Current human activities have led to the pollution of the environment, which has aroused as a global threat. In particular, the inefficient treatment of every-day products in wastewater treatment plants (WWTPs) is an important source of contaminants; especially when untreated sewage sludge and reclaimed water are valorised for agricultural porpoises. The present work describes the development of two bioremediation processes mediated with Trametes versicolor in order to treat different types of sludge. Additionally, one physical post-treatment has been proposed and studied so as to improve the final quality of a WWTP effluent. In both cases, the removal of pharmaceuticals (PhACs) in each stream has been assessed.

1. REMOVAL OF PHARMACEUTICALS
FROM WWTP STREAMS BY
BIOLOGICAL AND PHYSICAL
PROCESSES
Guillem
Llorens
Blanch
November
3th,
2016

2. !
Content
1. Introduc>on
2. Objec>ves
3. Removal
of
pharmaceu>cal
products
by
solid-­‐state
fermenta>on
4. Removal
of
pharmaceu>cals
in
bioslurry
systems
5. Post-­‐treatment
of
WWTP
effluent
6. Concluding
Remarks

3. !
INTRODUCTION

4. !
INTRODUCTION
1. Emerging
Pollutants
2. Bioremedia>on
3. Removal
techniques

5. !
INTRODUCTION
Water
pollu4on
The
introduc>on
of
a
foreign
substance
that
leads
to…
Quality
lost
Threat
for
the
environment
and
the
health
 Common
diseases
of
water
pollu>on:
– Waterborne
diseases
– Metal
poisoning
– Reproduc>ve
altera>ons
– Cancer
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S

6. !
INTRODUCTION
Emerging
Pollutants
(EPs)
 Synthe>c
and
natural
products
 Heterogenic
group
 Not
yet
regulated
 Detected
in
salt-­‐water,
freshwater,
wastewater,
sludges
and
soils
 Few
ng·∙L-­‐1
to
thousands
µg·∙L-­‐1
 Not
clearly
classificated
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S

7. !
INTRODUCTION
EPs
classifica4on
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S
Group
Abbrevia4on
Observa4ons
Pharmaceu>cal
products
PhACs
Prescribed
and
non-­‐prescribed
drugs
&
drug
abuse
substances
Personal
care
products
PCPs
Cosme>c
and
personal
hygiene
products
Endocrine-­‐disrup>ng
chemicals
EDCs
Natural
&
synthe>c
chemicals
Halogenated
compounds
PFCs
Surfactants,
lubricants,
paints
&
fire
retardants
Pharmaceu>cal
and
personal
care
products
PPCPs
PhACs
+
PCPs
Transforma>on
products
TPs
PPCPs
transformed
in
WWTPs

8. !
INTRODUCTION
Fate
and
distribu4on
Different
from
one
country
to
another
 Depends
on
the…
– Produc>on
– Consump>on
 EPs
reach
the
environment
through…
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S
Seasonal
fluctua>ons
Smart,
J.;
9
ways
guys
pee

9. !
INTRODUCTION
EPs
in
WWTPs
Plants
designed
to
remove
organic
maaer,
nutrients,
SS,
metals
and
pathogens
 EPs
goes
through
the
treatment
without
relevant
concentra>on’s
decrease
Why
is
that
happening?
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S

10. !
INTRODUCTION
EPs
in
WWTPs
 Not
tradi>onally
considered
as
pollutants
 Low
concentra>ons
 Wide
heterogenic
group
 Seasonal
fluctua>ons
 Detected
in
all
environmental
compartments
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S

11. !
INTRODUCTION
EPs
in
WWTPs.
Who
does
EPs
affect
the
environment?
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S
.
.
.
EPs
pollu>on
WWTP
Opera>onal
Consump>on
Hydrophobicity
Infiltra>ons
Sludge
Leachates

12. !
INTRODUCTION
EPs
in
WWTPs.
Valorisa4on
of
sewage
sludge
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S
Anaerobic
Diges>on
Biogas
Digestate
Liquid
liquor
Compos>ng
Compost
Land
applica>on
Sludge
40%
–
60%
of
OM
CH4
(60-­‐65%)
+
CO2
(30-­‐35%)
Stable,
nutrients
rich
&
pathogen
free
Nutrients,
structure,
water
infiltra>on,
porosity
&
erosion

13. !
INTRODUCTION
E
M
E
R
G
I
N
G
P
O
L
L
U
T
A
N
T
S
Wastewater
WT
Consump>on
Sludge
Condi>oning
Soil
WWTP
EPs
Hydrophilic
EPs
N
a
t
u
r
a
l
w
a
t
e
r
b
o
d
i
e
s
Hydrophobic
EPs

14. !
INTRODUCTION
Bioremedia4on
Elimina>on
of
pollutants
through
microorganisms
 In
contrast
to
physical
&
chemical
processes:
- Higher
opera>onal
>mes
- Lower
inputs
 Can
be
performed
in
situ
o
ex
situ
 Factors:
energy
source,
environmental
factors,
bioavailability
and
bioac>vity
B
I
O
R
E
M
E
D
I
A
T
I
O
N
,
what
is
it?

15. !
INTRODUCTION
Strategies
B
I
O
R
E
M
E
D
I
A
T
I
O
N
Natural
aSenua4on
Bios4mula4on
Bioaugmenta4on
Who?
Authochthonous
Authochthonous
Foreign
microorganism
What
is
supplied?
Nothing
(monitoring)
Bulking
material,
water,
nutrients
and/or
aera>on
Microorganism
+
bulking
material,
water,
nutrients
and/or
aera>on

16. !
INTRODUCTION
Fungal
remedia4on
1980s:
White-­‐rot
Fungi
(WRF)
- Basidiomycetes
- Filamentous
fungi
- Wood
decomposers
 Mineralisa>on
and
depolymerisa>on
of
lignin:
- Extracellular
enzyma>c
system
(LMEs)
- Intracellular
enzyma>c
system
(Cytochrome
P-­‐450)
B
I
O
R
E
M
E
D
I
A
T
I
O
N

17. !
INTRODUCTION
Trametes
versicolor
 Widely
distributed
in
the
environment
 Able
to
growth
in
solid
&
liquid
cultures
 Extracellular
enzyma>c
system
with
high
red-­‐
ox
capacity:
MnP,
LiP
and
Laccase
 Intracellular
enzyma>c
system:
Cytochrome
P450
B
I
O
R
E
M
E
D
I
A
T
I
O
N

18. !
INTRODUCTION
B
I
O
R
E
M
E
D
I
A
T
I
O
N
Trametes
versicolor
Liquid
matrix
Effluents
treatment
Solid
matrix
Sludge
and
soil
treatment
Synthe>c
dyes
Pharmaceu>cals
and
Personal
Care
Products
(PPCPs)
Endocrine-­‐Disrup>ng
Chemicals
(EDCs)
Polycyclic
Aroma>c
Hydrocarbons
(PAHs)
Brominated
flame
retardants
Chlorinated
solvents
(PCE,
TCE)
Air
pulsed
fluidized
bed
Biopile
(up)
and
bioslurry
(down)

19. !
INTRODUCTION
Removal
techniques
for
EPs
WWTPs
effluents
will
be
used
for
further
applica>ons,
its
necessary
to
remove
EPs
R
E
M
O
V
A
L
T
E
C
H
N
I
Q
U
E
S
 Fungal
bioremedia>on,
due
to
its
performance,
can
by
used:
- For
low
volumes/flows
- Where
is
produced
(in
situ)
or
specific
streams
of
WWTPs
 For
higher
volumes/flows

physical
process

20. !
INTRODUCTION
Removal
techniques
for
EPs
WWTPs
effluents
will
be
used
for
further
applica>ons,
its
necessary
to
remove
EPs
R
E
M
O
V
A
L
T
E
C
H
N
I
Q
U
E
S
Sludge

Water

Water
Slurry-­‐phase
bioreactor
Solid-­‐phase
bioreactor
Liquid
effluent
Adsorp>on

21. !
INTRODUCTION
Slurry-­‐phase
bioreactor
 Also
known
as
bioslurry
 For
pollutants
absorbed
in
solid
par>cles
 Ex
situ,
reactor:
- Suspension
of
a
solid
in
water
(5
–
40%
w/v)
- Agita>on
- Aera>on
 Indigenous
microorganisms
or
inoculated
 Addi>on
of:
nutrients,
neutralising
agents,
surfactants
and/or
co-­‐metabolites
 Monitored
parameters:
temperature,
pH
and
dissolved
oxygen
R
E
M
O
V
A
L
T
E
C
H
N
I
Q
U
E
S

22. !
INTRODUCTION
Solid-­‐phase
bioreactor
 Also
known
as
biopiles
 Ex
situ
or
in
situ
 Engineered
biological
process
in
order
to
mineralise
pollutants
- Sludge
mixed
with
bulking
material
- Complementary
systems:
aera>on,
irriga>on
and
leachate
collector
- Monitored
parameters:
moisture,
pH,
heat,
nutrients
and
oxygen
 Low
inputs
and
maintenance
(cost-­‐effec>ve)
R
E
M
O
V
A
L
T
E
C
H
N
I
Q
U
E
S

23. !
INTRODUCTION
Adsorp4on
techniques
 Removal
of
organic
and
inorganic
pollutants
 In
situ
or
ex
situ
 Adsorbents
can
be
natural
or
synthe>c
 Surface
phenomenon:
liquid
that
contains
a
solute
contacts
a
porous
solid
 Solutes
retained
in
the
porous
by
liquid-­‐solid
intermolecular
aarac>on
forces:
- Electrosta>c
aarac>on
- Chemical
mechanisms
- Physical
mechanisms
R
E
M
O
V
A
L
T
E
C
H
N
I
Q
U
E
S

24. !
OBJECTIVES

25. !
OBJECTIVES
AIM…
to
develop
novel
biological
and
physical
processes
to
remove
PPCPs
in
different
streams
of
WWTPs

26. !
OBJECTIVES
HOW?
 Finding
the
best
condi>ons
to
colonize
sewage
sludge
by
Trametes
versicolor
using
lignocellulosic
substrate
 Assessing
the
strategies
to
remove
PhACs
in
bioslurry
and
biopiles
systems
inoculated
with
Trametes
versicolor
under
non-­‐sterile
condi>ons
 Determining
the
applicability
of
the
anaerobic
diges>on
as
a
valorisa>on
method
for
the
fungal
biomass
 Studdying
the
microbial
communi>es
evolu>on
in
fungal
mediated
biopiles
 Inves>ga>ng
the
u>liza>on
of
low-­‐cost
sorbents
to
adsorb
EPs
as
post-­‐treatment
of
a
WWTP’s
effluent

27. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION

28. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
1. Introduc>on
2. Methodology
3. Results
 Total
drugs
removal
 Microbial
community
evolu>on

29. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
As
previously
men4oned…
 Sludge
is
an
inevitable
waste
of
any
WWTP
 Valorised
in
order
to
improve
agricultural
soils
(sludge
as
amendment)
 Tradi>onal
dewatering
techniques
and
stabiliza>on
methods
do
not
remove
all
EPs
 Must
be
treated
before
its
applica>on
I
N
T
R
O
D
U
C
T
I
O
N

30. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Trea4ng
sewage
sludge
Expensive
&
not
efficient
physicochemical
processes
has
been
proposed
to
remove
EPs
 In
contrast,
fungal
bioremedia>on:
sustainable
&
economical
- Fungi
degrade
a
wide
variety
of
compounds
- Minimum
maintenance
- Low
inputs
I
N
T
R
O
D
U
C
T
I
O
N

31. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Key
role
of
substrate…
 Fungal
colonisa>on
depends
on
it
 Must
act
as
bulking
agent
 Extensively
available
 Locally
produced
Economical
&
sustainable
I
N
T
R
O
D
U
C
T
I
O
N

32. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Agro-­‐industrial
waste
Forestry
by-­‐products
Valorised
waste
What
is
it?
Residues
from
growing,
processing
&
trea>ng
food
Non-­‐commercial
wood
by-­‐products
Green
waste:
food
&
small-­‐size
plant
wastes
Useful
because
of…
OM
(BOD
&
COD)
Structure
&
lignin
Nutrients,
OM
&
lignin
Examples:
Palm
fiber,
seeds
&
shells,
brewery
&
fish
ac>vi>es
Sawmill,
sawdust,
trimmings
&
bark
Compost
When
used
as
substrate…

LMEs

Removals

Structure

LMEs
 Removals

Structure

LMEs
≈
Removals

Structure
Agro-­‐industrial
waste
Forestry
by-­‐products
What
is
it?
Residues
from
growing,
processing
&
trea>ng
food
Non-­‐commercial
wood
by-­‐products
Useful
because
of…
OM
(BOD
&
COD)
Structure
&
lignin
Examples:
Palm
fiber,
seeds
&
shells,
brewery
&
fish
ac>vi>es
Sawmill,
sawdust,
trimmings
&
bark
When
used
as
substrate…

LMEs

Removals

Structure

LMEs
 Removals

Structure
Agro-­‐industrial
waste
What
is
it?
Residues
from
growing,
processing
&
trea>ng
food
Useful
because
of…
OM
(BOD
&
COD)
Examples:
Palm
fiber,
seeds
&
shells,
brewery
&
fish
ac>vi>es
When
used
as
substrate…

LMEs

Removals

Structure
I
N
T
R
O
D
U
C
T
I
O
N

33. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Different
substrates
can
be
used
for
fungal
biopiles…
but
not
all
work
as
bulking
material
 According
to
previous
studies
&
to
Valen>n
et
al.
(2009):
pine
bark
as
substrate
 Bark
of
Pinus
halepensis:
- The
most
common
tree
in
Catalonia
- The
lowest
economical
value
I
N
T
R
O
D
U
C
T
I
O
N

34. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
The
WWTP
of
El
Prat
de
Llobregat
 419,000m3·∙d-­‐1
wastewater
 2,000,000
equivalent
popula>on
I
N
T
R
O
D
U
C
T
I
O
N
Polymers

35. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Aim…
Rodríguez-­‐Rodríguez
(2014)
treated
dry
sludge
in
biopiles
with
a
non-­‐scalable
bulking
material:
wheat
straw
Determine
if
WWTP
sludge
could
be
treated
in
fungal
biopiles
systems
with
inexpensive
lignocellulosic
substrate
I
N
T
R
O
D
U
C
T
I
O
N

36. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
M
E
T
H
O
D
O
L
O
G
Y
PPPCs
at
real
concentra>ons
SUSBTRATE
DRY
SLUDGE
Trametes
versicolor
Total
PPCPs
removal
Laccase
ac>vity
Microbial
community
0
–
22
days
23
–
42
days
Triplicates
&
Sta>s>c
analysis

37. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Biopile
systems
Pre-­‐grown
fungus
onto
sterile
pine
barks
(7d)
+
non-­‐sterile
dry
sewage
sludge
 Enzyma>c
Ac>vity:
oxida>ve
state
of
T.versicolor
 Drugs
concentra>on:
removal
capaci>es
 Microbial
iden>fica>on:
communi>es
dinamics
R
E
S
U
L
T
S

38. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Maximum
enzyma>c
ac>vity
by
day
10:
0.007±0.002
U·∙g-­‐1
 Compared
to
other
substrates:
lower
degradability
 Degrada>on
pathway
involves:
metabolism,
co-­‐metabolism
&
detoxifica>on
mechanisms
R
E
S
U
L
T
S
ENZYMATIC
ACTIVITY

39. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
R
E
S
U
L
T
S
TOTAL
DRUGS
REMOVAL
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F:@G:=H7>$ /'&%$ *$ %'%,$ %..'..$ *$ %'..$ %..'..$ *$ %'..$ %..'..$ *$ %'..$
3*($')*&,
IJK$ )'&)$ *$ .'%/$ -,'8+$ *$ .'-,$ )8'(.$ *$ &'%8$ )8'(.$ *$ &'%8$
K=>?L@4?:6$ ,'.&$ *$ .'+%$ (%'&+$ *$ .'(8$ %+',%$ *$ .'%/$ +('&($ *$ .'+-$
E*B*>,$'5(+#)/$;,#.>,&"/+';)'$/+,+/F'$*.5,;)#)*.,>$(5;,
M7>N:OP4H:@$ (/'+8$ *$ %%'(8$ /.',-$ *$ .'/%$ +-'.($ *$ .'+-$ /&'%($ *$ .'/&$
DG4P<=?G=G:6$ 8.',&$ *$ ('8($ )('.%$ *$ .')($ ,+'--$ *$ .',+$ &(').$ *$ .',-$
!;@&"*#)$*&,>$(5;,
9:G=@4EP=>$ ,+'++$ *$ %.'-/$ -8',/$ *$ .'-($ %&'/&$ *$ .'8.$ (+'-+$ *$ .'(+$
A7PGP=@:67$ +8'+)$ *$ +'/&$ &8'&,$ *$ %'..$ &8'&,$ *$ %'..$ &8'&,$ *$ %'..$
Q@L417G:67$ +%'%-$ *$ )'(&$ )%'+($ *$ .')8$ ,8',&$ *$ .',($ ,+'%/$ *$ .',+$
R=P417G:67$ -8'%($ *$ +'+%$ ),'()$ *$ .'),$ ,&'(8$ *$ .',&$ ),'+,$ *$ .'))$
KP=H45467$ (-'&%$ *$ +'&,$ ,+'..$ *$ .',+$ +(',&$ *$ .'+-$ )('))$ *$ .')-$
S76@=N=1:67$ 8&'/-$ *$ %',8$ ('(-$ *$ .'.($
#
$ %/'/&$ *$ .'%,$
9"J$ +'.+$ *$ .'/&$
#, #,
.'-8$ *$ .'..$
0@=6H=E:67$ -',&$ *$ %'(%$ %..'..$ *$ %'..$ %..'..$ *$ %'..$ %..'..$ *$ %'..$
K4G=@$ -(.',&$ *$ %.('8/$ +/')&$ *$ .'+,$ -&'%)$ *$ .'+8$ //'-+$ *$ .'&/$
#
$T7>4<=@$64G$=??7??75U$N:6=@$346376GP=G:46$V=?$W:XW7P$GW=6$GW7$:6:G:=@$
19
drugs
at
0d
430.79
±
103.26
ng·∙g-­‐1
psychiatric
drugs
8
compounds
(42%)
Amount:
295.79
±
39.74
ng·∙g-­‐1
(69%)
5
drugs
Conjuga>on
process

40. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
R
E
S
U
L
T
S
TOTAL
DRUGS
REMOVAL
Re-­‐inoculated
144.51
±
4.06
ng·∙g-­‐1
66.45
±
0.96
%
Non-­‐re-­‐inoculated
218.92
±
49.71
ng·∙g-­‐1
49.18
±
0.52
%
Control
207.22
±
47.15
ng·∙g-­‐1
51.90
±
0.54
%

41. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Microbial
diversity
in
the
biopiles
system
assessed
by
- PCR-­‐DGGE
fingerprints
- Phylogene>c
affilia>ons
 Inoculated
cultures:
0d,
10d,
22d,
23d
&
42d
 Non-­‐inoculated
cultures:
0d,
22d
&
42d
R
E
S
U
L
T
S
MICROBIAL
COMMUNITY
EVOLUTION

42. PCR-­‐DGGE
fingerprints
of
fungal
popula4on
T.versicolor
T.versicolor

43. PCR-­‐DGGE
fingerprins
of
bacterial
popula4ons
Lysobacter
spp.

44. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
R
E
S
U
L
T
S
MICROBIAL
COMMUNITY
EVOLUTION
Fungal
community
11
different
fungi
detected
 Ini>al
>me
- T.versicolor:
predominant
band
(>99%)
for
inoculated
biopiles
- No
predominant
band
for
non-­‐inoculated
biopiles
 Fungal
community
came
from
substrate
 8
fungi
reported
to
degrade
EPs
 4
fungi
had
mycosta>c
capabili>es
Fungus
Order
Detected
at
4mes
(d)
Acremonium
sp.
Hypocreales
10
Pseudallescheria
ellipsoidea
Microascales
0
Peniophora
cinerea
Russulales
0
Rhodotorula
mucilaginosa
Sporidiobolales
0
Coriolopsis
galica
Polyporales
42

45. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
R
E
S
U
L
T
S
MICROBIAL
COMMUNITY
EVOLUTION
Fungal
community
11
different
fungi
detected
 Ini>al
>me
- T.versicolor:
predominant
band
(>99%)
for
inoculated
biopiles
- No
predominant
band
for
non-­‐inoculated
biopiles
 Fungal
community
came
from
substrate
 8
fungi
reported
to
degrade
EPs
 4
fungi
had
mycosta>c
capabili>es
Fungus
Order
Detected
at
4mes
(d)
Trichosporon
asahii
Tremellales
22
&
42
Wickerhamomyces
anomalus
Saccharomycetales
42
Meyerozyma
guilliermondii
Saccharomycetales
22
&
42
Meyerozyma
sp.
Saccharomycetales
42

46. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Bacterial
community
23
different
bacteria
detected
 Ini>al
>me:
organisms
from
Cloistridiales
order
(obligate
anaerobes)
 Predominant
organism:
Lysobacter
sp.
 First
days:
11
organisms
from
Bacillales
order
 Final
days:
Lysobacter
sp.,
Alcaligenes
sp.,
Salinimicrobium
sp.,
Pedobacter
bauzanensis
&
Brevibacterium
siliguriense
R
E
S
U
L
T
S
MICROBIAL
COMMUNITY
EVOLUTION

47. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
Global
community
Fungal
&
microbial
communi>es
disturbed
by
the
inocula>on
event
 Fungal
community:
- Inoculated:
3
prevalent
species,
1
lignocellulosic
decomposer,
2
with
mycosta>c
abili>es
&
all
with
pollutant
removal
abili>es
- Non-­‐inoculated:
2
prevalent
species,
both
with
inhibitory
systems
&
pollutant
removal
abili>es
 Bacterial
Community:
- Disturbed
adding
bulking
material,
water
&
O2
- Similar
evolu>on
of
inoculated
&
non-­‐inoculated
R
E
S
U
L
T
S
MICROBIAL
COMMUNITY
EVOLUTION

48. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
BY
SOLID-­‐STATE
FERMENTATION
 T.versicolor
improved
the
drugs
removal
from
sewage
sludge
in
biopiles
systems
with
pine
bark
as
substrate
under
non-­‐sterile
condi>ons
 T.versicolor
was
s>ll
in
the
biopiles
(at
least)
un>l
day
23
 Fungus’
re-­‐inocula>on
led
to
improved
removal
rates
 Addi>on
of
bulking
material
and
fungus
inoculum
changed
the
microbial
communi>es
 Similar
evolu>on
for
both
inoculated
and
non-­‐
inoculated
cultures
C
O
N
C
L
U
S
I
O
N
S

49. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS

50. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
1. Introduc>on
2. Methodology
3. Results
 T.versicolor
in
liquid
cultures
 Bioslurry
at
Erlenmeyer
scale
 Bioslurry
at
reactor
scale

51. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Membrane
Biological
Reactor
(MBR)
MBR
system
combines
a
suspended
biomass
reactor
with
a
filtra>on
process
(no
need
for
a
sealer)
 Interes>ng
way
to
improve
exis>ng
WWTPs:
technological
improvement
and
cost
reduc>on
 High
cellular
reten>on
>me
and
high
biomass
concentra>on:
promotes
the
biodegrada>on
of
organic
contaminants
 Two
configura>ons:
- Internal/submerged
configura>on
- External/side-­‐stream
configura>on
I
N
T
R
O
D
U
C
T
I
O
N

52. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
WWTP
of
Terrassa
Designed
to
treat
75,000
m3·∙d-­‐1
of
urban
and
industrial
wastewater
 MBR
with
internal
configura>on:
- Q:
7,200m3·∙d-­‐1
- TSS:
4-­‐5
g·∙L-­‐1
I
N
T
R
O
D
U
C
T
I
O
N

53. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Aim…
Determine
the
capacity
of
Trametes
versicolor
to
remove
drugs
from
raw
MBR
sludge
in
bioslurry
systems
Evaluate
the
valorisa4on
of
fungal
bioslurry’s
solids
in
an
anaerobic
digester
I
N
T
R
O
D
U
C
T
I
O
N

54. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
M
E
T
H
O
D
O
L
O
G
Y
T.v.
in
liquid
cultures
T.v.
in
bioslurry
at
Erlenmeyer
scale
T.v.
in
bioslurry
at
reactor
scale
Growth
&
removal
Laccase
Glucose
HZT
(10ppm)
9
days
Media
effect
&
(non-­‐)sterile
condiNon
Laccase
Glucose
HZT
(10ppm)
10
days
Spiked
drug
Drugs
removal
under
non-­‐
sterile
condiNon
Laccase
PPCPs
μorgs
15
days
Non-­‐spiked
drugs
Bioreactor
Anaerobic
Diges4on

55. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Liquid
cultures
M
E
T
H
O
D
O
L
O
G
Y
100mL
growing
medium
Trametes
versicolor
10ppm
HZT
130rpm
/
25ºC
/
9d
HZT
quan>fica>on
Laccase
ac>vity
Glucose
consump>on
Triplicates
&
Sta>s>c
analysis

56. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Bioslurry
in
Erlenmeyer
M
E
T
H
O
D
O
L
O
G
Y
100mL
Trametes
versicolor
10ppm
HZT
Only
for
spiked
130rpm
/
25ºC
HZT
quan>fica>on
Laccase
ac>vity
Glucose
consump>on
Spiked
Cultures
Defined
Glucose
No
nutrients
(sterile)
No
nutrients
(non-­‐sterile)
Non-­‐spiked
(No
nutrients)
sterile
non-­‐sterile
PPCPs
quan>fica>on
Microbial
analysis
Triplicates
&
Sta>s>c
analysis

57. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Bioslurry
at
reactor
scale
M
E
T
H
O
D
O
L
O
G
Y
Trametes
versicolor
115rpm
/
5d
PPCPs
quan>fica>on
Laccase
ac>vity
5L
raw
sludge
pH
control
Anaerobic
diges>on
Triplicates
&
Sta>s>c
analysis

58. !
Anaerobic
Diges4on:
BMP
Test
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
M
E
T
H
O
D
O
L
O
G
Y
Anaerobic
diges>on
VT:
1L
VW:
0.6L
[VS]:
3gVS·∙L-­‐1
Sta>c
Condi>ons
36ºC
Pressure
increment
Biogas
quan>fica>on
Triplicates
&
Sta>s>c
analysis

59. !
Anaerobic
Diges4on:
Inoculum
selec4on
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
M
E
T
H
O
D
O
L
O
G
Y
VT:
1L
VW:
0.6L
[VS]:
1.5gVS·∙L-­‐1
Sta>c
Condi>ons
36ºC
Pressure
increment
Biogas
quan>fica>on
AD
inoculums
Terrassa
Sabadell
Blanes
4.0
gCOD-­‐VFA·∙L-­‐1
(C2:C3:C4=
73:21:04
gCOD)
2mL·∙L-­‐1
nutrients
pH
=
7
Triplicates
&
Sta>s>c
analysis

60. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Liquid
cultures
Degrada>on
experiments
in
spiked
liquid
medium
cultures
were
carried
out
at
op>mal
T.versicolor
growth
condi>ons
 Assessment
of
fungus
capacity
to
grow
and
remove
spiked
drugs
in
liquid
medium
- Spiked
drug:
Hydrochlorothiazide
(HZT)
R
E
S
U
L
T
S

61. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Liquid
cultures
R
E
S
U
L
T
S
Removed:
45%
Adsorbed:
10%
Degraded:
35%

62. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Spiked
bioslurry
Once
the
removal
capaciPes
of
T.versicolor
in
liquid
medium
were
assessed…
Determine
the
effect
of
media
composi4on
on
the
degrada>on
of
spiked
HZT:
- MBR
sludge
- 3
mediums:
complete,
glucose
&
no-­‐nutrient
- Sterile
&
non-­‐sterile
condi>ons
R
E
S
U
L
T
S

63. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
R
E
S
U
L
T
S
Spiked
bioslurry
Medium

Complete
Glucose
No-­‐nutrient
(sterile)
No-­‐nutrient
(non-­‐sterile)
Raw
sludge
(non-­‐sterile)
Laccase
(max)
322
U·∙L-­‐1
(4d)
198
U·∙L-­‐1
(4d)
331
U·∙L-­‐1
(4d)
0
n.a.
Removal
13.8
%
71.4
%
69.1
%
93.2
%
94.1
%

64. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Spiked
bioslurry
 Medium
affects
the
degrada>on
of
HZT
R
E
S
U
L
T
S
Parameter
Value
pH
5.16
TSS
(g·∙L-­‐1)
3.98
±
0.04
VSS
(g·∙L-­‐1)
2.43
±
0.03
TC
(mg·∙L-­‐1)
181.789
±
4.72
TOC
(mg·∙L-­‐1)
74.348
±
5.20
TAN
(mg·∙L-­‐1)
42.9
±
0.04

65. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Spiked
bioslurry
 Medium
affects
the
degrada>on
of
HZT
 HZT
was
degraded
in
all
experimental
cultures:
- Highest
rate

systems
without
added
nutrients
 No-­‐nutrient
medium
was
selected
for
subsequent
experiments
R
E
S
U
L
T
S

66. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Non-­‐spiked
bioslurry
Once
the
operaPonal
condiPons
were
selected…
Non-­‐spiked
fungal
bioslurry
under
non-­‐sterile
condi>ons
at
Erlenmeyer
scale
 Assessment
of
fungus’
efficiency
to
eliminate
PPCPs
at
real
concentra4ons
 Evaluate
how
the
fungal
inocula>on
would
affect
the
autochthonous
microbial
popula4on
of
the
sludge
R
E
S
U
L
T
S

67. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
R
E
S
U
L
T
S
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LT?4D0:;>0! -6&(,! *! %,(,! .'(.!*!.(6! )5(5!*!.(6! .%(.!*!.(6!
!><&"*#)$*&,=$(4>,
=;92C4<?2:! )+6(5! *! %-(-! +%('!*!,(&! 5.(+!*!,(&! 5)(%*!,(&!
B01C2J2A;10! )--(%! *! .(.! &&('!*!,(-! 5('!*!,(-! #,
E4?230<2:! '5('! *! )(,! %,,(,!*!%(+! 55(,!*!%(+! -'(5!*!%(+!
=US! &&(+! *! )(%! 5+(,!*!,(5! #, #,
#?234>410! -5(%! *! %(-! %,,(,!*!,(5! %,,(,!*!,(5! %,,(,!*!,(5!
KC2132<;10! %5(-! *! %(&! %,,(,!*!,()! %,,(,!*!,()! .+(&!*!,()!
I/)#+J, %,V%6%(6! *! 6.5(%! !
38
PhACs
detected
at
ini>al
>me
(11
BQL)
[10,152
±
574
ng·∙L-­‐1]
5
PhACs
showed
nega>ve
elimina>on
rate

68. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
R
E
S
U
L
T
S
!"#$%#&'()*&#+,
-.*)*#+,&/.&'.)$#)*/.,
0,,12,3.45678
9,
:'%/;#+,<*'+=>,3?9,@
!
!"#$%&'()*+,-()./&)+
$#"*/(/#"-0+
!"#$%&'()*+,"#"1-()./&)+
$#"*/(/#"-0+
2#"1/"#$%&'()*+,"#"1
-()./&)+$#"*/(/#"-0+
A.#+4'>*&>,#.=,#.)*7*.B+#%%#)/$<,=$(4>,
"#$! %&'()! *! +(,! -+()!*!-(%! .'(.!*!-(%! #,
$/0123410! ''(,! *! 5()! 56(,!*!%(%! #, #,
78092:;14</01! ..(5! *! %(,! &.(&!*!.(6! &.(&!*!.(6! &.(&!*!.(6!
=4>0;10! -%(+! *! %(-! -+(&!*!,(%! %,,(,!*!,(%! +%()!*!,(%!
$?4<@</0123410! &(6! *! ,(-! .)(+!*!,(6! .)(+!*!,(6! .)(+!*!,(6!
$;?4A;82:! 6(%! *! ,(&! %,,(,!*!,(-! %,,(,!*!,(-! %,,(,!*!,(-!
A.)*"<C'$)'.>*;',
B2CD2?921! %-6()! *! %(+! -&(.!*!5(.!! ''(-!*!5(.! ''(-!*!5(.!
A.)"'+%*.)*&,
E0F2:;D4C! %,(.! *! ,(6! %,,(,!*!,(6! 5(%!*!,(6! -,(.!*!,(6!
A.)*7DE,
G21;9;>;10! -%(+! *! -(+! %,,(,!*!,()! %,,(,!*!,()! +)(5!*!,()!
F#+&*(%,F"#..'+,G+/&H'$>,
H;C9;230:! 5-('! *! ,(.! %,,(,!*!,(%! %,,(,!*!,(%! %,,(,!*!,(%!
I4?F0?2<2:;C! %'(.! *! ,(-! ,(,!*!%(&! .%(-!*!%(&! +()!*!%(&!
B0?2<2:;C! %&(5! *! ,(-! %,,(,!*!,(6! %,,(,!*!,(6! +,(.!*!,(6!
A.)*@*/)*&>,
=;<?4JC4A28;1! -.)&('! *! ))+(+! '+()!*!%(-! &%(,!*!%(-! 5&(6!*!%(-!
KLM! )+)%(5! *! %.-(&! ',(6!*!,('! &5(6!*!,('! 5%(6!*!,('!
73;9/?4:@8;1! 6+5(.! *! 5-(,! +'(&!*!,(&! +&(.!*!,(&! +)('!*!,(&!
NOM! %6'()! *! )(.! +6(,!*!,(&! +6(.!*!,(&! +)(.!*!,(&!
=C2?;9/?4:@8;1! .6(+! *! %(&! +6(+!*!,(-! %,,(,!*!,(-! 5)(5!*!,(-!
#GO! 5.()! *! '(6! +%(%!*!,(5! %,,(,!*!,(5! .'(.!*!,(5!
A.)*C+#)'+'),=$(4,
=C4<;>?4P0C! %%(%! *! ,(-! %,,(,!*!,(%! %,,(,!*!,(%! %,,(,!*!,(%!
F/.)$#>),%'=*(%,
Q4<?4:;>0! 5+,(6! *! 5-()! 5.(+!*!6(6!! %,,(,!*!6(6! %,,(,!*!6(6!
2*($')*&>,
RS#! 5,.(6! *! +(-! ++(%!*!%(%! -'(%!*!%(%! #,
LT?4D0:;>0! -6&(,! *! %,(,! .'(.!*!.(6! )5(5!*!.(6! .%(.!*!.(6!
!><&"*#)$*&,=$(4>,
=;92C4<?2:! )+6(5! *! %-(-! +%('!*!,(&! 5.(+!*!,(&! 5)(%*!,(&!
B01C2J2A;10! )--(%! *! .(.! &&('!*!,(-! 5('!*!,(-! #,
E4?230<2:! '5('! *! )(,! %,,(,!*!%(+! 55(,!*!%(+! -'(5!*!%(+!
=US! &&(+! *! )(%! 5+(,!*!,(5! #, #,
#?234>410! -5(%! *! %(-! %,,(,!*!,(5! %,,(,!*!,(5! %,,(,!*!,(5!
KC2132<;10! %5(-! *! %(&! %,,(,!*!,()! %,,(,!*!,()! .+(&!*!,()!
I/)#+J, %,V%6%(6! *! 6.5(%! !
3
removal
behaviours
Inoculated
cultures
led
to
lower
drug
concentra>on
(13
PhACs)
Inoculated
and
non-­‐
inoculated
cultures
got
the
same
final
drug
concentra>on
(8
PhACs)
Non-­‐inoculated
cultures
led
to
lower
drug
concentra>on
(2
PhACs)

69. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Non-­‐spiked
bioslurry
at
Erlenmeyer
scale
R
E
S
U
L
T
S
!"#$%#&'(&)"#*
+(#,%*-#,./01
2*
3#)&)(4*!"#$%#&'(&)"#*5*
67*
-#,./01
2*
+%8"9(4*:)%4;<*-=2*
!"#$%&'()*+
,-()./&)0+
!"#$%&'()*+
,1#"2
-()./&)0+
1#"2
/"#$%&'()*+
,1#"2-()./&)0+
3+4+566+ !"!#$%& '& ()(*$)& !#$(& %)$+& ),$(&
566+7+3+4+86+ )"($#& '& "*$-& **$(& !,$(& *,$(&
86+7+3+ ,*($!& '& (#$)& !"$!& -"$%& !"$(&
9#('&+ -(!*$!& '& -##$%& !"$%& %%$-& )#$,&

70. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Non-­‐spiked
bioslurry
R
E
S
U
L
T
S
Fungi
profile
 Ini>al
>me:
two
main
fungal
species:
F3
&
F4
 Final
>me:
microbial
diversity
of
the
mixture
increased
 All
fungal
bands
(except
F5)
correspond
to
unknown
fungi
Bacterial
profile
 More
bands
at
15d
Fungi
Bacteria

71. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Non-­‐spiked
bioslurry
at
reactor
scale
Once
it
was
proved
that
T.versicolor
could
degrade
PPCPs
at
real
concentraPons
from
MBR
sludge
without
external
nutrients
and
under
non-­‐
sterile
condiPons…
The
aim
was
to
eliminate
drugs
from
MBR
sludge
at
reactor
scale
and
use
the
resul4ng
biosolids
as
substrate
of
an
Anaerobic
Diges4on
R
E
S
U
L
T
S

72. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Bioreactor
Performance:
Fungus
ac4vity
R
E
S
U
L
T
S
 Evalua>on
of
the
fungus
state
through
the
laccase
ac>vity
 Maximum:
14.5
U·∙L-­‐1
at
day
2
 No
further
oxida>ve
poten>al
a{er
5d
Non-­‐spiked
bioslurry
at
reactor
scale

73. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Bioreactor
Performance:
PPCPs
removal
R
E
S
U
L
T
S
Non-­‐spiked
bioslurry
at
reactor
scale
Raw
MBR
sludge
Fungal
bioslurry
(5d)

74. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Bioreactor
Performance:
PPCPs
removal
R
E
S
U
L
T
S
Non-­‐spiked
bioslurry
at
reactor
scale
*
Removals
not
assesse
due
to
final
concentra>on
was
higher
than
ini>al
Total
removal:
40.0%

75. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Anaerobic
Bach
Assays:
inoculum
screening
R
E
S
U
L
T
S
Non-­‐spiked
bioslurry
at
reactor
scale
Terrassa
Sabadell
Blanes
!"#$%&%'(
)*+(,$$%'%&,+*-(
'*+.,"*(/'0(1234,
(
5*+.,"#6*"7$(8$+797+:(
;61<=123>6
??@
AB
>-
AB
C(
;0123>6??@
A
B
>-
AB
C!
!"##$%%$& !"#$%&%'"()% ("(()% ("((*(
'$($)"**& +","% ("-')% ("(#(%
+*$,"%& -$-"*!%&%-*"!#% ("()!% ("(*-%
,(
./+01+23,24/+%5,6718%&%82,+9,39%133/3%

76. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Anaerobic
Bach
Assays:
BMP
test
Once
the
inoculums
from
Sabadell’s
and
Blanes’
ADs
were
chosen,
the
biogas
produc>on
of
the
fungal
bioslurry
was
evaluated
R
E
S
U
L
T
S
Non-­‐spiked
bioslurry
at
reactor
scale
Inoculum
+
fungal
bioslurry
of
MBR
E
x
p
e
r
i
m
e
n
t
a
l
Inoculum
+
fungal
biomass
F
u
n
g
a
l
C
o
n
t
r
o
l
Inoculum
+
fungal
biomass
+
raw
MBR
sludge
S
l
u
d
g
e
C
o
n
t
r
o
l
VT:
1L
VW:
0.6L
[VS]:
3gVS·∙L-­‐1

77. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
R
E
S
U
L
T
S
Non-­‐spiked
bioslurry
at
reactor
scale
Sabadell
Blanes
Anaerobic
Bach
Assays:
BMP
test
Fungal
controls
showed
the
higher
biogas
produc>ons
Sludge
controls
and
experimental
cultures
had
similar
net
biogas
produc>on
un>l
day
10
(sta>onary
phase)
Some
inhibitory
product
was
produced
during
the
fungal
bioslurry

78. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
R
E
S
U
L
T
S
Non-­‐spiked
bioslurry
at
reactor
scale
Anaerobic
Bach
Assays:
BMP
test
 The
fungus
alone
(fungal
controls)
produced
high
amounts
of
biogas
 When
MBR
sludge
was
added
(sludge
controls)
the
produc>on
decreased
(increasing
complexity
of
the
matrix)
 The
biogas
produc>on
was
inhibited
or
slowed
with
the
biosolids
from
the
bioslurry
(experimental)
Culture
Sabadell*
Blanes*
Experimental
53.06
±
8.32
61.71
±
19.71
Fungal
Control
289.51
±
19.68
126.30
±
0.00
Sludge
Control
100.35
±
4.16
87.08
±
9.44
*
Net
accumulated
biogas
(mL)

79. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Anaerobic
Bach
Assays:
Total
PPCPs
removal
PhACs
at
real
concentra>on
was
monitored
for
AD
cultures
with
Blanes’
inoculum
in
order
to…
Evaluate
if
AD
process
improved
the
removal
of
drugs
R
E
S
U
L
T
S
Non-­‐spiked
bioslurry
at
reactor
scale

80. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
Anaerobic
Bach
Assays:
Total
PPCPs
removal
R
E
S
U
L
T
S
Non-­‐spiked
bioslurry
at
reactor
scale
!"#$"%&'!
(&)*)+,-
!"&./&*0+*)"&-
1&23456
78
!
9/#":+,-
+;*/0-
8)"<,%00=-1>7.
-
9/#":+,-1>7.
!
!"#$%&!$'()*+,$-$
./$
&01'()223$-$./$
"#$%&'()%*#! +,-+.!/!0-0,! 12-3!/!,-,! 00-4!/!,-,! +1-1!/!,-,-
5%)(%6%$#7'8#! 23-92!/!.-.9! 02-3!/!3-4- +- +
!
5:;7'<);=#:! 4-+.!/!3-33! ,33-3!/!3-3! +-
,33-3!/!3-,$
5;<#'8#! 4>-33!/!.-.9! 10-4!/!3-0! +.-3!/!3-0! 14-,!/!3-0!
?'%$#7%6! >3-4+!/!0-0,! 43-3!/!3-3- +- +
!
@;)%$#7%6! >+-19!/!0-0,! +3-+!/!,-+! 92-3!/!,-2! 19-3!/!,-2!
AB#8%$;8#! 0,-9.!/!0-0,! +- +- +
!
A');C'D%6! 3! +- +- +
!
A)%E%F*%*'8! ,,3-1,!/!0-0,! 4>-2!/!0-.
- +-
,0-,!/!0-.!
GH:&%6#*B;C%$;:#! 12-22!/!9-90! .4->!/!3-1! 42-3!/!3-1! ,33-3!/!3-1!
I;)%F#6'<#! +-3.!/!3-33!
+
! ,33-3!/!3-3! ,33-3!/!3-3!
!"#$%&'()"*$%+& ,-.-&/&-.0& ,1.,&/&-.2& 34.4&/&5.-&
+
!J#6;E%:!8;*!%FF#FF#<K!&'8%:!D;8D#8*)%*';8!L%F!B'=B#)!*B%8!*B#!'8'*'%:!
8
!5;8D#8*)%*';8!E%:H#F!/!F*%8<%)<!<#E'%*';8F!
.
!M));)F!#C7)#FF#<!%F!*B#!<#*#D*';8!:'6'*!;&!#%DB!D;67;H8<!<'E'<#<!(N!0&

81. !
REMOVAL
OF
PHARMACEUTICAL
PRODUCTS
IN
BIOSLURRY
SYSTEMS
 The
low
solids
content
of
the
sludge
makes
difficult
to
assess
whether
the
solid
or
the
liquid
were
treated
 MBR
sludge
can
be
treated
with
T.versicolor
at
laboratory
scale,
removing
a
wide
range
of
emerging
pollutants
 The
fungus
has
grown
under
non-­‐sterile
condi>ons
without
any
extra
nutrients
 Fungal
treatment
of
MBR
sludge
has
been
proved
as
an
adequate
pre-­‐treatment
prior
anaerobic
diges>on,
although
it
slowed
the
AD
process
C
O
N
C
L
U
S
I
O
N
S

82. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT

83. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
1. Introduc>on
2. Methodology
3. Results
 Substrate
Selec>on
 Total
drugs
removal
 Microbial
community
evolu>on

84. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Global
popula>on
increase
leads
to
a
growing
demand
of
water
 Groundwater
is
an
important
natural
resource
that
can
be
use
to
supply
water
for
municipal,
agricultural,
and
industrial
purposes
 Ar>ficial
recharge
methods
are
faster
than
natural
systems:
- Direct
aquifer
injec>on
systems:
to
put
water
directly
into
the
underground
water
basins
- Surface
spreading
recharge
systems:
to
replenish
aquifers
by
infiltra>on
I
N
T
R
O
D
U
C
T
I
O
N

85. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Soil-­‐Aquifer
Treatment
(SAT)
Surface
spreading:
reclaimed
wastewater
is
intermiaently
introduced
into
spreading
basins
 Water
percolates
across
the
ground
and
throughout
the
aquifer
 Water
quality
improvement
thanks
to
physical,
chemical,
and
biological
natural
processes
I
N
T
R
O
D
U
C
T
I
O
N

86. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Soil-­‐Aquifer
Treatment
(SAT)
I
N
T
R
O
D
U
C
T
I
O
N
Miotlinski
et
al.
2010

87. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
I
N
T
R
O
D
U
C
T
I
O
N
!
Alice
Springs
SAT
 Arid
Zone
Research
Ins>tute
(Alice
Springs,
NT,
Australia)
 Aquifer
recharge:
600ML·∙y-­‐1
 Recharge
area:
10,269m3
(5
basins)
 Infiltra>on
rate:
240mm·∙d-­‐1

88. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Adsorp4on
of
pollutants
The
mass
transfer
of
a
substance
from
a
liquid
to
a
solid’s
surface
 Ac>vated
carbon
is
the
universal
adsorbent,
but
it
is
expensive
 Local
materials
u>lized
as
inexpensive
sorbents:
Low-­‐cost
sorbents
(low
processing
&
abundant)
- NUA:
neutralised
used
acid
from
heavy
mineral
processing
- BIOCHAR:
charcoal
produced
by
the
pyrolysis
of
biomass
(eucalyptus)
I
N
T
R
O
D
U
C
T
I
O
N

89. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Aim…
Determine
if
it
was
possible
to
treat
a
WWTP
effluent
with
low-­‐cost
sorbents
in
order
to
use
it
in
a
soil-­‐aquifer
treatment
I
N
T
R
O
D
U
C
T
I
O
N

90. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
M
E
T
H
O
D
O
L
O
G
Y
Soil’s
adsorp>on
capacity
Amendment
ra>o
Removal
of
PPCPs
PhACs
adsorpNon
24
hours
Soil:
amendment
raNo
Adsorp>on
24
hours
Spiked
drugs
removal
Adsorp>on
21
days

91. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Soils
adsorp4on
capacity
&
Soil:amendment
ra4o
M
E
T
H
O
D
O
L
O
G
Y
Soil
from
Alice
Springs’
SAT
(basin
E)
0.1M
CaCl2
10rpm
12h
PhACs
10rpm
/
24h
PhACs
quan>fica>on
Adsorp>on
NUA
Biochar
Ra4os:
0.1,
0.5,
1,
2
&
5%
w/w
Triplicates
&
Sta>s>c
analysis

92. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Removal
in
amended
soils
M
E
T
H
O
D
O
L
O
G
Y
Soil
from
Alice
Springs’
SAT
(basin
E)
0.1M
CaCl2
10rpm
12h
PhACs
21d
PhACs
quan>fica>on
Removal
NUA
Biochar
1%
w/w
Triplicates
&
Sta>s>c
analysis

93. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Soil’s
adsorp4on
Determine
soil’s
natural
adsorp4on
capacity
of
PPCPs
under
experimental
condi>ons
 Spiked
drugs
selected
according
to
reclaimed
wastewater
characterisa>on
studies:

Ibuprofen

Carbamazipine

Sulfamethoxazole

Propranolol

Ketoprofen

Trimethoprim

Ofloxacin
R
E
S
U
L
T
S

94. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Soil’s
adsorp4on
 Kd:
equilibrium
constant
isoterm
of
adsorp>on
- Linear
rela>onship
between
sorbed
&
non-­‐sorbed
spices
- Aaenua>on
mechanisms
&
environmental
factors
are
considered
R
E
S
U
L
T
S

95. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Soil’s
adsorp4on
R
E
S
U
L
T
S
Compound
Kd
(mL·∙g-­‐1)
±
SD
Ketoprofen
1.21
±
0.03
Ibuprofen
4.24
±
0.02
Carbamazepine
2.25
±
0.05
Sulfamethoxazole
4.38
±
0.03
Propranolol
22.88
±
0.01
Trimethoprim
14.46
±
0.02
Ofloxacin
2487.9
±
0.4

96. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Selec4on
of
the
amendment
ra4o
Once
the
natural
adsorpPon
of
the
soil
was
determined…
Removal
of
PPCPs
was
studied
for
both
Biochar
and
NUA,
in
order
to
establish
the
best
ra4o
of
soil:amendment
 Ra>os:
0.1,
0.5,
1,
2
&
5
%
 Three
PhACs:
Sulfamethoxazole,
propranolol
&
trimethoprim
R
E
S
U
L
T
S

97. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
R
E
S
U
L
T
S
Trimethoprim
RaNo
(%)
NUA
+
Sand
NUA
+
Soil
Biochar
+
Sand
Biochar
+
Soil
0.1
2.92
±
0.44
13.42
±
0.35
0
3.12
±
0.17
0.5
*
10.37
±
0.33
*
1.31
±
0.58
1
0.37
±
0.30
13.42
±
0.23
0
2.09
±
0.11
2
0.88
±
0.39
9.45
±
0.09
0.43
±
0.32
1.46
±
0.49
5
0.48
±
0.21
11.60
±
0.03
*
1.83
±
0.10
*Kd
value
not
assessed,
measured
concentra>on
was
higher
than
the
ini>al
[mL·∙g-­‐1]
(±
standard
error)
Selec4on
of
the
amendment
ra4o

98. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
R
E
S
U
L
T
S
Propranolol
RaNo
(%)
NUA
+
Sand
NUA
+
Soil
Biochar
+
Sand
Biochar
+
Soil
0.1
2.69
±
0.04
18.44
±
0.12
8.74
±
0.30
14.06
±
0.27
0.5
5.23
±
0.04
18.38
±
0.03
11.48
±
1.02
24.52
±
0.27
1
3.76
±
0.29
17.53
±
0.04
17.11
±
0.04
15.81
±
0.07
2
5.47
±
0.13
17.68
±
0.10
39.39
±
0.13
28.54
±
0.14
5
9.59
±
0.29
40.17
±
0.04
39.81
±
0.31
33.42
±
0.07
[mL·∙g-­‐1]
(±
standard
error)
Selec4on
of
the
amendment
ra4o

99. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
R
E
S
U
L
T
S
Sulfamethoxazole
RaNo
(%)
NUA
+
Sand
NUA
+
Soil
Biochar
+
Sand
Biochar
+
Soil
0.1
*
4.83
±
0.09
34.70
±
0.08
29.62
±
0.02
0.5
*
3.76
±
0.07
36.97
±
0.00
35.80
±
0.01
1
*
4.53
±
0.00
38.85
±
0.04
44.88
±
0.03
2
*
4.63
±
0.04
56.30
±
0.04
68.35
±
0.04
5
0
4.55
±
0.04
181.74
±
0.14
181.04
±
0.14
*Kd
value
not
assessed,
measured
concentra>on
was
higher
than
the
ini>al
[mL·∙g-­‐1]
(±
standard
error)
Selec4on
of
the
amendment
ra4o

100. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Selec4on
of
the
amendment
ra4o
 Propranolol
and
trimethoprim
beaer
removed
with
biochar
 Sulfamethoxazole
beaer
removed
with
NUA
 Ra>o
of
1%
selected
for
further
experiments:
- All
the
tested
compounds
were
well
retained
- Higher
ra>o

Higher
opera>onal
costs
R
E
S
U
L
T
S

101. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Removal
of
PhACs
from
amended
soil
Once
the
amendment
raPo
was
selected…
The
removal
capacity
of
amended
soil
with
biochar
and
NUA
at
1%
was
determined
 Selected
PhACs:

Ofloxacin

Carbamazipine

Sulfamethoxazole

Propranolol

Ketoprofen

Trimethoprim

Ibuprofen
R
E
S
U
L
T
S
Constant
fluctuaNons
with
high
errors

102. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Removal
of
PhACs
from
amended
soil
R
E
S
U
L
T
S
Plots
a:
Biochar
Soil
Plots
b:
NUA
Sand
90.7%
77.7%
80.1%
77.8%

103. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Removal
of
PhACs
from
amended
soil
R
E
S
U
L
T
S
Plots
a:
Biochar
Soil
Plots
b:
NUA
Sand
100%
100%
97.6%
85.6%

104. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Removal
of
PhACs
from
amended
soil
R
E
S
U
L
T
S
Plots
a:
Biochar
Soil
Plots
b:
NUA
Sand
97.4%
91.4%
51.1%
51.7%
32.5%
30.6%

105. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
Removal
of
PhACs
from
amended
soil
In
general,
higher
removals
with
biochar,
but
no
sta>s>cal
differences
with
NUA
Biochar:
86.2%
NUA:
80.7%
 Removal
order
- Biochar:
OFX
>
TRM
=
PRN
>
CBZ
>
KTP
>
SMX
- NUA:
OFX
>
TRM
>
PRN
>
CBZ
=
KTP
>
SMX
R
E
S
U
L
T
S

106. !
POST-­‐TREATMENT
OF
WWTP
EFFLUENT
 Soil
could
not
adsorb
the
total
amount
of
PhACs
 High
removal
rates
for
both
NUA
and
biochar
as
amendments
a{er
21d
of
treatment
 A
ra>o
of
1%
(w/w)
of
amendment
was
enough
to
remove
the
selected
compounds
C
O
N
C
L
U
S
I
O
N
S

107. !
CONCLUDING
REMARKS

108. !
CONCLUDING
REMARKS
It
has
been
seen
that…
 Fungal
biopiles
can
be
made
of
forestry
by-­‐
products
to
treat
thermal
dried
sludge,
with
the
fungus
surviving
more
than
22d
and
accelera>ng
the
switch
of
the
microbial
popula>on
 MBR
sludge
can
be
treated
in
a
fungal
bioslurry
system,
and
the
resul>ng
biomass
can
be
energe>cally
valorised
 Low-­‐cost
sorbents
can
be
applyed
into
soil
in
order
to
treat
reclaimed
water
as
part
of
a
soil-­‐
aquifer
treatment

109. !
CONCLUDING
REMARKS
It
has
been
demonstrated
that…
 Different
WWTP’s
streams
can
be
biologically
and
physically
treated
in
order
to
remove
PhACs
 Fungal
mediated
systems,
both
liquid
and
solid,
enhanced
the
removal
of
PhACs
 Low-­‐cost
sorbents
can
improve
the
final
effluent
of
a
WWTP
by
removing
PhACs

110. !
CONCLUDING
REMARKS
Further
research
should
be
done
in
order
to…
 Increase
the
scale
of
the
biopiles
 Select
another
sludge
(rather
than
MBR
sludge)
 Study
the
PPCPs
adsorp>on
in
con>nuous
mode
and
with
real
reclaimed
water
 Search
new
ways
to
valorise
the
biomass
from
the
bioremedia>on
processess
 Establish
the
environmental
an
economical
feasibility
of
the
treatments

111. REMOVAL OF PHARMACEUTICALS
FROM WWTP STREAMS BY
BIOLOGICAL AND PHYSICAL
PROCESSES
Thanks
for
your
aaen>on!