Class Project 1 City Case StudiesIn this project student groups

Class Project 1: City Case Studies
In this project student groups will assess and compare
regulation and policy for a sample group of cities. The
foundation for this work is collection by student groups of all
key documents used in local land regulation in their case study
community. Groups of 2 students will participate in each case
study project. Student groups will be expected to collect and
assess all key documents used in local land regulation in their
case study community. The assignment will be divided into
modules, with sub-assignments handed out for each module.
The class will use the city of Boulder as a template and
baseline. Through guest lectures and document review we will
develop a thorough understanding of the Boulder case. This
assessment will focus on residential construction addressing
three development frameworks: (1) single-family residential
building; (2) multi-unit residential building or subdivision that
triggers land use or design review; and (3) multi-unit residential
structure or subdivision that triggers environmental review.
These three frameworks are intended to encompass core
regulatory problems of land development in this context, and
illustrate key issues of complexity, cost and conflict of
regulatory compliance for participants including both
developers and local governments. In addition, students will be
asked to collect (1) information on physical development
patterns, and (2) local media coverage of development politics.
A protocol for data collection will be handed out to students
when the project is assigned.
Assessment of case study documents will address the following
issues: (1) number and type of development regulations and
related application submittals; (2) analytical and evidentiary
requirements at each submittal step; and (3) process for review
of application submittals. Data will be assembled in a matrix for
each case study, and then evaluated against benchmarks derived
from further research and class discussions about the Boulder

county case. Time requirements for compliance with
development regulations will be identified. Students will write
up an evaluation of each case based on the matrix and
addressing key questions defined in the protocol. Students will
present their findings. Comparison between sites will occur
through in-class discussion.
Deliverables: (1) 8-page paper including matrix describing
regulations, policies and development patterns for case study
community, specifically referencing documents in the folder.
(2) Folder with all relevant documents uploaded to google drive
to made available to class. (3) 5-minute presentation to class
with power points.
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City Case Studies: Project Review and Conclusion to Case
Studies
In this project students will synthesize and conclude your
discussion of regulation and policy for your case study city. The
foundation for this work is collection of all key documents used
your city in local project review. Students are expected to
collect and assess all key documents used in project review.
This assignment is designed to help us explore key issues of
complexity, cost and conflict of regulatory compliance for
participants including both developers and local governments.
The class will use the city of Boulder as a template and baseline
including the following five steps.
1. Step 1. Compilation of key documents in a google document,
to be created by you, and with the url included in the beginning
of the assignment.
2. Step 2. Evaluation against the benchmark of the city of
Boulder, available at the bottom of this assignment sheet.
3. Step 3. Prepare a two-page paper describing and synthesizing
the work you’ve done on your own and in class about the city
plan, zoning code and project review process.

Include at least the following.
1. What aspects of project review, and comp plan and zoning
code, are emphasized in your city? Based on your knowledge of
your city, why do you think these aspects are emphasized?
code, stand out or seem most interesting to you? Why are they
significant?
code, are missing or need more emphasis? Why?
4. Step 4. An interview will be conducted with a local planner
in your city to further explore both project review and your
earlier work on planning and zoning. Reread the plan, zoning
code and project review process, so you can talk about them
easily.
Include at least the following questions in your interview. We’ll
discuss further in class.
1. are any aspects of the comp plan, zoning code and project
review process have been discussed as needing updates,
modification? Why?
2. What are the time requirements for compliance with different
stages of project review? Are these realistic?
3. What aspects of project review receive the most complaints?
Are there aspects that you think could be simplified?
4. Step 5. Add two more pages to your paper describing the
results of the interview.
Deliverables: (1) 4-page paper including additional checklist
comparison describing regulations, policies and development
patterns for case study community, specifically referencing
documents in the folder. (2) Folder with all relevant documents
uploaded to google drive to made available to class. (3) possible
3-minute presentation to class with power points.
Due: Nov 10, midnight, submittal on canvas.
City of Boulder: Project Review
This is a synopsis of administrative review and land use review
for the City of Boulder. Admin review and land use review are

described below, in the Background section below.
The Documents section below describes all the relevant
documents to be used in various section of both admin and land
use review. The Application itself is at the end of this document
below. In this assignment, please compare the project review
process in your case study city – information requirements,
number of steps, complexity - to what the city of Boulder
requires.
1. Background
Administrative Review
From the website. "Administrative Reviews check that proposed
projects are compliant with city development standards and
codes.
Administrative applications are due at 10 a.m. every other
Monday and are reviewed on two-week cycles. View the 2020
submittal calendar (Links to an external site.) .
Search for the "Administrative Review Application Form" in
the Applications & Forms Database (Links to an external
site.) to find a complete list of application requirements and
fees for the projects below."
· Accessory Dwelling Units or Detached Dwelling Units
· Parking Reductions
· Setback Variances
· Solar Exceptions
· Antenna for Wireless Telecommunications
· Change of Address or Street Name
· Cooperative Housing Unit
· Group Home Facility
· Landscape Standards Modification
· Minor Modification of an Approved Discretionary Review
Plan
· Non-Conforming Uses
· Public Utility Easement Vacation
· Residential and Congregate Care Facilities

· Small Restaurants and Taverns
· Temporary Outdoor Entertainment
· Site Access
· Small Recycling Collection Facilities
· Substitution of Restaurant Use
· Solar Access Permit
· Zoning Verification
Land Use Review
From the web site. "All proposed changes to land uses,
including annexation, site review, use review and preliminary
plat, require Land Use Review. If you are planning to apply for
a building permit and your project falls into one of the project
categories listed below, you will need to go through the land
use review process before applying for a building permit.
Land Use Review applications are due at 10 a.m. on specified
Mondays. Check the 2020 submittal calendar (Links to an
external site.) for due dates. Land Use Review applications are
reviewed on three-week cycles.
Search for the "Land Use Review Application" in
· Annexation Information
· Annexation/Initial Zoning
· Annexation Feasibility Study
· Concept Plan Review and Comment
· Minor Subdivision
· Outside City Utility Permit
· Preliminary Plat for Subdivision
· Rezoning
· Site Review
· Use Review
· Extension of Development Approval - Planning Board (Section
9-2-12, BRC)
· Vacation Review of Street, Alley and Access Easements

2. Documents
· Accessory Dwelling Unit (ADU) (Links to an external
site.) Attachment to Admin. Review application, includes ADU
application requirements and review criteria
· Administrative Parking Deferral (Links to an external
site.) Attachment to Admin. Review application, includes
parking deferral requirements and review criteria
· Administrative Parking Reduction (Links to an external
parking reduction requirements and review criteria
· Administrative Review Application Form (Links to an external
site.) Main application used to apply for all Administrative
Review requests. Attachments to this form are available for
each administrative review type.
· Administrative Setback Variance (Links to an external
setback variance criteria
· Administrative Solar Access Exception (Links to an external
site.) Attachment to Admin. Review application, includes solar
access exception criteria
· Christmas Tree Vendor Information/Sign Regulation
Sheet (Links to an external site.) Attachment to Admin. Review
application, includes site standards and sign regulations for
Christmas tree sales lots
· Community Garden Form (Links to an external site.) For land
or rooftops gardened by a group of people that do not reside on
the property

· Cooperative Housing Units (Links to an external
definition of co-op housing and review criteria
· Day Care Center (Links to an external site.) Attachment to
Admin. Review application, includes definition of day care
center and review criteria
· Detached Dwelling Units with Two Kitchens (Links to an
external site.) Attachment to Admin. Review application,
includes review criteria for detached dwelling units with two
kitchens
· Gasoline Service Station (Links to an external
definition of gasoline service station and review criteria
· Group Home Facility (Links to an external site.) Attachment
to Admin. Review application, includes definition of group
home facility and review criteria
· Minor Modification (Links to an external site.) Attachment to
Admin. Review application, includes review criteria for minor
modifications to previously approved discretionary reviews
(e.g. PUD's, Site Reviews)
· Religious Assembly (Links to an external site.) Attachment to
Admin. Review application, includes definition of religious
assembly and review criteria
· Residential Care Facility (Links to an external
definition of residential care facility and review criteria
· Section 9-3.2-14, "Two Detached Dwellings on a Single

Lot" (Links to an external site.) Attachment to Admin. Review
application, includes review criteria for two detached dwellings
on a single lot
· Temporary Sales or Outdoor Entertainment (Links to an
external site.) Administrative Review application for events
with temporary sales or outdoor entertainment
· Vacation Review of Utility and Drainage Easements (Links to
an external site.) Attachment to Admin. Review application,
includes review criteria for vacating utility and drainage
easements
· Wireless Telecommunications Facilities (Links to an external
definition of wireless facility and review criteria
· Zoning Confirmation for BLA (Links to an external
confirmation form and checklist for Beverage Licensing
Authority
Land Use Review
· Annexation / Initial Zoning (Links to an external
site.) Attachment to Land Use Review Application. Includes
information about the annexing property to the City of Boulder
and designating initial zoning.
· Annexation Feasibility Study (Links to an external
site.) Attachment to Land Use Review Application
· Changes to BVCP Land Use Designations (Links to an
external site.) Attachment to Land Use Review Application
· Concept Plan Review & Comment (Links to an external
description of concept plan review and comment (conceptual

plan review prior to site review) and review criteria.
· Form-Based Code (FBC) Review (Links to an external
· Inclusionary Housing Compliance (Links to an external
site.) Preliminary determination of Inclusionary Housing
Compliance
· Inclusionary Housing Summary (Links to an external
site.) Requirements for meeting the Inclusionary Housing
Ordinance
· Land Use Review Application Form (Links to an external
site.) Main application form used to apply for all land use
review requests. Includes list of application requirements and
fee schedule. Attachments to this form are available for each
development review type.
· Land Use Review Project Fact Sheet (Links to an external
site.) Basic project facts for Land Use Review
· LUR Process Summary (Links to an external site.) Land Use
Review and Technical Document Submittal Summary
· Minor Subdivision (Links to an external site.) Attachment to
Land Use Review Application
· Outside City Utility (Links to an external site.) Attachment to
Land Use Review Application. Includes review criteria for city
utility service to out-of-city properties.
· Persons of Interest Form (Links to an external site.) Persons of
Interest Form
· Pre-Application Meetings (Links to an external site.) Brochure

describing the first optional step of the development review
process, pre-application meetings. Includes meeting schedule
form.
· Preliminary Plat for Subdivision (Links to an external
review criteria and application requirements for prel iminary plat
for subdividing lots.
· Rezoning Review (Links to an external site.) Attachment to
Land Use Review Application. Includes review criteria for
rezoning from one zoning designation to another.
· Site Review (Links to an external site.) Attachment to Land
Use Review Application. Includes review criteria and
application requirements for site review, generally for
requesting city code variations.
· Use Review (Links to an external site.) Attachment to Land
Use Review Application. Includes review criteria to allow
specific use review uses in certain zoning districts.
· Vacation Review (Links to an external site.) Attachment to
Land Use Review Application. Includes requirements for
vacating easements or city owned Right-of-Way.
· Vested Rights Option Form & Waiver (Links to an external
site.) Include this form (if required) with your Land Use Review
application materials. See specific variance criteria and
informational attachment for the type of review for which you
are applying.
3. Application
https://www-
static.bouldercolorado.gov/docs/PDS/forms/100_adr_application
.pdf

See discussions, stats, and author profiles for this publication
at: https://www.researchgate.net/publication/331522524
Degradation of diesel oil by soil bacteria in shake fl ask system
using food
waste amendments
Article in Malaysian Applied Biology · March 2019
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Deciphering the Proteolytic Activity of Trypsin-like Serine
Proteases and the Expression Levels of Serine Protease
Inhibitors Genes in Regulating Coagulation of
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Malaysia. View project

Ze As
2 PUBLICATIONS 0 CITATIONS
SEE PROFILE
Noraznawati Ismail
Universiti Malaysia Terengganu
SEE PROFILE
Fazilah Ariffin
Universiti Malaysia Terengganu
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* To whom correspondence should be addressed.
Malays. Appl. Biol. (2019) 48(1): 35–41
DEGRADATION OF DIESEL OIL BY SOIL BACTERIA IN
SHAKE
FLASK SYSTEM USING FOOD WASTE AMENDMENTS
GAN SIK ZE1, NORAZNAWATI ISMAIL2 and FAZILAH
ARIFFIN1*
1School of Fundamental Science, Universiti Malaysia
Terengganu,
21030 Kuala Nerus, Terengganu, Malaysia
2Institute Marine Biotechnology, Universiti Malaysia
Terengganu,
21030 Kuala Nerus, Terengganu, Malaysia
*E-mail: [email protected]

Accepted 4 January 2019, Published online 20 March 2019
ABSTRACT
Diesel oil contains compounds that can cause harm to humans
and the environment. Hence, biodegradation method is an
alternative way to reduce the pollution caused by diesel oil. The
aim of this study is to determine the diesel oil degradation by
soil bacteria amended with food wastes in the flasks system. It
also aims to determine the food wastes such as sugarcane
bagasse and fishbone to enhance the biodegradation of diesel
oil. The degradation analysis was performed in an enrichment
culture flask containing soil, diesel oil with the addition of food
waste. The degradation analysis was carried out for 42 days
at 30°C at 150 rpm. The bacteria was isolated and identified
based on colony morphology and biochemical tests. Five
potential
diesel oil-degrading bacteria were preliminary identified as
Pseudomonas fluorescens, Pseudomonas aeruginosa, Klebsiella
species, Shewanella putrefaciens and Bacillus cereus. Diesel oil
degradation compound was analyzed using Gas
Chromatography - Mass Spectrometry. Four compounds namely
styrene, ethanol, 2-butoxy, benzene, 1-ethyl-2, 3-dimethyl
and benzene 1-ethyl-2, 3-dimethyl showed degradation by
bacteria amended with food wastes. The results of this study
demonstrate the potential use of food wastes such as sugarcane
bagasse and fish bone as substrates for enhancing the
remediation of hydrocarbon contaminated soil.
Key words: Degradation, diesel oil, soil bacteria, shake flask
INTRODUCTION
Polycyclic Aromatic Hydrocarbons (PAH) pollution
is one of the most serious problems affecting the
world nowadays. The widespread of the PAHs in

the environment may cause negative effect on
human health such as hamper body defense
system, stimulate cancer growth and can cause
death (Mumtaz & George, 1996). Analysis done
by Agency for Toxic Substances and Disease
Registry shows that 17 groups of PAHs have bad
consequences on human health (Buha, 2011).
Therefore, to reduce the amount of PAHs in the
environment, a bioremediation method was
developed. Bioremediation is considered as
promising environmental friendly treatment to
remediate hydrocarbon compounds. It has the
capability to reduce the toxic substance to non-toxic
substance as the end product. Besides that, the
technique is safe and give the complete
mineralization of pollutant without affecting the
natural ecosystem.
PAHs pollution in the environment can be
reduced using PAHs-degrading bacteria. However,
bioremediation has some drawbacks such as the low
abundance, diversity and activity of indigenous
hydrocarbon degrading bacteria and also their slow
growth rates. Therefore, food waste composting was
developed as a cost-effective and highly efficient
tool in biodegradation of hydrocarbon pollution site.
This is one of the methods by adding valuable
nutrient contain in the food waste such as nitrogen,
phosphorus, and carbon to their environment and to
enhance the rate of PAHs degradation. Besides
increasing PAH decomposition rate, the food waste
accumulation in the environment can be reduced.
The objective of this study is to determine the diesel
oil degradation by soil bacteria with amended with
food wastes in the flasks system. It is also to

determine the potential food wastes such as
36 DEGRADATION OF DIESEL OIL BY SOIL BACTERIA
sugarcane bagasse and fishbone for enhanced
biodegrading of diesel oil.
MATERIALS AND METHODS
Sample Collection
The soil sample was collected from diesel oil
contaminated soil at motorcycle service in Kuala
Terengganu, Malaysia. The soil sample was
collected from 10 cm subsurface of the soil using a
sterile spatula. The temperature and pH of the soil
was measured. Diesel oil was collected at a petrol
station area in Kuala Terengganu. Food wastes such
as sugarcane bagasse and fishbone were collected
from canteens at Universiti Malaysia Terengganu.
The food wastes were dried at 90°C and blended to
make a powder.
Enrichment of diesel oil-degrading bacteria
The enrichment culture (EC) composed of
22.5 g of soil sample and 15 ml of diesel oil. Then,
flasks C and C1 were filled with 112.5 ml of mineral
salt medium (MSM) whereas the other flasks were
filled with 105 ml of MSM. MSM consisted of
the following composition: 0.2 gL-1 of MgSO, 0.02
gL-1 of CaCl, 1.0 gL-1 of KHPO, 1.0 gL-1 of KHPO,
1.0 gL-1 of NHNO and 0.05 gL-1 FeCl. The flasks
were then left undisturbed for 2 days. After 2 days

of incubation, 7.5 gL-1 of sugarcane bagasse powder
was added into flasks A and A1 while the same
amount of fishbone powder was added into flasks B
and B1. Flasks C and C1 served as control. The
enrichment culture was shaken at 150 rpm (Abioye
et al., 2010).
Diesel oil extraction from medium
The diesel oil was extracted at every 7 days
intervals for 42 days. The enrichment culture (EC)
was suspended with n-hexane (1:1 ratio). The
suspension was homogenized for 1 hour and then
centrifuged at 150 rpm for 5 minutes. The upper
layer of supernatant obtained was resuspended with
1:1 ratio of supernatant to n-hexane. The funnel was
shaken gently by inverting. The upper phase was
collected and evaporated using a rotary evaporator.
The diesel oil extracted was stored at 4°C for further
analysis.
Enumeration of diesel oil-degrading bacteria
EC was collected every 7 days for the
enumeration of diesel oil- degrading bacteria. The
EC was serially diluted and 0.1 ml of diluted sample
was transferred to nutrient agar (NA) plates using the
spread plate method. The CFU/ml of sample was
calculated using the formula below:
number of colonies
Total number of bacteria =
(dilution factor) x (actual volume plated)
Screening for diesel oil biodegradation

The extracted diesel oil was diluted to 1 ppm.
The diluted samples were filtered using 0.45 µm
PTFE membrane filter before injecting into the GC-
MS (QP2010 ultra). The analysis conditions for the
GC-MS are shown in Table 1.
The degradation percentage of the diesel oil
compound was calculated using the modified
formula (Deng et al., 2014):
Table 1. Analysis condition of GC-MS
GC-MS GCMS-QP2010 ultra
[GC]
Column Rtx–5MS 0.25 x 30 m df=0.25 µm
Inlet mode Splitless
Vaporizing chamber temperature
Column oven temperature 50°C (1 min) 300°C (5 min)
Carrier gas Helium
Control mode Constant linear velocity 35.5 cm/sec
High-pressure injection 150kPa (1.00min)
Purge flow rate 3.0 mL/min
Injection rate 1 µl
[MS]
Interface temperature 300°C
Ion source temperature 200°C
Data sampling time 1.00 min
Measurement mode Scan
Mass range m/z 50-600
Event time 0.05 sec

DEGRADATION OF DIESEL OIL BY SOIL BACTERIA 37
W1 – W5
DR = x 100%
W1
Where:
DR = Diesel oil degradation rate
W1 = Diesel-oil Concentration in Week 1
W5 = Diesel-oil Concentration in Week 5
Identification of diesel oil-degrading bacteria
The diluted samples were transferred to oil agar
(OA) and incubated for 24 hours at 30°C. The
bacterial colonies with the clear clearing zones on
OA were isolated and sub cultured on nutrient agar.
Further sub-culturing of bacteria was conducted until
pure isolates was were obtained. The isolated
bacteria were identified using Gram staining
technique and the cell morphology of bacterium was
recorded. The bacterial species was further
identified using BBL Crystal Identification Kit
(Fisher Scientific, USA) as per the manufacturer’s
instructions. A single pure bacteria colony was
suspended in an inoculum fluid tube and then filled
into the target area of the base. The inoculum was
rolled gently along the tracks and the panel, closed
with a lid and incubated at 37°C. The isolated
bacterium was identified using BBL Crystal MIND.
RESULTS
Effect of the addition of food waste into the flask

system on the growth of bacteria
The bacterial growth curves for all the flasks
show the same pattern, which steadily increase and
then gradually decrease, after reaching their
respective peaks (Figure 1). The flask containing
sugarcane bagasse (SB) show the highest growth
rate at week 3 compared to the flask system
containing fishbone (FB). All the flask systems show
approximately the same viable cell count at week 1
and week 2. The bacterial count of the flask system
containing SB slightly increases from 2.89 x 109
CFU/ml at week 1 to 1.58 x 1011 CFU/ml at week 3
and the bacterial count of the flask system without
any food waste increases from 2.70 x 109 CFU/ml at
week 1 to 4.95 x 1010 CFU/ml at week 3. On the
other hand, the bacterial count of the flask system
containing fishbone slightly increases from 2.81 x
109 CFU/ml at week 1 to 9.55 x 1011 CFU/ml at week
3 (Figure 1).
Effect of addition of food waste on pH value in
flasks system
The pH of the shake flask containing fish bone
and sugarcane bagasse were dropped at week 5 to
pH 5.75 and 5.96 respectively, whereas pH for
control did not show significant changes during the
incubation period (Figure 2).
Identification of diesel oil- degrading bacteria
Physiological characteristics in Table 2 shows
that there were four Gram-negative bacteria and one

Gram-positive bacterium that had been identified.
Isolate 2 and Isolate 3 showed negative result for
oxidase test while all isolates showed negative result
for indole test. All of the isolated bacteria are white
in colour with a circular form. These isolated
bacteria had an entire margin and bacillus shape.
The elevation of Isolate 2, Isolate 3, Isolate 4, and
Isolate 5 are raised, whereas Isolate 1 is convex.
Fig. 1. Bacteria count (CFU/ml) in the flask system containing
sugarcane bagasse, fishbone and control
after 5 weeks of incubation.
Fig. 2. pH value in the flask system containing sugarcane
bagasse, fishbone and control after 5 weeks of incubation.
Table 2. Colony morphology and biochemical activities of
isolated diesel oil -degrading bacteria
Characteristics
Gram- Oxidase Indole Colony Morphology
stain test test Color Form Margin Elevation Shape
Isolate 1 – + – White Circular Entire Convex Bacillus
Isolate 2 – – – White Circular Entire Raised Bacillus
Isolate 3 – – – White Circular Entire Raised Bacillus
Isolate 4 – + – White Circular Entire Raised Bacillus
Isolate 5 + ND ND White Circular Entire Raised Bacillus
– = negative result; + = positive result; ND – Not Detected.

Table 3. Diesel oil -degrading bacteria preliminary identified
by BBL Crystal Identification Kit
Isolated bacteria Bacteria Identification
1 Pseudomonas fluorescens
2 Klebsiella sp.
3 Shewanella putrefaciens
4 Pseudomonas aeruginosa
5 Bacillus cereus
Table 3 shows the isolated bacteria identified
using the BBL Crystal Identification Kit. It shows
that, five potential diesel oil-degrading bacteria
known as Pseudomonas fluorescens, Klebsiella
species, Shewanella putrefaciens, Pseudomonas
aeruginosa, and Bacillus cereus has been pre-
liminarily identified.
Effect of the addition of food waste on the
degradation rate of the diesel oil compounds
In this study, it was demonstrated that the flask
containing sugarcane bagasse shows the highest
degradation rate for compounds styrene and ethanol,
2-butoxy but has the lowest degradation rate for
benzene, 1-ethyl-2, 3-dimethyl. However, the flask
containing fishbone shows the highest degradation
rate for compound benzene, 1-ethyl-2, 3-dimethyl
and the lowest degradation rate for compound
styrene.
DISCUSSION
According to Figure 1, the counts of diesel oil-
degrading bacteria shows a normal bacterial growth

curve, which consists of four phases. It begins with
the lag phase, followed by the log phase, stationary
phase and it ends with a decline phase. During week
1 and week 2 of incubation, the bacteria are trying
to adapt to the new environment. Then, the bacterial
growth was increased exponentially in week 3 as it
had been adapted to the environment. At this stage,
bacterial cells are most metabolically active and are
able to utilize the diesel oil in the medium. However,
the bacterial growth was decreased at week 4 and
week 5 due to the limited nutrient supply, which is
a decrement in diesel oil concentration and
accumulation of toxic wastes produced by the
bacteria in the medium. Unlike the bacteria in the
flask system containing sugarcane bagasse and
fishbone, bacteria in the flask system without food
waste grew slower because there was no additional
nutrient compared to flask system containing food
waste. This finding was supported by Agarry (2018),
who stated that amendments of organic and
inorganic nutrients might enhance microbial
growth, which can promote the efficiency of
hydrocarbon degradation.
The amendments of food waste in the
enrichment medium promoted the growth of the
bacteria population. This is because the sugarcane
bagasse and fishbone provide valuable nutrients for
the bacteria to grow (Ibrahim et al., 2016). In terms
of nutrient composition, sugarcane bagasse is made
up of 40% cellulose, 35% hemicellulose and 15%

lignin, which contain a carbon source for bacteria
to get energy (Alves Da Costa et al., 2015).
Fishbone also has a high content of natural calcium
and phosphorus (Malde et al., 2010). Therefore,
with the amendments of the food waste in the
enrichment medium, it may enhance the biological
activity of bacteria, which is a necessary nutrient
for biodegradation activities by the bacteria.
Normally, the efficiency for bacteria in undergoing
biodegradation is affected by the conditions that are
favorable for the bacteria. In this study, organic
matter content from food waste supplies enough
nutrients for the bacteria to grow. Therefore, it may
contribute to the efficiency of diesel oil degradation
as well as some minerals needed for enhancing the
biodegradation activities (Das & Chandran, 2011).
Based on Figure 2, the graph shows a slight
decrease in pH after week 3 because the bacteria had
reached their decline phase. This stage is attained
due to the depletion of nutrients and accumulation
of metabolic waste products by the bacteria in the
media. This condition facilitates the bacteria to
move on to the death phase. Besides, bacteria will
utilize the compounds in the diesel fuel as a carbon
source for growth. Thus, it will produce metabolite
byproducts after utilizing the compound. This
metabolite byproduct, metabolic wastes, dead
bacteria and toxic materials will cause the
environment to be acidic.
In this study, the isolated diesel oil -degrading
bacteria were preliminary identified are Pseudo-
monas fluorescens, Klebsiella species, Shewanella
putrefaciens, Pseudomonas aeruginosa and

Bacillus cereus. These identified bacteria have
utilized the compounds in diesel oil as sole source
of carbon and energy source for growth. Potential
oil degraders such as Pseudomonas fluorescens
have shown the properties of high biodegradability
of an aliphatic hydrocarbon and low as well as slow
biodegradation of aromatic hydrocarbon (Sepic
et al., 1997). Apart from that, Pseudomonas
aeruginosa is well-known for its degradation ability.
Various studies showed that P. aeruginosa is able
to degrade crude oil and oil sludge approximately
by 70% (Zhang et al., 2005; Das & Chandran, 2011;
Al-Wasify & Hamed, 2014). Klebsiella species
shows ability to degrade toluene, xylene,
naphthalene, and nonane (Rodrigues et al., 2009).
It is believed that Klebsiella species isolated in this
study also have the capability to degrade diesel oil.
However, Shewanella putrefaciens have been
reported as crude oil degrader in coastal seawater and
sea ice (Das & Das, 2015). Bacillus cereus is a Gram-
positive bacterium that is widely distributed
environmentally. Bacillus species have been
reported to be capable of utilizing phenanthrene,
anthracene, and pyrene as carbon sources (Dou et
al., 2010; Fazilah et al., 2016; Rao et al., 2015).
The percentage of diesel oil biodegradation in
the three flask systems is shown in Figure 3. The
degradation rate of styrene, ethanol-2-butoxy, and
benzene, 1-ethyl-2, 3-dimethyl for flask system
without food wastes is 5%, 10.7% and 9.5%
respectively. For the flask system containing
sugarcane bagasse, the degradation rate of styrene,
ethanol-2-butoxy, and benzene, 1-ethyl-2, 3-
dimethyl is 12.8%, 18.4%, and 0.1% respectively.
While, for the degradation rates of flask system

containing fishbone for styrene, ethanol-2-butoxy,
and benzene, 1-ethyl-2, 3-dimethyl is 4.2%, 14.1%,
and 13.9% respectively. The flask system
containing the food waste has increased the
degradation rate of styrene, ethanol-2-butoxy, and
benzene, 1-ethyl-2, 3-dimethyl. However, not all the
compounds show the same variations. For styrene,
the flask system containing sugarcane bagasse
enhances the degradation of styrene by 8.6%, but
the flask system containing the fishbone has
decreased the degradation rate of styrene by 0.8%.
This is due to the concentration of nutrients such
as calcium and phosphorus provided by the fishbone
is too high for Pseudomonas fluorescens, which
eventually inhibits the degradation of the styrene
(Beltrametti et al., 1997; Das & Chandran, 2011;
Lang et al., 2016). The degradation rate of ethanol,
2-butoxy for the flask system containing food waste
is higher than the flask system without the food
waste. This shows that sugarcane bagasse and
fishbone have enhanced the degradation rate of
Fig. 3. Degradation percentage (%) of the compounds in diesel
oil (styrene, ethanol,
2-butoxy, and benzene, 1-ethyl-2, 3-dimethyl-) in the flask
containing sugarcane bagasse
and fishbone and flask without food waste (control).
ethanol, 2-butoxy by 7.7% and 3.4% respectively.
Then, the degradation rate of benzene, 1-ethyl-2, 3-
dimethyl in the flask system containing sugarcane
bagasse is inhibited. However, the flask system

containing fishbone shows enhancement of 4.4%
for the degradation rate of benzene, 1-ethyl-2, 3-
dimethyl. This is because the sugarcane bagasse
powder is easily absorbed by the bacteria
responsible for degrading benzene, 1-ethyl-2, 3-
dimethyl. Hence, it becomes the competitor for the
bacteria to utilize it as a carbon source for growth.
CONCLUSION
The results of this study shows the addition of food
waste to the flask system enhanced the growth rate
of bacteria and increased the degradation rate of
the compounds in diesel oil. Four compounds such
as styrene, ethanol, 2-butoxy, benzene, 1-ethyl-2,
3-dimethyl and benzene 1-ethyl-2, 3-dimethyl
showed degradation by bacteria with amended with
food wastes. Five potential diesel oil-degrading
bacteria have been identified as Pseudomonas
fluorescens, Pseudomonas aeruginosa, Shewanella
putrefacians, Klebsiella species and Bacillus cereus.
These preliminary results highlighted the potential
application of sugarcane bagasse and fish bone
for enhancing the biodegradation of diesel oil
contaminated soil.
ACKNOWLEDGMENTS
The authors would like to thank Universiti Malaysia
Terengganu (UMT) for supporting this research
through Tabung Penyelidik Muda (TPM) Grant.
REFERENCES
Abioye, P.O., Abdul Aziz, A. & Agamuthu, P. 2010.
Enhanced biodegradation of used engine oil in

soil amended with organic wastes. Water, Air,
and Soil Pollution, 209(1-4): 173-179.
Agarry, S.E. (2018). Evaluation of the effects of
inorganic and organic fertilizers and activated
carbon on bioremediation of soil contaminated
with weathered crude oil. Journal of Applied
Sciences and Environmental Management,
22(4): 587-595.
Al-Wasify, R.S. & Hamed, S.R. 2014. Bacterial
biodegradation of crude oil using local isolates.
International Journal of Bacteriology, 2014:
1-8.
Alves Da Costa, D., Lucas De Souza, C., De Oliveira,
E., Saliba, S., Da, J. & Carneiro, C. 2015. By-
products of sugarcane industry in ruminant
nutrition. International Journal of Agronomy
and Agricultural Research, 3: 1-9.
Beltrametti, F., Marconi, A.M., Bestetti, G.,
Colombo, C., Galli, E., Ruzzi, M. & Zennaro, E.
1997. Sequencing and functional analysis of
styrene catabolism genes from Pseudomonas
fluorescens ST. Applied and Environmental
Microbiology, 63(6): 2232-9.
Buha, A. 2011. Polycyclic aromatic hydrocarbon.
Retrieved October 27, 2016 from www.toxi
p e d i a . o r g / d i s p l a y / t o x i p e d i a / P o l y c y c l i c
Aromatic Hydrocarbons

Das, N. & Chandran, P. 2011. Microbial degradation
of petroleum hydrocarbon contamina nts: an
overview. Biotechnology Research Inter-
national. 941810.
Das, N. & Das, D. 2015. Strategies for remediation
of polycyclic aromatic hydrocarbons from
contaminated soil-an overview. Journal of
Critical Reviews, 2(1): 20-25.
Deng, M.C., Li, J., Liang, F.R., Yi, M., Xu, X.M.,
Yuan, J.P. & Wang, J.H. 2014. Isolation and
characterization of a novel hydrocarbon-
degrading bacterium Achromobacter sp. HZ01
from the crude oil-contaminated seawater at the
Daya Bay, Southern China. Marine Pollution
Bulletin, 83(1): 79-86.
Dou, J., Ding, A., Liu, X., Du, Y., Deng, D. & Wang,
J. 2010. Anaerobic benzene biodegradation by
a pure bacterial culture of Bacillus cereus under
nitrate reducing conditions. Journal of Environ-
mental Sciences (China), 22(5): 709-15.
Fazilah, A., Darah, I. & Noraznawati, I. 2016.
Bioremediation of phenanthrene by mono-
cultures and mixed culture bacteria isolated
from contaminated soil. International Journal
of Biological, Biomolecular, Agricultural, Food
and Biotechnological Engineering, 10(9): 495-
498.
Ibrahim, M.H., Quaik, S. & Ismail, S.A. 2016.
Introduction to Organic Wastes and Its
Management. In Prospects of Organic Waste

Management and the Significance of Earth-
worms Springer, Cham, 1-21.
Lang, S., Tarayre, C., Destain, J., Delvigne, F.,
Ongena, M. & Thonart, P. 2016. The effect of
nutrients on the degradation of hydrocarbons
in mangrove ecosystems by microorganisms.
International Journal of Environmental
Research, 10(4): 583-592.
Malde, M.K., Graff, I.E., Siljander-Rasi, H.,
Venalainen, E., JulShamn, K., Pedersen, J.I. &
Valaja, J. 2010. Fish bones – a highly available
calcium source for growing pigs. Journal of
Animal Physiology and Animal Nutrition, 94(5):
66-76.
Mumtaz, M.M. & George, J.D. 1996. Toxicological
profile for polycyclic aromatic hydrocarbon.
U.S. Department of Health and Human Services,
1-487.
Rao, M.P., Rajithasri, A.B., Sagar, K. & Satyaprasad,
K. 2015. Isolation of polycyclic aromatic
hydrocarbons degrading PGPR from a oil
products contaminated site in Hyderabad,
Telangana State, India. International Journal
of Pharma and Biosciences, 6(4): 305-317.
Rodrigues, D.F., Sakata, S.K., Comasseto, J.V.,
Bicego, M.C. & Pellizari, V.H. 2009. Diversity
of hydrocarbondegrading Klebsiella strains
isolated from hydrocarbon contaminated
estuaries. Journal of Applied Microbiology,
106(4): 1304-1314.

Šepic
�
, E., Bricelj, M. & Leskovšek, H. 1997.
Biodegradation studies of polyaromatic
hydrocarbons in aqueous media. Journal of
Applied Microbiology, 83(5): 561-568.
Zhang, G., Wu, Y., Qian, X. & Meng, Q. 2005.
Biodegradation of crude oil by Pseudomonas
aeruginosa in the presence of rhamnolipids.
Journal of Zhejiang University Science B, 6(8):
725-730.
�
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with power points.

with power points.
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City Case Studies: Project Review and Conclusion to Case
Studies
In this project students will synthesize and conclude your
discussion of regulation and policy for your case study city. The
foundation for this work is collection of all key documents used
your city in local project review. Students are expected to
collect and assess all key documents used in project review.
This assignment is designed to help us explore key issues of
complexity, cost and conflict of regulatory compliance for
participants including both developers and local governments.
The class will use the city of Boulder as a template and baseline
including the following five steps.
1. Step 1. Compilation of key documents in a google document,
to be created by you, and with the url included in the beginning
of the assignment.
2. Step 2. Evaluation against the benchmark of the city of
Boulder, available at the bottom of this assignment sheet.
3. Step 3. Prepare a two-page paper describing and synthesizing
the work you’ve done on your own and in class about the city
plan, zoning code and project review process.
Include at least the following.

code, are emphasized in your city? Based on your knowledge of
your city, why do you think these aspects are emphasized?
code, stand out or seem most interesting to you? Why are they
significant?
code, are missing or need more emphasis? Why?
4. Step 4. An interview will be conducted with a local planner
in your city to further explore both project review and your
earlier work on planning and zoning. Reread the plan, zoning
code and project review process, so you can talk about them
easily.
Include at least the following questions in your interview. We’ll
discuss further in class.
1. are any aspects of the comp plan, zoning code and project
review process have been discussed as needing updates,
modification? Why?
2. What are the time requirements for compliance with different
stages of project review? Are these realistic?
3. What aspects of project review receive the most complaints?
Are there aspects that you think could be simplified?
4. Step 5. Add two more pages to your paper describing the
results of the interview.
Deliverables: (1) 4-page paper including additional checklist
comparison describing regulations, policies and development
patterns for case study community, specifically referencing
documents in the folder. (2) Folder with all relevant documents
uploaded to google drive to made available to class. (3) possible
3-minute presentation to class with power points.
Due: Nov 10, midnight, submittal on canvas.
City of Boulder: Project Review
This is a synopsis of administrative review and land use review
for the City of Boulder. Admin review and land use review are
described below, in the Background section below.

The Documents section below describes all the relevant
documents to be used in various section of both admin and land
use review. The Application itself is at the end of this document
below. In this assignment, please compare the project review
process in your case study city – information requirements,
number of steps, complexity - to what the city of Boulder
requires.
1. Background
From the website. "Administrative Reviews check that proposed
projects are compliant with city development standards and
codes.
Administrative applications are due at 10 a.m. every other
Monday and are reviewed on two-week cycles. View the 2020
submittal calendar (Links to an external site.) .
Search for the "Administrative Review Application Form" in
· Accessory Dwelling Units or Detached Dwelling Units
· Parking Reductions
· Setback Variances
· Solar Exceptions
· Antenna for Wireless Telecommunications
· Change of Address or Street Name
· Cooperative Housing Unit
· Group Home Facility
· Landscape Standards Modification
· Minor Modification of an Approved Discretionary Review
Plan
· Non-Conforming Uses
· Public Utility Easement Vacation
· Residential and Congregate Care Facilities
· Small Restaurants and Taverns

· Temporary Outdoor Entertainment
· Site Access
· Small Recycling Collection Facilities
· Substitution of Restaurant Use
· Solar Access Permit
· Zoning Verification
Land Use Review
From the web site. "All proposed changes to land uses,
including annexation, site review, use review and preliminary
plat, require Land Use Review. If you are planning to apply for
a building permit and your project falls into one of the project
categories listed below, you will need to go through the land
use review process before applying for a building permit.
Land Use Review applications are due at 10 a.m. on specified
Mondays. Check the 2020 submittal calendar (Links to an
external site.) for due dates. Land Use Review applications are
reviewed on three-week cycles.
Search for the "Land Use Review Application" in
· Annexation Information
· Annexation/Initial Zoning
· Annexation Feasibility Study
· Concept Plan Review and Comment
· Minor Subdivision
· Outside City Utility Permit
· Preliminary Plat for Subdivision
· Rezoning
· Site Review
· Use Review
· Extension of Development Approval - Planning Board (Section
9-2-12, BRC)
· Vacation Review of Street, Alley and Access Easements

· Cooperative Housing Units (Links to an external
definition of co-op housing and review criteria
· Day Care Center (Links to an external site.) Attachment to
Admin. Review application, includes definition of day care
center and review criteria
· Detached Dwelling Units with Two Kitchens (Links to an
external site.) Attachment to Admin. Review application,
includes review criteria for detached dwelling units with two
kitchens
· Gasoline Service Station (Links to an external
definition of gasoline service station and review criteria
· Group Home Facility (Links to an external site.) Attachment
to Admin. Review application, includes definition of group
home facility and review criteria
· Minor Modification (Links to an external site.) Attachment to
Admin. Review application, includes review criteria for minor
modifications to previously approved discretionary reviews
(e.g. PUD's, Site Reviews)
· Religious Assembly (Links to an external site.) Attachment to
Admin. Review application, includes definition of religious
assembly and review criteria
· Residential Care Facility (Links to an external
definition of residential care facility and review criteria
· Section 9-3.2-14, "Two Detached Dwellings on a Single
Lot" (Links to an external site.) Attachment to Admin. Review

application, includes review criteria for two detached dwellings
on a single lot
· Temporary Sales or Outdoor Entertainment (Links to an
external site.) Administrative Review application for events
with temporary sales or outdoor entertainment
· Vacation Review of Utility and Drainage Easements (Links to
an external site.) Attachment to Admin. Review application,
includes review criteria for vacating utility and drainage
easements
· Wireless Telecommunications Facilities (Links to an external
definition of wireless facility and review criteria
· Zoning Confirmation for BLA (Links to an external
confirmation form and checklist for Beverage Licensing
Authority
Land Use Review
· Annexation / Initial Zoning (Links to an external
information about the annexing property to the City of Boulder
and designating initial zoning.
· Annexation Feasibility Study (Links to an external
· Changes to BVCP Land Use Designations (Links to an
external site.) Attachment to Land Use Review Application
· Concept Plan Review & Comment (Links to an external
description of concept plan review and comment (conceptual
plan review prior to site review) and review criteria.

· Form-Based Code (FBC) Review (Links to an external
· Inclusionary Housing Compliance (Links to an external
site.) Preliminary determination of Inclusionary Housing
Compliance
· Inclusionary Housing Summary (Links to an external
site.) Requirements for meeting the Inclusionary Housing
Ordinance
· Land Use Review Application Form (Links to an external
site.) Main application form used to apply for all land use
review requests. Includes list of application requirements and
fee schedule. Attachments to this form are available for each
development review type.
· Land Use Review Project Fact Sheet (Links to an external
site.) Basic project facts for Land Use Review
· LUR Process Summary (Links to an external site.) Land Use
Review and Technical Document Submittal Summary
· Minor Subdivision (Links to an external site.) Attachment to
Land Use Review Application
· Outside City Utility (Links to an external site.) Attachment to
Land Use Review Application. Includes review criteria for city
utility service to out-of-city properties.
· Persons of Interest Form (Links to an external site.) Persons of
Interest Form
· Pre-Application Meetings (Links to an external site.) Brochure
describing the first optional step of the development review

process, pre-application meetings. Includes meeting schedule
form.
· Preliminary Plat for Subdivision (Links to an external
review criteria and application requirements for preliminary plat
for subdividing lots.
· Rezoning Review (Links to an external site.) Attachment to
Land Use Review Application. Includes review criteria for
rezoning from one zoning designation to another.
· Site Review (Links to an external site.) Attachment to Land
Use Review Application. Includes review criteria and
application requirements for site review, generally for
requesting city code variations.
· Use Review (Links to an external site.) Attachment to Land
Use Review Application. Includes review criteria to allow
specific use review uses in certain zoning districts.
· Vacation Review (Links to an external site.) Attachment to
Land Use Review Application. Includes requirements for
vacating easements or city owned Right-of-Way.
· Vested Rights Option Form & Waiver (Links to an external
site.) Include this form (if required) with your Land Use Review
application materials. See specific variance criteria and
informational attachment for the type of review for which you
are applying.
3. Application
https://www-
static.bouldercolorado.gov/docs/PDS/forms/100_adr_application
.pdf

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