Removal of pharmaceuticals from WWTP streams by biological and physical processes

REMOVAL OF PHARMACEUTICALS FROM WWTP STREAMS BY BIOLOGICAL AND PHYSICAL

PROCESSES

Guillem Llorens Blanch November 3th, 2016

! 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

!

INTRODUCTION

!

INTRODUCTION

1. Emerging Pollutants 2. Bioremedia>on 3. Removal techniques

!

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

EMERGING POLLUTANTS

!

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

EMERGING POLLUTANTS

!

INTRODUCTION

EPs classifica4on EMERGING POLLUTANTS

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

!

INTRODUCTION

Fate and distribu4on Different from one country to another  Depends on the… – Produc>on – Consump>on

 EPs reach the environment through…

EMERGING POLLUTANTS

Seasonal fluctua>ons

Smart, J.; 9 ways guys pee

!

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?

EMERGING POLLUTANTS

!

INTRODUCTION

EPs in WWTPs  Not tradi>onally considered as pollutants  Low concentra>ons  Wide heterogenic group  Seasonal fluctua>ons  Detected in all environmental compartments

EMERGING POLLUTANTS

!

INTRODUCTION

EPs in WWTPs. Who does EPs affect the environment? EMERGING POLLUTANTS

.

. .

EPs pollu>on

WWTP Opera>onal

Consump>on

Hydrophobicity

Infiltra>ons

Sludge

Leachates

!

INTRODUCTION

EPs in WWTPs. Valorisa4on of sewage sludge EMERGING POLLUTANTS

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

!

INTRODUCTION

EMERGING POLLUTANTS

Wastewater

WT

Consump>on

Sludge Condi>oning

Soil

WWTP EPs

Hydrophilic EPs

Natural

water bodies

Hydrophobic EPs

!

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

BIOREMEDIATION

, what is it?

!

INTRODUCTION

Strategies BIOREMEDIATION

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

!

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)

BIOREMEDIATION

!

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

BIOREMEDIATION

!

INTRODUCTION

BIOREMEDIATION

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)

!

INTRODUCTION

Removal techniques for EPs WWTPs effluents will be used for further applica>ons, its necessary to remove EPs

REMOVAL TECHNIQUES

  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

!

INTRODUCTION

Removal techniques for EPs WWTPs effluents will be used for further applica>ons, its necessary to remove EPs

REMOVAL TECHNIQUES

Sludge

Water

Water

Slurry-‐phase bioreactor

Solid-‐phase bioreactor

Liquid effluent Adsorp>on

!

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

REMOVAL TECHNIQUES

!

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)

REMOVAL TECHNIQUES

!

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

REMOVAL TECHNIQUES

!

OBJECTIVES

!

OBJECTIVES

AIM…

to develop novel biological and physical processes to remove PPCPs in different streams

of WWTPs

!

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

! REMOVAL OF PHARMACEUTICAL PRODUCTS BY SOLID-‐STATE FERMENTATION

!

REMOVAL OF PHARMACEUTICAL PRODUCTS BY SOLID-‐STATE FERMENTATION

1.  Introduc>on 2.  Methodology 3.  Results

 Total drugs removal  Microbial community evolu>on

!


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

INTRODUCTION

!


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

INTRODUCTION

!


Key role of substrate…  Fungal colonisa>on depends on it  Must act as bulking agent  Extensively available  Locally produced

Economical & sustainable

INTRODUCTION

!


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


Forestry by-‐products



Non-‐commercial wood by-‐products

Useful because of… OM (BOD & COD) Structure & lignin


ac>vi>es

Sawmill, sawdust, trimmings & bark



LMEs   Removals Structure




Useful because of… OM (BOD & COD)


ac>vi>es



INTRODUCTION

!


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

INTRODUCTION

!


The WWTP of El Prat de Llobregat  419,000m3·∙d-‐1 wastewater  2,000,000 equivalent popula>on

INTRODUCTION

Polymers

!


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

INTRODUCTION

!


METHODOLOGY

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

!


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

RESULTS

!


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

RESULTS

ENZYMATIC ACTIVITY

!


RESULTS

TOTAL DRUGS REMOVAL

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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

!


RESULTS

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 %

!


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

RESULTS

MICROBIAL COMMUNITY EVOLUTION

PCR-‐DGGE fingerprints of fungal popula4on

T.versicolor T.versicolor

PCR-‐DGGE fingerprins of bacterial popula4ons

Lysobacter spp.

!


RESULTS


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

!


RESULTS


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

!


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

RESULTS


!


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

RESULTS


!


 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

CONCLUSIONS

! REMOVAL OF PHARMACEUTICAL PRODUCTS IN BIOSLURRY SYSTEMS

!

REMOVAL OF PHARMACEUTICAL PRODUCTS IN BIOSLURRY SYSTEMS


 T.versicolor in liquid cultures  Bioslurry at Erlenmeyer scale  Bioslurry at reactor scale

!


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

INTRODUCTION

!


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

INTRODUCTION

!


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

INTRODUCTION

!


METHODOLOGY

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

!


Liquid cultures

METHODOLOGY

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

!


Bioslurry in Erlenmeyer

METHODOLOGY

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

!


Bioslurry at reactor scale

METHODOLOGY

Trametes versicolor 115rpm / 5d

PPCPs quan>fica>on

Laccase ac>vity

5L raw sludge

pH control

Anaerobic diges>on Triplicates

& Sta>s>c analysis

!

Anaerobic Diges4on: BMP Test


METHODOLOGY 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

!

Anaerobic Diges4on: Inoculum selec4on


METHODOLOGY 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

!


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)

RESULTS

!


Liquid cultures

RESULTS

Removed: 45% Adsorbed: 10% Degraded: 35%

!


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

RESULTS

!


RESULTS

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 %

!


Spiked bioslurry  Medium affects the degrada>on of HZT

RESULTS

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

!


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

RESULTS

!


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

RESULTS

!


RESULTS

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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(%! !

38 PhACs detected at ini>al >me (11 BQL)

[10,152 ± 574 ng·∙L-‐1] 5 PhACs showed

nega>ve elimina>on rate

!


RESULTS

!"#$%#&'()*&#+,-.*)*#+,&/.&'.)$#)*/.,

0,,12,3.456789,

:'%/;#+,<*'+=>,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)

!


Non-‐spiked bioslurry at Erlenmeyer scale

RESULTS

!"#$%#&'(&)"#*+(#,%*-#,./012*

3#)&)(4*!"#$%#&'(&)"#*5*67*

-#,./012*

+%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+ ,*($!& '& (#$)& !"$!& -"$%& !"$(&9#('&+ -(!*$!& '& -##$%& !"$%& %%$-& )#$,&

!


Non-‐spiked bioslurry

RESULTS

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

!


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

RESULTS

!


Bioreactor Performance: Fungus ac4vity

RESULTS

 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

!


Bioreactor Performance: PPCPs removal

RESULTS


Raw MBR sludge

Fungal bioslurry (5d)

!


Bioreactor Performance: PPCPs removal

RESULTS


* Removals not assesse due to final concentra>on was higher than ini>al

Total removal: 40.0%

!


Anaerobic Bach Assays: inoculum screening

RESULTS


Terrassa Sabadell

Blanes !"#$%&%'( )*+(,$$%'%&,+*-(

'*+.,"*(/'0(1234,(

5*+.,"#6*"7$(8$+797+:(;61<=123>6??@AB>-ABC(

;0123>6??@A

B>-ABC!!"##$%%$& !"#$%&%'"()% ("(()% ("((*(

'$($)"**& +","% ("-')% ("(#(%+*$,"%& -$-"*!%&%-*"!#% ("()!% ("(*-%

,(./+01+23,24/+%5,6718%&%82,+9,39%133/3%

!


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

ESULTS


Inoculum + fungal

bioslurry of MBR

Experimental Inoculum +

fungal biomass

Fungal Control

Inoculum + fungal

biomass + raw MBR sludge

Sludge Control

VT: 1L VW: 0.6L

[VS]: 3gVS·∙L-‐1

!


RESULTS


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

!


RESULTS


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)

!


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

RESULTS


!


Anaerobic Bach Assays: Total PPCPs removal

RESULTS


!"#$"%&'!(&)*)+,-

!"&./&*0+*)"&-1&2345678!

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&

!


 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

CONCLUSIONS

!

POST-‐TREATMENT OF WWTP EFFLUENT

!POST-‐TREATMENT OF WWTP EFFLUENT


 Substrate Selec>on  Total drugs removal  Microbial community evolu>on


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

INTRODUCTION


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

INTRODUCTION


Soil-‐Aquifer Treatment (SAT)

INTRODUCTION

Miotlinski et al. 2010


INTRODUCTION

!

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


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)

INTRODUCTION


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

INTRODUCTION


METHODOLOGY

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


Soils adsorp4on capacity & Soil:amendment ra4o

METHODOLOGY

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


Removal in amended soils

METHODOLOGY

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


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

RESULTS


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

RESULTS


Soil’s adsorp4on

RESULTS

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


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

RESULTS


RESULTS

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


RESULTS

Propranolol


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




RESULTS

Sulfamethoxazole


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




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

RESULTS


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

RESULTS

Constant fluctuaNons with high errors


Removal of PhACs from amended soil

RESULTS

Plots a: Biochar Soil Plots b: NUA Sand

90.7% 77.7% 80.1% 77.8%



RESULTS


100% 100% 97.6% 85.6%



RESULTS


97.4% 91.4%

51.1% 51.7%

32.5% 30.6%


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

RESULTS


 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

CONCLUSIONS

!

CONCLUDING REMARKS

!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

!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

!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

REMOVAL OF PHARMACEUTICALS FROM WWTP STREAMS BY BIOLOGICAL AND PHYSICAL

PROCESSES

Thanks for your aaen>on!

Environment

Removal of pharmaceuticals from WWTP streams by biological and physical processes