Treatment of simulated textile wastewater with aerobic granular sludge

100 Pages·2016·2.64 MB·English

Treatment of simulated textile wastewater with aerobic
granular sludge: efficiency and detoxification potential

João Pedro Viana da Silva

Master in Biotechnology

Supervisor: Dra. Ana Cristina Anjinho Madeira Viegas and Dra. Nídia Dana Mariano
Lourenço de Almeida

Examination Committee
Chairperson: Dra. Leonilde de Fátima Morais Moreira
Supervisor: Dra. Ana Cristina Anjinho Madeira Viegas
Member of the Committee: Dra. Helena Maria Rodrigues Vasconcelos Pinheiro

November 2016
Acknowledges:

A very special thanks to both Dra. Ana Cristina Anjinho Madeira Viegas and Dra. Nídia Dana Mariano
Lourenço de Almeida for the opportunity to perform this work, the patience and the support
demonstrated towards me.
Also, must be thanked the FCT – Fundação para a Ciência e a Tecnologia for partially financing this
work through the UID/BIO/04565/2013 project.
And lastly but no less important, to my friends and family who have given me strength to live and work
with both energy and a smile in my heart.

“You may have a fresh start any moment you choose, for this thing that we call 'failure' is not the falling
down, but the staying down.” Mary Pickford
“Then there is a still higher type of courage - the courage to brave pain, to live with it, to never let
others know of it and to still find joy in life; to wake up in the morning with an enthusiasm for the day
ahead.” Howard Cosell

i
Abstract
:

Textile wastewaters are resistant to aerobic treatment and characterised by high organic loads and dyes
with a relevant environmental impact. Previous works have reported a successful biodecolourization of
textile wastewaters containing azo dyes, by using aerobic granular sludge (AGS) technology in
sequencing batch reactors (SBR). The present study compares the performance, at both efficiency and
detoxification levels, of two AGS SBRs with different hydrodynamic regimens in the treatment of
simulated textile wastewater containing the azo dye acid red 14 (AR14). Similar colour and chemical
oxygen demand (COD) removal yields (80%) were obtained for both SBRs, yet higher sludge retention
time was achieved in the slower feeding rate, SBR2. Under dye and organic loading shocks, both SBRs
maintained high colour removal yields, while COD removal yields dropped to 70% due to a lower
anaerobic COD consumption. The detoxification potential of both systems, determined with yeast-based
assays, pointed to a better performance of SBR2 and all of the SBR1 samples were significantly more
toxic. Thus, SBR2 appeared better suited for the treatment of this simulated wastewater. Residual AR14
and the major stable biodegradation product, 4-amino-1-naphthalenesulfonic acid (4A1NS), were not
directly responsible for the toxicity of the samples. Statistical analysis pointed to the existence of
correlations between the levels of two additional unidentified metabolites present in the SBR2 samples
and the cytotoxicity data. Overall results suggest the development of a more diverse microbial
community in the SBR2 able to biodegrade to less harmful metabolites the dye present in the textile
wastewaters.

Key words: Textile wastewater; Azo dye; aerobic granular sludge; sequencing batch reactor;
detoxification; yeast-based toxicity assays

ii
Resumo:

As águas residuais têxteis são resistentes ao tratamento aeróbico e são caracterizadas pela elevada
carga orgânica e presença de corantes, tendo um elevado impacto ambiental. A descoloração destas
águas contendo corantes azo já tinha sido alcançada em reator sequencial descontinuo (SBR) e com
biomassa granular. Este estudo compara o desempenho, em termos de eficiência e de potencial de
destoxificação, de dois SBRs com regimes hidrodinâmicos distintos no tratamento de uma água residual
artificial têxtil contendo o corante azo acid red 14 (AR14). Em ambos, os rendimentos de remoção de
cor e COD foram semelhantes (80%), contudo o SBR2, com a velocidade de alimentação mais baixa,
atingiu uma idade de lamas superior. O aumento das cargas de corante e orgânica não influenciou a
capacidade de remoção de cor, mas reduziu a capacidade de remoção de COD final. O potencial de
destoxificação obtido com os dois bioreatores foi determinado com base em ensaios de toxicidade com
levedura e revelou o melhor desempenho do SBR2 face ao SBR1, cujas amostras foram
significativamente mais tóxicas. Demonstrou-se que os compostos AR14 e o seu metabolito estável 4-
amino-1-naftaleno-sulfonato (4A1NS), não são responsáveis pela toxicidade detetada. A análise
estatística indicou a existência de dois metabolitos presentes nas amostras do SBR2 que apresentam
correlação com os resultados de toxicidade. Estes resultados sugerem o desenvolvimento de uma
comunidade microbiana capaz de degradar em metabolitos menos nocivos o corante presente nesta
água residual.

Palavras chave: Água residual têxtil; corante azo; grânulos aeróbicos; Reator sequencial descontinuo;
destoxificação; ensaios de toxicidade em levedura.

iii
Index:

Acknowledges: ..........................................................................................................................................i
Abstract..................................................................................................................................................... ii
Resumo: .................................................................................................................................................. iii
1. Introduction .......................................................................................................................................... 1
1.1. Textile industry and textile wastewaters ....................................................................................... 1
1.1.1. Composition and environmental impact ................................................................................ 1
1.1.2. Azo dyes ................................................................................................................................ 4
1.2. Textile wastewater treatment ....................................................................................................... 9
1.2.1. Physical-chemical treatments ................................................................................................ 9
1.2.2. Biological treatments ........................................................................................................... 10
1.3. Aerobic granular sludge (AGS) .................................................................................................. 14
1.4. Assessment of the toxicity of azo dye wastewaters ................................................................... 15
1.5. Saccharomyces cerevisiae and Caenorhabditis elegans: Relevance in toxicological studies of
xenobiotics ......................................................................................................................................... 17
1.6. Thesis aims and outline .............................................................................................................. 19
2. Materials and methods ...................................................................................................................... 20
2.1. Bioreactor setup and operation .................................................................................................. 20
2.2. Synthetic textile wastewater ....................................................................................................... 21
2.3. Sampling ..................................................................................................................................... 22
2.4. Analytic methods ........................................................................................................................ 22
2.4.1. Total suspended solids and volatile suspended solids ....................................................... 22
2.4.2. Sludge volume index and sludge retention time ................................................................. 23
2.4.3. Chemical oxygen demand removal, colour removal and pH ............................................... 23
2.4.4. High performance liquid chromatography ........................................................................... 24
2.5. Assessment of the toxicity of the collected samples .................................................................. 24
2.5.1. Control solutions and sample processing............................................................................ 24
2.5.2. S. cerevisiae microplate susceptibility assay ...................................................................... 25
2.5.3. S. cerevisiae gene expression assay .................................................................................. 25
2.5.4. C. elegans reproduction assay ............................................................................................ 26
2.5.5. Preliminary work in toxicity assays ...................................................................................... 26
2.5.6. Statistical analysis of toxicity data ....................................................................................... 27
3. Results and discussion ...................................................................................................................... 27
3.1. SBR operation ............................................................................................................................ 27
3.1.1. Biomass properties .............................................................................................................. 28
3.1.2. SBR performance ................................................................................................................ 30
3.1.3. Cycle characterization ......................................................................................................... 32
3.2. Toxicity assessment of the samples from operation period I ..................................................... 38
3.2.1. Sample processing and preliminary studies ........................................................................ 38
iv
3.2.2. Effects on yeast growth ....................................................................................................... 41
3.2.3. Effects on the transcript levels of the yeast gene GRE2 ..................................................... 44
3.2.4. Effects on C. elegans reproduction ..................................................................................... 46
3.2.5. Wastewater detoxification potential and AR14 metabolite profiles ..................................... 48
3.3. Toxicity assessment of the samples from operation period II .................................................... 53
4. Final discussion and future work ....................................................................................................... 58
5. Bibliography ....................................................................................................................................... 60
6. Annexes: ............................................................................................................................................ 70
Annex I – Toxicity reports on both mono and acid azo dyes ............................................................. 70
Annex II – Chromatograms from samples collected from SBRs during the period I ......................... 80
Annex III – Chromatograms from samples collected from SBRs during the period II ....................... 86

Figure index:

FIGURE 1 - MANUFACTURING TEXTILE PROCESSING LINE REPRESENTED WITH BOTH MAIN RELEVANT
CHARACTERISTICS AND MAJOR POLLUTANTS PRESENT IN THE WASTEWATER GENERATED (ADAPTED FROM
VERMA ET AL. (2012)). .................................................................................................................................. 2
FIGURE 2- SCHEMATIC REPRESENTATION OF THE EFFECT OF TEXTILE WASTEWATER DISCHARGE INTO THE
ENVIRONMENT (ADAPTED FROM VERMA ET AL. (2012)). .............................................................................. 3
FIGURE 3- CHEMICAL STRUCTURE OF AR14. ........................................................................................................ 5
FIGURE 4 - SCHEMATIC DIAGRAM SHOWING INTERACTIONS OF ENVIRONMENTAL FACTORS (EXTRINSIC: OXYGEN,
LIGHT, MOISTURE, ETC.; AND INTRINSIC: MICROBIAL ENZYMES, ETC.) WITH THE AZO DYES (CHEMICAL
STRUCTURE) AND THEIR (BIO)DEGRADATION PRODUCTS, WHICH ALTOGETHER DETERMINE THE
ENVIRONMENTAL FATE AND ECOLOGICAL CONSEQUENCES OF AZO DYES. THE SOLID LINES REPRESENT
PATHWAYS LEADING TO TOXIC METABOLITES, WHEREAS DOTTED LINES REPRESENT PATHWAYS LEADING
TO NON-TOXIC ENVIRONMENTALLY SAFE METABOLITES (ADAPTED FROM RAWAT ET AL. (2016)). .............. 6
FIGURE 5- CHEMICAL STRUCTURES OF THE AZO DYE AR14 AND OF THE TWO AROMATIC AMINES FORMED
DURING THE AZO BOND REDUCTION REACTION, 1-NAPHTHOL-2-AMINO-4-SULFONIC ACID (1N2A4S) AND
4-AMINO-1-NAPHTHALENESULFONIC ACID (4A1NS) (ADAPTED FROM FRANCA ET AL. (2015)). ................. 9
FIGURE 6 -- SCHEMATIC OF SBR OPERATION, WITH THE FILLING, REACTION, SETTLING AND DRAWING WHICH CAN
BE FOLLOWED BY AN IDLE PHASE. ................................................................................................................ 14
FIGURE 7 - TYPES OF TOXICITY TESTING METHODS USED IN THE ASSESSMENT OF THE HAZARD POSED BY AZO
DYES AND THEIR DEGRADATION PRODUCTS THAT MAY BE PRESENT IN TREATED WASTEWATERS (ADAPTED
FROM SOLÍS ET AL. (2012)). ........................................................................................................................ 16
FIGURE 8 - TSS MEASUREMENTS IN THE MIXED LIQUOR OF SBR1 (▪, FULL BLACK LINE) AND SBR2 (●, FULL
GREY LINE) AND IN THE DISCHARGED EFFLUENT OF SBR1(▫, DASH BLACK LINE) AND SBR2 (○, DASH GREY
LINE) DURING PERIOD I (68 UNTIL 103 OPERATIONAL DAY) AND PERIOD II (162 UNTIL 169 OPERATIONAL
DAY). ............................................................................................................................................................. 29
FIGURE 9 - SLUDGE RETENTION TIME (SRT) CALCULATED FOR SBR1 (▪, FULL BLACK LINE) AND SBR2 (●, FULL
GREY LINE) DURING PERIOD I (68 UNTIL 103 OPERATIONAL DAY) AND PERIOD II (162 UNTIL 169
OPERATIONAL DAY). ..................................................................................................................................... 29
FIGURE 10 – TOTAL DISSOLVED COD REMOVAL YIELDS FOR SBR1 (▪, FULL BLACK LINE) AND SBR2 (●, FULL
GREY LINE) DURING PERIOD I, OPERATION DAYS FROM 68 TO 103 (FEED: 500MG O2/L AS COD, 71.5
MGNH4CL/L AND 20 MG AR14/L); AND PERIOD II, OPERATION DAYS FROM 162 TO 173 (DAY 162 WITH
FEED: 500MG O2/L AS COD, 71.5 MGNH4CL/L AND 60 MG AR14/L; DAYS 167 AND 169 WITH FEED:
1500MG O2/L AS COD, 214.5 MGNH4CL/L AND 60 MG AR14/L). ............................................................ 31
v
FIGURE 11 - COLOUR REMOVAL YIELDS FOR SBR1 (▪, FULL BLACK LINE) AND SBR2 (●, FULL GREY LINE),
CALCULATED FROM ABSORBANCE READINGS AT THE WAVELENGTH OF 515 NM, DURING PERIOD I,
OPERATION DAYS FROM 68 TO 103 (FEED: 500MG O2/L AS COD, 71.5 MGNH4CL/L AND 20 MG AR14/L),
AND PERIOD II, OPERATION DAYS FROM 162 TO 173 (DAY 162 WITH FEED: 500MG O2/L AS COD, 71.5
MGNH4CL/L AND 60 MG AR14/L; DAYS 167 AND 169 WITH FEED: 1500MG O2/L AS COD, 214.5
MGNH4CL/L AND 60 MG AR14/L). .............................................................................................................. 32
FIGURE 12 – TOTAL DISSOLVED COD, IN FULL LINE, AND COLOUR, IN DASH LINE, REMOVAL PROFILES OF
OPERATIONAL DAY 75 IN BOTH SBR1 (▪) AND SBR2 (●). THE BLACK VERTICAL LINE MARKS THE ONSET OF
AERATION. SBR’S FEEDING SOLUTION WITH 500MG O2/L AS COD, 71.5 MG NH4CL/L AND 60 MG
AR14/L. ....................................................................................................................................................... 33
FIGURE 13 – TOTAL DISSOLVED COD, IN FULL LINE, AND COLOUR, IN DASH LINE, REMOVAL PROFILES OF
OPERATIONAL DAY 82 IN BOTH SBR1 (▪) AND SBR2 (●). THE BLACK VERTICAL LINE MARKS THE ONSET OF
AERATION. SBR’S FEEDING SOLUTION WITH 500MG O2/L AS COD, 71.5 MG NH4CL/L AND 60 MG
AR14/L. ....................................................................................................................................................... 34
FIGURE 14 - TOTAL DISSOLVED COD, IN FULL LINE, AND COLOUR, IN DASH LINE, REMOVAL PROFILES OF
OPERATIONAL DAY 89 IN BOTH SBR1 (▪) AND SBR2 (●). THE BLACK VERTICAL LINE MARKS THE ONSET OF
AERATION. SBR’S FEEDING SOLUTION WITH 500MG O2/L AS COD, 71.5 MG NH4CL/L AND 60 MG
AR14/L. ....................................................................................................................................................... 34
FIGURE 15 - TOTAL DISSOLVED COD, IN FULL LINE, AND COLOUR, IN DASH LINE, REMOVAL PROFILES OF
OPERATIONAL DAY 162 IN BOTH SBR1 (▪) AND SBR2 (●). SBR’S FEEDING SOLUTION WITH 500MG O2/L
AS COD, 71.5 MG NH4CL/L AND 60 MG AR14/L. THE BLACK VERTICAL LINE MARKS THE ONSET OF
AERATION. .................................................................................................................................................... 35
FIGURE 16 - TOTAL DISSOLVED COD, IN FULL LINE, AND COLOUR, IN DASH LINE, REMOVAL PROFILES OF
OPERATIONAL DAY 169 IN BOTH SBR1 (▪) AND SBR2 (●). SBR’S FEEDING SOLUTION WITH 1500MG O2/L
AS COD, 214.5 MG NH4CL/L AND 60 MG AR14/L. THE BLACK VERTICAL LINE MARKS THE ONSET OF
AERATION. .................................................................................................................................................... 36
FIGURE 17 – REPRESENTATIVE PH PROFILES ALONG THE REACTION PHASE OF THREE TREATMENT CYCLES OF
SBR1, NAMELY CORRESPONDING TO THE DAY 75 (FULL LINE, 500MG O2/L AS COD, 71.5 MG NH4CL/L
AND 20 MG AR14/L), DAY 162 (DASH LINE, 500MG O2/L AS COD, 71.5 MG NH4CL/L AND 60 MG
AR14/L) AND DAY 169 (DOT LINE, 1500MG O2/L AS COD, 214.5 MG NH4CL/L AND 60 MG AR14/L). THE
BLACK VERTICAL LINE MARKS THE ONSET OF AERATION. ............................................................................ 37
FIGURE 18 - REPRESENTATIVE PH PROFILES ALONG THE REACTION PHASE OF THREE TREATMENT CYCLES OF
SBR2, NAMELY CORRESPONDING TO THE DAY 75 (FULL LINE, 500MG O2/L AS COD, 71.5 MG NH4CL/L
AND 20 MG AR14/L), DAY 162 (DASH LINE, 500MG O2/L AS COD, 71.5 MG NH4CL/L AND 60 MG
AR14/L) AND DAY 169 (DOT LINE, 1500MG O2/L AS COD, 214.5 MG NH4CL/L AND 60 MG AR14/L). THE
BLACK VERTICAL LINE MARKS THE ONSET OF AERATION. ............................................................................ 37
FIGURE 19 – MASS CONCENTRATION OF AR14 DETERMINED BY HPLC IN THE SOLUTION WWFEED SUBJECTED
TO THREE DIFFERENT STERILIZATION PROCEDURES. FILTRATION THROUGH STERILE WHATMAN™
PURADISC FILTERS WITH 0.2µM PORE; FILTRATION THROUGH STERILE PALL® ACRODISC/SUPOR FILTERS
WITH 0.2µM PORE AND AUTOCLAVED AT 121°C FOR 20 MINUTES. ............................................................. 39
FIGURE 20 – MASS CONCENTRATION OF 4A1NS DETERMINED BY HPLC IN THE SOLUTION WWAMINE
SUBJECTED TO THREE DIFFERENT STERILIZATION PROCEDURES. FILTRATION THROUGH STERILE
WHATMAN™ PURADISC FILTERS WITH 0.2µM PORE; FILTRATION THROUGH STERILE PALL®
ACRODISC/SUPOR FILTERS WITH 0.2µM PORE AND AUTOCLAVED AT 121°C FOR 20 MINUTES. ................ 39
FIGURE 21 – EFFECT ON THE YEAST GROWTH INHIBITION RATIO OF THE SAMPLES COLLECTED AT THE ONSET
OF THE REACTION PHASE (WWFEED) AND THE WWAMINE SOLUTION IN COMPARISON WITH FEED
SOLUTION WITHOUT DYE (WWCONTROL). ERROR BARS REPRESENT ±1 STANDARD DEVIATION. MEANS
FOR WWFEED AND WWAMINE WERE NOT SIGNIFICANTLY DIFFERENT FROM WWCONTROL (TUCKEY´S
TEST; P > 0.999). ......................................................................................................................................... 42
FIGURE 22 – EFFECT ON THE YEAST GROWTH INHIBITION RATIO OF SAMPLES COLLECTED FROM THE SBR1 AT
THE END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF AERATION (WWEFFLUE) FOR THE
OPERATION DAYS 76, 83 AND 90, IN COMPARISON WITH THE FEED SOLUTION WITHOUT DYE
(WWCONTROL). ERROR BARS REPRESENT ±1 STANDARD DEVIATION. “●” DENOTES MEAN VALUES
SIGNIFICANTLY DIFFERENT FROM THE CONTROL WWCONTROL (TUKEY´S TEST; P < 0.0001). ................. 43
vi
FIGURE 23 – EFFECT ON THE YEAST GROWTH INHIBITION RATIO OF SAMPLES COLLECTED FROM SBR2 AT THE
END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF AERATION (WWEFFLUE) FOR THE
OPERATION DAYS 76, 83 AND 90, IN COMPARISON WITH THE FEED SOLUTION WITHOUT DYE
(WWCONTROL). ERROR BARS REPRESENT ±1 STANDARD DEVIATION. “●” DENOTES MEAN VALUES
SIGNIFICANTLY DIFFERENT FROM THE CONTROL WWCONTROL (TUKEY´S TEST; P < 0.0001). ................. 43
FIGURE 24 – TOXICITY ASSESSMENT BASED ON INCREASED EXPRESSION OF THE YEAST STRESS-INDICATOR
GENE GRE2 OF THE SAMPLES COLLECTED AT THE ONSET OF THE REACTION PHASE (WWFEED) AND THE
FEEDING SOLUTION WITH THE STABLE AROMATIC AMINE 4A1NS INSTEAD OF THE AR14 (WWAMINE), IN
COMPARISON WITH THE FEED SOLUTION WITHOUT DYE (WWCONTROL). ERROR BARS REPRESENT ±1
STANDARD DEVIATION. ................................................................................................................................. 44
FIGURE 25 - TOXICITY ASSESSMENT BASED ON INCREASED EXPRESSION OF THE STRESS YEAST INDICATOR
GENE GRE2 OF SAMPLES COLLECTED FROM SBR1 AT THE END OF THE ANAEROBIC REACTION
(WWANAER) AND THE END OF AERATION (WWEFFLUE) FOR THE OPERATION DAYS 76, 83 AND 90 IN
COMPARISON WITH FEED SOLUTION WITHOUT DYE (WWCONTROL). ERROR BARS AND “ND” REPRESENT,
RESPECTIVELY, ±1 STANDARD DEVIATION AND NOT DETERMINED. ............................................................. 45
FIGURE 26 - TOXICITY ASSESSMENT BASED ON INCREASED EXPRESSION OF THE STRESS YEAST INDICATOR
GENE GRE2 OF SAMPLES COLLECTED FROM SBR2 AT THE END OF THE ANAEROBIC REACTION
(WWANAER) AND THE END OF AERATION (WWEFFLUE) FOR THE OPERATION DAYS 76, 83 AND 90 IN
COMPARISON WITH FEED SOLUTION WITHOUT DYE (WWCONTROL). ERROR BARS REPRESENTS ±1
STANDARD DEVIATION. ................................................................................................................................. 46
FIGURE 27 – EFFECTS ON REPRODUCTION OF C. ELEGANS BRISTOL N2 OF SAMPLES COLLECTED FROM SBR1
AT THE END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF THE AERATION (WWEFFLUE)
FROM THE OPERATION DAYS 76 AND 83, IN COMPARISON WITH THE FEED SOLUTION WITHOUT DYE
(WWCONTROL). ERROR BARS REPRESENT ±1 STANDARD DEVIATION....................................................... 47
FIGURE 28 - EFFECTS ON REPRODUCTION OF C. ELEGANS BRISTOL N2 OF SAMPLES COLLECTED FROM SBR2
AT THE END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF THE AERATION (WWEFFLUE)
FROM THE OPERATION DAYS 76 AND 83, IN COMPARISON WITH THE FEED SOLUTION WITHOUT DYE
(WWCONTROL). ERROR BARS REPRESENT ±1 STANDARD DEVIATION....................................................... 47
FIGURE 29 - MASS CONCENTRATION OF AR14 (DESCENDANT DIAGONAL STRIPES) AND 4A1NS (ASCENDANT
DIAGONAL STRIPES) IN THE SAMPLES COLLECTED FROM SBR1, IN THE OPERATIONAL DAYS 76, 83 AND
90, AT THE END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF AERATION (WWEFFLUE)
DETERMINED BY HPLC (SPECTROPHOTOMETRIC DETECTION AT 220 NM USING A LICHROCART
PUROSPHER STAR RP-18E COLUMN). “N” REPRESENTS A MASS CONCENTRATION VALUE EQUAL TO
ZERO. ............................................................................................................................................................ 49
FIGURE 30 - MASS CONCENTRATION OF AR14 (DESCENDANT DIAGONAL STRIPES) AND 4A1NS (ASCENDANT
DIAGONAL STRIPES) IN THE SAMPLES COLLECTED FROM SBR2, IN THE OPERATIONAL DAYS 76, 83 AND
90, AT THE END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF AERATION (WWEFFLUE)
DETERMINED BY HPLC (SPECTROPHOTOMETRIC DETECTION AT 220 NM USING A LICHROCART
PUROSPHER STAR RP-18E COLUMN). “N” REPRESENTS A MASS CONCENTRATION VALUE EQUAL TO
ZERO. ............................................................................................................................................................ 49
FIGURE 31 – AREA OF RELEVANT PEAKS DETECTED IN CHROMATOGRAMS OF THE SAMPLES COLLECTED FROM
SBR1 IN OPERATIONAL DAYS 76, 83 AND 90, AT THE END OF THE ANAEROBIC REACTION (WWANAER)
AND THE END OF AERATION (WWEFFLUE). PEAK Α1 (PURPLE BARS), PEAK Β1 (ORANGE BARS) AND PEAK
Γ1 HAD A RETENTION TIME OF 22, 27 AND 29 MINUTES, RESPECTIVELY. SPECTROPHOTOMETRIC
DETECTION AT 220 NM USING A LICHROCART PUROSPHER STAR RP-18E COLUMN. ........................... 51
FIGURE 32 - AREA OF RELEVANT PEAKS DETECTED IN CHROMATOGRAMS OF THE SAMPLES COLLECTED FROM
SBR2 IN OPERATIONAL DAYS 76, 83 AND 90, AT THE END OF THE ANAEROBIC REACTION (WWANAER)
AND THE END OF AERATION (WWEFFLUE). PEAK Α2 (PURPLE BARS), PEAK Β2 (ORANGE BARS) AND PEAK
Γ2 HAD A RETENTION TIME OF 22, 27 AND 29 MINUTES, RESPECTIVELY. SPECTROPHOTOMETRIC
DETECTION AT 220 NM USING A LICHROCART PUROSPHER STAR RP-18E COLUMN. ........................... 52
FIGURE 33 – EFFECT ON THE YEAST GROWTH INHIBITION RATIO OF THE SAMPLES COLLECTED AT THE ONSET
OF THE REACTION PHASE DURING THE PERIOD II AND THE FEED WITH A COD OF 1500 MG O2/L, 214.5 MG
NH4CL/L AND WITHOUT DYE (WWCONTROL3) IN COMPARISON WITH FEED SOLUTION WITHOUT DYE
(WWCONTROL). FEEDING SAMPLES DURING OPERATIONAL DAYS 159 TO 165 (WWFEED+) WITH COD OF
500MG O2/L, 71.5 MG NH4CL/L AND 60 MG AR14/L AND 165 TO 173 (WWFEED3) WITH COD OF
vii
1500MG O2/L, 214.5 MG NH4CL/L 60 MG AR14/L. ERROR BARS REPRESENT ±1 STANDARD DEVIATION.
MEANS FOR WWFEED+, WWCONTROL3 AND WWFEED3 WERE NOT SIGNIFICANTLY DIFFERENT FROM
WWCONTROL (TUCKEY´S TEST; P > 0.999). .............................................................................................. 53
FIGURE 34 – EFFECT ON THE YEAST GROWTH INHIBITION RATIO OF SAMPLES COLLECTED FROM SBR1 AT THE
END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF AERATION (WWEFFLUE) FOR THE
OPERATION DAYS 165 AND 173, IN COMPARISON WITH THE FEED SOLUTION WITHOUT DYE
(WWCONTROL). SBR’S FEEDING SOLUTION AT THE OPERATIONAL DAY 165 WITH 500MG O2/L AS COD,
71.5 MG NH4CL/L AND 60 MG AR14/L; AND AT THE DAY 173 WITH 1500MG O2/L AS COD, 214.5 MG
NH4CL/L AND 60 MG AR14/L. ERROR BARS REPRESENT ±1 STANDARD DEVIATION. ............................... 54
FIGURE 35– EFFECT ON THE YEAST GROWTH INHIBITION RATIO OF SAMPLES COLLECTED FROM SBR2 AT THE
END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF AERATION (WWEFFLUE) FOR THE
OPERATION DAYS 165 AND 173, IN COMPARISON WITH THE FEED SOLUTION WITHOUT DYE
(WWCONTROL). SBR’S FEEDING SOLUTION AT THE OPERATIONAL DAY 165 WITH 500MG O2/L AS COD,
71.5 MG NH4CL/L AND 60 MG AR14/L; AND AT THE DAY 173 1500MG O2/L AS COD, 214.5 MG NH4CL/L
AND 60 MG AR14/L. ERROR BARS REPRESENT ±1 STANDARD DEVIATION. ............................................... 55
FIGURE 36 - MASS CONCENTRATION OF AR14 (DESCENDANT DIAGONAL STRIPES) AND 4A1NS (ASCENDANT
DIAGONAL STRIPES) IN THE SAMPLES COLLECTED FROM SBR1, IN THE OPERATIONAL DAYS 165 AND 173,
AT THE END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF AERATION (WWEFFLUE)
DETERMINED BY HPLC (SPECTROPHOTOMETRIC DETECTION AT 220 NM USING A LICHROCART
PUROSPHER STAR RP-18E COLUMN). ...................................................................................................... 56
FIGURE 37 - MASS CONCENTRATION OF AR14 (DESCENDANT DIAGONAL STRIPES) AND 4A1NS (ASCENDANT
DIAGONAL STRIPES) IN THE SAMPLES COLLECTED FROM SBR2, IN THE OPERATIONAL DAYS 165 AND 173,
AT THE END OF THE ANAEROBIC REACTION (WWANAER) AND THE END OF AERATION (WWEFFLUE)
DETERMINED BY HPLC (SPECTROPHOTOMETRIC DETECTION AT 220 NM USING A LICHROCART
PUROSPHER STAR RP-18E COLUMN). ...................................................................................................... 56
FIGURE 38 - AREA OF RELEVANT PEAKS DETECTED IN CHROMATOGRAMS OF THE SAMPLES COLLECTED FROM
SBR1 IN THE OPERATIONAL DAYS 165 AND 173, AT THE END OF THE ANAEROBIC REACTION (WWANAER)
AND THE END OF AERATION (WWEFFLUE). PEAK Α1 (PURPLE BARS), PEAK Β1 (ORANGE BARS) AND PEAK
Γ1 HAD A RETENTION TIME OF 22, 27 AND 29 MINUTES, RESPECTIVELY. SPECTROPHOTOMETRIC
DETECTION AT 220 NM USING A LICHROCART PUROSPHER STAR RP-18E COLUMN. ........................... 57
FIGURE 39 - AREA OF RELEVANT PEAKS DETECTED IN CHROMATOGRAMS OF THE SAMPLES COLLECTED FROM
SBR2 IN THE OPERATIONAL DAYS 165 AND 173, AT THE END OF THE ANAEROBIC REACTION (WWANAER)
AND THE END OF AERATION (WWEFFLUE). PEAK Α2 (PURPLE BARS), PEAK Β2 (ORANGE BARS) AND PEAK
Γ2 HAD A RETENTION TIME OF 22, 27 AND 29 MINUTES, RESPECTIVELY. SPECTROPHOTOMETRIC
DETECTION AT 220 NM USING A LICHROCART PUROSPHER STAR RP-18E COLUMN. ........................... 58
FIGURE 40 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 76 (76 WWANAER). ............................................................................................................................. 80
FIGURE 41 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 76 (76 WWEFFLUE). ............................................................................................................................ 80
FIGURE 42 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 76 (76 WWANAER). ............................................................................................................................. 81
FIGURE 43 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 76 (76 WWANAER). ............................................................................................................................. 81
FIGURE 44 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 83 (83 WWANAER). ............................................................................................................................. 82
FIGURE 45 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 83 (83 WWEFFLUE). ............................................................................................................................ 82
FIGURE 46 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 83 (83 WWANAER). ............................................................................................................................. 83
FIGURE 47 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 83 (83 WWEFFLUE). ............................................................................................................................ 83
FIGURE 48 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 90 (90 WWANAER). ............................................................................................................................. 84
FIGURE 49 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 90 (90 WWEFFLUE). ............................................................................................................................ 84
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FIGURE 50 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 90 (90 WWANAER). ............................................................................................................................. 85
FIGURE 51 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 90 (90 WWEFFLUE). ............................................................................................................................ 85
FIGURE 52 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 165 (165 WWANAER). ......................................................................................................................... 86
FIGURE 53 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 165 (165 WWEFFLUE). ........................................................................................................................ 86
FIGURE 54 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 165 (165 WWANAER). ......................................................................................................................... 87
FIGURE 55 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 165 (165 WWEFFLUE). ........................................................................................................................ 87
FIGURE 56 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 173 (173 WWANAER). ......................................................................................................................... 88
FIGURE 57 – CHROMATOGRAM OBTAIN FROM SBR1 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 173 (173 WWEFFLUE). ........................................................................................................................ 88
FIGURE 58 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF ANAEROBIC PHASE ON THE OPERATIONAL
DAY 173 (173 WWANAER). ......................................................................................................................... 89
FIGURE 59 – CHROMATOGRAM OBTAIN FROM SBR2 AT THE END OF AERATION PHASE ON THE OPERATIONAL
DAY 173 (173 WWEFFLUE). ........................................................................................................................ 89
Tables index:
TABLE 1 - CHEMICAL AND PHYSICAL CHARACTERISTIC OF WASTEWATER GENERATED AT DIFFERENT STAGES OF
THE TEXTILE INDUSTRY PROCESSING (ADAPTED FROM DASGUPTA ET AL. (2015)). ................................. 3
TABLE 2 - SUPPLEMENT 8 OF THE REGULATION CE 1907/2006 OF 18 OF DECEMBER OF 2006. LIST OF TOXIC
AND CARCINOGENIC AROMATIC AMINES GENERATED BY REDUCTIVE CLEAVAGE OF AZO DYES FORBIDDEN
IN THE EUROPEAN UNION. .................................................................................................................. 8
TABLE 3 - SOME EXAMPLES OF RESULTS FROM PHYSICAL AND CHEMICAL TREATMENTS OF SYNTHETIC TEXTILE
INDUSTRY WASTEWATERS CONTAINING AZO DYES. ............................................................................. 10
TABLE 4 - ADVANTAGES AND DISADVANTAGES OF THE DIFFERENT STRATEGIES TYPES OF INTEGRATED
ANAEROBIC-AEROBIC BIOREACTORS. ................................................................................................. 12
TABLE 5 – SOME RESULTS FROM BIOLOGICAL TREATMENTS OF SYNTHETIC TEXTILE INDUSTRY WASTEWATERS
CONTAINING AZO DYES. .................................................................................................................... 13
TABLE 6 -SUMMARY OF THE OPERATING CONDITIONS FOR BOTH SBR1 AND SBR2. .................................... 21
TABLE 7 – SLUDGE VOLUME INDEX (SVI) VALUES FROM SBR1 AND SBR2 MEASURED AFTER 5 MIN SETTLING
(SVI5) AND AFTER 30 MIN SETTLING (SVI30). ................................................................................... 30
TABLE 8 – PERCENTAGE OF DECOLOURIZATION OF THE WWFEED SOLUTION, MEASURED AS ABSORBANCE AT
515NM (THE WAVELENGTH OF MAXIMUM ABSORBANCE OF THE AR14, IN THE VISIBLE REGION), BY S.
CEREVISIAE UPON 14 HOURS’ INCUBATION AT 30°C. SUPPLEMENTATION WITH YPD WHEN PROVIDED WAS
REPRESENTED WITH + YPD. ............................................................................................................. 40
TABLE 9 - PERCENTAGE OF DECOLOURIZATION OF THE WWFEED SOLUTION, MEASURED AS ABSORBANCE AT
515NM (THE WAVELENGTH OF MAXIMUM ABSORBANCE OF THE AR14, IN THE VISIBLE REGION), BY E. COLI
ALONE AND WITH C. ELEGANS UPON 72 HOURS’ INCUBATION AT 20°C. ................................................ 41
TABLE 10 – PEARSON CORRELATION BETWEEN THE VALUES OF THE YEAST GROWTH INHIBITION RATIO AND THE
MASS CONCENTRATIONS OF AR14 AND 4A1NS IN THE SAMPLES COLLECTED FROM BOTH SBR UNITS’. 50
ix