FINAL YEAR PROJECT

THE EFFECT OF COSMOBALL PLASTIC MEDIA

CONFIGURATION ON WASTEWATER TREATMENT

By : Muhamad Asri bin Mohd Arif (174823)

Supervisor : Prof Dr Azni bin Idris

 CHAPTER 1

INTRODUCTION

1.1 INTRODUCTION
 Wastewater treatment becomes more and more advance nowadays.

There are always research that been done to develop a better

wastewater treatment. Today, cosmoball that has been introduced by

Professor Dr. Azni bin Haji Idris from Universiti Putra Malaysia (UPM)

which was started on 1990 and has become popular and well known

among industries. The technology developed by Prof Dr. Azni is also

known as Biofil system which can treat river or any other

contaminated water. The best feature of this Biofil which that it does

not use any chemical which is parallel to the vision of Green

Engineering of UPM itself. However, we as UPM researchers are still

pursuing to increase the adaptability and the effectiveness of the

Biofil.

1.2 BACKGROUND OF STUDY

Recent issues that have become a major concern on the cosmoball

treatment are the low self - sustainability characteristic. Thus, it

causes high maintenance cost which is not favourable by the

consumers. Hence, the study of the cosmoball configuration on how

the cosmoball is arranged is investigated to determine its effects

towards the treatment efficiency.

1.3 STATEMENT OF PROBLEM

There are several issues that have come up with the cosmoball in the

maintenance aspect. One of it is that after a certain period of

cosmoball usage the surface of the cosmoball will be filled by layers of

sludge that consists of bacteria. The main problem is when there is

accumulation of sludge it limits the availability of bacteria on the

surface of the cosmoball which reduce the efficiency of decomposition

process. Besides that, eventhough that the sludge inside the

cosmoball can be washed away with water but with large quantity of
 cosmoball due to packing arrangement the cleaning process will be

easier said than done. Even though there several approach to wash

the sludge from comsmoball such as using water jet, but it is costly

and wasteful. So, is there a solution for these problems?

1.4 RESEARCH OBJECTIVES

i. To identify a better configuration of the cosmoball which are

applicable in most types of condition by comparing to the

previous configuration.
ii. To determine optimize the cosmoball quantity in a specific

volume in order to reduce cost.

1.5 ASSUMPTIONS AND HYPOTHESES

There are several assumptions and hypotheses that should be taking

into account such as the following:

i.

1.6 SIGNIFICANCE OF STUDY

Cosmoball has contributed huge development in wastewater treatment

technology as it approaching towards the Green Engineering. Thus, in

depth research in the configuration aspect might be important to

improve the cosmoball to become a whole lot more improve compared

to the cosmoball available today.

1.7 SCOPE AND LIMITATION OF STUDY

Even though the study is to focus on the configuration of the

cosmoball itself, there is limitation due to the available shape of

cosmoball commercialized which only available in only one shape and

has a fixed dimension. The only things that can be modified are that it

can be separated into two semi sphere shape of cosmoball and how it

could be arranged. There are several techniques of arrangement that

may be applied to the cosmoball configuration but the effectiveness is

not yet being assured. Hence, trial and error method might be the best
 solution to find a better cosmoball with high self sustainability

characteristics.

1.8 STRUCTURE OF THESIS

1.9 SUMMARY
 CHAPTER 2

LITERATURE REVIEW

2.1 Wastewater and Its Treatment

Wastewater treatment is the only existent process in which you cannot

control the inputs, but have to create a defined output. Treating

wastewater is vital. Less than 3% of the worlds total water supply is

available as fresh water, and salt water is costly to desalinate. Due to

this fact, water that was previously used for cleaning, bathing, and

drinking will eventually end up back in a river or lake for it to be

consumed once again (Ellis, 2004). Nature has the ability to manage

and clean certain amounts of waste and pollution in water through its

use of microorganisms and algae. In the past few decades, however,

the amount of waste that is disposed each year has increased,

consequently affecting our environment in negative ways. Natures

inability to sufficiently treat the increasing amount of waste in bodies of

water is one of the reasons why wastewater treatment plants have

become a focal path in securing our future water supply. Wastewater

treatment plants are essential to manage and clean the effluent from

industrial, agricultural, and domestic sources. Treated wastewater is

discharged to lakes and rivers and is eventually reused, so monitoring

the chemical and bacteriological composition is crucial in guaranteeing

public health (Ellis, 2004). The treatment of wastewater is also

imperative to the protection of many aquatic species. Wastewater can

have low dissolved oxygen conditions, as well as excess nutrients such

as nitrogen and phosphorous. Under these conditions the ecosystem is

prone to develop a syndrome, in response to human activities, known

as eutrophication. Eutrophication refers to excessive plant growth

(mostly algae), which leads to a change in habitat from the lack of

oxygen present in bodies of water. Figure 1 below provides a

description of the process of eutrophication (Cloern, 2007).

Figure 1: Process and Effects of Eutrophication (Cloern, 2007)

2.1.1 Wastewater Components

Wastewater is composed of many organic and inorganic

materials, chemicals, and nutrients, among others. The

composition of wastewater varies every day. Components in

the wastewater can differ depending on location, season, and

time of day. There is no way to control the radical changes in

wastewater, given that they depend on many factors. The

composition of wastewater is roughly 99.9% water and 0.1%

total organic and inorganic matter (Olutiola, 2010).

Microorganisms
Many types of microorganisms can be preset in wastewater.

The most common are: bacteria, viruses, protozoa and

helminthes. Most of these microorganisms can be considered

harmless, given that they are present in the human body, but

there are some that might cause disease. In order to get rid

of disease causing microorganisms, the wastewater must go

through rigorous processes such as filtering and chlorination

(Jimenez et al., 2008).

Organic Material
The organic composition of wastewater is approximately

50% proteins, 40% carbohydrates, 10% fats and oils, and

trace amounts of priority pollutants and surfactants (Ellis,

2004). The organic material present in wastewater includes

detergents, pesticides, fats and oils. These substances can be

toxic and must be eliminated from wastewater. Within the

organic material, biodegradable organic material also resides.

Biodegradable organic material refers to all waste that will

naturally degrade with time. This material is comprised of

wood, plant matter, paper, and human and animal waste.

Even though it does not present a direct risk to the

environment, it is a way in which microorganisms are

transferred. Many microorganisms are introduced to the

water in this manner, and since water composition needs to

be monitored to prevent diseases, organic matter needs to be

removed (Henze, 2002).

Basic Nutrients
Basic nutrients present in wastewater are most commonly

nitrogen, phosphorous and ammonia. These nutrients can

cause depletion of oxygen, i.e., eutrophication when they are

present in excess. As seen in Figure 1, the nutrients are

capable of causing significant damage to the environment,

due to the changes they impart on the ecosystem (Henze,

2002).

Metals and Inorganic Material

Most of the metals and inorganic materials found in

wastewater are part of industrial waste. Some of these

metals and inorganic materials present in water are mercury,

lead, cadmium, nickel and hydrogen sulphide. The amounts

of these components that is present in wastewater greatly

surpass the trace amounts that humans and animals need for

health purposes. Wastewater is eventually reused due to its

scarcity, so these components need to be eliminated. They

can cause corrosion, and be toxic to the environment as well

as humans (Henze, 2002). Table 1, shows the components

present in wastewater and their effect on the environment.

 Table 1: Components in Wastewater (Henze, 2002)

2.1.2 Overview Wastewater Treatment

There are many ways in which wastewater is treated,

primarily because many factors need to be taken into

consideration prior to the treatment of sewage. As previously

mentioned, the location and time of the year can drastically

affect the composition of wastewater. For example, the area

in which the water is treated and where the water will be

discharged affects its cleaning process. If the water is to be

discharged in a river, the chemical and bacteriological

content needs to be less than if it were to be used for

irrigation of fields. Wastewater can be treated for different

purposes, some of which are for agricultural and potable use.

The treatment of water for potable standards is very rigorous,

while the treated water for agricultural use can have traces of

organic matter and solid waste.

Wastewater treatment is generally divided in three or four

main stages, which represent the degree to which the water

is treated. These stages are: preliminary treatment, primary

treatment, secondary treatment, and tertiary or advanced

treatment. Within these stages there are different steps and

methods that can be employed to treat the water (Prescod,

1992).

Preliminary Treatment
Preliminary treatment is used to eliminate large, solid objects

that are often present in wastewater. Although this not an

essential step in the cleaning process, treatment plants

typically employ it as a precaution to prevent any damage

these objects may cause to machinery in subsequent stages.

The most common methods utilized in this stage are

screening and grit removal.

Screening is the first step in the preliminary treatment of the

wastewater. The water flows through a series of screens

which filters out large, medium, and small objects from the

liquid. Usually three different mesh sizes are used. The mesh

size of the largest grid is about 5.0 centimeters, and it is used

to remove large objects, such as branches and cloth. The

mesh sizes of the second and third grids range between 0.5

to 3.0 centimeters, and 1.5 to 3.0 millimeters, respectively

(Hogan, 2010). The grids are cleaned regularly, either

manually or sometimes by using an automated process.

The next step in the preliminary process is known as grit

removal. This process consists of separating sand, gravel,

and any food particles present in the wastewater. There are

different types of grit chambers, some of which are aerated;

others just have a specific flow pattern. The turbulence,

either from flow or air, settles the heavy components in the

water, while light organic particles are suspended. The

settled grit is then removed by a pumping system that takes

it out of the chamber (EPA, 2004). Using this grit removing

process will decrease the chances of damaging any type of

mechanical system that may be used in the following steps

(Vesilind, 2003).

None of the steps in preliminary treatment involve chemical

or biological improvement of the wastewater. This is one of

the reasons why some treatment plants, especially small

ones, do not include this stage in their process, but include

screening as the first step in their primary treatment instead

(Hogan, 2010).

Primary Treatment
The primary treatment of wastewater is the stage in which

the majority of organic and inorganic material, as well as

contaminants in the water, are removed. Solids are removed

by sedimentation, while oils and grease are removed by

skimming (Hogan, 2010). Approximately 25% to 50% of the

incoming biochemical oxygen demand (BOD), 50% to 70% of

the total suspended solids (SS), and 65% of the oil and

grease are removed during primary treatment (Prescod,

1992).

In this stage the wastewater is subjected to a process known

as coagulation flocculation, which leads to the settling of the

solids. Since the particles in the wastewater are fairly small, a

coagulant is added to the water to simplify the removal

process. The purpose of this chemical is to attract the

particles that are present to make them clump together; this

simplifies their removal.

Figure 2: Steps Followed During the Primary Treatment (The

Water Treatments, 2008)

Figure 2 shows the process that the wastewater follows in the

primary treatment stage. First, the coagulant is added. Then,

the water is rapidly mixed for a short period of time to ensure

dispersion of the coagulant. After rapid mixing, the speed of

the mixer is lowered to allow the particles clump. Once flocs

are formed, they either settle to the bottom of the tank, or

rise to the top. The bottom flocs are removed as sludge, while

the top ones are removed as scum, using a continuously

moving scrapper (Vesilind, 2003).

Secondary Treatment
Secondary treatment of wastewater employs biological

processes and the use of microorganisms to rid the water of

any organic compounds that may still be present. This stage

simulates what actually happens in nature, when

microorganisms break down organic wastes. There are three

main approaches that can be used in this stage: fixed film,

suspended film and lagoon systems (Mancl, 2009).

Fixed Film
In a fixed film system, microorganisms grow on rocks, sand,

or plastic, to create a film. They grow on these surfaces by

feeding off the organic matter and nutrients in the

wastewater that flows over them. There are three main fixed

film systems that are commonly used today: trickling filters,

rotating biological contactors, and sand filters (Mancl, 2009).

Suspended Film
Suspended film systems consist of suspending the

microorganisms in the wastewater. While in the water, they

absorb the organic waste and nutrients around them, which

allows them to grow and reproduce to form micro-colonies.

These micro-colonies settle as sludge, which is then removed

and either reused in the process by being re-suspended, or

treated in a sludge treatment process. Some examples of

suspended film are: activated sludge, extended aeration, and

sequential batch reactor systems.

Lagoon Systems
Lagoon systems are settling ponds in which the wastewater is

retained for an extended period of time. During this time,

microorganisms degrade the organic compounds present in

the water.

The systems used in secondary treatment completely rely on

natures ability to clean the wastewater through natural

resources. The use of sunlight, algae, oxygen, and

microorganisms is essential (Mancl, 2009). The result of

effective primary plus secondary treatment is removal of

about 90% of the organic matter, virtually all pathogens, and

most solids. Between 10% and 20% of the nitrogen is also

automatically removed because the decomposer bacteria

require this much for their own growth (The Sound Book,

2010). After these three treatment stages, the water is

sufficiently clean for use in non-potable applications. These

include: agricultural uses, non-potable household uses, and

irrigation of parks.

Advanced Treatment
Advanced wastewater treatment refers to any steps that are

utilized to raise the quality of water obtained from primary

and secondary treatments. Problems such as color, odor, and

taste are dealt with in this stage. The water is disinfected

using either one or a combination of techniques such as

chlorination, UV disinfection, and ozonation. Nutrients like

nitrogen and phosphorous are also sometimes removed

(Hogan, 2010).

Chlorination is the step in which chlorine is added to the

water as a disinfectant to eradicate any bacteria left behind

from the previous steps. UV disinfection consists of exposing

the treated water to radiation by UV light. The UV light

penetrates through an organisms cell wall and disrupts its

genetic material, impeding any reproduction. Ozonation is the

use of ozone gas (O3) to disinfect and improve the color and

odor in the water (Prescod, 1992).

An increasing number of wastewater treatment plants are

incorporating this last stage in their processes. The water

exiting at this stage in the treatment can be used for portable

 applications, which means the water can come in contact

with humans. Figure 3 depicts all of the stages used in

wastewater treatment processes, prior to releasing the water

to the environment.

Figure 3: Stages in the Wastewater treatment Process

(Sewage Treatment, 2009)

2.2 Biofilms

A biofilm is a community of microorganisms that develops in moist

environments. The organic and inorganic materials that comprise a biofilm

can range from decaying products in wastewater, to the millions of species

of microorganisms in a lake (Deibel et al., 2003). A biofilm population can

be made up of as little as one bacterial species, and is not limited to any

particular amount of species. Dental plaque, for example, is one type of

biofilm that is comprised of over 500 bacterial species (MSU Center for

Biofilm Engineering, 2008).

The varying species in biofilms, such as bacteria, fungi, algae, yeasts, and

protozoa, form clusters by producing an extracellular polymeric

substance, also known as EPS. The cells are held together by these

sugary molecular strands (EPS) and develop complex three-dimensional,

resilient communities. These clusters of microorganisms excrete a glue-

like substance, which allows them to adhere to a surface of almost any

type (MSU Centre for Biofilm Engineering, 2008). Where a combination of

moisture and nutrients exists on a surface, it is likely to find a biofilm

there. Biofilms have the ability to grow on natural materials above and

below ground, on metals, plastics, medical implant materials, and plant

and body tissue. Over time, a biofilm in the appropriate environment will

grow and become strongly attached to the surface it lives on (Deibel et al.,

2003).

Biofilms vary in size, depending on their surrounding conditions. A healthy

biofilm can grow to be many inches thick in an environment with the

proper nutrients and a steady temperature. To synthetically develop a

sustainable biofilm, constant replenishing of nutrients is necessary (MSU

Center for Biofilm Engineering, 2008).

2.2.1 The Life of Biofilms

Behaviors and survival strategies of bacterial colonies significantly

surpass the capabilities of individual bacteria. For example,

microbial biofilms are naturally tolerant of antibiotic doses up to

1,000 times greater than doses that kill free-swimming, planktonic

bacteria (MSU Center for Biofilm Engineering, 2008). The picture in

Figure 4, illustrates the five stages in biofilm development (Cogen et

al., 2004):
1. Initial attachment
2. Irreversible attachment
3. Maturation 1
4. Maturation 2
5. Dispersal

Figure 4: Stages of Biofilm Development (Cogen et al., 2004)

Initial Attachment
Initial attachment occurs when a material surface is exposed to an

aqueous medium (e.g., water, blood) and is coated by polymers

from that medium. This is known as the conditioning layer; it is

organic, forms within minutes of exposure, and continues to grow

for several hours. The cells that comprise the conditioning film

attach quickly and efficiently to hydrophobic, nonpolar surfaces,

such as plastics. In addition, initial attachment and colonization

increase as surface roughness increases. This is due to the larger

surface area on rougher surfaces (Donlan, 2002). Planktonic

microorganisms instinctually attach themselves to this conditioning

layer to become part of the developing biofilm (Cogen et al., 2004).

Irreversible Attachment
The second stage, irreversible attachment, begins once the

microorganisms start producing the extracellular polymeric

substance, EPS, which represents the house of the biofilm cells

(Flemming et al., 2007). The polyhydroxyl groups in EPS anchor the

bacteria in the biofilm to the surface through hydrogen bonding

(Kjelleberg et al., 2007). At this time, the microbes can no longer

move away from the surface. It has been determined that mature

biofilms are anchored to their place until the final stage of growth

(Kolari et al., 2001).

The Cell Matrix

The EPS is comprised of a wide variety of proteins, glycoproteins,

glycolipids, and extracellular DNA. The biopolymers in the

extracellular polymeric substance are highly hydrated and form a

matrix that holds the biofilm together and retains water. This matrix

interacts with the environment to provide nutrients for biofilm

organisms. Micro-colonies are formed within the EPS during the

maturation stages of development. The colonies are separated by

pores and water channels, which function as pathways for nutrients

and oxygen to flow through, and nourish the biofilm (Flemming et

al., 2007).

Maturation
During the maturation stages, the biofilms main goal is to grow in

three dimensions. This is achieved by picking up debris in the

surrounding environment, such as sand, and by recruiting new

planktonic bacteria. The biofilm also grows in these stages through

reproduction, which occurs regularly in the micro-colonies. Some

biofilms can grow to be several inches thick, as seen in Figure 5,

which is a photograph of a streambed in Yellowstone National Park

in the United States. This particular biofilm is heavily populated by

green algae (MSU Center for Biofilm Engineering, 2008).

 Figure 5: Streambed in Yellowstone National Park (MSU Center for

Biofilm Engineering, 2008)

Dispersal
The final stage of development is dispersal, or shedding of cells.

This stage is sometimes referred to as expansion. While many

bacterial cells disperse from biofilms by passive processes, bacterial

biofilms can also experience active dispersal events in which

sessile, matrixencased biofilm cells convert to free-swimming

planktonic bacteria (Webb, 2007). Quorum sensing, commonly

referred to as cell-to-cell signaling, is essential to biofilm dispersal

(Donlan, 2002). Through quorum sensing, bacterial populations will

activate dispersion only when they are able to sense that their

population is numerous enough to make it advantageous. For

example, one study observed a cellular division of labor through

quorum sensing, where one group of cells stayed attached to the

surface and made nutrients available to the second group, which

reproduced and released daughter cells to the surrounding water

(MSU Center for Biofilm Engineering, 2008).

Quorum Sensing
An illustration of quorum sensing can be seen in Figure 6.

Figure 6: Quorum Sensing Depiction (Cunningham et al., 2011)

This image compares the cell-to-cell signaling abilities of planktonic

bacteria to the bacteria in biofilms. Planktonic bacteria and biofilm

bacteria both secrete chemical signals, or homoserine lactones

(HSLs). The difference is that the population of bacteria in a biofilm

is denser than that of planktonic bacteria, so the signals are more

concentrated, and thus, more effective (Cunningham et al., 2011).

Biofilm dispersal is not always intentional; passive dispersion, such

as erosion and sloughing, can occur due to hydrodynamic or shear

forces in the aqueous environment. Layers of biofilm may be broken

off due to natural abrasion, nutrient deprivation, and/or

hydrodynamic forces caused by the velocity of the liquid (Webb,

2007). Regardless of which way the dispersal occurs, a layer of cells

(the conditioning layer) remains on the surface for the growth

process to continue, or start over. It is important to note that the

higher the shear force, the denser the biofilm will be. This means

that a biofilm created under high shear conditions will be more

compact and less porous than one created under low shear
 conditions (Celmer et al., 2008). Furthermore, a less porous biofilm

will be less susceptible to natural erosion.

2.3 Biofilm Applications

The awareness of biofilms has increased enormously in recent years due

to the impact of biofilms on natural and industrial systems, as well as

human health. Biofilms cost the U.S. billions of dollars every year in

energy losses, equipment damage, product contamination, and medical

infections (MSU Center for Biofilm Engineering, 2008). The situations in

which biofilms exist can range from contamination of catheters, to

adhesion to rocks in steams, to corrosion of pipes in the oil industry.

Because both disease-causing and non-disease-causing bacteria are

incorporated into biofilms, food and medical industries have to be kept

biofilm-free (Deibel et al., 2003). The formations of biofilms cause

contamination or corrosion of equipment because of their physical

properties, which make them resistant to anti-microbial agents (Cogen et

al., 2004).

2.3.1 Biomedical Device Industry

An example of contamination caused from biofilms is found in the

biomedical device industry. It is not uncommon for biofilms to grow

on medical devices and implants within the human body. These

organisms typically originate from the skin of a patient or health

care worker, or tap water to which the device is exposed. Such

medical devices include urinary catheters, central venous catheters,

prosthetic heart valves, and artificial hip prosthesis (Kokare et al.,

2008).

The organisms most commonly isolated from catheter biofilms are

S. epidermidis, S. aureus, C. albicans, P. aeruginosa, and K.

pneumonia. They gain access to the catheter by migrating

externally from the skin along the exterior catheter surface or

internally from the catheter port. It has been observed that

colonization and biofilm formation can occur within 3 days of

inserting the catheter, but biofilm formation on internal surfaces of

the catheters is more likely to be present for those that remain in

place for longer periods of time (Donlan, 2001).

According to the National Institutes of Health, more than 65% of all

microbial infections are caused by biofilms (Proal, 2008). Specific to

urinary catheters, biofilms that develop will infect the patient and

result in a urinary tract infection. This is more likely to happen in an

open system, where the catheter drains into an open collection

center, than a closed system, where the catheter empties into a

plastic bag. Time is also a variable; essentially all patients who have

a urinary catheter for more than 30 days get infected with a UTI

(Kokare et al., 2008).

2.3.2 Water Systems

Although biofilms sometimes have harmful effects, they also offer

huge potential for certain applications, such as bioremediating

hazardous waste sites, biofiltering municipal and industrial water

and wastewater, and forming biobarriers to protect soil and

groundwater from contamination (MSU Center for Biofilm

Engineering, 2008). When used in engineered systems of

wastewater treatment, biofilms are often beneficial.

Some of these systems include trickling filter systems, modified

lagoons, and specialized systems for nutrient or waste removal.

Biofilm based treatment systems are advantageous because the

microbial communities are resistant to changing environmental

conditions, which makes them resilient to variation in toxicity

concentrations (Clark Ehlers et al., 2012).

Heavy metals such as copper, lead, and zinc are typical pollutants

in freshwater and wastewater. A study was performed by Teitzel et

al. to examine the effects of these heavy metals on biofilm and

planktonic P. aeruginosa. A rotating-disk biofilm reactor was used in

this experiment, and it was determined that biofilms can be up to

600 times more resistant to heavy metal stress than free-swimming

cells. By binding metal ions to the EPS matrix, biofilm is capable of

eliminating heavy metals from the surrounding liquid (Teitzel et al.,

2003).

CHAPTER 3

METHODOLOGY
3.0 Analytical Testing Techniques
Various techniques were applied in order to test the efficiency of biofilms

in wastewater. These were used to monitor the development of the

biofilm, as well as its ability to cleanse the wastewater. The nylon mesh

and plastic membrane frames were scanned daily to observe the biofilms

growth. Additionally, water samples were taken daily to test for changes

occurring in the water. The following experiments were designed to

determine the effect of biofilms on wastewater.

3.0.1 Total Organic Carbon (TOC) Test

The purpose of the total organic carbon (TOC) test was to determine

the concentration of organic carbon in the reactor tank each day of

the experiment. By testing for the existence of organic carbon in the

liquid, a conclusion can be drawn on the biofilms ability to function

properly. In theory, a biofilm should oxidize the organic components

in wastewater, which consist of organ carbon, causing the TOC

results to decrease over time.

The TOC machine measures the organic carbon in the water by

mixing the water sample with a strong acid. This acid reacts with

the inorganic carbon, which is converted into CO2 and is evaporated

from the liquid. The organic matter is left behind. Combustion takes

place in order to convert the organic carbon into water and CO2.

The CO2 is then transferred to an infrared analyzer, where the

adsorption wavelength of CO2 is read, and the concentration of

organic carbon can be obtained. Results are given in milligrams of

organic carbon per liter of water (mg/L) (LAR, 2008).

3.0.2 Ammonium Test

The ammonium test determines the concentration of ammonium

present in the wastewater sample. The method used for this test is

known as Nesslerization. The Nessler reagent is an aqueous solution

made of sodium hydroxide, potassium iodide, and mercuric iodide.

This reagent is added to a diluted sample of wastewater after a

mineral stabilizer and polyvinyl alcohol; the mineral stabilizer is

added to reduce cloudiness in the samples. A spectrophotometer is

then used to read the absorbance of ammonium present in the

sample (Tissue, 2000). A calibration curve is prepared with a sample

of pure distilled water, to which the reagent, stabilizer, and

polyvinyl alcohol are also added.

The intensity of the color can be correlated with the ammonium

concentrations present in the wastewater. A bright, yellow color

implies a low concentration of ammonium, while dark, brown

implies a high concentration. The concentration of ammonium in

the wastewater can be directly correlated to biofilm health. High

concentrations of ammonium suggest the biofilm is not growing

properly, while low concentrations suggest the biofilms are healthy,

and efficiently treating the wastewater.

In theory, the ammonium concentration of wastewater in which

biofilms are growing should decrease. This is because a certain

amount of the microorganisms that comprise biofilms have nitrifying

properties. Nitrifying microorganisms oxidize ammonium in the

wastewater, converting it to nitrite. These nitrites are then further

oxidized to nitrates. Two different groups of bacteria work on each

of these reactions. Nitrosomonas, the most common ammonium-

oxidizing bacteria, work on the first reaction in which ammonium is

converted to nitrite: NH3 + O2 NO2- + 3H+ + 2e-. Nitrobacters

work on the second reaction in which nitrite is converted to nitrate:

NO2- + H2O NO3- + 2H+ + 2e- (Environmental Protection

Agency, 2002).

3.0.3 Ultraviolet-Visible Spectroscopy

The amount of nitrates and the chemical oxygen demand (COD) in a

wastewater sample was determined through the use of ultraviolet-

visible spectroscopy. In this qualitative testing technique, a beam of

light passed through the given sample and measured the reflected

wavelength. The range of wavelengths used for the wastewater

samples was 190nm to 600nm. The beam of light (in the UV

regions), excited the particles in the sample for each wavelength.

After the sample was subjected to every wavelength in the

determined range, a graph was displayed. The graph showed peaks

at certain wavelengths, indicating the presence of certain

compounds with which the wavelength was associated.

For wastewater, there was one absorbency wavelength found in UV-

Vis spectroscopy that was important: 254nm. The absorbance at

254nm was correlated with the COD in the sample. The COD

measured all the oxidizable matter present in the wastewater.

Theoretically, as the biofilm treats the wastewater, the COD will

decrease because there is less oxidizable matter in the wastewater,

thus less need for oxygen.

On the contrary, nitrate concentrations, in theory, should increase

over time. This is because the biofilms are oxidizing ammonium,

therefore, creating nitrate. To find the nitrate concentration in the

sample, the second derivative of the absorbencies was used. Within

the range of 220 to 240nm, the highest value of the second

derivatives corresponding to the specified wavelength range was

isolated. These maximum values are directly proportional to the

nitrate concentration. Using the following equation, the nitrate

concentration in each sample was found (Pons, 2011):

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