treatment of wine distillery wastewaters by high rate anaerobic digestion and submerged ...

133 Pages·2008·0.99 MB·English

TREATMENT OF WINE DISTILLERY WASTEWATERS BY HIGH
RATE ANAEROBIC DIGESTION AND SUBMERGED MEMBRANE
SYSTEMS

A thesis submitted in fulfilment for the degree of

DOCTOR OF PHILOSOPHY

Of

RHODES UNIVERSITY

By

Xolisa Lorraine Melamane

December 2006
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ABSTRACT
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Experiences in treating wine distillery wastewaters (WDWs) contribute to the field of oenology
as many oenologists are concerned with the selection, efficiency and economy of their
wastewaters. Wine distillery wastewaters are strongly acidic, have high chemical oxygen
demand (COD), high polyphenol content and are highly variable. Primary attention was
focussed on sustainable biological treatment of raw wine distillery wastewater (RWDW) and
fungally pre-treated wine distillery wastewater (FTWDW) by energy-efficient high rate
anaerobic digestion (AD). This study also explored the development of a novel dual-stage
anaerobic digestion ultrafiltration (ADUF) process, using a ceramic submerged membrane
bioreactor (SMBR) in the treatment of both RWDW and FTWDW. The first stage was for the
selection of microorganisms that were able to treat the toxic pollutants from WDWs. It was
operated at a high feed-to-microorganism ratio. The second stage, a secondary digester, was
operated like a typical membrane bioreactor at a low feed-to-microorganism ratio to sustain a
stable efficient population for a long period. The characteristics of RWDW were as follows: pH
3.83, 15 000 mg/l soluble COD (COD) and 5229 mg/l of phenols. After pre-treatment of
s
RWDW with Trametes pubescens, starting parameters for FTWDW were as follows: pH 6.7,
7000 mg/l soluble COD (COD ) and 1440 mg/l of phenols. During operation of a high rate
S
anaerobic digester for RWDW treatment, K HPO was required for buffering the digester.
2 4
Volatile fatty acid concentrations were <300 mg/l throughout the study, indicating degradation
of organic acids present. Mean COD removal efficiency for the 130 day study was 87 %, while
S
the mean polyphenol removal efficiency was 85 %. Addition of 50 mg/l Fe3+ increased the
removal efficiencies of COD to 97 % and of polyphenols to 99 %. High removal efficiencies
S
of COD and polyphenols were attributed to the addition of macronutrients and micronutrients
S
that caused pH stability and stimulated microbial activity. The COD removal efficiency of
S
high rate anaerobic digestion of FTWDW reached 99.5%. During FTWDW digestion, pH
buffering was achieved using K HPO . A combination of a SMBR and a secondary digester
2 4
was tested for the treatment of RWDW and FTWDW during a 30 day study. Results for
RWDW showed that pH buffering was achieved by dosing the feed stream with CaCO and
3
K HPO . Buffering proved to be significant for optimum performance of the system in removal
2 4
of soluble COD , and volatile fatty acids (VFAs). Different batches of RWDW used for feeding
S
the reactor had variable compositions with respect to concentrations of nitrates, ammonium and
total phenolic compounds. Ammonium accumulated in the secondary digester after 14 days of
system operation, indicated the time required for the establishment of anaerobic conditions in
the system. Dosing of the SMBR treating FTWDW with CaCO and K HPO buffered the pH;
3 2 4
ii
this proved significant for optimum performance of the system in removal of COD . The
S
system eliminated an average of 86 (± 4) % of COD present in the FTWDW. The residual
S
COD levels in the effluent were approximately 400 mg/l, significantly lower than the
S
concentrations observed when treating RWDW, indicating that fungal pre-treatment might have
provided additional nutrients for removal of recalcitrant components of the wastewater. The
resulting effluent was rich in nitrates and phosphates and might be used as a fertiliser.
Alternatively, a membrane process, such as reverse osmosis (RO) or nanofiltration (NF) could
be applied to raise the water quality to meet the levels required for reuse. Biomass samples
were obtained from the four treatment systems and population shifts characterization using
phospholipids fatty acids (PLFA) and 16S rRNA analysis to provide an indication of
limitations within the microbial population. The values of the concentrations of the individual
PLFAs detected in the samples indicated that ten bacterial species were present, with the GC
content of the 16S rRNA increasing from 1 to 10. Analysis of denaturing gradient gel
electrophoresis DGGE data indicated that the composition of the archeal community changed
the consortia used for both RWDW and FTWDW treatment. Changes in band intensities
indicated the presence of different components of the archeal communities. The results were
not conclusive in terms of species identity as cloning, sequencing and phylogenetic analyses
were not performed, but they did indicate microbial population shifts and species diversity for
high rate anaerobic digestion. The results also confirmed prevalence of relatively few species
during operation of SMBRs for treatment of RWDW and FTWDW, which suggested that the
microorganisms that survived were either tolerant of toxic components of RWDW and
FTWDW or they were able to remove polyphenols.

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TABLE OF CONTENTS
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CONTENT PAGE NUMBER

ABSTRACT ii
TABLE OF CONTENTS iv
LIST OF FIGURES vii
LIST OF TABLES ix
LIST OF ABBREVIATIONS x
LIST OF ABBREVIATED UNITS xii
LIST OF PUBLICATIONS xiii
ACKNOWLEDGEMENTS xiv
CHAPTER 1: INTRODUCTION AND SYNOPSIS 1
1.1 Introduction 1
1.2 Synopsis of the study 5
CHAPTER 2: LITERATURE REVIEW 8
2.1 Wastewater generation in distillation industries 8
2.2 Wastewaters characteristics 9
2.3 Wine distillery wastewater disposal and use 10
2.4 Traditional treatment practices 14
2.5 Current treatment and disposal options 15
2.6 Biological treatment 17
2.7 Membrane bioreactors in the treatment of distillery wastewaters 22
2.8 Concluding remarks 28
2.9 Interim summary and conclusions 29
CHAPTER 3: OBJECTIVES OF THE STUDY 30
3.1 Characterization of RWDW and FTWDW 30
3.2 Treatment of RWDW and FTWDW in high rate anaerobic digester 30
3.3 Treatment of RWDW and FTWDW in dual-stage anaerobic submerged
membrane bioreactors 30
3.4 Population shifts characterization using PFLA analysis and 16S rRNA 31
CHAPTER 4: HIGH RATE ANAEROBIC DIGESTION OF RWDW 32
4.1 Introduction 32
4.2 Materials and Methods 33
iv
4.2.1 Set-up of a high rate anaerobic digester 33
4.2.2 Performance of a high rate anaerobic digester during treatment of RWDW 34
4.3 Results and Discussion 35
4.4 Interim conclusions 42
CHAPTER 5: TREATMENT OF RWDW USING SMBR 43
5.1 Introduction 43
5.2 Materials and Methods 44
5.2.1 Biological wastewater treatment system 44
5.2.2 Sample analysis 45
5.3 Results and Discussion 46
5.4 Interim conclusions 55
CHAPTER 6: TREATMENT OF FTWDW USING SMBR 57
6.1 Introduction 57
6.2 Materials and Methods 57
6.2.1 Biological wastewater treatment system 57
6.3 Results and Discussion 59
6.4 Interim conclusions 65
CHAPTER 7: HIGH RATE ANAEROBIC DIGESTION OF FTWDW 66
7.1 Introduction 66
7.2 Materials and Methods 67
7.2.1 Set-up of a high rate anaerobic digester 67
7.2.2 Sample analysis 68
7.2.3 Performance of a high rate anaerobic digester during treatment of FTWDW 68
7.3 Results and Discussion 69
7.4 Interim conclusions 72
CHAPTER 8: POPULATION DIVERSITIES IN ANAEROBIC DIGESTERS
AND SUBMERGED MEMBRANE SYSTEMS 73
8.1 Introduction 73
8.2 Materials and Methods 75
8.2.1 Sample collection and lyophillization 75
8.2.2 Phospholipid fatty acids extraction, transesterification
and analyses 75
8.2.3 Deoxyribonucleic acid isolation 78
8.2.4 Polymerase chain reaction amplification 78
8.2.5 Denaturing gradient gel electrophoresis analyses 78
8.3 Results and Discussion 79
v
8.3.1 Phospholipid fatty acids results evaluation 79
8.3.2 Denaturing gradient gel electrophoresis results evaluation 84
8.4 Interim conclusions 87
CHAPTER 9: GENERAL DISCUSSION 88
CHAPTER 10: CONCLUSIONS AND RECOMMENDATIONS 95
10.1.1 Characterization of RWDW and FTWDW. 95
10.1.2 Treatment of RWDW and FTWDW in high rate anaerobic digester. 95
10.1.3 Treatment of RWDW and FTWDW in dual-stage anaerobic SMBRs. 95
10.1.4 Population shifts characterization using PFLA analysis and 16S rRNA. 96
10.2 Recommendations for Future Work 96
CHAPTER 11: REFERENCES 98

APPENDICES 111

APPENDIX A: PARAMETERS MONITORED DURING HIGH RATE
ANAEROBIC DIGESTION OF RWDW. 111

APPENDIX B.1: PARAMETERS MONITORED DURING TREATMENT
OF RWDW USING SMBR. 113

APPENDIX B.2: OTHER PARAMETERS MONITORED DURING TREATMENT
OF RWDW USING SMBR. 114

APPENDIX C.1: PARAMETERS MONITORED DURING TREATMENT OF
FTWDW USING SMBR. 115

APPENDIX C.2: OTHER PARAMETERS MONITORED DURING
TREATMENT OF FTWDW USING SMBR. 116

APPENDIX D: PERFORMANCE OF FUNGAL UNIT FOR PRE-TREATMENT
OF RWDW. 117

APPENDIX E: PARAMETERS MONITORED DURING HIGH RATE ANAEROBIC
DIGESTION OF FTWDW. 118

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LIST OF FIGURES
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Figure 2.1: Configurations of a solid-liquid MBR: A. External MBR B. Submerged MBR 25
(Stephenson et al., 2000).
Figure 4.1: The high rate anaerobic digester made of PVC. A peristaltic pump (Watson 33
Marlow 501RL2) was used to pump in the feed at C, with rubber tubing of 3.2 cm
diameter. The digester had 15 l capacity and a height of 36.09 cm. A = supernatant outlet,
B = sampling outlet, C = feed inlet, D = inlet for nitrogen gas, E = unused port, F = sludge
inlet and G = stirrer bar to provide mixing.
Figure 4.2: Anaerobic digester after settling. 35
Figure 4.3: Volatile fatty acids concentration and pH in the digester during the trial. 37
CaCO (2000 mg/l) was added to buffer the pH value of the system from day 62 to day 73.
3
From days 73 to 84, this was replaced with a mixture of CaCO and K HPO (1 g/l).
3 2 4
K HPO (1 g/l) was used to buffer the system from day 84 until day 130 as indicated by
2 4
the vertical dotted lines.
Figure 4.4: Removal efficiency of COD and polyphenols in the anaerobic digester as a 38
S
function of the time of incubation, with dotted lines marking addition of 50 mg/l Fe3+
between days 86 and 92.
Figure 4.5: Nitrate and ammonium concentrations measured in the supernatant during 39
RWDW anaerobic digestion.
Figure 4.6: Phosphate and total suspended solids concentrations measured in the 41
supernatant during WDW anaerobic digestion.
Figure 5.1: The biological wastewater treatment system used in the study, showing the 45
flow paths between reactors A through D. A = influent tank, B = SMBR,
C = permeate balancing tank, D = secondary digester.
Figure 5.2: pH of the supernatant in the individual reactors of the bioreactor system as 47
function of time of operation. A = influent tank, B = SMBR, C = permeate balancing tank,
D = secondary digester. The first dotted line on day 12 indicates the time when extra Deleted: indicated
CaCO was added as buffer and the second dotted arrow on day 22 when Reactor B was
3
inoculated using sludge from Reactor D.
Figure 5.3: pH of the supernatant in the individual reactors of the bioreactor system as 48
function of time of operation from B = SMBR and D = secondary digester. The dotted
lines on days 2 and 12 indicate the period when the pH was <7.4 and VFA concentration
was at its highest.
Figure 5.4 Volatile fatty acid concentrations in reactors B and D of the bioreactor system 48
as function of time of operation. A = influent tank, B = SMBR, C = permeate balancing
tank, D = secondary digester. The dotted lines on days 2 and 12 for reactors B and D
bracket the period when the pH was <7.4 and [VFA] was highest.
vii
Figure 5.5 Removal efficiency of COD in the SMBR and the total system as a function of 50
S
time. SMBR = reactor B, total = final effluent from secondary digester (reactor D). The
first dotted line on day 16 and the second on day 30 for both reactor B and D bracket the
period when COD removal efficiency began to stabilize and then increased slightly.
S
Figure 5.6 The total concentration of phenolic compounds in the bioreactor system 51
expressed in phenol equivalents as a function of time. A = influent tank, B = SMBR, C =
permeate balancing tank, D = secondary digester. The dotted lines indicate the beginning
and end of the period when phenol levels became very low.
Figure 5.7: Concentration of nitrates in the system as function of time. A = influent tank, 54
B = SMBR, C = permeate balancing tank, D = secondary digester. The dotted lines on days
14 and 30 bracket the period when nitrate levels became very low.
Figure 5.8: Concentration of ammonium in the system as function of time. A = influent 55
tank, B = SMBR, C = permeate balancing tank, D = secondary digester. The first dotted
line on day 14 and the second on day 30 for reactor A, B, C and D bracket the period when
ammonium levels became very low.
Figure 6.1: The bioreactor used for fungal pre-treatment of wastewater with floating 58
Trametes pubescens mycelial balls. The working volume of the bioreactor was 102 l with
the following dimensions: height 2.3 m (of which 1.2 m was the lower, V-shaped part), and
a square 0.5 × 0.5 m cross-section. Two ports at the bottom were used for aeration (for
both oxygenation and mixing) and for withdrawal of samples. (Adapted from Strong and
Burgess, in press).
Figure 6.2: Prevailing pH of the bulk liquid in the individual reactors of the experimental 60
system as function of time of operation of the bioreactor system. A = influent tank,
B = SMBR, C = permeate balancing tank, D = secondary digester.
Figure 6.3: Total VFA concentrations in reactor B (SMBR) and reactor D (secondary 61
digester) as function of time.
Figure 6.4: Removal efficiency of COD in the SMBR and over the total treatment system 62
S
as a function of time.
Figure 6.5: The total concentration of phenolic compounds in the treatment system 63
expressed in phenol equivalents as a function of time operation. A = influent tank,
B = SMBR, C = permeate balancing tank, D = secondary digester.
Figure 6.6: Concentration of nitrates (6a,(cid:204) (cid:162)) and ammonium (6b, (cid:88) (cid:165)) in the system as 64
function of incubation time. A = influent tank, B = SMBR, C = permeate balancing tank,
D = secondary digester.
Figure 7.1: pH and VFA concentration as a function of time during anaerobic digestion. 71
Figure 7.2: The total concentration of phenolics and COD as a function of time during 71
s
anaerobic digestion.
Figure 8.1: Denaturing gradient gel electrophoresis profiles of DNA extracted during 86
treatment of RWDW and FTWDW.

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LIST OF TABLES
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Table 2.1: Chemical characteristics of distillery wastewaters. 11
Table 2.2: Performance levels of anaerobic digestion of wine distillery wastewaters. 24
Table 4.1: Incremental changes in digester feed strength. 34
Table 4.2: Characterization of digester influent and effluent. 36
Table 7.1: Characteristics of FTWDW and RWDW. 69
Table 8.1: Results of the PLFA analyses of the anaerobic fermentation of RWDW. 80
Table 8.2: Results of the PLFA analyses of the anaerobic fermentation of FTWDW. 81
Table 8.3: Results of the PCA analysis of RWDW PLFA data. 82
Table 8.4: Results of the PCA analysis of FTWDW PLFA data. 82
Table 8.5: Identification of samples in Figure 8.1, above. 86
Table 9.1: DWAF (1996) guidelines for irrigation water quality targets. 91
Table 9.2: Mean characteristics of effluents from high rate anaerobic digestion and SMBR 94
(secondary digester) systems treating RWDW and FTWDW.
Table B.1: Parameters monitored during treatment of RWDW using SMBR. 113
Table B.2: Other parameters monitored during treatment of RWDW using SMBR. 114
Table C.1: Parameters monitored during treatment of FTWDW using SMBR. 115
Table C.2: Other parameters monitored during treatment of FTWDW using SMBR. 116
Table D.1: Results for COD, total phenol, colour changes and laccase activities for the four 117
white rot fungi in raw and PVPP-treated wastewater.

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LIST OF ABBREVIATIONS
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AD Anaerobic digestion
ADUF Anaerobic digestion ultrafiltration
BOD Biochemical oxygen demand in milligrams per litre
BOD Five day test for biochemical oxygen demand in milligrams per litre
5
C Carbon
COD Chemical oxygen demand
COD Soluble chemical oxygen demand
S
COD Total chemical oxygen demand
T
DNA Deoxyribonucleic acid
DGGE Denaturing gradient gel electrophoresis
DWAF Department of Water Affairs and Forestry
EC Electrical conductivity
et al And others
EMBR External membrane bioreactor
FAMEs Fatty acid methyl esters
FAU Formazine attenuation units
F/M Food to microorganism
FTWDW Fungally pre-treated wine distillery wastewater
GC\MS Gas chromatography\mass spectrophotometry
HRT Hydraulic retention time
MBR Membrane bioreactor
MCRT Mean cell residence time
MF Microfiltration
MLSS Mixed liquor suspended solids
MS Mixed solids
MSS Mixed suspended solids
MW Molecular weight
N Nitrogen
NO - Nitrates
3
P Phosphorus
PCA Principal component analysis
PCR Polymerase chain reaction
x