THESIS APPROVAL

GRADUATE SCHOOL, KASETSART UNIVERSITY

Master of Engineering (Environmental Engineering)

DEGREE

Environmental Engineering Environmental Engineering

FIELD DEPARTMENT

TITLE: Nutrient Removal Enhancement by a Continuous Biocarrier- Anoxic/

Aerobic-MBR Process from Synthetic Domestic Wastewater

NAME: Miss Patchaya Ruchirattanawarakorn

THIS THESIS HAS BEEN ACCEPTED BY

THESIS ADVISOR

( Associate Professor Chart Chiemchaisri, D.Eng. )

THESIS CO-ADVISOR

( Associate Professor Wilai Chiemchaisri, D.Tech.Sc. )

THESIS CO-ADVISOR

( Professor Chia-Yuan Chang, Ph.D. )

DEPARTMENT HEAD

( Associate Professor Chart Chiemchaisri, D.Eng. )

APPROVED BY THE GRADUATE SCHOOL ON

DEAN

( Associate Professor Gunjana Theeragool, D.Agr. )

 THESIS

NUTRIENT REMOVAL ENHANCEMENT BY A CONTINUOUS

BIOCARRIER-ANOXIC/AEROBIC-MBR PROCESS FROM
SYNTHETIC DOMESTIC WASTEWATER

PATCHAYA RUCHIRATTANAWARAKORN

A Thesis Submitted in Partial Fulfillment of

the Requirements for the Degree of
Master of Engineering (Environmental Engineering)
Graduate School, Kasetsart University
2013
 Patchaya Ruchirattanawarakorn 2013: Nutrient Removal Enhancement by a
Continuous Biocarrier–Anoxic/Aerobic-MBR Process from Synthetic Domestic
Wastewater. Master of Engineering (Environmental Engineering), Major Field:
Environmental Engineering, Department of Environmental Engineering. Thesis
Advisor: Associate Professor Chart Chiemchaisri, D.Eng. 87 pages.

The possibility in using a continuous biocarrier-A/O-MBR process in place of

A2O-MBR process for treating synthetic domestic wastewater was studied for its
performance in terms of organic and nutrient removal. The continuous biocarrier-A/O-
MBR process has two compartments including an anoxic and an oxic biological
reactor integrated with membrane filtration. The system was operated for 260 days in
5 different stages (1) anoxic (sponge)-MBR with 3Q recirculation rate (2) extra
alkalinity (NaHCO3) addition (3) anoxic (new sponge model) and pH (NaOH)
adjustment (4) 2Q recirculation rate (5) adjustment of COD/N ratio to 7. The results
showed that TCOD, SCOD and ammonia removal was higher than 99%. The TN and
TP removal of 85% and 81% can be achieved. The concentration of ammonia in MBR
tank, nitrite and nitrate in anoxic tank were almost non-detected, which indicated that
this operating condition can reach complete nitrification and high efficiency of
denitrification. The sponge addition in the anoxic tank helped reducing membrane
fouling and the results suggested that sustainable operation up to 55 days was
maintained without any membrane cleaning.

/ /
Student’s signature Thesis Advisor’s signature
 ACKNOWLEDGEMENTS

I would like to extend my gratitude to all those who gave me hope for the
success of this thesis. I would like to give special thanks to the Department of
Environmental Engineering (Kasetsart University, Thailand) and the Department of
Environmental & Science (Chia Nan University of Pharmacy and Science, Taiwan)
for giving me the financial support for dual master degree program.

I am deeply indebted to my advisor Associated Professor Chart Chiemchaisri

and co-advisor Professor Dr. Chia-Yuan Chang, Associate Professor Wilai
Chiemchaisri who give me help, suggestions and encouragement all the time of
research and for writing this thesis. Also I would like to thank the committee
members Assistant Professor Monthon Thanuttamavong, Professor Dr. Chettiyappan
Visvanathan, Hong-Thih Lai Ph. D. and Cheng-di Dong Ph. D. for good comments to
improve my thesis.

Special thanks to my colleagues in Taiwan and Thailand who assisted me in

my research works for all their help, support, interest and valuable hints.

Especially, I would like to give my thanks to my parents, who help me in

everything and always give me the greatest love, willpower and financial support
until the completion of this study.

Patchaya Ruchirattanawarakorn
September, 2013
 i

TABLE OF CONTENTS

Page

TABLE OF CONTENTS i
LIST OF TABLES ii
LIST OF FIGURES iii
LIST OF ABBREVIATIONS v
INTRODUCTION 1
OBJECTIVES 5
LITERATURE REVIEW 7
MATERIALS AND METHODS 33
Materials 35
Methods 39
RESULTS AND DISCUSSION 46
CONCLUSIONS AND RECOMMENDATIONS 74
Conclusions 74
Recommendations 75
LITERATURE CITED 76
CURRICULUM VITAE 87
 ii

LIST OF TABLES

Table Page

1 Typical composition of untreated domestic wastewater 8

2 Definition of the various terms used to define various nitrogen species 11
3 Domestic wastewater phosphorus contribution by different sources
in a household 20
4 Advantages and disadvantages of MBR configurations 29
5 Process removal performances under the different combined
MBR systems 30
6 Sponge Characteristics 35
7 Membrane Characteristics 37
8 Characteristics of Synthetic Domestic Wastewater 38
9 Stages of study at different operating conditions 42
10 Parameters and analysis methods 43
11 The range, mean value and highest removal efficiency of TCOD
and SCOD at 5 stages under different operation conditions 48
12 The range, mean value and highest removal efficiency of ammonia
and total nitrogen of 5 stages under different operation conditions 51
13 The range, mean value and highest removal efficiency of total
phosphorus at 5 stages under different operation conditions 62
14 Average MLSS concentration in suspended termed and attached
termed in anoxic tank. 64
15 Average value of COD volumetric loading rate for 5 stages 69
 iii

LIST OF FIGURES

Figure Page

1 Nitrogen transformation in biological treatment processes 12

2 Model of metabolism of phosphate accumulating organism PAOs under
(a) anaerobic metabolism (b) aerobic/anoxic metabolism 25
3 MBR configurations (a) side stream MBR and (b) submerged MBR 28
4 Biocarrier-anoxic/aerobic-MBR Experimental Plans 34
5 Sponge Material Applied to Anoxic Tank 36
6 Hollow Fiber Membrane Applied to Aerobic Tank 36
7 Schematic of the lab-scale biocarrier-anoxic/aerobic-MBR system 40
8 Biocarrier-anoxic/aerobic-MBR system 41
9 TCOD variation and removal during the experiment 48
10 SCOD variation and removal during the experiment 49
11 Relationship between COD removal and MLVSS in anoxic tank 49
12 Ammonia removal profile during the study period 50
13 Total nitrogen removal profile during the study period 51
14 Relationship between alkalinity and ammonia removal efficiency 53
15 Nitrification and denitrification during the period of study 53
16 Variation of pH on denitrification 55
17 Variation of ORP on denitrification 56
18 Variation of pH on nitrification 56
19 Variation of ORP on nitrification 57
20 Effect of COD/N ratio on total nitrogen removal 58
21 Continuous denitrification with different COD/N Ratio in anoxic tank 59
22 Relation of MLSS, MLVSS with total nitrogen removal 59
23 Nitrogen concentration in permeate 60
24 Phosphorus removal profile during the study period 62
25 Average MLSS in anoxic tank during experimental period 64
 iv

LIST OF FIGURES (Continued)

Figure Page

26 Variation of MLSS in this experimental period 65

27 Variation of MLVSS in this experimental period 66
28 Variation of F/M ratio in this experimental period 67
29 Variation of COD volumetric loading rate in this experimental period 69
30 Average flux through experimental period 71
31 Variation of TMP and COD/N ratio through experimental period 73
 v

LIST OF ABBREVIATIONS

CAS = conventional activated sludge

COD/N = COD to nitrogen ratio
COD = chemical oxygen demand
DO = dissolved oxygen
EPS = extracellular polymeric substance
HRT = hydraulic retention time
F = permeate flux (L/m2.h)
M = meter
m/s = meter per second
m3/s = cubic meter per second
mg/L = milligram per liter
MBR = membrane bioreactor
A/O-MBR = anoxic/aerobic-membrane bioreactor
MF = Microfiltration
MLSS = mixed liquor suspended solids
MLVSS = mixed liquor volatile suspended solids
NaOCl = sodium hypochlorite
NF = Nanofiltration
-N = ammonium nitrogen
-N = nitrite nitrogen
-N = nitrate nitrogen
PVDF = polyvinylidene fluoride
SCOD = soluble chemical oxygen demand
TCOD = total chemical oxygen demand
TDS = total dissolved solids
TMP = transmembrane pressure
 vi

LIST OF ABBREVIATIONS (Continued)

TN = total nitrogen
SRT = solid retention time
A2O = anaerobic-anoxic-oxic
Aer. = aerobic
Ana. = anaerobic
Ano. = anoxic
BNR = biological nutrient removal
Eff. = effluent
Effi. = efficiency
Inf. = influent
SBR = sequencing batch reactor
UCT = University of Cape Town
Org-N = organic nitrogen
TP = total phosphorus
ORP = oxidation-reduction potential rate
NaOH = sodium hydroxide
 1

NUTRIENT REMOVAL ENHANCEMENT BY A CONTINUOUS

BIOCARRIER-ANOXIC/AEROBIC-MBR PROCESS FROM
SYNTHETIC DOMESTIC WASTEWATER

INTRODUCTION

Domestic wastewater contains considerable amount of nutrient and organic

matter. The principal sources of domestic wastewater in a community are the
residential areas and commercial districts. Other important sources include
institutional and recreational facilities. It can intensify water eutrophication if not
adequately treated before discharging into surface waters. Eutrophication is a major
water quality problem worldwide that causes algal blooms that stimulate the growth
of algae, other photosynthetic microorganisms such as toxic cyanobacteria and
disrupts the normal functioning of aquatic ecosystems. Due to the very limited
regional environmental capacity of the receiving water, there can be an increase in
rigorous standards for effluent discharge in the coming future. Removal of nitrogen
and phosphorus from domestic wastewater is one of the key strategies in preventing
eutrophication. Reducing nutrient input to nearby waters, along with other technical
innovations, is an obligatory mission for both wastewater treatment plants (WWTPs)
engineers and the neighboring communities (Zhang et al., 2011).

Over the past several decades, biological nutrient removal (BNR) processes
have been widely studied, which have proven that biological nutrient removal process
is very efficient for removing nitrogen and phosphorus, as well as chemical oxygen
demand (COD) to prevent eutrophication due to their economic advantages compared
with chemical treatment methods. The BNR systems developed include the
sequencing batch reactor (SBR), the University of Cape Town (UCT) system, the
Bardenpho process and the Anaerobic/Anoxic/Oxic (A2O) system, etc. Amongst
these processes, the most commonly used process is the A2O process (Peng et al.,
2006).
 2

The continuous-flow A2O process is simultaneous removal of carbon,

nitrogen and phosphorus, usually the biological nutrient removal process is connected
with anaerobic, anoxic and aerobic (A2O) treatment zone. It is a single-sludge
suspended growth system. In this system, denitrifiers perform the major role in
nitrogen removal and phosphate-accumulating organisms (PAOs) are responsible for
enhanced biological phosphorus removal (EBPR). Both of these microorganisms
require carbon source (COD) to independently carry out the reaction. Accordingly,
the availability of COD is often an essential limiting factor when simultaneous
removal of nitrogen and phosphorus is attempted. For nitrogen removal,
denitrification occurs only under anaerobic or anoxic condition. During
denitrification, the biological reductions of nitrate (NO3-) to nitrogen gas (N2) done by
heterotrophic bacteria and the nitrifying liquid from aerobic tank is to be return to
anaerobic or anoxic tank. The nitrification is conversion of ammonium (NH4) to
nitrate nitrogen. Nitrification is a two-step process. Bacteria Nitrosomonas convert
ammonia and ammonium to nitrite. Second step is bacteria Nitrobacteria conversion
of nitrite to nitrate. Nitrification occurs only under aerobic conditions at dissolved
oxygen (Peng and Zhu, 2006). For phosphorus removal under anaerobic conditions,
volatile fatty acids (VFA) are taken up from the liquid phase and stored as PHB. At
the same time an energy carrier is released during the construction of PHB from
glycogen. The energy required comes from the hydrolysis of polyphosphate and the
subsequent formation of ATP. It results in an increase of orthophosphate
concentration in solution. Under anoxic or aerobic condition, the stored PHB will be
oxidized to CO2. The polyphosphate accumulating organisms (PAOs) grow and
restocked with polyphosphate from orthophosphate and glycogen. This results in a
decrease of orthophosphate concentration in solution (Comeau et al, 1986).

Membrane bioreactors (MBRs) are generally not restricted in their biological

process design, and many biological process designs have been used in combination
with membrane technology to create MBR systems. Biological process integrated
with membrane separation is one of the fastest growing technologies. It is a new
generation of hybrid techniques. The concept of MBR systems consists of utilizing a
bioreactor and microfiltration as one unit process for wastewater treatment thereby
 3

replacing, and in some cases supplementing, the solids separation function of

secondary clarification and effluent filtration. The membrane bioreactor (MBR)
system have small footprint, produce consistent effluent quality even in varying
influent condition, and provide effective treatment for high organic levels in
wastewater. MBR allows the biological process to be operated at long sludge ages.
High mixed liquor suspended solids (MLSS) concentrations and long SRT support
massive process advantage including stable operation, complete nitrification, and low
bio-solids production. High MLSS concentration also reduce biological volume
requirement. Moreover, the membrane tank could provide highly space efficient solid
separation (Malamis and Andreadakis, 2009).

In this way, the exclusive performance of membrane filtration and biological

nutrient removal was succeeded. An anaerobic, anoxic and aerobic-membrane
bioreactor (A2O-MBR) system was applied in laboratory scale by Yeom et al. (2009)
in which the performance of nitrogen and phosphorus removal efficiencies are 60-
65% and 74-82% respectively. An effective operation was adopted in this study to
complete several functions in one way including carbon and nitrogen removals,
phosphorus recovery, sludge reduction as well as water reuse. It is obvious that a good
performance on phosphorus removal was achieved as well as the recovery of
phosphorus from sludge was accomplished in this study. However, the nitrogen
removal of this process was not high enough. It seems to indicate that the operating
condition was advantageous to phosphorus removal, and not with nitrogen. In
addition, the carbon source consume in anaerobic/anoxic unit for phosphorus removal
should be higher than that of the unit for nitrogen removal. Theoretically, the COD/P
ratio for phosphorus removal generally is in the range of 40-50 (Reddy and Movva,
1998), which is much higher than the COD/N ratio of 7-10 for nitrogen removal (Choi
et al., 2008). The high COD/P ratio was easy to be achieved in Yeom’s (2009) study
since they sent back the thermal digested sludge to anaerobic unit to supply the carbon
source. However, it’s difficult for some kinds of wastewater to have a high COD/P
ratio without sludge return. Also, thermal digestion of sludge is an energy cost
process for most wastewater treatment plant.
 4

In this study, hybrid MBR system was tried to find an alternative related to
have the function for carbon and nutrient removal. In Wang’s (2009) previous study,
an anoxic/aerobic-membrane bioreactor (A/O-MBR) was conducted to estimate the
performance and the result showed a better removal efficiencies of COD was over
94%, ammonia-nitrogen, total nitrogen and total phosphorus removal efficiencies
were 92%, 78% and 64% respectively. The research result shown that even without
anaerobic process the simultaneous removal of nutrient could be done and the high
removal efficiencies were achieved. Several advantages of A/O-MBR could be
involved including smaller footprint, operating cost and maintenance reduction.
Hence, on the basis of economical factor, biocarrier-anoxic/aerobic-MBR consists of
anoxic/aerobic-membrane bioreactor (A/O-MBR) with nitrate liquid recycle and
sponge addition as bio-carrier in anoxic zone was use in this study for organic and
nutrient removal in wastewater treatment.
 5

OBJECTIVES

1. To examine the possibility of biocarrier-A/O-MBR process for simultaneous

removing organic and nutrient from wastewater.

2. The different operating conditions, alkalinity, pH, sludge recycling rate and
COD/N ratio, were assessed in this study. This study tried to identify the themes of
these four parameters and enhance the system efficiency for the biological nutrient
removal in this biocarrier-A/O-MBR process.

3. To study the possible mechanism and efficiency of nutrient removal in

biocarrier-A/O-MBR process by use a synthetic domestic wastewater.

4. To use biocarrier (sponge) to increase the treatment performance including

denitrification and try to reduce the membrane fouling in this biocarrier-A/O-MBR
process.
 6

Scope of this study

The scope of the study is to investigate and conduct experiments with

synthetic domestic wastewater under ambient conditions by using a biocarrier-
anoxic/aerobic-MBR process with sponge addition in to the anoxic tank which
different operating conditions, alkalinity, pH, sludge recycling rate and COD/N ratio.
Furthermore, the mechanism of nitrogen as the nitrogen concentrations in the anoxic
and aerobic compartment were analyzed to find the detailed removal pathways of
nitrogen and enhance performance of removal organic and nutrient by a biocarrier-
anoxic/aerobic-MBR process was evaluated.
 7

LITERATURE REVIEW

1. Domestic wastewater

Domestic wastewater refers to waste water that is discarded from households,

hotel, commercial and institutional facility in urbanized area. Also referred to as
sanitary sewage, such water contains a wide variety of dissolved and suspended
impurities. It amounts to a very small fraction of the sewage by weight. But it is large
by volume and contains impurities such as organic materials and plant nutrients that
tend to rot. The typical composition of untreated domestic wastewater is shown in
Table 1. The main organic materials are food and vegetable waste, plant nutrient
come from chemical soaps, washing powders, etc. Domestic wastewater is also very
likely to contain disease causing microbes. In general, domestic wastewater includes
black water (fecal sewage) and gray water (wastewater from dishwashers, washing
machines, sinks and baths). Black water accounts for 32.5% of domestic wastewater,
while gray water accounts for 67.5% (Metcalf and Eddy, 2004). Thus, disposal of
domestic wastewater is a significant technical problem.

Today, many people dump their garbage into streams, lakes, rivers, and seas,
thus making water bodies the final resting place of cans, bottles, plastics, and other
household products. The various substances that we use for keeping our houses clean
add to water pollution as they contain harmful chemicals. In the past, people mostly
used soaps made from animal and vegetable fat for all types of washing. But most of
today’s cleaning products are synthetic detergents and come from the petrochemical
industry. Most detergents and washing powders contain phosphates, which are used to
soften the water among other things. These and other chemicals contained in washing
powders affect the health of all forms of life in the water. When fresh water is
artificially supplemented with nutrients, it results in an abnormal increase in the
growth of water plants. This is known as eutrophication. The discharge of waste from
urban communities into water bodies generally stretches the biological capacities of
aquatic systems. Excess nutrients cause the water body to become choked with
organic substances and organisms. When organic matter exceeds the capacity of the
 8

microorganisms in water that break down and recycle the organic matter, it
encourages rapid growth, or blooms of algae. When they die, the remains of the algae
add to the organic waste already in the water, eventually, the water becomes deficient
in oxygen. Anaerobic organisms (those that do not required oxygen to live) then
attack the organic wastes, releasing gases such as methane and hydrogen sulphide,
which are harmful to the oxygen requiring (aerobic) forms of life (Metcalf and Eddy,
2004). The result is a foul smelling, waste filled body of water.

Eutrophication can produce problems such as bad taste and odor as well as
green scum algae. Also the growth of rooted plants increases, which decreases the
amount of oxygen in the deepest water of the lake. It also leads to the death of all
forms of life in the water bodies. The effects of water pollution are not only
devastating to people but also to animals such as fish, and birds. Polluted water is
unsuitable for drinking, recreation, agriculture, and industry. It diminishes the
aesthetic quality of lakes and rivers. More seriously, contaminated water destroys
aquatic life and reduces its reproductive ability. Eventually, it is a hazard to human
health. Construction of public underground sewerage systems is the key to
minimizing domestic water pollution.

Table 1 Typical composition of untreated domestic wastewater

Concentration
Contaminants Unit Low Medium High
strength strength strength
Solids, Total mg/L 390 720 1230
(TS)
Dissolved solids, mg/L 270 500 860
total (TDS)
Suspended solids, mg/L 120 210 400
total (TSS)
Settleable solids mg/L 5 10 20
 9

Table 1 (Continued)

Concentration
Contaminants Unit Low Medium High
strength strength strength
Biochemical mg/L 110 190 350
oxygen demand,
5-d, 20°C (BOD,
20°C)
Total organic mg/L 80 140 260
carbon (TOC)
Chemical oxygen mg/L 250 430 800
demand (COD)
Chlorides mg/L 30 50 90
Sulfate mg/L 20 30 50
Nitrogen mg/L 20 40 70
(total N)
Organic mg/L 8 15 25
Free ammonia mg/L 12 25 45
Nitrites mg/L 0 0 0
Nitrates mg/L 0 0 0
Phosphorus (TP) mg/L 4 7 12
Organic mg/L 1 2 4
Inorganic mg/L 3 5 8
Oil and grease mg/L 50 90 100
Volatile organic µg/L ˂100 100-400 >400
compound
(VOCs)

Source: Metcalf and Eddy (2004)

 10

2. Nitrogen

The element nitrogen is significant to the growth of microorganisms, plants

and animals, which is known as nutrients or biostimulants. Nitrogen is the most
important nutrient because it is the essential building block in the synthesis of protein,
and nitrogen data will be required to estimate the treatability of wastewater by
biological process. Insufficient nitrogen can necessitate the addition of nitrogen to
make the waste treatable. Nutrient requirements for biological waste treatment which
control for algal growth in the receiving water is necessary, removal and letdown of
nitrogen in wastewater before discharge may be desirable.

2.1 Forms of nitrogen

The chemistry of nitrogen is complicated, because of the several oxidation

states that nitrogen can assume and the changes of the oxidation state can be brought
about by living organisms. The oxidation state changes brought about by bacteria can
be either positive or negative depending upon whether aerobic or anaerobic conditions
slam. The oxidation states of nitrogen are summarized below (Sawyer et al, 1994).

-III 0 I II III IV V
NH3 N2 N2O NO N2O3 NO2 N2O5

The most common and important forms of nitrogen in wastewater and their
corresponding oxidation state in the water/soil environment are ammonia (NH3, -III),
ammonium (NH4+, -III), nitrogen gas (N2, 0), nitrite ion (NO2─, +III), and nitrate ion
(NO3─, +V). The oxidation state of nitrogen in most organic compounds is -II. Total
nitrogen, as reported in Table 2, is comprised of organic nitrogen, ammonia, nitrite,
and nitrate.
 11

Table 2 Definition of the various terms used to define various nitrogen species

Form of nitrogen Abbrev. Definition

Ammonia gas NH3 NH3

Ammonium ion NH4+ NH4+

Total ammonia nitrogen TANa NH3 + NH4+

Nitrite NO2─ NO2─

Nitrate NO3─ NO3─

Total inorganic nitrogen TINa NH3 + NH4+ + NO2─ +

NO3─
Total Kjeldahl nitrogen TKNa Organic N + NH3 + NH4+

Organic nitrogen Organic Na TKN - (NH3 + NH4+)

Total nitrogen TNa Organic N + NH3 + NH4+

+ NO2─ + NO3─

a
All species expressed as N

Source: Metcalf and Eddy (2004)

2.2 Sources of nitrogen

The major source of nitrogen compounds are

2.2.1 The nitrogenous compound of plant and animal source. Ammonia

came from the distillation of bituminous coal such as nitrogen got from decayed plant
material (Metcalf and Eddy, 2004; Sedlak, 1991).
 12

2.2.2 Sodium nitrate (NaNO3) is found mainly in mineral precipitates in

Chile and in the fertilizer found in seabird rookeries (Metcalf and Eddy, 2004; Sedlak,
1991).

2.2.3 Atmospheric nitrogen. The production of nitrogen from the

atmosphere is term fixation. Because fixation is a biologically mediated process and
because NaNO3 precipitates are relatively rare, most sources of nitrogen in
soil/groundwater are of biological origin (Metcalf and Eddy, 2004; Sedlak, 1991).

2.3 Nitrogen transformations in biological treatment processes

The nitrogen transformations that may occur in biological treatment

systems are showed in Figure 1.

Figure 1 Nitrogen transformation in biological treatment processes

Source: Metcalf and Eddy (2004); Sedlak (1991)

 13

From Figure 1, organic nitrogen present in raw wastewater may be

transformed to ammonia through bacteria decomposition of proteinaceous matter and
hydrolysis of urea. In any biological treatment system some bacteria growth always
takes place. Since nitrogen constitutes 12 to 13 percent of cell dry mass, some
ammonia nitrogen will be assimilated in newly formed cells. Depending on the
treatment process and the loading condition, cell autoxidation and lysis will also
occur. Hence part of the ammonia used for cell synthesis will be returned to the liquid
through lysis and autoxidation. The remaining assimilated nitrogen can be removed
from the system in the net growth, or wasted biological sludge.

Under appropriate conditions, ammonia nitrogen can be oxidized in a two-

step process to form nitrates. This process, called nitrification, is carried out by two
groups of microorganisms (nitrifiers) in the presence of oxygen. Finally, nitrates may
be transformed to nitrogen gas through a process called denitrification. This
transformation is accomplished by denitrifying microorganisms in the absence of
oxygen. An organic carbon source is required for denitrification to occur. The
nitrogen gas formed escapes to the atmosphere. A residual of nondegradable soluble
organic nitrogen of about 1 mg N/L will remain in the effluent.

2.4 Available nitrogen removal options

Nitrogen entering a biological treatment system in the organic or ammonia

form can be either removed or transformed to another form. Removal of nitrogen is
obtained by assimilation and by conversion to nitrogen gas through nitrification and
denitrification (Metcalf and Eddy, 2004; Sedlak, 1991).

2.5 Nitrogen removal by assimilation

Since nitrogen is an essential constituent of microbial cells, any net

growth of biomass that is removed from the waste stream will cause some nitrogen
removal. The amount of nitrogen that can be removed by this mechanism is limited by
the amount of net growth, which in turn depends on the carbonaceous organic content
 14

of the wastewater and the system’s operating conditions. Since the nitrogen content of
microbial cells is approximately 12.5 percent (on a dry weight basis). The nitrogen
removal by assimilation is limited to approximately 2 to 5 percent of the raw
wastewater BOD, depending on operating conditions. The percent nitrogen removal in
the treatment of domestic wastewater may range from 8 to 20 percent. This removal
mechanism may become quite significant in wastewaters having relatively high
concentrations of BOD, such as in some industrial wastewaters or municipal
wastewater with a large industrial contributor. Net growth should be maximized (by
increasing organic loading) in order to maximize assimilative nitrogen removal
(Metcalf and Eddy, 2004; Sedlak, 1991).

2.6 Nitrification and denitrification process

2.6.1 Nitrification process plays an important role in removal of ammonia

nitrogen. During nitrification process, ammonia is biologically oxidized to nitrate
(with nitrite intermediate substance) with autotrophic bacteria such as Nitrosomonas
and Nitrobacter which carry out the reaction in two steps in aerobic tank (Spagni and
Marsili-Libelli, 2009). Nitrification process is the oxidation of nitrogen compounds in
which nitrogen acts as an electron donor and oxygen as electron acceptor.
Stoichiometry of biological nitrification follows as

Nitroso-bacteria:

2 NH 4  3O2  2 NO2  4H   2H 2O (1)

Nitro-bacteria:

2 NO2  O2  2 NO3 (2)

 15

Total oxidation reaction:

NH 4  2O2  NO3  2H   H 2 O (3)

Base on total oxidation reaction, the oxygen required for complete

oxidation of ammonia is 4.33 g O2/g N oxidized with 3.22 g O2/g used for nitrite
production and 1.11 g O2/g NO2 oxidized (Werzernak and Gannon, 1967). The
amount of alkalinity required to carry out the reaction each g of ammonia nitrogen (as
N) converted, 0.16 g of new cells are formed, 7.07 g of alkalinity as CaCO3 are
removed, and 0.08 g of inorganic carbon are utilized in the formation of new cells
(Metcalf and Eddy, 2004; Sedlak, 1991).

2.6.2 Factors affecting nitrification

a) pH: Nitrification is pH-sensitive and rates decline significantly at

pH values below 6.8. Optimal nitrification rates occur at pH values in the 7.5 to 8.0
range. A pH of 7.0 to 7.2 is normal used to maintain reasonable nitrification rates
(Metcalf and Eddy, 2004; Sedlak, 1991).

b) Toxicity: Nitrification organisms are sensitive to a wide range of

organic and inorganic compounds and at concentrations well below those
concentrations that would affect aerobic heterotrophic organisms. In many case
toxicity may be sufficient to kill the nitrifying bacteria and significantly reduced
nitrification rates. Nitrification has been shown to be good indicators of the presence
of organic toxic compounds at low concentrations (Blum and Speece, 1991).
Compounds that are toxic include solvent organic chemicals, amines, proteins,
tannins, phenolic compounds, alcohols, cyanates, ethers, carbamates and benzene
(Hockenbury and Grady, 1977; Sharma and Ahlert, 1977).
 16

c) Metal: Metals have shown to completely inhibit ammonia

oxidation at 0.25 mg/L nickel, 0.25 mg/L chromium and 0.10 mg/L copper (Skinner
and Walker, 1961).

d) Un-ionized ammonia: Nitrification is also inhibited by un-ionized

ammonia (NH3) or free ammonia and un-ionized nitrous acid (HNO2). The inhibition
effects are dependent on the total nitrogen species concentration, temperature and pH.
At 20°C and pH 7.0, the NH4-N concentrations at 100 mg/L and 20 mg/L may initiate
inhibition of NH4-N and NO2-N oxidation, respectively, and NO2-N concentration at
280 mg/L may initiate inhibition of NO2-N oxidation (U.S. EPA., 1993).

e) Temperature: Optimal temperature for nitrification reaction is

range 28-36 °C (Metcalf and Eddy, 2004; Sedlak, 1991).

f) SRT: Requiring much longer SRT values for nitrifying

organisms. Typical design SRT values may range from 10 to 20 days at 10 °C and 4
to 7 days at 20 °C. At elevated temperatures, the relative kinetics of NH4-N and NO2-
N oxidation change, and NO2-N will accumulate at lower SRT (Metcalf and Eddy,
2004; Sedlak, 1991).

g) DO: Nitrification rates increase up to DO concentration of 3 to 4

mg/L. At low DO concentration (˂ 0.50 mg/L) where nitrification rates are greatly
inhibited, the low DO inhibition effect has been shown to be greater for Nitrobacter
than for Nitrosomonas (Metcalf and Eddy, 2004; Sedlak, 1991).

2.6.3 Denitrification process is the biological conversion of nitrate

nitrogen to more reduced forms such as N2, N2O and NO with heterotrophic bacteria
such as Pseudomonas and Bacillus in anoxic tank (You and Chen, 2008). Fu et al.,
(2009) and Ahn et al., (2003) found that denitrification was the rate limiting step for
total nitrogen removal in combined process. Denitrification involves biological
oxidation of many organic substrates in wastewater treatment in which nitrate acts as
 17

electron acceptor instead of oxygen (Metcalf and Eddy, 2004). Stoichiometry of

biological denitrification follows as

C10 H19O3 N  10 NO3  5N 2  10CO2  3H 2O  NH 3  10OH  (4)

In heterotrophic denitrification reaction, one equivalent of alkalinity

is produced per equivalent of NO3-N reduced, which equates to 3.57 g of alkalinity
(as CaCO3) production per g of nitrate nitrogen reduced. As a general rule, (Barth et
al., 1968) estimated that 3.7 g of COD is needed per gram of NO3 reduced. However,
the actual value will depend on the system operating condition and the type of
electron donor used for denitrification. From a steady stage COD balance it can be
shown that the COD removed is oxidized or accounted for in cell growth, which 0.45
g of new cells are produced per g of NO3 reduced.

2.6.4 Factors affecting denitrification

a) DO: Dissolved oxygen can inhibit nitrate reduction by repressing

the nitrate reduction enzyme. A dissolved oxygen concentration of 0.2 mg/L and
above has been reported to inhibit denitrification for a Pseudomonas culture (Skerman
and Macrae, 1957; Terai and Mori, 1975).

b) pH: No significant effect on the denitrification rate has been

reported for pH between 7.0-8.0, while Dawson and Murphy (1972) showed a
decrease in the denitrification rate as the pH was decrease from 7.0 to 6.0.

c) Temperature: Denitrification rate reduced at low temperature due

to low dissolved oxygen at high temperature.

d) Carbon source: Type of carbon source has effect to denitrification

rate due to high denitrification rate can occur for the readily biodegradable organics
such as methanol.
 18

3. Phosphorus

3.1 Phosphorus compound

Phosphorus is a normal part of the environment. It is a nutrient used by

organisms for growth. It occurs in form of phosphates in natural water, wastewater,
containing rocks and as the excretory and decay products of plants and animals. In the
natural world phosphorous is never encountered in its pure form, but only as
phosphates, which consists of a phosphorous atom bonded to four oxygen atoms. This
can exists as the negatively charged phosphate ion (PO43-), which is how it occurs in
minerals, or as organophosphates in which there are organic molecules attached to
one, two or three of the oxygen atoms. Phosphates come from a variety of sources
including agricultural fertilizers, domestic wastewater, detergents, industrial process
wastes and geological formations.

3.2 Chemical forms of phosphate

The usual forms of phosphorous found in aqueous solutions include:

3.2.1 Orthophosphate, for example, Trisodium phosphate (Na3PO4),

Disodium phosphate (Na2HPO4), Monosodium phosphate (NaH2PO4), Diamonium
phosphate ((NH4)2HPO4), are available for biological metabolism without further
breakdown.

3.2.2 Polyphosphate or Condensed phosphate, for example, Sodium

hexametaphosphate (Na3(PO4)6), Sodium tripolyphosphate (Na5P3O10), Tetrasodium
pyrophosphate (Na4P2O7). It included those molecules with two or more phosphorus
atom, oxygen atoms, and, in some case hydrogen atoms combined in a complex
molecule. The polyphosphate are dehydrated phosphate by undergo hydrolysis in
aqueous solution and revert to the orthophosphate forms, this hydrolysis is usually
quite slow. The rate of reaction depended on temperature, low pH and higher rate in
wastewater.
 19

3.2.3 Organic phosphate, for example, Nucleic acid, Phospholipids, Sugar

phosphate. The organically bound phosphorus is usually of minor importance in most
domestic waste, but it can be an important constituent of industrial wastes and
wastewater sludge.

3.3 Source of phosphorus in wastewater

Under normal water flows, roughly two-thirds of the total phosphorus load
to lakes and rivers comes from nonpoint sources.

3.3.1 Synthetic detergent and household cleaning product

Phosphorus in the wastewater comes from toilet wastes as well as

bath, laundry, and kitchen wastewater. Toilet wastes contribute 30%-70% of P in the
form of nucleic acids and adenosine tri–phosphate excreted in feces. Detergent
(laundry) and dish soaps (kitchen and sinks) used in the household contribute another
30%-70% of P (Wilhelm et al., 1994). The U.S. EPA (U.S.EPA, 2002) estimates that
each person in the United State contribute 2.7 grams of total P to wastewater each day
as shown in Table 3. Of this total P, toilet wastewater contributes the highest amount
(59%), followed by bath, sinks, and appliances (37%), and garbage disposals (4%).
 20

Table 3 Domestic wastewater phosphorus contribution by different sources in a

household.

Source of Phosphorus Contribution Contribution

(grams/person/day) (% of total)
Garbage disposal (kitchen) 0.1 4%
Toilet 1.6 59%
Bath, sinks, and 1.0 37%
appliances
Total phosphorus 2.7

Source: U.S.EPA (2002)

3.3.2 Pasture, waste materials and agricultural

The primary pasture, waste materials and agricultural nonpoint

source pollutants are nutrients (particularly nitrogen and phosphorus), sediment,
animal wastes, pesticides, and salts. It enters to surface water through direct surface
runoff or through seepage to ground water that discharges to a surface water outlet.
Various farming activities result in the erosion of soil particles. The sediment
produced by erosion can damage fish habitat and wetlands and, in addition, often
transports excess agricultural chemicals resulting in contaminated runoff. This runoff
in turn affects changes to aquatic habitat such as temperature increases and decreased
oxygen. Phosphorus loading contributed by runoff from pastures and cropland is
largest source of nonpoint phosphorus on a statewide basis.

3.3.3 Industrial and commercial uses

Phosphorus has many applications to the agriculture and farm

industries. In particular, phosphorus is used extensively in fertilizers as concentrated
phosphoric acid. Phosphorus is used in the production of special glasses (sodium
 21

lamps), as a cleaning agent (trisodium phosphate), and as a water softener (for the
prevention of pipe and boiler tube corrosion on boilers). Phosphorus is also used in
the production of phosphor bronze and steel.

3.4 Phosphorus removal

Phosphate removal from wastewater involves the incorporation of

phosphate into a particulate form (suspended solids) and then the removal of the
suspended solids. The types of suspended solids into which phosphate can be
incorporated are either biological (micro-organisms) or chemical (sparingly soluble
metal phosphate precipitates). The physical removal and subsequent processing of
these phosphate-containing solids should be accomplished without allowing
significant release of phosphate into liquid streams.

3.4.1 Chemical phosphorus removal

Chemical precipitation of phosphate usually becomes necessary

when the phosphorus discharge criteria are low. Chemical process for phosphate
removal commonly rely on the formation of sparingly soluble orthophosphate that
can be removed by solids separation process either together with raw sludge and/or
waste biological solids or separately. Phosphate precipitation process can be classified
according to their location in the process stream (Sedlak, 1991).Phosphate
precipitation is achieved by the addition of the salts of one of three metals that form
sparingly soluble phosphates. These are calcium (Ca (II)), iron (either ferriciron, Fe
(III), or ferrous iron, Fe (II), and aluminum (Al (III)) (Metcalf and Eddy, 2004). The
salt most commonly employed as shown below.

a) Lime (Ca (OH)2). Lime dosage is more influenced by alkalinity

than phosphorus concentration; the pH must be raised to 10.5 for phosphorus removal
to occur. The amount of lime required is approximately 1.5 times the alkalinity
concentration in mg/L. The chemical equations for lime removal of phosphorus are
given below.
 22

Ca(OH)2 HCO CaCO H2O (5)

5Ca2 OH- HPO 2-

Ca5(OH)(PO ) H2O (6)

b) Aluminum salts. Aluminum is commercially available in five

forms: (Al2 (SO4)3, sodium aluminate (Na2Al2O4), poly-aluminum chloride (PAC),
aluminum chloride (AlCl3), aluminum chlorohydrate. Theoretically, 0.87 pound of
aluminum removes one pound of phosphorus (as P). The different aluminum forms
consume differing amounts of alkalinity. Alum uses approximately 0.5 mg/L of
alkalinity for each mg/L of aluminum added. Aluninum chloride uses 1 mg/L. PAC
uses almost no alkalinity. The optimum pH is 6.5. A simplified chemical equation
illustrating aluminum precipitation of phosphorus is given below

Al PO - Al(PO ) (7)

Al HCO AlOH (8)

c) Iron salt. Iron is commercially available in forms: ferric chloride

(FeCl3), ferric sulfate (Fe2 (SO4)3), ferrous sulfate (FeSO4) and ferrous chloride
(FeCl2). Pickle liquor, a waste product of the steel industry, containing ferrous iron in
either a sulfuric or hydrochloric acid solution, also is used as a source of iron for
phosphate precipitation. All are corrosive and therefore must be carefully handled.
The basic reactions involved in the precipitation of phosphorus with iron are as
follows

Ferric chloride (FeCl3):

-
eCl PO ePO Cl- (9)

Ferrous chloride (FeCl2):

-
3 eCl2 2PO e (PO )2 Cl- (10)
 23

Ferrous sulfate (FeSO4):

2-
eSO 2PO -
e (PO )2 SO (11)

Theoretically, 1.8 pounds of iron is required to remove one pound of phosphorus (as
P). However, to achieve low phosphorus concentration, much more is required.
Approximately 1 mg/L of alkalinity is consumed for each mg/L of iron added; as a
result, the wastewater pH drops approximately 0.1 per 10 mg/L of iron added. Iron
works over a wide pH range. Iron salt solutions contain some trace metals: up to 75-
100 mg/L depending on the product.

3.4.2 Biological phosphorus removal

Phosphorus removal is generally done to control eutrophication

because phosphorus is a limiting nutrient in most freshwater systems. Treatment plant
effluent discharge limits have range from 0.10 to 2.0 mg/L of phosphorus depending
on plant location and potential impact on receiving waters. The principal advantages
of biological phosphorus removal are reduced chemical costs and less sludge
production as compared to chemical precipitation. In the biological removal of
phosphorus, the phosphorus in the influent wastewater is incorporated into cell
biomass, which after that is removed from the process a result of sludge wasting.
Phosphorus accumulating organism (PAOs) is promote to grow and consume
phosphorus in systems that use a reactor condition that provide PAOs with a
competition advantage more other bacteria. The reactor configuration is consisted of
an anaerobic tank that is positioned before the activated-sludge aeration tank. The
anaerobic tank is mixed with the return sludge and influent wastewater.

a) Process occurring in the anaerobic zone

Normally heterotrophic bacteria in activated-sludge treatment the

typical phosphorus composition is 1.5 to 2.0 %. However, many bacteria are able to
store phosphorus in their cells in the form of energy-rich polyphosphates, resulting in
 24

phosphorus content as high as 20 to 30 % by dry weight. In the anaerobic zone

acetate, propionic and glucose are produced by fermentation of biodegradable soluble
chemical oxygen demand (bsCOD) which is dissolved degradable organic matter that
can be adsorb easily by biomass. The PAOs adsorb acetate and produced intracellular
polyhydroxybutyrate (PHB) storage products by using energy available from stored
polyphosphates. Some of glycogen contained in the cell is also used. PHB is produced
44 w/w% by usage of acetate. At the same time with the acetate uptake is the release
of orthophosphate (O-PO4), as well as magnesium, potassium and calcium cations. It
results in an increase of orthophosphate concentration in solution (Metcalf and Eddy,
2004; uomeau et al., 1986; U.S.EPA, 1987; hiswanath et al., 1988). The PHB
content in the PAOs increases while the polyphosphate decreases. The reaction of
phosphorus release in anaerobic zone is shown in below.

Fermentation PAO
Acetate PHB
Substrate
Propionic PHV (12)

b) Processes occurring in the aerobic/anoxic zone

In the aerobic phases of the process, the organisms, in the

presence of dissolved oxygen (anoxic phase presence of nitrate is electron acceptor in
place of oxygen by different group of bacteria), convert the stored PHB to CO2, water,
and increased cell mass. The stored of PHB is metabolized and some glycogen is
produced. This process providing energy from oxidation and carbon for new cell
growth. The energy is used form polyphosphate bounds in cell storage so that soluble
orthophosphate (O -PO 4 ) is removed from solution and incorporated into
polyphosphate within the bacteria cell. The reactions of phosphorus up take in aerobic
and anoxic zones are shown in equation 9 and 10 respectively (uomeau, 1986). The
new biomass with high polyphosphate storage explains for phosphorus removal. The
stored phosphorus is removed from the biotreatment reactor for disposal with the
waste sludge.
 25

PH O2 nergy ew cell H2 O CO2 (13)

-
PH O nergy ew cell H2O CO2 (14)

The mechanism that occurring in the anaerobic and aerobic/anoxic zones are shown
on Figure 2.

(a) (b)

Figure 2 Model of metabolism of phosphate accumulating organism PAOs under (a)

anaerobic metabolism (b) aerobic/anoxic metabolism

Source: smolders (1994)

3.5 Phosphorus storing microorganisms

Fuhs and Chen (1975); Buchan (1983) and Malnou et al. (1984) identified
the organism associated with phosphorus removal as Acinetobacter. These bacteria
are short, chubby, gram negative rods with a size of 1-1.5 µm. They present in pairs,
 26

short chains or clusters. These bacteria are known to prefer simple substrates. Other
bacteria are species of Pseudomonas and Aeromonas. Pseudomonas present to be
responsible for biological phosphorus uptake and Aeromonas present to be important
for complete fermentation and VFA production (Brodish and Joyner, 1983; Brodish,
1985). Due to many biological phosphorus removal systems have relationship with
nitrification and denitrification, the capability for phosphorus storing microorganisms
to reduce nitrate is an important issue. However, many researches have observed
phosphorus uptake in anoxic zone concurrent with nitrate reduction. The organism in
anoxic zone associated with phosphorus removal as some species of Pseudomonas
and Aeromonas (Laurraine et al., 1985).

3.6 Factors affecting biological phosphorus removal performance

3.6.1 DO concentration: System performance is not affected by DO as

long as the aerobic zone DO concentration is higher than 1.0 mg/L at pH lower than
6.5 that sufficient phosphorus uptake occurs provided the aerobic detention time is
long enough (Sedlak, 1991).

3.6.2 Temperature: Many research reported that phosphorus removal was

not affected by temperature as low as 10°C (Sell et al., 1981; Kang et al, 1985).
However, phosphorus removal capacity does not seem to be affected at low
temperature operations, Shapiro et al. (1967) showed that the specific phosphorus
release rate for batch activated sludge sample increased 5 times as temperature
increased from 10 to 30 °C. This means that more time may be required in aerobic
zone at low temperature for complete fermentation and/or substrate uptake to occur.
Also have some research show high phosphorus removal efficiency at higher
temperature in the range 15 to 33 °C (Helmer and Kunst, 1998; Mamais and Jenkins,
1992).

3.6.3 pH: The study on effects of pH show that higher efficiency biological
phosphorus removal occur at pH 7.5 to 8.0, which the maximum specific growth rate
of Acinetobacter was higher 42% at pH 8.5 compare with 7.0 (Groenestijn and
 27

Deinema, 1985). Liu et al. (1996) showed the maximum specific phosphorus release
rate in anaerobic zone is 80 mg P/g VSS at pH 7.1 to 8.6. The effect of pH on the
specific phosphorus uptake rate in aerobic zone, which Tracy and Flammino (1985)
reported that high specific phosphorus uptake rate occur at pH 6.6 to 7.4 and all
activity was lost at pH lower than 5.2.

3.6.4 Cations: The cations associated with phosphorus storage must be

available, which Wentzel et al. (1989) suggested molar ratio for Mg, K, and ua to
phosphorus are 0.71, 0.50 and 0.25 respectively. The reported by sedlak (1991) show
that influent phosphorus concentration 10 mg/L the concentration of Mg, K and ua
would be required are 5.6, 6.3 and 3.2 mg/L respectively. As phosphate storage are
0.28, 0.26 and 0.09 mole/mole of phosphorus respectively.

3.6.5 SRT (Solids retention time): The biological phosphorus removal,

which phosphorus is removed from the biotreatment reactor for disposal with the
waste sludge. Fukase et al., (1982) study in anaerobic-aerobic system treating
municipal wastewater show BOD: P removal ratio increased 19 to 26 as SRT
increased from 4.3 to 8 days and phosphorus content of the activated sludge decreased
from 5.4 to 3.7%. These result indicated that long SRT need more amount BOD
removal as low effluent phosphorus concentration.

4. Membrane processes

4.1 Introduction to Membrane Processes

4.1.1 Microfiltration (MF) is a pressure-driven process with microporous

membranes for separating particles from solid/liquid suspensions (Kwon et al., 2000).

4.1.2 Ultrafiltration (UF) is a pressure driven membrane process of

importance to the separation and concentration of colloid suspensions, which include
clay particle as well as proteins and other macromolecules (Bacchin et al., 1996).
 28

4.1.3 Nanofiltration (NF) is also used in a wide range of drinking water

treatment and wastewater reclamation. NF membranes can reject smaller size
molecule that cannot be removed by MF and UF membranes. However it needs lots of
energy consumption during the operation (Choi et al., 2002).

4.1.4 Reverse Osmosis (RO) is increasingly being used for desalination of

brackish water and in wastewater (Kumar et al., 2007). Water and solute transport in
RO membranes has been described in terms of nonporous models where the
membrane is assumed to have no pores with all water and solute transport occurring
by diffusion (Spiegler et al., 1996).

4.2 Membrane Bioreactor

The MBR system was first introduced in 1970 for treatment of sanitary
wastewater, and consisted of a suspended-growth biological reactor combined with a
membrane unit process into a single process. The operation of MBR can be
categorized into two type, the first one is side-stream MBR and the second one is
submerged MBR, both of them are shown in Figure 3. The advantages and
disadvantages of both membrane configuration types are summarized in Table 4 as
well.

(a) (b)

Figure 3 MBR configurations (a) side stream MBR and (b) submerged MBR
 29

Table 4 Advantages and disadvantages of MBR configurations

MBR configuration Advantages Disadvantage

Side-stream MBR  Small footprint  Aeration limitations
 Complete solids  Membrane fouling
removal from effluent  Membrane costs
 Effluent disinfection  High operating costs
 High loading rate  High pumping cost (60-
capability 80% of total costs)
 Combined COD, solids  High cleaning
and nutrient removal in Requirement
a single unit  Process complexity
 Low/zero sludge
production
 Rapid start up
 Sludge bulking not a
problem
Submerged MBR  Small footprint  Susceptible to
 Feed-forward control of membrane fouling
O2 demand  High aeration cost
 Less frequent cleaning
required
 Lower operating costs
 Low liquid pumping
costs (28% of total
costs)
 Low energy
consumption
 30

4.3 Aerobic and Anaerobic MBRs

Membrane bioreactor system can operate either aerobic or anaerobic

condition of microorganism cultures. This system can be effective remove organic
pollutant from high contaminant wastewater. Recently, aerobic submerged membrane
bioreactor has been studied with respect to better effluent quality and lower sludge
production when compare to conventional activated sludge process.

The one advantage of MBRS is that it allowed HRT and SRT to be

independent of each other. Combined MBRS has been proved effective removal
organic pollutants from both municipal wastewater and industrial wastewater as
shown in Table 5.

Table 5 Process removal performances under the different combined MBR systems

Parameter Performance
(% removal)
Process
SRT HRT COD/N Return COD NH4+ TN
(days) (days) ratio Q
Anoxic/Aerobic Infinite 1.5 9.30 - 96.20 86.30 83.00
MBR
Anoxic/Aerobic Infinite 15 - 3 96.21 97.58 72.19
MBR
Anoxic/Aerobic 33.3 10 (hrs) 8.40 1, 3, 5, 92.00 - 89.00
MBR 10

Source: Fu et al., (2009); Wang et al., (2005); Tan and Ng, (2008)
 31

4.4 Membrane operation parameters

Transmembrane pressure is a driving force used to evaluate permeated

flux in MBR system.

TMP (15)
F
 ( Rf  Rm)

Where, F = flux (L/m2-hr), TMP = Transmembrane pressure (Pa), Rf = Fouling

resistance (m-1), Rm = Intrinsic membrane resistance (m-1), μ = viscosity of sludge
(Pa.s)

Jeison and Van Lier (2007) determined the membrane resistance and critical flux by:

TMP
Rm  (16)
J

Where, Rm = resistance due to membrane + resistance due to fouling and stable cake
formation, η = water viscosity (Pa.s), J = Flux (L/m2-hr).

Another fouling equation following form

PT
J (17)
Rt

Where, Rt = Rm+Rc+Rf, ΔPT = the tranmembrane pressure (Pa), η = the viscosity of

the permeate (Pa.s), Rt = the total resistance (m-1), Rm = Intrinsic membrane resistance
(m-1), Rc= Cake resistance formed by cake layer deposited over membrane surface, R f
= Resistance cause by pore plugging and/solute adsorption onto the membrane pore
and surface

Flux optimization for fouling control was pursued since 1980, and the
relationship between TMP and flux utilized to reduce excessive fouling in MBR. The
 32

use of constant flux and TMP rise is useful to monitor fouling in complex fluids and
applied to many MBR systems. Normally step flux-TMP test was used to decide the
optimum flux. When the TMP is no longer stable and its value increases abruptly
which indicates that there is rapid accumulation of foulants on membrane surface. The
flux associated to the point where there is a rapid change in TMP is usually referred as
critical flux.

The critical flux depends on the back transport provided by the cross flow
or turbulence generated by imposed liquid flow and/or bubbling as well as the specific
solute–membrane interactions, which are affected by charge and hydrophobicity.
Solute size also plays an important role in determining the mechanism of back
transport whether it is diffusive or inertial lift (Belfort and Nagata, 1985). High local
concentrations of sludge that promote local aggregation due to concentration
polarization will also determine the cohesiveness of the foulant layer. Recently the
evaluation of critical flux in sponge submerged MBR indicated that suspended sponge
could significantly reduce the membrane fouling and enhance sustainable flux and
also could achieve higher quality effluent with total organic carbon removal
efficiency over 95% in all cases (Guo et al., 2008).
 33

MATERIALS AND METHODS

The experimental plan of this system was operated in continuous biocarrier-

anoxic/aerobic-MBR process. The continuous biocarrier-A/O-MBR process was
applied from Yeom’s (2009) study which consists of anoxic tank with sponge addition
and oxic tank combine with membrane. The experimental was separated into 5 stages
different operating conditions with alkalinity, pH, sludge recycling rate and COD/N
ratio that shown in the Figure 4.

The analytical parameters in this experiment such as COD, TN, TKN, NO3--N,
NO2--N, NH4+-N, TP, MLSS, MLVSS, turbidity, pH, alkalinity, ORP etc. were
measured to evaluate system performance and removal efficiencies of organic and
nutrients.
 34

Laboratory-scale biocarrier - A/O - MBR study

Stage1 Stage 2 Stage 3 Stage 4 Stage 5

Operate with Operate with Operate with Operate with Operate with
Anoxic Anoxic Anoxic Anoxic Anoxic
(sponge) (sponge) (New model (New model (New model
MBR with MBR with sponge) sponge) sponge)
3Q 3Q MBR with MBR with MBR with
recirculation recirculation 3Q 2Q 2Q
recirculation recirculation recirculation

COD, TN, Extra

NO3--N, alkalinity Extra Extra Extra
NO2--N, (NaHCO3) alkalinity alkalinity alkalinity
NH4-N, addition (NaHCO3) (NaHCO3) (NaHCO3)
TKN, TP, addition addition addition
SS, VSS,
Turbidity, COD, TN,
pH, ORP, NO3--N, pH (NaOH) pH (NaOH) pH (NaOH)
Alkalinity, NO2--N, adjustment adjustment adjustment
etc. NH4-N,
TKN, TP,
SS, VSS, COD, TN, COD, TN, Adjust
Turbidity, NO3--N, NO3--N, COD/N ratio
pH, ORP, NO2--N, NO2--N, to 7
Alkalinity, NH4-N, NH4-N,
TKN, TP, TKN, TP, COD, TN,
etc.
SS, VSS, SS, VSS, NO3--N,
Turbidity, Turbidity, NO2--N,
pH, ORP, pH, ORP, NH4-N,
Alkalinity, Alkalinity, TKN, TP,
etc. etc. SS, VSS,
Turbidity,
pH, ORP,
Alkalinity,
etc.

Figure 4 Biocarrier-anoxic/aerobic-MBR Experimental Plans

 35

Materials

The biocarrier –A/O-MBR process consist of anoxic with sponge addition and
oxic tank with membrane separation. The characteristic of sponge was shown in Table
6 and Figure 5. In the oxic zone, diffusers at the bottom were used to generate
oxygen. The solid liquid separation happened in oxic tank with 9 membrane module
as characteristic of membrane was shown in Table 7 and Figure 6. The total working
volume was 18 L including an anoxic tank 6 L and oxic tank 12 L. The synthetic
domestic wastewater was use in this study, which was controlled flow rate by liquid
level sensor.

1. Sponge and Membrane Characteristics

1.1 Sponge

Table 6 Sponge Characteristics

Type Sponge
Size 1.0 cm x 1.0 cm x 1.0 cm
Shape Cubic
Surface area 1 cm2
Volume 1 cm3
 36

Figure 5 Sponge Material Applied to Anoxic Tank

1.2 Membrane

Figure 6 Hollow Fiber Membrane Applied to Aerobic Tank

 37

Table 7 Membrane Characteristics

Type Polyvinylidene fluoride (PVDF)

Membrane length 15 cm
Pore size 0.1 µm
Radius 0.48 mm
Total membrane 9 modules
Total Area of membrane 0.141 m2
Brand Tianjin Polytechnic University, China

2. Sludge Seeding

The sludge sampled from a domestic wastewater treatment plant located at

An-Ping Tainan, Taiwan was seeded into the reactor of this study. The initial
concentration of sludge was 5,600 mg/L and acclimatized in a laboratory scale during
start-up period of this study.

3. Characteristics of Synthetic Domestic Wastewater

Wastewater used in this study was a synthetic domestic wastewater which

was basically composed of a mixed carbon source, macro nutrients (N and P).an
alkalinity control (NaHCO3) and a microelement solution. The composition contained
(L-1) 210 mg glucose, 200 mg NH4Cl, 220 mg NaHCO3, 22-34 mg KH2PO4,
microelement solution (0.19 mg MnCl2 . 4H2O, 0.0018 mg ZnCl2 . 2H2O, 0.022 mg
CuCl2 . 2H2O, 5.6 mg MgSO4 . 7H2O, 0.88 mg FeCl3 . 6H2O, 1.3 mg CaCl2 . 2H2O).
 38

Table 8 Characteristics of Synthetic Domestic Wastewater

Parameter Range Mean value

Temperature (°C) 24~29 27±1
pH 6.10~8.57 7±1
ORP (mv) (-463)~(-202) (-324)±59
DO (mg/L) 0~0.9 0.35±0.24
Conductivity (µs/cm) 1101~2900 2098±606
Turbidity (NTU) 64~144 100±25
Color (ADMI) 5~26 13±5
Alkalinity (mg CaCO3/L) 260~435 356±55
SS (mg/L) 200~600 428±114
TDS (mg/L) 533~1800 1138±428
TP (mg/L) 25~37 32±4
TCOD (mg/L) 178~382 237±69
TN (mg/L) 48~61 54±3
Org-N (mg/L) Non-detected Non-detected
NH4+-N (mg/L) 48~58 53±3
NO2—N (mg/L) 0~0.19 0.01±0.03
NO3—N (mg/L) 0~3.96 1±0.88

The composition of synthetic domestic wastewater that shown in Table

8 was measured immediately after prepares wastewater. Which this experimental was
prepare synthetic domestic wastewater every 3 day with ambient temperature.
 39

Methods

1. Laboratory-scale biocarrier A/O-MBR system

The experimental system was schematized in Figure 7.The total working

volume of the Biocarrier-A/O-MBR system was 18 L. It has two compartments
including an anoxic bioreactor (6 L) and an oxic biological reactor with integrated
membrane for solid-liquid separation (12 L).

The synthetic wastewater was fed into the system at a flow rate of 40 mL/min.
The influent was controlled by a liquid level sensor in the oxic tank to maintain an
average HRT of 9 hours. The influent was fed directly to anoxic tank and flow to oxic
tank by overflow. The HRT of anoxic and oxic was 3 and 6 h, respectively. The
system was provided an internal recycle of 3Q or 2Q from oxic tank to anoxic tank
depending on the experimental condition. In the oxic zone, diffusers in the bottom of
the reactor were used to generate air bubbles for providing oxygen need of
microorganisms and hydrodynamic scouring effect to reduce membrane fouling.
Dissolved oxygen (DO) concentration was controlled at least at a level of 3.5 mg/L.
The solid liquid separation happened in oxic tank with 9 membrane modules (hollow
fiber membrane is made from Polyvinylidene fluoride, PVDF). The characteristic of
membrane is presented in Table 6. The suction pump was operated with 5 min on and
1 min off. The cubic shape commercial sponge with size of 1ˣ1ˣ1 cm3 and 10%
volume fraction of anoxic bioreactor was added into the anoxic tank in this study as a
biocarrier to improve denitrification and organic removal, reduce membrane fouling.
Sludge was withdrawn from the system to maintain sludge retention time (SRT) of 20
days.

The system was operated at ambient temperature. This study consisted of 5

stages shown in Table 9, (1) anoxic (sponge)-MBR with 3Q recirculation rate (2)
anoxic (sponge)-MBR with 3Q recirculation rate and extra alkalinity (NaHCO3)
addition (3) anoxic (new model sponge)-MBR with 3Q recirculation rate, extra
alkalinity (NaHCO3) addition and pH (NaOH) adjustment (4) anoxic (new model
 40

sponge)-MBR with 2Q recirculation rate, extra alkalinity (NaHCO3) addition and pH

(NaOH) adjustment (5) anoxic (new model sponge)-MBR with 2Q recirculation rate,
extra alkalinity (NaHCO3) addition, pH (NaOH) adjustment and adjusted COD/N
ratio to 7.

Figure 7 Schematic of the lab-scale biocarrier-anoxic/aerobic-MBR system

(1; synthetic wastewater tank, 2; influent pump, 3; anoxic tank, 4; sponge,

5; stirrer, 6; MBR tank, 7; aerator, 8; membrane module, 9; level sensor,
10; pressure gauge, 11; permeate pump, 12; sludge return pump)
 41

Figure 8 Biocarrier-anoxic/aerobic-MBR system

 42

Table 9 Stages of study at different operating conditions

Stages 1 2 3 4 5 Note
Conditions
Sponge
(Model No.1)

Surface area =
√ √
510 cm2

New model sponge

(Model No.2)

Surface area =
√ √ √
3600 cm2

Recirculation rate 3Q 3Q 3Q 2Q 2Q Q = 40 ml/min

Influent alkalinity 280 385 385 385 385 Add extra

(mg uauO3/L) alkalinity
(NaHCO3)
Influent 6.5 6.8 8 8 8 Adjust pH
pH (NaOH)

Influent 4 4 4 4 7 Add extra

COD/N ratio carbon source
 43

2. Analytical Determinations

The analysis of water qualities to determine treatment performance of

biocarrier-A/O-MBR process will be performed according to the parameters specified
in Table 10.

Table 10 Parameters and analysis methods

Parameter Equipment Method Method

Number
pH pH Meter (YSI - -
Environmental pH 100)

Temperature pH Meter (YSI - -

Environmental pH 100)

Turbidity Turbidity Meter (Hach - -

2000)
ORP pH Meter (YSI - -
Environmental pH 100)
TKN Kjedahl Method Taiwan & USEPA W451.51A
Standard Method
NO2--N Spectrophotometer Taiwan & USEPA W448.51B
(CT Chrom-Tech Model Standard Method
number CT-2800)
NO3--N Spectrophotometer Taiwan & USEPA W419.51A
(CT Chrom-Tech Model Standard Method
number CT-2800)
NH4-N Spectrophotometer Taiwan & USEPA W448.51B
(CT Chrom-Tech Model Standard Method
number CT-2800)
 44

Table 10 (Continued)

Parameter Equipment Method Method

Number
Color Spectrophotometer Taiwan & USEPA NIEA
(Thermo Scientif Model Standard Method W223.51B
Genesys 20)
Total Spectrophotometer Taiwan & USEPA NIEA
Phosphorus (CT Chrom-Tech Model Standard Method W427.53B
number CT-2800)
Suspended Filtration, Evaporation Taiwan & USEPA NIEA
Solid Standard Method W210.57A
Volatile Filtration, Volatilization Taiwan & USEPA Standard
Suspended Standard Method Method 209C
Solid
Total Dissolved Evaporation Taiwan & USEPA NIEA
Solid Standard Method W210.57A
Alkalinity Titration Taiwan & USEPA NIEA
Standard Method W449.00B
COD Close Reflux Method Taiwan & USEPA NIEA
Standard Method W517.51B
Permeate Flux Flow Metre - -
Transmembrane Pressure Gauge - -
Pressure

All samples for the determination of total kjehldahl nitrogen (TKN), nitrite
nitrogen (NO2--N), nitrate nitrogen (NO3--N), ammonia nitrogen (NH4-N), total
phosphorus (TP) and color will be filtered by using a 0.45 µm membrane filter (accept
for alkalinity) and followed by spectrophotometric analysis (CT Chrom-Tech Model
number CT-2800 used for TKN, NO2--N, NO3- -N, NH4-N, TP and Thermo Scientif
 45

Model Genesys 20 used for color) in accordance to Taiwan Environmental Protection

Agency (EPA) standard method.

3. Membrane Cleaning

Membrane cleaning was required when TMP was increased up to 30 kPa. The
procedure of membrane cleaning was commenced by disconnecting the suction lines
from the membrane modules, and then the membranes were taken out from the reactor
in order to remove the cake layer on membrane surface by shaking in a 5 L plastic
cylinder which contains reverse osmosis water (RO water). After that dilute sodium
hypochlorite (NaOCl) 10% to 3% by RO water and the membranes were submerged
in a chemical cleaning containing 3% sodium hypochlorite (NaOCl) for 6 hours. After
6 hours the membranes modules were taken to remove cleaning solution by rinsing
with RO water.
 46

RESULTS AND DISCUSSION

In this study, the biocarrier-A/O-MBR system was operated for 260 days and
synthetic domestic wastewater was used as the feed wastewater. The characteristic of
synthetic domestic wastewater was shown in the section of material and method. The
system was operated at ambient temperature. This study consisted of 5 stages which
contained different operation conditions and SRT 20 of days, HRT of 9 hrs were
controlled. The performance of the system as well as removal efficiency of TCOD,
SCOD, total ammonia, total nitrogen and total phosphorus were evaluated.

1. COD removal efficiency

The chemical oxygen demand (uOD) is used as a measure of the oxygen

equivalent of the organic matter content of a sample that is susceptible to oxidation by
a strong chemical oxidant. In this study have measure TCOD and SCOD which
TCOD was without filtration and SCOD was filtrate through 0.45 µm pore diameter
membrane filter separates dissolved from suspended forms of organic matter. The
influent and effluent TCOD, SCOD, as well as their removals during the experimental
period are shown in Figures 9 and 10. The average, range and highest removal
efficiency of TCOD and SCOD in 5 stage are shown in Table 11.

The average influent of TCOD and SCOD of stage 1-4 was 206 mg/L and 88
mg/L, respectively, then raised up to 376 mg/L and 245 mg/L at stage 5 (day 183) by
adding extra carbon source (glucose) for adjusting the COD/N ratio. The TCOD and
SCOD removals of stage 1 ranged at 88-98% and 78-97%. It is obvious that the
removal efficiency of stage 1 fluctuated during the start-up period and the
performance was unstable. The removals became more stable at stage 2 and 3 since
extra alkalinity was added at stage 2 and pH was adjusted for improving nitrification
and denitrification at stage 3. The TCOD and SCOD removals of stage 2 and 3 ranged
at 91-100% and 80-100%. At stage 4, TCOD and SCOD removals ranged at 92-100%
and 80-99% but a decrease trend of both was observed from the Figures 9 and 10 due
to the reduction of sludge recycling rate from 3Q to 2Q for improving the
 47

denitrification in anoxic tank. The abrupt change of sludge recycling rate from 3Q to
2Q induced less supply of nitrate to anoxic tank, however, the same organic level in
influent at this stage was fed. It meant that an unbalance electron transmitting between
organic substance and nitrate occurred in anoxic tank, so the residual organic would
be accumulated in permeate to have lower COD removal at this stage. The COD
removal efficiencies increased at stage 5 with the increase of COD/N ratio to 7 in the
influent. The range of TCOD and SCOD removals were 98-100% and 96-100%. At
this stage, the sludge recycling rate kept the same value of 2Q as stage 4, however,
both TCOD and SCOD removals increased in this stage.

There are two possibilities to explain this phenomenon according to

experimental data of this study and the suggestion of previous study conducted by
Zou et al., (2012). First of all, the increase of influent organic level could increase the
growth rate of microorganisms in the system, in fact, the level of MLVSS in anoxic
tank at this stage increased from 1400 mg/L to around 4800 mg/L as shown in the
Figure 11. By this way, it could be speculated that the quantity of microorganisms
increased in this stage, especially for heterotrophic bacteria such as denitrifiers and
DNPAOs. Actually, 30% increase of TN removal and an increase of P removal were
obtained at this stage. Based on the results from Zou’s (2012) study , the increase of
COD concentration in the influent can enhance the speed adsorption of the soluble
organic onto the organism, which accelerates the first removal step of organic matter,
and carbon source was used for bacteria growth and accompanied by denitrification
improvement as well as the decrease of nitrate accumulation rate.

In this study, the permeate COD concentrations were averagely lower than 10
mg/L at stage 5. It showed that most of the COD was removed in anoxic tank and the
whole system provided a stable performance of organic removal and a stable effluent
quality. The residual of COD in permeate may be due to the impact on formation of
EPS which the chemical structure of polymeric substances secreted by the cells into
the environment is diversified. The EPS compounds belong to such different classes
of macromolecules as polysaccharides, proteins, nucleic acids and glycoproteins
(Sutherland, 2001; Branda et al., 2005). However these compounds are slowly
 48

biodegradable chemical oxygen demand (SBCOD) which can be considered as one of

the main factors contributed to membrane fouling.

Table 11 The range, mean value and highest removal efficiency of TCOD and SCOD
at 5 stages under different operation conditions

Mean value of Highest removal Range removal

removal efficiency (%) efficiency (%)
Stage
efficiency (%)
TCOD SCOD TCOD SCOD TCOD SCOD
1 94±3 88±7 98 97 88-98 78-97
2 96±3 91±6 99 98 91-99 80-98
3 97±2 91±6 ˃99 ˃99 93-100 84-100
4 95±3 88±7 ˃99 99 92-100 80-99
5 99±1 98±1 ˃99 ˃99 98-100 96-100

Figure 9 TCOD variation and removal during the experiment

 49

Figure 10 SCOD variation and removal during the experiment

Figure 11 Relationship between COD removal and MLVSS in anoxic tank

 50

2. Nitrogen removal efficiency

Figures 12, 13 and Table 12 illustrate ammonia and total nitrogen (TN)
removals in this biocarrier-A/O-MBR system. Ammonia removal of stage 1 fluctuated
from 40% to 99% and TN removal was quite unstable from (-40) % to 82% since the
system was at start-up period. In the second stage, the average ammonia and total
nitrogen removal efficiencies was 91% and 41%, respectively, the increase tend could
be due to the addition of extra alkalinity for the improvement of nitrification in MBR
tank.

Figure 12 Ammonia removal profile during the study period

 51

Figure 13 Total nitrogen removal profile during the study period

Table 12 The range, mean value and highest removal efficiency of ammonia and
total nitrogen of 5 stages under different operation conditions

NH4 Removal efficiency Total nitrogen removal

(%) efficiency (%)
Stage
Range Highest Mean Range Highest Mean
value value
1 40-99 99 71±17 (-40)-82 82 13±33
2 73-99 99 91±8 20-74 74 41±20
3 88-100 ˃99 97±4 19-41 41 30±6
4 90-100 ˃99 97±4 26-57 57 39±10

5 95-100 ˃99 99±2 58-85 85 80±8

The nitrification is a two steps reaction: ammonia is firstly oxidized to nitrite

by Nitrosomonas bacteria and nitrite is then oxidized to nitrate by Nitrobacteria
 52

bacteria, during the nitrification, 7.14 mg/L of alkalinity as CaCO3 is consumed for
each mg/L of ammonia nitrogen oxidized. For denitrification, alkalinity is generated
at 3.57 mg/L for each mg/L of nitrogen reduced (Sedlak, 1991). As shown in Figures
14 and 15, the average influent alkalinity of stage 1 was 280 mg/L then raised up to
386 mg/L at stage 2-5 by adding extra alkalinity (NaHCO3). The rang alkalinity in
anoxic tank at stage 1 was very low at 60 mg/L to 176 mg/L, and then kept increase
tend from 124 mg/L to 500 mg/L at stage 2 and 3 which extra alkalinity was added at
stage 2 as well as modified sponge model and pH control were introduced at stage 3.
Alkalinity kept stable at stages 4 and 5 ranged from 470 mg/L to 560 mg/L, which
return sludge was changed from 3Q to 2Q at stage 4 and added extra carbon source at
stage 5.

After extra alkalinity was added, 20% increase of ammonia removal was
observed and the ammonia in MBR tank became low, meanwhile, the average level
was observed at 0.98 mg/L in permeate. On the other hand, nitrate in anoxic tank and
MBR tank increased from 6.48 to 37.86 mg/L and 6.54 to 38.44 mg/L, respectively.
It indicates that alkalinity was not enough for ammonia to be oxidized to nitrate and
the amount of alkalinity consumed in nitrification is greater than alkalinity generated
in denitrification at stage 1. At stage 4-5 alkalinity generated from denitrification was
greater than alkalinity consumed in nitrification resulting in a complete nitrification. It
indicated that alkalinity in the system was enough for nitrification. The alkalinity
from denitrification met the requirement of nitrification at stages 4-5 could be due to
the influent COD was higher than that of former stages. The complete denitrification
found at high influent COD concentration led to low nitrate concentration in
permeate, and alkalinity produced in denitrification could make up for compensated
alkalinity consumed in nitrification. The residual alkalinity was higher than 40 mg/L
to insure adequate buffering and for complete nitrification required (Sedlak, 1991).
 53

Figure 14 The relationship between alkalinity and ammonia removal efficiency

Figure 15 Nitrification and denitrification during the period of study

At stage 3, a modified sponge model was induced to increase the surface area
as well as the attached microorganisms. The results indicated that the biomass-riched
sponge in anoxic tank apparently enhanced the treatment performance and reduced
membrane fouling.
 54

The other condition adjusted at this stage was pH value. When alkalinity was
produced during denitrification, solution pH was also elevated, instead of being
depressed in nitrification. However, it was reported that no significant effect on the
denitrification rate when pH is in between 7.0 and 8.5, while showed a decrease in the
denitrification rate as the pH was decrease from 7.0 to 6.0. For nitrification, pH is
sensitive and rate declines significantly at pH value blow 6.8. The optimum
nitrification rates occur at pH values in the range of 7.5 to 8.5 (Sedlak, 1991).

Figure 16 showed the effect of denitrification on pH, which average pH during

denitrification in anoxic was 6.3, 7 and 8 at stages 1, 2 and stage 3-5, respectively.
The pH value increased to 7 at stage 2 could be due to the addition of extra alkalinity
for improving nitrification. At stages 3-5, solution pH increased to 8 was due to
NaOH addition and resulted in enhancing the performance of denitrification and
nitrification.

However, ORP of anoxic tank was observed to be reduced gradually from 66

mV at stage 1 to -196 at stage 3 and kept decrease trend at stages 4 and 5 as shown in
Figure 17. ORP decrease is expected due to the absence of oxygen coupled with mass
substrate consumption which induced reduction reaction in this process (Vollertsen
and Hvitved-Jacobsen, 2002). In this case, nitrate was electron acceptor in
denitrification process when nitrate decreased which resulted in further decrease in
ORP. It indicated that good denitrification occurred at low ORP with sufficient carbon
source present, while low carbon source (indicated by high ORP) caused the failure of
denitrification. The average concentration of nitrite and nitrate in anoxic tank in stage
3 was 0.29 mg/L and 32.6 mg/L. Actually, nitrite was almost non-detected at stage 3,
it means that nitrite was not accumulated in the anoxic tank, however, nitrate in
anoxic tank was still high at an average of 32.6 mg/L. The average of total nitrogen
removal efficiency kept stable at 30% at stage 3.

Figure 18 shows the effect of pH on nitrification which average pH at stage 1

was 5.88 and then increased to 7.3 by adding extra alkalinity for improving
nitrification at stage 2. At stages 3, 4 and 5, pH in MBR tank was controlled by NaOH
 55

in the average of 8.13 to enhance the performance of denitrification and nitrification.

The result showed that the removal efficiencies of ammonia kept stable at 97%
(stage3). The average concentration of ammonia and nitrate in MBR were 0.64 mg/L
and 35.48 mg/L, respectively. It means that nitrification was almost complete.

During nitrification, ORP values increases due to the presence of aeration in

this phase while oxygen is an oxidizing agent. As shown in Figure 19, ORP values
fluctuated form 50 mV to 263 mV at stage 1 because this stage is an unstable stage.
The ORP values became more stable in the range of 99-167 mV at stage 2-5, while
ammonia concentration became low. It shows that high nitrification reaction can
occur as high ORP value, which has been reported from previous studies that
nitrification failed when ORP values were lower than 50 mV and low ORP
corresponded to poor effluent quality (Hamamoto et al., 1997; Chapentier et al., 1998;
Lo et al., 1994; Holman and Wareham, 2003; Chen et al., 2002; Kim et al., 2004).

Figure 16 Variation of pH on denitrification

 56

Figure 17 Variation of ORP on denitrification

Figure 18 Variation of pH on nitrification

 57

Figure 19 Variation of ORP on nitrification

In order to improve the denitrification of this system, sludge recycling rate

was changed from 3Q to 2Q at stage 4 and then average COD/N ratio was raised from
4 to 7 at stage 5. As shown in Figure 12, ammonia removal decreased from 100% to
90% at stage 4 and then increased more than 99% again at stage 5 after changing
COD/N ratio. The decrease of ammonia removal at stage 4 may be due to the change
of sludge recycling rate, which returned less nitrate from oxic tank to anoxic tank. In
addition, more residual carbon source from anoxic to MBR tank could essentially
result in the inhibition of growth rate and biological-activity of ammonifying bacteria
and nitrifying bacteria (Wu et al., 2008; Nootong and Shieh, 2008; Osaka et al.,
2008).

The result obtained from this study also indicated that the effect of COD/N
ratio on system performance was significant, especially on TN removal, as shown in
Figure 20, the TN removal increased significantly from 26% (stage 4) to 85% (stage
5). Figure 21 also shows the nitrogen concentration in the anoxic tank after change
COD/N ratio from 4 to 7, nitrite kept stable at 0.33 mg/L, nitrate decreased from 34 to
non-detected level (almost 0 mg/L) and total nitrogen decrease from 38.5 to 13.71
mg/L. On the other hand, nitrate in MBR tank decrease from 38 to 5 mg/L. The
 58

complete denitrification in anoxic tank could be accomplished due to the increase of

COD /N ratio because some carbon was used in new biomass formation, which is
shown in Figure 22 that MLSS and MLVSS were increased from 4300 mg/L to 7600
mg/L and 1400 mg/L to 4800 mg/L, respectively. The new biomass can improve
denitrification reaction and enhance total nitrogen removal efficiency.

The incomplete denitrification and accumulation of nitrate during stages 1 to 4

could be due to the shortage of carbon source which is insufficient for the denitrifiers
bacteria to grow in the anoxic tank. The results obtained from this study showed that
the COD/N ratio of influent significantly affects the growth of microorganisms
(heterotrophic denitrifiers) since denitrifiers utilized carbon source as electron donor
and nitrate as electron acceptor (Bliss and Barnes, 1983; Tiedje, 1988; Wiesmann,
1994). The concentration of nitrogen in permeate was shown in Figure 23 that
organic-N, Nitrite-N and ammonia-N are very low and almost non-detected level
(almost 0 mg/L). The total nitrogen also decreased from 38 mg/L to 8.5 mg/L, of
which the residual nitrate was the only nitrogenous compound in permeate.

Figure 20 Effect of COD/N ratio on total nitrogen removal

 59

Figure 21 Continuous denitrification with different COD/N Ratio in anoxic tank

Figure 22 Relation of MLSS, MLVSS with total nitrogen removal

 60

Figure 23 Nitrogen concentration in permeate

3. Phosphorus removal efficiency

In previous studies, many researchers tried to find a way to simultaneously

remove nitrogen and phosphorus in an A2O process (Yeom et al., 2009; Peng et al.,
2006; Ma et al., 2005). Which polyphosphate-accumulating organisms (PAOs) take
up carbon sources while releasing orthophosphate in the anaerobic phase and store
them in the form of polyhydroxybutyrate (PHB), using the energy produced through
the hydrolysis of intracellular polyphosphate. In the subsequent aerobic phase, PAOs
grow and take up orthophosphate by using the store PHB as the carbon and energy
source (Zeng et al., 2004) However, many conflicts between N and P removals could
be seen in A2O process such as SRT, carbon competition and sludge recycling
operation etc.

This study tried to find a way to conduct a sponge unit to replace anaerobic
and anoxic units with a sponge anoxic and integrate an aerobic MBR unit to enhance
the system performance. Figure 24 and Table 13 shows total phosphorus removal
 61

efficiency of the biocarrier-A/O-MBR process in this study. It is clear that the

removal efficiency of total phosphorus at stage 1 fluctuated significantly in the range
of (-68)-35%. At stages 2 and 3, the highest removal efficiency was 23 and 44%,
respectively. The poor removal could be due to the high residual nitrate in anoxic
tank, which average of nitrate concentration in the anoxic tank in stage 2 and 3 shown
25.31 and 32.60 mg/L. The high level of nitrate can play as an inhibitor for
phosphorus release in anoxic zone resulting in low phosphorus up-take in the oxic
zone.

For improving phosphorus release in anoxic zone (reduce residual nitrate

concentration) the reduction of sludge recycling from 3Q to 2Q was carried out in
stage 4. The results of this stage showed that there was no negative removal efficiency
and the average removal of phosphorus was not high around 22%. Meanwhile, nitrate
in anoxic tank decreased from 34 mg/L to 5 mg/L at stage 4. It indicated that
reduction of nitrate in anoxic tank can improve phosphorus removal (more
phosphorus release in anoxic zone and up take in oxic zone). On the other hand, the
COD/N ratio of 4 at stage 4 was much lower than those of other studies and this
probably the main reason for poor phosphorus removal since the carbon was
insufficient for PAOs to compete with denitrifying bacteria to accomplish the
phosphorus release. The phosphorus removal efficiency often decreased when the
available organic carbon content is low (Morling, 2001). For that, the extra carbon
was added into the system to increase the COD/N ratio from 4 to 7 at stage 5.
Afterward, the removal efficiency of phosphorus increased from 14% to 81% and
nitrate concentration in the anoxic tank become low at 0.2 mg/L. Also the total
phosphorus concentration in permeate decreased from 30 mg/L to around 6.5 mg/L.

With a high COD/N ratio, denitrification was found to be almost complete and
also more amount of phosphorus release was observed in the system, it means the
residual carbon free from denitrification can meet the necessity of phosphorus release
that take up by PAOs in the anoxic phase. The poor removal efficiency of phosphorus
in previous stage could be due to shortage of carbon source, which caused by organic
matter in influent was consumed to remove nitrogen by denitrification rather than
 62

phosphorus removal. Hence, the biological phosphorus removal process was

prohibited by insufficient available organic carbon source. It indicated that high
COD/N ratio can improve total phosphorus removal efficiency, while, low COD/N
ratio resulting in low removal efficiency.

Figure 24 Phosphorus removal profile during the study period

Table 13 The range, mean value and highest removal efficiency of total phosphorus
at 5 stage under different operation conditions

Total phosphorus removal efficiency (%)

Stage
Range Highest Mean value
1 (-68)-35 35 (-9)±31
2 (-20)-23 23 3±11
3 (-11)-44 44 15±17
4 2-22 22 15±6
5 14-81 81 56±26
 63

4. Sludge characterization

During start up period, biomass with an initial MLSS concentration of 5700

mg/L was acclimatized to synthetic domestic wastewater. MLSS and MLVSS of the
system were determined regularly and a SRT of 20 days was controlled. Sludge color
of both anoxic and MBR tank appeared as khaki-brown. Biomass in anoxic tank
presented in two forms, both suspended and attached growth. The biomass was
entrapped into the sponge cubes and apparently, there was almost no biofilm observed
on the media surface. Certain volume of sponge was taken out from the tanks to
measure biomass. Sponge was squeezed and rinsed with water thoroughly in order to
get all biomass out of the sponge. An average MLSS mass in suspended and attached
forms in anoxic tank at stages 1-5 is shown in Figure 25 and Table 14.

From this result, it was shown that the average MLSS in suspended termed has
decrease tend from 27.13 g (stage 1) to 15.60 g (stage 4) and increase to 21.82 g at
stage 5. The decrease trend may be due to shortage of carbon source which carbon
source is food for microorganism growth, it directly effects function microorganism
population. At the same time, the increase at stage 5 was due to the addition of extra
carbon source to improve denitrification. However, the extra carbon source can
produce new microorganism and improve growth rate of microorganism. Also
average MLSS in attached termed increase from 11.58 g (stage1) to 13.77 g (stage2)
and then kept stable until stage 4 and increase again to 15.42 g at stage 5.
 64

Table 14 Average MLSS concentration in suspended termed and attached termed in

anoxic tank.

MLSS (g) MLSS (g)

Stage Suspended Attached Total MLSS (g)
termed termed
1 27.13±6.94 11.58±2.19 38.71±7.09
2 20.93±6.41 13.77±4.30 34.70±6.33
3 16.98±4.49 13.97±2.96 30.95±4.64
4 15.60±1.92 14.10±1.76 29.70±2.93
5 21.82±5.17 15.42±1.84 37.24±5.61

Figure 25 Average MLSS in anoxic tank during experimental period

Figures 26 and 27 showed the variation of MLSS and MLVSS during the
experimental period. The MLSS in the anoxic tank shown in Figures 26 is the
summation of suspended and attached termed. At stage 1, there was a variation of
MLSS concentration in anoxic and MBR tank that fluctuated from 4467 mg/L to 8833
mg/L and from 2133 to 5213 mg/L, respectively. The difference in MLVSS
 65

concentration in anoxic and MBR tank is from 2200 to 4800 mg/L and 1267 to
4160mg/L, respectively. The instability in this period is observed since it is the start
up of the system. At stage 2-4, MLSS and MLVSS in anoxic and MBR tank became
more stable, the range of MLSS and MLVSS in anoxic were 3867 mg/L to 7600 mg/L
and 1200 mg/L to 4100 mg/L, respectively. In the MBR, the range of MLSS and
MLVSS were 1200 mg/L to 3400 mg/L and 1000 mg/L to 3400 mg/L, respectively.
At stage 5 both the MLSS and MLVSS concentration in the anoxic and MBR tank
increased as the COD/N ratio also increased from 4 to 7, because high COD loading
results in an increase in microbial formation. At this stage, the MLSS concentration in
the anoxic and MBR tank increases from 4300mg/L to 7600 mg/L and from 1200
mg/L to 4600 mg/L, respectively, and the MLVSS concentration in the anoxic and
MBR tank also increases from 1400 mg/L to 4800 mg/L and 1200 mg/L to 4200
mg/L, respectively.

Figure 26 Variation of MLSS in this experimental period

 66

Figure 27 Variation of MLVSS in this experimental period

5. Volumetric loading rate and F/M ratio

The F/M ratio or food to microorganism is defined as the rate of COD applied
per unit volume of mixed liquor. Also volumetric loading rate is defined as the
amount of COD and NH4 applied to the system volume per day. The variations of
F/M ratio and volumetric loading rate are shown in Figure 28 and 29. The average
volumetric loading rate is shown in Table 15
 67

Figure 28 Variation of F/M ratio in this experimental period

The ratio of food to microorganism (F/M ratio) refers to the balance between
the food supply and the mass of microorganism in the system. As shown as Figure 28,
the highest F/M ratio in the anoxic and MBR tank at stages 1-4 were 0.16 g SCOD/g
VSS.d and 0.16 g SCOD/g VSS.d, respectively. At stage 5, the F/M ratio in the
anoxic tank increased up to 0.27 g SCOD/g VSS.d and then decreased gradually to
around 0.15 g SCOD/g VSS.d, while the F/M ratio in the MBR was around 0.03-0.13
g SCOD/g VSS.d. An increase of F/M ratio in the anoxic tank in the beginning at
stage 5 may be due to high organic loading from extra carbon source addition and
then F/M ratio keep decreasing due to the increase of microorganism. Typically, MBR
runs at lower F/M ratio than conventional activated sludge (CAS) process (0.4-0.6 g
COD/g MLVSS day ) as shown the preferred F/M ratio range in MBR in a typical
domestic MBR plant is approximately a third to a half of that in CAS, 0.1-0.3 g
COD/g MLVSS day (MBR design, 2011).

As compared with the other studies, F/M ratio in this study was lower than
other studies, however, the result showed high removal efficiency at stage 5 was
 68

obtained as the removal of SCOD, NH4+ and total nitrogen was 98%, 99 % and 80%,
respectively. The F/M ratio in anoxic tank became slight higher than that of MBR
tank at stage 4 and then much higher at stage 5. In fact, the highest of COD and
nitrification at stage 4 was 99% and more than 99 %, respectively. It indicated that the
system have good performance on COD removal and nitrification. However, the
nitrogen removal at stage 4 was only 39% but 80% at stage 5. The low efficiency of
nitrogen removal at stage 4 may be due to insufficient of carbon source and the higher
nitrification can occur in stage 5 even higher COD/N ratio due to most of carbon
source was consume in the anoxic tank. It indicated that anoxic tank is flexible and
has high capability in this system.

In fact, MLVSS in anoxic and MBR tank increased from 2000 to 4800 mg/L
and 2000 to 4000 mg/L at stage 5, an increase of microorganism resulted in higher
nutrient removal. It indicated that high F/M ratio can enhance performance and the
removal efficiency of this system. On the other hand, increase of F/M ratio at stage 5
basically would cause serious membrane fouling which could shorten filtration
duration. The F/M ratio can affect the accumulation of fine particles and soluble
microbial product in the supernatant, in addition, F/M ratio was also reported to
positively affect the bound EPS, thus lead to fouling (Trussell et al., 2006).
 69

Figure 29 Variation of COD volumetric loading rate in this experimental period

Table 15 Average value of COD volumetric loading rate for 5 stages

Volumetric loading rate (kg COD /m3.d)

Stage
Overall Anoxic MBR
1 0.27±0.07 0.14±0.08 0.10±0.05
2 0.27±0.06 0.18±0.08 0.09±0.06
3 0.24±0.04 0.12±0.07 0.09±0.06
4 0.22±0.03 0.17±0.06 0.06±0.03
5 0.75±0.04 0.47±0.19 0.18±0.03

Figure 29 and Table 15 show the average volumetric loading rate (VLR) of
COD. The volumetric loading rate of overall, anoxic and MBR at stages 1-4 was
around 0.22-0.27 kg/m3.d, 0.12-0.18 kg/m3.d and 0.06-0.10 kg/m3.d, respectively and
up to 0.75 kg/m3.d, 0.47 kg/m3.d and 0.18 kg/m3.d, respectively at stage 5. The
increase of volumetric loading rate at stage 5 was due to the addition of extra carbon
source for improving denitrification at this stage.
 70

The results show that the volumetric loading rate in the MBR tank was very
low and the organic carbon was almost removed in anoxic tank, also, the increase of
organic loading rate led to a higher denitrification rate. Actually, increase of extra
carbon source at stage 5 resulted in increasing volumetric loading rate from 0.22 to
0.75 kg/m3.d and removal efficiency, which removal efficiency of SCOD, NH4+ and
total nitrogen was shown 88 to 98%, 97% to 99 % and 39 to 80% respectively.

At the same time, the average MLVSS in anoxic and MBR tank at stage 5 also
increase from 2000 to 4800 mg/L and 2000 to 4000 mg/L, respectively. It might be a
result of the synthesis of heterotrophic biomass, as expected at high organic loadings.
Some report also suggested that nitrogen removal decreased dramatically with a
decrease of the organic loading rate in the influent due to an organic limitation in
heterotrophic denitrification (Choi et al., 2008; Rene et al., 2008). The present study
has demonstrated that biocarrier-A/O-MBR process completely accomplish the
removals of organic and nitrogen compounds despite different the organic loading in
the range of 0.22-0.75 kg/m3.d. Furthermore, this biocarrier-A/O-MBR process can
produce a stable and high quality permeate with SCOD and NH4 lower than 10 mg/L
and 0.3 mg/L, respectively. Finally, this study found that an appropriate COD/N ratio
is a criteria for the simultaneous removal carbon, nitrogen and phosphorus.

6. Membrane fouling

In order to investigate the fouling behavior, the change in TMP with operation
time was monitored at constant flux. During the experiment, membrane flux was kept
at an imposed value of 17 L m-2 h-1. As shown in Figure 30, at stage 1 and the
beginning of stage 2, the average flux is quite unstable and fluctuated from 12 to
17.42 L m-2 h-1. However, the flux became more stable at the end of stage 2 and was
kept stable until stage 5 with an average flux of 16.68 L m-2 h-1
 71

Figure 30 Average flux through experimental period

The approach for fouling control in this study is a continuous aeration to scour
the membrane surface and intermittent suction-relaxation operation of the membrane
(1 min rest for every 5 min of operation). As presented in Figure 31, for stage 1 and
the beginning of stage 2, the TMP normally increased rapidly every 10 days, which
reached about 40 kPa. Once the TMP was up to 40 kPa, the membrane module was
taken out from the bioreactor for chemical cleaning. Chemical cleaning of the
membrane was carried out with sodium hypochlorite solution (NaOCl). After the
chemical cleaning, the membrane filtration capability was recovered. The frequency
of chemical cleaning membrane was high at this stage. In the end of second stage,
membrane had a longer filtration of around 21 days.

At stage 3, a change of sponge model was made in anoxic tank, on which the
new model of sponge provided more surface area for the increase microorganism
attachment. Sponge has been considered as a reasonable attached growth media
because it can act as a mobile carrier for active biomass resulting in an improved
organic and nutrients removal as well as reduces fouling of the membrane (Guo et al.,
2009; Chae et al., 2004; Ngo et al., 2006; Psoch and Schiewer, 2006). Figure 32
shows that a chemical cleaning interval of 48 days was observed between stage 3 and
stage 4. It was found that a longer filtration of membrane occurred after the change of
 72

new sponge model. The longer filtration may be due to microorganism type adhesive
on the surface attach on sponge more than surface of membrane, which at this stage
provide more surface area of sponge from 510 cm2 to 3600 cm2 lead to increase of
microorganism attach to surface of sponge and lower attach on membrane surface. It
implicated that sponge cubes with size 1 cm3 can be considered as a media of choice
in attach growth MBR for effective organic and nutrients removal and prolong
filtration duration keeping in view its economic viability. The sustainable operation
without any cleaning, whatever water or chemical, up to 55 days was maintained at
stage 4. Even though in this stage sludge recycling rate was changed from 3Q to 2Q to
improve the denitrification in the anoxic tank. It means that the reduction of sludge
flow rate from MBR tank to anoxic tank did not cause an accumulation of sludge in
MBR tank, which can led to membrane fouling. The first 41days of stage 4, TMP
values kept at a low level (less than 10 kPa). However, TMP began to increase rapidly
after 41 days and reached about 37 kPa around day 55. The interval became short
again at stage 5 due to the change of COD/N ratio from 4 to 7. It is obvious that the
increase of COD/N ratio thereby increased growth rate of microorganism and EPS in
the system. In fact, the level of MLVSS in MBR tank at this stage, increased from
1200 mg/L to around 4200 mg/L and probably EPS also increased under high glucose
content in the medium.

EPS is an organic compound that is secreted by cells. The characteristic of

EPS is slime, which makes it easy to attach and be adsorbed on membrane surface
after that block membrane pores finally cause severe fouling. Many investigations
have showed that the accumulations of EPS in MBR tank leads on formation of EPS
which can be considered as main factor that contributed to membrane fouling (Yuan
et al., 2008). The average filtration run in this period was observed to be 18 days.
These results indicate that the increase of COD/N ratio can possibly increase
membrane fouling, however, further study could be conducted in the future to realize
the real mechanism.
 73

Figure 31 The Variation of TMP and COD/N ratio through experimental period
 74

CONCLUSIONS AND RECOMMENDATIONS

Conclusions

In this study, a biocarrier-A/O-MBR process was investigated for treating

synthetic domestic wastewater. It was conducted to evaluate the operating different
conditions for organic and nutrient removals. The operation was continued for 260
days. Base on the results of the system, the findings of this research can be
summarized as follows.

1. The poor removal efficiency of ammonia and total nitrogen posed serious
problem during start up period because several reasons, such as shortage of carbon
source to anoxic tanks, low alkalinity supplement of MBR tank and low biomass
concentration in both reactor. From experiment results, it can be concluded that an
external carbon source and an alkalinity addition can improve the removal efficiency
of the biocarrier-A/O-MBR system.

2. The results showed that the highest SCOD, TN and TP removals of more
than 99%, 85% and 81% can be achieved under a specific operating mode (sludge
recycling rate of 2Q, influent alkalinity 385 mg CaCO3/L and COD/N ratio of 7), with
complete nitrification and without nitrite accumulated in the system.

3. Increased COD/N ratio resulted in high MLSS level played an important

role in membrane fouling and pore blocking in this study. Many investigations have
showed that the accumulations of EPS in MBR tank can be considered as main factors
contributed to membrane fouling, which can speculate that EPS may be one of the
factors of membrane fouling in this study.

4. Sponge addition could improve effective organic and nutrients removal,

sustainable flux and reduce membrane fouling. It can longer membrane filtration
duration and keeping in view its economic viability that can be observed in this study.
 75

5. Stable and excellent permeate was obtained with the average COD ˂ 10
mg/L, NH4+- ˂ 0.2 mg/L, T ˂ 11 mg/L, TP ˂ .5 mg/L, no color, no odour and
free of SS. This indicates the stability of MBR technology for produce high permeate
quality.

6. The results indicated that the biocarrier-A/O-MBR process developed by

this study can replace A2O-MBR system and it shown that even without anaerobic
process and lower recirculation rate (2Q), the removal of COD was complete, total
nitrogen and total phosphorus removal efficiency could be achieved up to 80%.
Comparing with another research of A2O-MBR system, the COD, total nitrogen and
phosphorus removal efficiency was 94%, 60% and 74% respectively, the performance
of this biocarrier-A/O-MBR process is much better than A2O-MBR system. It also
indicated that biocarrier-A/O-MBR system has high organic and nitrogen removals,
smaller footprint, less operating and maintenance costs.

Recommendations

1. EPS study should be applied in the research for finding the relation with
COD/N ratio and the mechanism of membrane fouling.

2. The stage 5 should be extended to reach a stable stage. Actually, TN and TP

removal kept increase and probably the higher removal efficiency could be achieved
in the coming future.

3. Attached microorganisms on sponge should be studied in this research for

finding growth rate of microorganism on sponge and realizing the relationship
between attached biomass and pollutants removal.
 76

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 87

CURRICULUM VITAE

NAME : Ms. Patchaya Ruchirattanawarakorn

BIRTH DATE : August 14, 1984

BIRTH PLACE : Bangkok, Thailand

EDUCATION : YEAR INSTITUTE DEGREE/DIPLOMA

2008 Kasetsert Univ. B.Eng. (Chemical
Engineering)
2013 Kasetsert Univ. M.Eng. (Environmental
Engineering)

POSITION/TITLE : Sales Engineer

WORK PLACE : Process Technology and Services Co.,Ltd.