Chapter 1

INTRODUCTION

Ever since man started to live in communities, we have been throwing things away.
-William Rathje
Production of solid waste is inevitable. A report released by the World Bank (WB) in
2011 states that municipal solid waste that will be generated by Philippine cities will go up
by 165 percent to 77,776 tons per day from 29,315 tons as a consequence of a projected 47.3percent hike in urban population by 2025. As we are constantly growing in number, the
allotted land areas as reservoirs for our waste generation are diminishing. Some of them, such
as the case of Irisan dumpsite, have even reached their limits. This results to a scenario where
solid waste in landfills stays longer and becomes more concentrated leading to aggravated
environmental problems that require immediate action.
In the recently concluded National Solid Waste Management Commission (NSWMC)
policy forum for the amendment of the Ecological Solid Waste Management Act of 2000
(RA 9003) and its Implementing Rules and Regulations last September 23 and 24, 2014, a
major concern on landfill leachate treatment was raised due to the lack of attention given to
the topic despite its adverse effects to the ecosystem. Contamination of groundwater,
emission of methane gas to the atmosphere, and the proliferation of disease-bearing animals
and microorganisms all these to name a few.
By definition, landfill leachate is the liquid that drains or leaches from a landfill. It
varies widely in composition considering the age of the landfill and the type of waste that it
contains. It usually includes both dissolved and suspended material. In a landfill that receives

a mixture of municipal, commercial, and mixed industrial waste, but excludes significant
amounts of concentrated specific chemical waste, landfill leachate may be characterized as a
water-based solution of four groups of contaminants: (1) dissolved organic matter, (2)
inorganic macro components, (3) heavy metals (Pb, Cd, Cu, Hg), and (4) xenobiotic organic
compounds such as halogenated organics (Kjeldsen et al., 2002).
Currently, landfill leachate treatment can be one or a combination of biological,
chemical, and physical processes. Biological treatment removes organics and nitrogen
through processes such as activated sludge, trickling filters, and nitrification or
denifrification. Chemical processes are used to control pH, precipitate metals, and remove
some organics by oxidation. Suspended matter can be removed by the physical processes of
sedimentation and filtration (Tchobanoglous, et.al., 1993). These treatment technologies can
be used to treat leachate for discharge into surface waters, or as pre-treatment prior to
transport to a municipal wastewater treatment facility for further treatment. All of these
management options for leachate treatment require energy input and will increase the landfill
operation and maintenance cost with no additional benefit.
There is substantial energy in leachate that is currently wasted or lost in treatment
processes and these come from waste organic matter and revolutionalize landfill leachate
treatment. One of the most potentially interesting technologies for the production of electrical
energy from organic matters is use the use of Microbial Fuel Cell or MFC a device that
converts chemical energy to electrical energy by the catalytic reaction of microorganisms
(Divetiya, 2014). Capturing this energy would provide a new source of electrical power.
Application of MFC technology for waste treatment was first suggested by early
researchers and was demonstrated for food processing wastewater in 1983. The range of

organic compounds that can be metabolized by chemotropic microorganisms (and their

enzymes) is extremely large so almost any waste stream may be utilized in an MFC (Caye et.
al., 2008). Among the possible sources of potential bacteria for an MFC is a landfill leachate
due to its microorganism diversity. Currently, the most studied exoelectrogens belong to the
-, -, -, and -proteobacteria (e.g., Geobactersulfurreducens, Geobactermetallireducens,
She- wanellaoneidensis, Escherichia coli, Rhodopseu- domonaspalustris) (Bond and Lovley,
2003; Min et al., 2005a; Ringeisen et al., 2006; Qiao et al., 2008; Xing et al., 2008); while
some non- proteobacteria (e.g., Geothrixfermentans (Bond and Lovley, 2005) and yeasts
(e.g., Saccharomyces cerevisiae (Walker and Walker, 2006)) are also capable of exocellular
electron transfer.
In an MFC, the oxidation of an electron donor compound is physically separated from
the terminal electron acceptor. When microorganisms consume a substance under aerobic
conditions, they produce carbon dioxide and water. However, when oxygen is not present,
they produce carbon dioxide, protons, and electrons. The electrons pass from one part of the
cell called the anode through a circuit that includes an external resistance load, then to a
cathode and finally to the terminal electron acceptor contained in the cathode chamber.
Typically, the anode compartment is separated from the cathode compartment by a proton
exchange membrane (PEM) or cation exchange membrane (CEM). Hydrogen ions pass from
the anode compartment through the membrane to the cathode compartment and combine with
oxygen to form water.

By diverting electron flow from microbial respiration to the

electrodes, MFCs convert chemical energy to electrical energy commonly referred to as

bioelectricity generation.
An understanding of the basic interactions inside the MFC as they affect

characteristics of the substrate is essential. These interactions can explain how the system
functions as a treatment process. Since reduction-oxidation reactions occur when bacteria
consume the organic compounds in a Microbial Fuel Cell, reduction in physico-chemical
parameter concentrations such as biochemical oxygen demand (BOD) and chemical oxygen
demand (COD) are critical to be measured. The effectiveness of mitigating heavy metal
concentration is also observed. Recording pH is also necessary.

Though leachate composition may vary widely, three types of leachates can be
defined according to landfill age (Baig et al.).
Table No. 1 Leachate Classification
Leachate type

Young

Intermediate

Stabilised

Landfill age year

<5

5-10

>10

pH

<6.5

>7.5

COD g/l

>20

3-15

<2

BOD/COD

>0.3

0.1-0.3

<0.1

Organic matter

70-90% VFA

20-30%VFA

HMW

Nitrogen

100-2000 mg/l TKN

100-2000 mg/l TKN

100-2000 mg/l TKN

Metals g/l

<2

<2

Source: (Baig et al.)

The BOD is the amount of dissolved oxygen required to meet the metabolic needs of
aerobic microorganisms in water that is rich in organic matter, such as sewage (Crites, 2000).
The COD, meanwhile, represents the number of electrons contained in a compound,
expressed as the mass of oxygen required to accept the electrons as the compound is

completely oxidized. The power output from an MFC is directly related to the electrons
captured from the substrate (Caye et al., 2008).

THEORETICAL AND CONCEPTUAL FRAMEWORK

This is an experimental research which investigates the application of Microbial Fuel
Cell in bioelectricity generation and the treatment of landfill leachate from Urdaneta City,
Pangasinan.
A microbial fuel cell is a bioreactor that converts chemical energy in organic
compounds (wastewater) to electrical energy. In an MFC, microorganisms oxidize organic
matter to generate electrons and protons. The electrons are transferred through various
methods to the anode (as an electron acceptor). These electrons are then conducted over a
resistor to the aerobic cathode where oxygen is reduced and meets with protons to produce
water molecules. Microorganisms carry out the conversion under anaerobic conditions
(Ganesh, 2012).
The overall reaction is the breakdown of the substrate to carbon dioxide and water
with a simultaneous production of electricity as a by-product.
In this experiment, a laboratory scale mediator less two chamber cell is used. The
system is composed of an anode and cathode chamber separated by a membrane permeable
to protons. The anode compartment is the anaerobic region where anaerobic bacteria are
located, while the cathode compartment is aerobic. The membrane separation helps to
maintain these conditions for each compartment, yet allows a charge transfer between the
electrodes. Electrodes are placed in each chamber to facilitate the electron transfer process.
Electrons and protons are produced through the oxidation of organic matter. The electrons are

transferred from the anode electrode, in the anode compartment, and travel through a wire
and resistor to the cathode electrode. Here, the electrons join with the protons which have
diffused through the membrane from the anode compartment, and oxygen to form water. A
catholyte solution must be used to facilitate this reaction.
One part of the fuel cell is the anode. Often, the anode is composed of graphite,
carbon paper or carbon cloth (Clauwaert et. al., 2007). The anodic chamber is filled with the
carbon substrate that the microbes metabolize to grow and produce energy. In this study,
leachate was used as the anode because of its microbial diversity.
Another component of MFC is the cathode. It completes the circuit of the cell by
transferring electrons to a high-potential electron acceptor. The cathode chamber is
commonly filled with a conductive media to facilitate oxygen reduction reaction. Thus, brine
solution was used in this study since it is safe, cheap and readily available. To complete the
cathode chamber, an oxidizing reagent needs to be introduced to maintain the abiotic
environment. Oxygen is the preferred oxidizing agent since it simplifies the operation of the
cell. It is supplied with the aid of an aeration pump that lets the air flow passively inside the
cathode chamber.
To determine the effectiveness of the microbial fuel cell, the power produced was
determined. The voltage of the cell and the resistance of the resistor were measured. With
these, the power generated was computed using the formula Power = (Voltage)2/(Resistance).
Leachates are known for its microbial diversity. It has bacteria, which includes
aerobic, coliform and fecal coliform, psychrophilic and mesophilic bacteria, and sporeforming bacteria (Matejczyk, et.al, 2011). Microorganisms, such as bacteria, are responsible
for decomposing organic waste. When organic matter such as dead plants, leaves, grass

clippings, manure, sewage, or even food waste is present in a water supply, the bacteria will
begin the process of breaking down this waste. If there is a large quantity of organic waste in
the water supply, there will also be a lot of bacteria present working to decompose this waste.
In this case, the demand for oxygen will be high so the BOD level will be high. As the waste
is consumed or dispersed through the water, BOD levels will begin to decline. The 5-Day
BOD test was used which involves the dilution of the samples to a volume of 300mL in a
stoppered bottle. The dissolved oxygen of the diluted samples was measured before and after
incubation of five days.
Chemical Oxygen Demand represents the number of electrons contained in a
compound, expressed as the amount of oxygen required to accept the electrons as the
compound is completely oxidized. A chemical oxidation reduction involves the breaking of
chemical bonds and the removal of electrons. The electrons are transferred from the
contaminant to the oxidant. The contaminant is in turn oxidized and the oxidant, the electron
acceptor, is reduced. In this study, initial and final COD concentrations were measured using
HACH colorimeter.
To survive under metal-stressed conditions, bacteria have evolved several types of
mechanisms to tolerate the uptake of heavy metal ions. These mechanisms include the efflux
of metal ions outside the cell, accumulation and complexation of the metal ions inside the
cell, and reduction of the heavy metal ions to a less toxic state. The complex structure of
microorganisms implies that there are many ways for the metal to be taken up by the
microbial cell. The biosorption mechanisms are various and are not fully understood. They
may be classified according to the dependence on the cell's metabolism, which are
metabolism dependent and non metabolism dependent. Also, they may be classified

according to the location where the metal removed from solution is found: extra cellular
accumulation/precipitation,

cell

surface

sorption/precipitation;

and

intracellular

accumulation. To test the ability of microorganisms to reduce heavy metal concentration of

the leachate sample, the initial and final concentrations of heavy metals were measured using
Atomic Absorption Spectroscopy (AAS).\

PROBLEM STATEMENT AND HYPOTHESES

To serve as a guide in achieving the objectives of this research, the following
questions are formulated and must be answered at the end of this work:
01. What is the percent reduction of BOD, COD, Pb, Cu, Cd after the treatment?
a. Microbial Fuel Cell
b. Via Precipitation
02. Is there a significant difference in the characteristics (BOD, COD, Pb, Cu, Cd) of
landfill leachate after the treatment between the MFC and via precipitation?
Hypothesis: There is a significant reduction of BOD, COD and heavy metal
concentration before and after the leachate treatment.
03. Is there a significant difference in Chemical Oxygen Demand reduction with
the following variation in load resistance in the Microbial Fuel Cell: 50,
100, and 150?
Hypothesis: There is a significant difference in the Chemical Oxygen
Reduction with varying load resistance (50, 100, and 150) in the
Microbial Fuel Cell.
04. What is the trend of Bioelectricity produced with respect to time?

I
N
P
U
T

P
R
O
C
E
S
S

Raw Material:
- Landfill Leachate
- Brine Solution
Salt Bridge
- Agar
- Sodium Chloride
Fuel Cell
- PET bottle
- PVC pipe
- Copper wire
- Carbon brush

Collection of Landfill Leachate

Formulation of Agar for Salt Bridge
Assembly of Fuel Cell
Determination of initial and final
BOD, COD and heavy metal
concentration

O
U
T
P
U
T

Microbial Fuel Cell

Figure No. 1 Paradigm of the Study

SIGNIFICANCE OF THE STUDY

This research aims to establish that MFC is an effective treatment of landfill leachate
and a sustainable energy source. With the current issues pertaining to solid waste
management and the incessant quest for alternative renewable energy sources, the greatest
significance of this work is that it introduces an integrated function of Microbial Fuel Cell as
a landfill leachate treatment system associated with bioelectricity generation utilizing a
sample from the Engineered Sanitary Landfill of Urdaneta City, Pangasinan which currently
caters the garbage of nearby areas in Northern Luzon that includes Baguio City (Camilo and
PEZA), Bautista, Mangatarem, Mapandan, Pozzorubio, Sual and the city of Urdaneta itself.

SCOPE AND DELIMITATION

This study covers the analysis of the landfill leachate characteristics such as BOD,
COD and Heavy Metal concentration utilizing a sample from the Engineered Sanitary
Landfill of Urdaneta City, Pangasinan. The study does not include outside inoculation of
bacteria due to the presence of mixed bacterial community in the leachate. The heavy metal
concentration reduction is limited to lead (Pb), cadmium (Cd), and copper (Cu). For the
cathode compartment and PEM, the salt concentrations are based from related literatures.
For each problem and parameter being investigated on this study, three trials were
conducted since it is the lowest number of trials that is allowed for experimentation. The
observation of the performance of the MFC is limited to every six-hour reading for five days
under room temperature.

10

CHAPTER 2
RESEARCH DESIGN AND METHODOLOGY

Research Design and Methodology

This research study adopted the experimental research design that would add
foundation for similar undertakings in the future. The design approach was employed for it
is known to be the most reliable, intellectual and accurate approach. Experimental research
involves the control or manipulation of conditions for the purpose of discovering the
influence of one or more factors upon a condition. The study would demonstrate the viability
of constructing an inexpensive microbial fuel cell as a leachate treatment other than the
conventional methods and toproduce electricity from raw material other than the
conventional sources.
The researchers conducted this experiment to establish the extent of the leachate
treatment andits potential as a renewable source of energy through a microbial fuel cell. The
researchers gathered leachate from the Engineered Sanitary Landfill of Urdaneta City,
Pangasinan. Samples were immediately brought to the Environmental Research Laboratory
and conducted the necessary test to determine its physico-chemical properties. Leachate
samples were then transferred to its respective compartments in the fuel cell and the
generated electricity in terms of volts measured every 6 hours for 5 days.
Data Gathering Tools
For gathering data, the researchers performed an experiment utilizing the different
resources found in the Environmental Research Laboratory (ERL) of Saint Louis University.
The researchers also supplemented the data with information from scholar articles in the

11

internet, related literature and research materials about Microbial Fuel Cell (MFC) and
related books about leachate treatment process.
To support the validity of the research, the researchers performed an experiment
using several trials in varying the resistance used for each sample. Statistical methods, such
as T-test, Analysis of Variance (ANOVA) were also used to analyse the collected data.
Data Gathering Procedure
A. Collection of Raw Materials
The leachate sample was obtained from the Engineered Sanitary Landfill of Urdaneta
City, Pangasinan. The sample was put into a cooler to ensure freshness.

It was then

transported to the laboratory for immediate tests.

B. Leachate characterization
Leachate characterization was conducted before and after the leachate treatment
process in Microbial Fuel Cell. Analyses for pH, Biochemical Oxygen Demand (BOD) and
Chemical Oxygen Demand (COD) were conducted at Environmental Research Laboratory
(ERL) of Saint Louis University. Meanwhile, AAS for heavy metals were conducted at
Philex Mines.
1. Biochemical Oxygen Demand (BOD) Measurement
10 mL of pure leachate sample was transferred in an empty BOD bottle. The BOD
bottle was filled with distilled water up until the opening of the bottle and was immediately
covered with glass stopper. The bottle was inverted 10 times to mix the leachate sample and
the distilled water. The stopper was removed and 6 mL of the mixture was discarded. 2 mL
of alkaline iodide solution and 2 mL of manganous sulphate solution were added. The bottle
was then covered with the stopper, inverted 20-30 times and flocs were allowed to settle. 2

12

mL of concentrated sulphuric acid was added, and the bottle was then inverted 20-30 times to
allow mixing. 203 mL of the solution was measured and transferred to a 250 mL Erlenmeyer
flask. The solution was titrated using sodium thiosulphate as the titrant until the color of the
solution turns to pale yellow. Enough amount of starch solution was added such that the
color of the solution turned to blue. Titration was continued until the solution became
colorless. Afterwards, the volume of the sodium thiosulphate consumed was recorded. This
served as the initial dissolved oxygen in mg/L.
10 mL of the leachate sample was transferred to an empty BOD. The BOD bottle was
filled with dilution water up until the near opening of the bottle, and was then covered with
glass stopper. The bottle was covered with cap to ensure that there was no air entering. The
bottle was stored in an incubator at 20 C and analyzed after 5 days.
2. Chemical Oxygen Demand (COD) Measurement
10 mL of the leachate sample was transferred in a beaker and was diluted with 100 mL
distilled water. Afterwards, 2 mL of the diluted solution was transferred in the COD vial and
inverted gently several times to allow mixing. The vials were placed in the COD digester
and heated to 150 C for 2 hours. The vials were then allowed to cool to 120 C or less and
the chemical oxygen demand was measured using HACH colorimeter.
3. Heavy Metal Measurement
Leachate samples were digested as follows: 50 mL of well-mixed sample was transferred
to a 250 mL beaker and heated using a hot plate with a temperature of 200-300 until the
volume was reduced to 15 20 mL. Concentrated nitric acid (15 mL) was added and the
sample was heated for 30 minutes. Afterwards, 15 mL of concentrated hydrochloric acid was
added and the sample was further heated for 30 minutes. The solution was then allowed to

13

cool in a water bath for 15-20 minutes.

The digested sample was then transferred

quantitatively with deionized water to a 100 mL volumetric flask and diluted to volume.
After the digestion, samples were brought to Philex Mines for analysis using Atomic
Absorption Spectroscopy (AAS).
C. Microbial Fuel Cell Construction
1. Construction of Anode and Cathode Compartments
Gatorade bottles (500 ml) were collected and used as anode and cathode compartments.
Circular holes were made into the bottom half of the Gatorade bottles about 2 inches in
diameter using a heated red hot tansan (crown). Sandpaper was used to smoothen the edges
of the holes made. These bottles were washed with mild soap and water.
2. Preparation of Electrode
For every cell, two copper wires were cut into 6-inches long. Each of the copper
wires was soldered into carbon brushes (SM-56). The cover of the Gatorade bottle was
drilled to make a hole to which the sponge and the copper wire with the carbon brush was
inserted for the anode compartment. For the cathode compartment, a tiny hole in the center
of the sponge was used to hold the copper wire so it freely hung to the saltwater solution.
3. Preparation of Proton Exchange Membrane (Salt bridge)
Moldex PVC pipes were used to hold the salt bridge. The pipes were cut into 5
inches long using saw. 3 pipes were cut for the fuel cells.
In preparation of the agar solution, 500 ml of distilled water was heated in a beaker
using a beaker using a Bunsen burner. Twenty grams of laboratory grade Agar-Agar powder
was put into the beaker then stirred. When the solution started to boil, 10 grams of salt was
added to the mixture. The solution was stirred continuously until it became viscous. The

14

pipes were placed in another 1000 ml beaker. The solution of salt and agar was then poured
into this beaker filling up the pipes. These were cooled to room temperature until the agaragar hardened.
4. Preparation of Saltwater Solution
125 grams of Sodium Chloride (NaCl) crystals was weighed using a digital balance
then dissolved into 1 liter of distilled water until a solution was formed. The solution was
transferred into a plastic bottle. For a set-up of 3 cells, one liter of salt solution was needed
5. Fuel Cell Assembly
After the preparation of the materials, the fuel cell was assembled. The salt bridge
was inserted to the holes of the Gatorade bottles, connecting the two compartments. A glue
gun was used to cover the spaces of the salt bridge and the hole to ensure no leakage to
occur. The anode compartment was for the leachate. Meanwhile, the cathode was for the
salt solution. The electrodes were inserted after placing the bacteria and the salt solution.

D. Testing the Potential of Leachate to Generate Power

Considering three trials consisting of three sample set-ups, 225 ml of leachate was
used for this experiment. The leachate was allowed to have contact with the salt bridge and
the electrode. 225 mL of salt solution was then poured into each of the cathode compartment
of each MFC. This amount was just enough to fill half of the bottle. The electrode was then
fully immersed into the salt solution.
After this, a multitester was used to measure and monitor the voltage and current
produced of each MFC in 6-hour intervals for five days. These data were then recorded.

15

Treatment of Data
After the experimentation , the data will be subjected to computational and statistical
analysis.
The amounts of reduction for different parameters (BOD, COD, Pb, Cu, Cd) will be
expressed in terms of percentages.

% =

100

( Equation 2.1)

To determine the significant difference in BOD, COD, pH, Pb, Cd, Cu between the
control and the Microbial Fuel Cell, the results will be statistically treated using T-test. The
equation given will be used to determine the difference needed for viability of treatment.

(Equation 2.2)

A line graph will be used to establish the relationship and identify the trend between
the amount of power produced with time. The time will be the abscissa while power will be
the ordinate of the graph.
In the determination of the existence of a significant difference in the COD reduction
of the varied resistance of the Microbial Fuel Cell, the results are statistically treated using
One-way Analysis of Variance (ANOVA) and Tukeys Honest Significant Difference (HSD)
Test.

16

The F-Test through the One-Way Analysis of Variance (ANOVA) is used to

determine the significance of the differences among the COD reduction obtained using varied
resistance of Microbial Fuel Cell. The formulas utilized are given by the one-way ANOVA
table below:
Table 2-1
One-way ANOVA Table

SOURCE OF

DEGREES OF

SUM OF

MEAN

VARIATION

FREEDOM

SQUARES

SQUARE

Between Groups

k-1

SSC

S12=SSC/k-1

Within Groups

N-k

SSE

Total

N-1

SST

SST =

SSC=

SSE=SST-SSC
Where:
k= no.of treatment
N=total sample in the whole treatment
n=total sample per treatment

17

S2 =SSE/k-1

S12
S22

SS=sum of squared deviations

F=degrees of freedom
X=sample being analysed

The Tukeys Honest Significant Difference (HSD) Test was used to determine the
best resistance for COD reduction in Microbial Fuel Cell. The test statistic was computed the
following formula:

Where:
Q= value from THSD table
MSwithin= Mean Square within Groups from ANOVA Table
n= number of trials per treatment

To resolve the problems of the study, the researchers carefully and systematically
presented the data in tables which are supported by computations. Descriptions and
interpretations. It was through these means that the researchers were able to come up with
analyses and interpretations showing the relationship of the findings to the problem.

18

19