " # g= " # G6779H G H" D G H" GE H B # ?2 / ! $##" 6:RM66R# " E#" E " # " # # G6779H 8 ! G H 8 ! k # # ! /5" :94RM:9R5# " ># '# G:RR:H # / $40" 396M395# " # E#" % !" Z# " %# )# E# )# G6779H * $ B # ? 04" 99;M929# " # " ># '# G:RR;H " # ! $5" :67RM:6:6# " A# f #" " # # >" ># '# G:R4:H ) B # # $ 0$" 693M625# " ># %# Z " # # G6777H % 6O 6O # ! "/" 963M96R# E " '# %# # # E ! " 8# G6777H 8 B q) /" :3M62#

E ! " 8# G:RR7H ! ! # E ! " 8# # 9M5# 8 ' " ?B ' # E ! " 8# G677:H $ M B $ # 2 14" 679M6:;# E ! " 8# G6779H # # ! # # 5M:6# ) B 8E $" # E ! " 8# % " P# '# G:RR5H 8 # ! # $$" 695M657# E ! " 8# ) M " %# # G:RR5H 8 # $ ! 0"" 3R3M47;# E ! " 8#" ) " %# % " P# '# G:RR9H # ! ! 0$" 46;M46R# f " %# ?# " # G677;H 8 B # ! $ "0G5H" 263M25:#

( ' 8 7 / !( !" G " " " " H" M " " " ! = $ # " = ! # * " ! G 979 µ)H# ! " M! " # G:6 1 H" ! G92 µΜH " # #M " # * ! 4RS# 3 _{> % = ) :47 G67:7H 923M959}

0,$, -* = M M $ # * G " $ " H $ T " M GE ! 677:H# G 6776 H# ' " " M! 25Q " G) 6774H ! G) 677RH" M # M " M " G ! H " " $ # ! " 1 G 1 H G ! " H" 1 G 6776 T ) 6774H# ) " " " ! # N %" $ G H" " #

# " % T %" G8 :RR2T = * $ :RRRT P 677:T g= g= 6779T E 6779T ) * 6772T D 6773H# " % G " ! H G :R49H G # #" H # 0 " % $ " G 6772H# * $ G " " " H G% :RR:T * :RR9T A :RR9H# % $ G 1 HT " G :RR5T 6776 T 6779H# ' $ T $ " $ ;O 9O _{# *} " GD Z :R33T = * $ :RRRH# " = GN 677:T f 677;T A 677RH# ' G6774H G " " D " P H T G H %' ( " $ # * % 6O" " %

" G% 6775H M G E 6772H $ # % " ! # ' $ M T ! ! G 6779T ) 677RH# !" $ M! ! # * ! $ = " M M # 0,", 2#6#:# " !" D N GD N H :9;2 # M " G 6" 6#6#:H#

2#6#6# # " G6777 " 5 H 97 ) * G) !H # M " 25 Q " # !" G 9H# M! " $ # 2#6#9# & ! G 9" 9#6#9H# 2#6#2# ) / " ) 0 " " # * = " % " $ " G " D " P H" G " " " H # ) G % :RR4H# # * " M M G H G0 R;M7R" 8 " H" Z 3725 % # # D M M G H G " 8 " H ) % 6776 %

# * G B " " $ H" % " = " " % 9#7# # * = " 7#25µ = # i 7#25µ ! 8 6# " Z%₆ 0₂G) !H # * 7#25 µ = # " % # * G* H G H *0 M5777 G = H# * G*0 H * # * M ! 277 GD !" *" H" # :777 1 G) !H 7#5 ) %D09 = # " :77 1 T " # # ;O _{G = H#} * $ ! #

2#6#5# # * G " : 6H" " ! GD09H6" D 6" P 6 GD09H9" ) % G:7 )H" % ;#7# :" G M_{" D0} 9 M ₀ 2 6M_{H # )} % $ " " " " = G :RRR T :RRR H# 2#6#;# ) D (?O G 2#5H G ) 6779H# ) ) D (?O " # " G H " G = * $ :RRRT ) 6772T ) 677RH G) 6772H ) D (?O# * $ G 6O _D6O P 6O_{H * G 9H#}

2#6#3# ! * B # & % ;#7 D 0%# * " 7#25µ = # f " :6 1 " ! 577 !" :57 " 65 Q # 97 " " # Z G 9H 97 # $ " T $ ! G :SH !# % # 2#6#4# * $ " " G ^ ;H#

0,/, 2#9#:# # 0 / * " # " = # " " " G* 2#:HT G M 69;1R4H # G H " # % " # * " " ! # " # #M G M :R42H # * * 2#: ! M # * # * $ G694 EH % G6#7H 9O G $ :R32H" G* 2#:H# * " ! G " = H" " #

% ( 0,$, M = # *

G M :R42H G M 69;1R4H

#

US-EPA Portuguese law

pH (units) 6.0-9.0 2.0 Eh (mV) 238 TC (mg/l) 11.5 TOC (mg/l) 10.7 IC (mg/l) 0.8 Cl- (mg/l) 90.5 F- (mg/l) 12 NO3- (mg/l) 50 7.8 SO42- (mg/l) 2000 374 PO43- (mg/l) 0.08 Cu2+ (mg/l) 3.38 1.0 0.5 (µM) 53.1 16 8 Ni2+ (mg/l) 3.98 2.0 17.8 (µM) 67.8 34 303 Zn2+ (mg/l) 2.61 NS 0.8 (µM) 39.9 NS 12 Total Cr (mg/l) 2.77 2.0 1.2 (µM) 53.3 38 23 Cr6+ (mg/l) 0.1 <0.06 (µM) 2 <1

Wastewater limit discharge criteria _{Industrial effluent}

" * 2#: G6#55 ) 9#4R )" H" " ! G7#979 )H# * ! " " " !" G H " ;O 9O Gf ? 677;H# * 2#: G H " " G $ H ! # 2#9#6# 0 / * " " B ( ZT ( $ Z % # * B %" $ " $ G H" # * " # * ( " " G " " H Gf 677RH# * " $ " " % # 9" M

! G " D P H" !" G) 677RH# " % ;#7 $ % # $ % # N % 5#7 R#7 G8 :RR2H" % 5 M ; 6O_" 6O_" 6O_{" D}6O_{" P} 6O 9O _{G> :RR:T} 8 :RR2T = * $ :RRRT g= g= 6779T ) * 6772H# % ;#7" " G $ " " " $ M H" G $ H G H G) 6772H# " ( # 0 " % ; % G* 2#:H % ; # " c# # 9O _{d" % "} $ G) 6772H" ! # % " % $ G # # D P H $ G) 6772H# * & G H " # * " =

" ! G* 2#6H# " " $ " )6O G #H # " % 6T " G0%H6O ; ,< & G0%H9 * 2#6# $ " " 2#:# % ( 0,", " Z" " ! = " % ;#7" % 5#7e#

Ligand Cr(OH)2+ _Cu2+ _Ni2+ _Zn2+ _Reference

Biomass 3.56 Ferraz et al. (1999)

3.99 3.34 3.59 Machado et al. (2009) Cl- _3.77a) _0.20b) _0.41b) _0.40b) F- _8.86a) _1.80b) _1.40b) _1.30b) NO3- 2.30a) 0.50b) 0.40b) 0.40b) HCO3- _ -3.56c) _ _ 1.80b) _2.10b) _1.50b) SO42- 2.38d) 2.36b) 2.30b) 2.34b) H2PO4- 6.42a) _ _ _ Herrero et al. (2005) log K e " ) ? B H G0%H6O_{O ? O %}O _{? O %60} H ) O ? )?

H 6O_{O % 0} 9M 09O %O H G0%H6O_{O 02}6M _{0% 02} $ " " 2#:# B + Mn+ _MB (s) 1 + + F-MF(aq) 5 6 SO4 2-MSO4 (aq) 3 MNO 3(aq) NO3 - + + C l-MCl (aq) 2 4 H + + M HP_O 4 (a_q) H 2PO 4 -+ + H CO 3 -MC_O 3 (a_q) + H + 7 B + Mn+ _MB (s) 1 + + F-MF(aq) 5 6 SO4 2-MSO4 (aq) 33 MNO 3(aq) NO3 - + + C l-MCl (aq) 22 4 H + + M HP_O 4 (a_q) H 2PO 4 -+ + H CO 3 -MC_O 3 (a_q) + H + 7 44 H + + M HP_O 4 (a_q) H 2PO 4 -+ + H CO 3 -MC_O 3 (a_q) + H + 77 + 0,$# M # ) B ) O_# 8 G HB 8# " $ # N G :H# * G 6 M ;H # * * 2#6 " " ! = " G H # * " " $ # " )% 09" ) " ) )D09 " 6" 9" 2 5" G 2#:H" $ T " 1 ! 1 = G :H 1 1 1 "

# * 2#6 = G H # 0 " ! # * ! G ;H# % " # * :#5 1 " * 2#:" G9327 1 H" 62S " # #" S ! G 2#6HT G932 1 H" ! 95S" ::S G 2#6H# % ;O 9O _# * ! ! G 2#6H" " $ " ;O 9O_{# 8 " !} $ %T $ 637 E % 6#5 ;O 9O G $ :R32H# * G H G* 2#6H"

" $ G 2 3" H" " G 2#:H# 0 20 40 60 80 100 N ic ke l s pe ci es (% ) 3.9x 10-3_M (374 mg/l) 3.9x 10 -2_M (3740 mg/l) Sulphate concentration 0 20 40 60 80 100 N ic ke l s pe ci es (% ) 3.9x 10-3_M (374 mg/l) 3.9x 10 -2_M (3740 mg/l) Sulphate concentration + 0,"# ! :#5 1 D N :9;2# ! G HT ! G HT ! G ! H# * ! G) D (?OHT # " " ! = ! B VV ≅ P V D G Z * 2#6H G = * $ :RRRT ) 677RH# * M $ T " " " G8 :RR2H# * " " Gc8 dH Gs c)dH $ " # % "