CHLORDANE AND DIELDRIN CONTAMINATION AND BIODEGRADATION IN

ALA WAI CANAL SEDIMENT

A THESIS SUBMITTED TO THE GRADUATE DIVISION OF THE

UNIVERSITY OF HAWAIʻI AT MĀNOA IN PARTIAL FULFILLMENT

OF THE REQUIREMENTS FOR THE DEGREE OF

MASTER OF SCIENCE

IN

CIVIL ENGINEERING

MAY 2012

By

Cindy M.Y. Lee

Thesis Committee:

Tao Yan, Chairperson

Roger Babcock

Michelle Teng

i

ACKNOWLEDGEMENTS

This thesis would have not been possible without the encouragement, guidance, advice,

and assistance of several individuals. First, I would like to thank my advisor, Dr. Tao

Yan for his mentorship and involvement with this thesis. I would like to thank my

Graduate Committee members, Dr. Roger Babcock and Dr. Michelle Teng for their

valuable advice about my thesis. I would like to thank Mr. Joseph Lichwa and Ms.

Bunnie Yoneyama for their assistance with the laboratory equipment. I would like to

thank the current and previous members of Dr. Yan’s research group for their assistance

in the lab. Also, I would like to thank my supervisors and co-workers at Pearl Harbor

Naval Shipyard & IMF for their encouragement and advice. Finally, I would like to

thank my parents and brothers for their assistance and constant support.

ii

TABLE OF CONTENTS

Acknowledgements…………………………………………………………………… i

List of Tables…………………………………………………………………………. iv

List of Figures………………………………………………………………………… v

CHAPTER 1 INTRODUCTION……………………………………………………... 1

1.1 Chlordane and Dieldrin Contamination………………………………………. 1

1.2 Urbanization and Stream Modifications……………………………………… 2

1.3 Evaluating Potential Effects Associated with Locations Contaminated with

Chlordane and Dieldrin……………………………………………………….. 3

1.4 Treatment Efforts for Locations Contaminated with Chlordane and Dieldrin.. 4

1.5 Objectives……………………………………………………………………... 5

CHAPTER 2 HORIZONTAL AND VERTICAL DISTRIBUTION OF

CHLORDANE AND DIELDRIN IN ALA WAI CANAL SEDIMENT…………….. 6

2.1 Introduction…………………………………………………………………… 6

2.2 Methods……………………………………………………………………….. 8

2.2.1 Sample site and Collection……………………………………………… 8

2.2.2 Laboratory Analysis…………………………………………………….. 10

2.2.2.1 Sample Preparation……………………………………………... 10

2.2.2.2 Extraction……………………………………………………….. 11

2.2.2.3 Cleanup and Concentration……………………………………... 12

2.2.2.4 Calibration and Analysis………………………………………... 13

2.3 Results………………………………………………………………………… 16

2.3.1 Concentration Profiles of Sediment Cores……………………………… 16

2.3.2 Comparison of the Concentration of Surface Sediments to Aquatic Life

Protection Guidelines…………………………………………………… 18

2.4 Discussion…………………………………………………………………….. 20

CHAPTER 3 DETERMINE WHETHER OR NOT CHLORDANE CAN BE

BIODEGRADED IN ALA WAI CANAL SEDIMENT……………………………... 25

3.1.Introduction…………………………………………………………………… 25

3.2.Methods……………………………………………………………………….. 27

3.2.1 Sample Site and Collection……………………………………………... 27

3.2.2 Culture Preparation and Sampling……………………………………… 28

3.2.3 Extraction and Cleanup…………………………………………………. 29

3.2.4 Calibration and Analysis………………………………………………... 30

3.3 Results………………………………………………………………………... 33

3.4 Discussion…………………………………………………………………….. 34

iii

CHAPTER 4 CONCLUSIONS AND RECOMMENDATIONS…………………… 35

4.1 Conclusions…………………………………………………………………... 35

4.2 Recommendations……………………………………………………………. 36

Appendix A…………………………………………………………………………... 39

Appendix B…………………………………………………………………………... 43

Appendix C…………………………………………………………………………... 44

References……………………………………………………………………………. 67

iv

LIST OF TABLES

Table 1.1: Types of Stream Modifications…………………………………………... 3

Table 2.1: Summary of Analytical Methods………………………………………… 10

Table 3.1: Set Up of the Laboratory Microcosm Experiments………………………. 28

Table A.1: Dry Weights of Sediment Collected for Set A…………………………... 39

Table A.2: Dry Weights of Sediment Collected for Set B…………………………... 40

Table A.3: Concentration, % Moisture, and % Recovery of Sediment Collected for

Set A…………………………………………………………………………………. 41

Table A.4: Concentration, % Moisture, and % Recovery of Sediment Collected for

Set B…………………………………………………………………………………. 42

Table B.1: Years Structures Built in the Pālolo, Mānoa, McCully/Mōʻiliʻili, and

Makiki Areas………………………………………………………………………… 43

Table B.2: Number of Units in Structures in the Pālolo, Mānoa, McCully/Mōʻiliʻili,

and Makiki Areas…………………………………………………………………….. 43

Table C.1: Chlordane Concentration of Samples Taken from Microcosms…………. 44

Table C.2: pH of Samples Taken from Microcosms………………………………… 55

v

LIST OF FIGURES

Figure 2.1: Areas Surrounding the Ala Wai Canal…………………………………... 9

Figure 2.2: Sediment Core Collection Sites…………………………………………. 10

Figure 2.4: Site 1 Concentration Profile……………………………………………... 16

Figure 2.5: Site 2 Concentration Profile……………………………………………... 16

Figure 2.6: Site 3 Concentration Profile……………………………………………... 17

Figure 2.7: Site 4 Concentration Profile……………………………………………... 17

Figure 2.8: Site 5 Concentration Profile……………………………………………... 17

Figure 2.9: Chlordane Concentration at the Surface for Each Site and the Probable

Effect Level (PEL) of Chlordane…………………………………………………….. 19

Figure 2.10: Dieldrin Concentration at the Surface for Each Site and the Probable

Effect Level (PEL) of Dieldrin………………………………………………………. 19

Figure 3.1: Sediment Core Collection Site…………………………………………... 27

Figure 3.2: Decachlorobipheyl Calibration Curve…………………………………... 31

Figure 3.3: Chlordane Calibration Curve……………………………………………. 32

Figure 3.4: Chlordane Concentration of Samples Taken from Microcosms………… 33

Figure C.1: pH of Samples Taken from Microcosms………………………………... 46

Figure C.2: Chromatographs of Samples Taken on January 14, 2012………………. 47

Figure C.3: Chromatographs of Samples Taken on February 4, 2012………………. 52

Figure C.4: Chromatographs of Samples Taken on March 10, 2012………………... 57

Figure C.5: Chromatographs of Samples Taken on March 31, 2012………………... 62

1

CHAPTER 1 INTRODUCTION

1.1 Chlordane and Dieldrin Contamination

Chlordane and dieldrin are organochlorine pesticides, chlorine-containing

compounds, that were used worldwide for residential and agriculture applications to

control termites and other insects. Chlordane was produced in 1947 and was sold as an

insecticide for agricultural applications and termiticide for underground treatment around

the foundation of structures [30, 33]. Dieldrin was produced in 1948 and was also sold as

an insecticide and termiticide [27, 32].

Chlordane and dieldrin were banned in the 1970s for agricultural applications and

1980s for all other uses due to the harmful effects on aquatic life, wildlife, and humans

[4]. Chlordane and dieldrin are persistent, toxic, and can bioaccumulate in the food chain

[27, 30]. Even when chlordane and dieldrin are no longer used, bottom sediments and

marine biota display significant concentrations of organochlorine compounds since

organochlorine compounds take years to deplete from the environment [4].

Organochlorine compounds are commonly transported to water sources through soil

erosion and runoff [4]. Organochlorine compounds are hydrophobic, do not dissolve well

in water and bind strongly to sediment, making the compounds hard to deplete from the

environment [4].

Sediments are analyzed for organochlorine compounds since “sediment serves as

both a source and a removal mechanism for contaminants to and from the stream, and as

a means of contaminant transport downstream” [4]. Sediment also provides a habitat for

benthos [4]. Benthos ingest the organochlorine compounds in bottom sediments and fish

and birds then eat the benthos, which leads to the accumulation of organochlorine

2

compounds in the fatty tissues of species in the food chain [4]. Also, marine biota are

analyzed to determine the presence of organochlorine compounds and contamination

levels since the accumulation of organochlorine compounds are harmful to aquatic life

and to those who consume them [4]. Bottom sediments and marine biota contain higher

concentrations of organochlorine compounds than the surrounding water since

organochlorine compounds are hydrophobic and bioaccumulative, which increases the

chances of detecting organochlorine compounds when analyzing sediment and tissue [4].

1.2 Urbanization and Stream Modifications

As the population increases more land is used to make room for residential,

industrial, commercial, and recreation areas. Also, land is needed to built roads,

freeways, and highways for these areas. In order to have more land for these

development projects, streams are modified for these projects. Stream modifications

include lined channels, blocked or filled in channels, elevated culverts, extended culverts,

and revetments [18, 38]. Also, vegetation near the stream maybe removed for these

stream modifications. Clearing of land for construction near streams or removal of

streamside vegetation can promote soil erosion and increase runoff to streams, which

contributes to chlordane and dieldrin contamination.

3

Table 1.1: Types of Stream Modifications [18, 38]

Type of Stream Modification Description

Lined Channel Artificial channel where the natural banks and stream

bed have been replaced, usually with concrete.

Blocked or Filled In Channel Part of the original channel is blocked.

Elevated Culvert Relatively short (usually less than 60 meters) conduit

structures that are usually found under highways. An

artificial waterfall is created with these structures

since they are built above the water level

immediately downstream.

Extended Culvert Similar to an elevated culvert but longer and usually

found in residential areas.

Revetment Either one or both sides of the banks are reinforced

but the channel isn’t reinforced.

Vegetation Removed –

Channel Realigned

The stream is realigned and the vegetation on the

banks is removed.

1.3 Evaluating Potential Effects Associated with Locations Contaminated with

Chlordane and Dieldrin

Guidelines have been established for chlordane and dieldrin to determine what

concentration of chlordane and dieldrin will cause adverse biological effects [4]. The

Canadian Sediment Quality Guidelines (CSQG) for aquatic life is used to determine if

aquatic life expose to chlordane and dieldrin in sediment are affected by these

contaminants [4, 31]. The CSQG for aquatic life were established by conducting

sediment toxicity tests on various aquatic life such as mollusks, shrimp, and sea urchin

and oyster larvae to assess the biological significance of contamination levels found in

sediments and comparing the tests to contamination levels found in sediments and data

pertaining to the characteristics of the sediments [31]. The probable effect level (PEL) is

the “level above which adverse effects are expected to occur frequently” meaning “more

than 50% adverse effects occurs” when aquatic life is exposed to sediment above this

4

level [31]. The PEL is 8.87 µg/kg dry weight for chlordane and 6.67 µg/kg dry weight

for dieldrin [4, 31].

1.4 Treatment Efforts for Locations Contaminated with Chlordane and Dieldrin

Because chlordane and dieldrin are still found in the environment and a potential

health hazard, studies have been done to find ways to deplete chlordane and dieldrin.

Chlordane and dieldrin undergo aerobic or anaerobic biodegradation in the water column

and in the sediment, respectively. Aerobic biodegradation involves the breakdown of

chlordane and dieldrin by microbes when oxygen is present [25], whereas anaerobic

biodegradation occurs when non-oxygen electron acceptors are utilized [28]. Chlordane

and dieldrin are hydrophobic and have low solubility levels, making it hard for microbes

to get to these in the sediment so that they can be degraded. Aerobic degradation has

been reported by Cuozzo et al. (2012) for chlordane and Matsumoto et al. (2009) for

dieldrin [8, 13]. Anaerobic degradation has been reported by Hirano et al. (2007) for

chlordane and Chiu et al. for dieldrin (2005) [7, 10]. From the studies reported, suitable

conditions are needed for degradation of chlordane and dieldrin to occur, which depends

on the contamination levels, temperature, and carbon and energy source [2, 7, 8, 10, 13,

14, 20]. Therefore, by determining the degradation microbes that are available in the

environment and discovering the right conditions and then optimizing the conditions for

the microbes will help the microbes to degrade chlordane and dieldrin.

5

1.5 Objectives

The objectives are (1) investigate the contamination levels of chlordane and

dieldrin both horizontally (i.e. at different locations of the Ala Wai Canal) and vertically

(i.e. at different depth in the sediment) in Ala Wai Canal sediment, and (2) determine

whether or not chlordane in Ala Wai Canal sediment can be biodegraded. Objective 1

was accomplished by collecting sediment core samples at various locations along the Ala

Wai Canal and the concentrations of chlordane and dieldrin were quantified for each

sampling location to formulate a concentration profile. Objective 2 was accomplished by

performing laboratory microcosm experiments with Ala Wai Canal sediment spiked with

chlordane.

6

CHAPTER 2 HORIZONTAL AND VERTICAL DISTRIBUTION OF

CHLORDANE AND DIELDRIN IN ALA WAI CANAL SEDIMENT

2.1 Introduction

The Ala Wai Canal was constructed in 1921-1928 by Walter F. Dillingham’s

Hawaiian Dredging Company to drain the “mosquito infested wetlands” and to “create

land suitable for development into commercial and residential real estate” [15].

Apartments and condominiums surround the Ala Wai Canal and the banks of the Ala Wai

Canal are landscaped with coconut palms and flowers. The Ala Wai area is used for

recreation with bike paths, canoes, kayaks, a golf course, and an illuminated sidewalk for

running and walking. However, there are signs posted near the Ala Wai Canal informing

people not to swim in the Ala Wai Canal and that the fish and shellfish in the Ala Wai

Canal are contaminated. Three streams, Pālolo, Mānoa, and Makiki, were altered to

make room for residential, commercial, and recreation areas and transports trash and

contaminates that drains into the Ala Wai Canal. Due to a low flow rate, the Ala Wai

Canal cannot naturally filter and clean the water so pesticides, toxic chemicals, heavy

metals, and trash accumulate in the Ala Wai Canal [17].

As part of the U.S. Geological Survey’s National Water-Quality Assessment

(NAWQA) program, stream bed sediment and fish samples were collected during the fall

of 1998 from six streams on Oahu to determine the occurrence of organochlorine

pesticides [4]. One of the sampling sites was Mānoa Stream. The chlordane

concentration in the sediment was 300 µg/kg dry weight [4]. The chlordane

concentration in the fish tissue was 950 µg/kg wet weight and 700 µg/kg wet weight [4].

The dieldrin concentration in the sediment was 150 µg/kg dry weight [4]. The dieldrin

7

concentration in the fish tissue was 850 µg/kg wet weight and 1250 µg/kg wet weight [4].

The concentration of chlordane and dieldrin in the sediment exceeded the Canadian

Sediment Quality Guidelines (CSQG) for the protection of aquatic life, 8.87 µg/kg for

chlordane and 6.67 µg/kg for dieldrin [4, 31]. The concentration of chlordane and

dieldrin in the fish tissue exceeded the New York State Department of Environmental

Conservation (NYSDEC) guidelines for the protection of fish eating birds and mammals,

500 µg/kg for chlordane and 120 µg/kg for dieldrin [4, 31]. Since the Mānoa Stream is

contaminated with chlordane and dieldrin, the Ala Wai Canal will also be contaminated

since the contents of the Mānoa Stream are transported to the Mānoa-Pālolo Drainage

Canal and then to the Ala Wai Canal.

There is a concern about chlordane and dieldrin because of the harmful effects to

humans, other species, and the environment. The fish and the shellfish in the Ala Wai

Canal are unsafe to eat and may have developed deformities due to living in a

contaminated environment and ingesting chlordane and dieldrin or eating benthos that

ingest chlordane and dieldrin in the sediment. When the Ala Wai Canal was dredged, a

Mantis shrimp (Odontodactylus Scyllarus) that was much larger than the normal size was

pulled from the sediment and weighted in at “1.35 pounds and 15 inches” [9].

Before the Ala Wai Canal was contaminated, people used to fish and catch

shellfish in the Ala Wai Canal. Today, most people know not to consume the fish and

shellfish in the Ala Wai Canal, but first-generation immigrants who may not speak or

speak little English, go to the Ala Wai Canal to catch crabs since they may not know the

hazards of consuming contaminated crabs from the Ala Wai Canal [9]. People continue

to use the canal for recreational purposes, in spite of the presence of contaminants.

8

Therefore, it is important the concentration levels of harmful contaminants, such as

chlordane and dieldrin, are accurately determined. This can be accomplished by

analyzing sediment since from sediment, the sources of contamination and where these

contaminants are being transported can be determined and trends can be discovered on

how these contaminants are being degraded naturally. With the up-to-date concentration

data, especially if the levels are high and exceeds the aquatic life guideline, sound

regulatory actions and warning mechanisms can be put in place to warn people about the

harmful health effects of consuming contaminated fish and shellfish and the hazardous

potentials associated with paddling in the Ala Wai Canal.

This study is focused on investigating the contamination levels of chlordane and

dieldrin both horizontally (i.e. at different locations of the Ala Wai Canal) and vertically

(i.e. at different depth in the sediment) in Ala Wai Canal sediment. This was

accomplished by collecting sediment cores along and near the Ala Wai Canal and

analyzing the layers of the sediment core for chlordane and dieldrin.

2.2 Methods

2.2.1 Sample Site and Collection

The Ala Wai Canal is approximately 2 miles (3.22 km) long, that receives water

and sediment from the Mānoa-Pālolo Drainage Canal and Makiki Stream and empties to

the Ala Wai Harbor. Besides, collecting runoff from the Pālolo, Mānoa, and Makiki

areas, the Ala Wai Canal also collects storm water runoff from the Kapahulu,

McCully/Mōʻiliʻili, Ala Moana, and Waikīkī areas (Figure 2.1).

9

Figure 2.1: Areas Surrounding the Ala Wai Canal [34]

Sediment samples were collected on November 24, 2010 and November 25, 2010

at five transects along or near the Ala Wai Canal (Figure 2.2). Each transect had two

sampling sites where a sediment core was collected. At each sample site there was a

floating dock used by paddlers. A sediment core was collected at opposite ends of the

floating dock, one on the right side and the other on the left side. Sediment cores 2

inches (5.08 cm) in diameter and 8 inches to 20 inches (20.32 cm – 50.8 cm) in length

were collected using an AMS multi-stage sludge and sediment core sampler (American

Falls, Idaho, U.S.A). The sediment cores were capped and placed in an ice cooler with

ice packs. The sediment cores were taken to the laboratory and were frozen in the freezer

at -20 °C.

10

Figure 2.2: Sediment Core Collection Sites [34]

2.2.2 Laboratory Analysis

The samples were prepared, extracted, cleaned, concentrated, and analyzed per

Environmental Protection Agency (EPA) Methods (Table 2.1).

Table 2.1: Summary of Analytical Methods

Type of Analytical Method Method

Pressurized Fluid Extraction SW-846 Method 3545A

Florisil Cleanup SW-846 Method 3620C

Fraction Concentration Appendix A to Part 136 Method 608

Gas Chromatography SW-846 Method 8081B

2.2.2.1 Sample Preparation

The samples were prepared using SW-846 Method 3545A [37]. The frozen

sediment cores were thawed and then removed from the plastic liner. The sediment cores

were sectioned at 2 inch (5.08 cm) intervals using a clean stainless steel knife. Any water

ALA WAI CANAL

Mānoa-Pālolo Drainage Canal

Drainage Canal

Makiki Stream

11

found was removed from the sediment sample and discarded. The sediment samples

were dried in a hood for 48 hours to 72 hours on hexane-rinsed aluminum foil. Foreign

objects such as twigs, rocks, rope, and fish bones were removed from the sediment

samples and discarded. The sediment samples were grinded using a ceramic mortar and

pestle and then strained through a strainer to remove any remaining foreign objects.

Approximately 5 grams of ground sediment was weighed for extraction. Each sample

that will be extracted was mixed with the surrogate, 2 mL of decachlorobiphenyl at 250

µg/L, and was set aside so that the spiking solvent, acetone, would evaporate. An

additional 5 grams of ground sediment was weighed into a tared aluminum crucible and

dried overnight at 103 °C to 105 °C to calculate the percent dry weight.

2.2.2.2 Extraction

The samples were extracted using SW-846 Method 3545A [37]. An extraction

cell was prepared for each spiked sediment sample. A 20 mm disposable glass fiber filter

was placed in the bottom of a 5 mL cell and then weighed. The spiked sediment sample

was added to the cell and then weighted. The difference of the cell with sample and the

empty cell was used to calculate the weight of spiked sediment sample. Drying agent,

diatomaceous earth, was added to fill up the cell and was mixed with the spiked sediment

sample with a clean stainless steel spatula.

The spiked sediment samples were extracted with acetone and hexane (1:1, v/v)

using a DIONEX ASE 200 Accelerated Solvent Extractor (Sunnyvale, California, U.S.A)

with the following operating conditions. The oven temperature was set to 100 °C and the

pressure was set to 1500 psi to 2000 psi. The static time was set to 5 minutes. The

12

flushing volume was set to 60% of the cell volume. The cell was purged with nitrogen

and the purge time and pressure was set to 60 seconds and 150 psi.

After extraction, the volume of each extract was measured. The extracts were

stored in the freezer at -20 °C until cleanup.

2.2.2.3 Cleanup and Concentration

The samples were cleaned using SW-846 Method 3620C [37]. A column

containing 20 grams of activated Florisil and 20 grams of anhydrous sodium sulfate on

top of the Florisil was prepared for each cleanup. One-tenth (1/10) of the total extract

volume was used for cleanup. The columns were pre-eluted with 60 mL of hexane and

the elutes were discarded. The extract was added to the column and 1 mL of hexane was

added twice to rinse the column after the extract was added. The columns were eluted

with two fractions containing ethyl ether and hexane. Fraction 1 contained 200 mL of

ethyl ether and hexane (6:94, v/v). Fraction 2 contained 200 mL of ethyl ether and

hexane.

The factions were concentrated using Method 608 [24]. The fractions were

concentrated in a hood using a Kuderna-Danish (K-D) apparatus on a hot water bath and

then further concentrated in a hood using a nitrogen evaporator. Three or four boiling

chips were placed in the 500 mL K-D flask with a 10 mL concentrator tube. The fraction

was poured into the K-D flask with a concentrator tube and a three-ball Snyder column

was attached to the K-D flask. The Snyder column was pre-wetted with 1 mL of hexane.

The K-D apparatus was placed on a water bath set to 85 °C. The water temperature was

increased and the placement of K-D apparatus in the water bath was adjusted to complete

the concentration. When the volume reached to approximately 5 mL, the K-D apparatus

13

was removed from the water bath. When the K-D apparatus was cooled, the Snyder

column was removed and the K-D flask was rinsed with hexane. The fraction was placed

in a 5 mL vial for further concentration. The fraction was concentrated to 1 mL using an

N-EVAP™

III Nitrogen Evaporator with an OA-SYS™

Heating System (Berlin,

Massachusetts, U.S.A). The 1 mL of fraction was placed in a 2 mL gas chromatography

(GC) vial containing a cap with silicone/PTFE septa. The volume of each fraction was

adjusted with 1 mL of hexane. The fractions were stored in the freezer at -20 °C until

analysis.

2.2.2.4 Calibration and Analysis

The concentration of chlordane and dieldrin in the samples were determined using

SW-846 Method 8081B [37]. Chlordane and dieldrin in sediment was analyzed using a

Hewlett Packard 5890 Series II Gas Chromatograph equipped with an electron capture

detector (GC-ECD) (Palo Alto, California, U.S.A) with the following operating

conditions. The oven temperature was initially held at 100 °C for 2 minutes, ramped

from 100 °C to 160 °C at 15 °C/min, and then ramped from 160 °C to 270 °C at 5

°C/min. The carrier gas was helium (ultra high purity) and the pressure was set to 60 psi.

The makeup gas for the detector was nitrogen and the pressure was set to 25 psi. The

detector temperature was set to 300 °C. The GC-ECD also had an Agilent GC

AutoSampler Controller and 6890 Series Injector (Santa Clara, California, U.S.A) to

inject 2 µL of sample. The injector temperature was set to 225 °C. Also, a J&W DB-

XLB column of 30 m × 0.533 mm × 150 µm size was used (Folsom, California, U.S.A).

Chlordane and dieldrin concentrations were quantified using calibration standards

and were corrected by surrogate recoveries, ranging from 0.45% to 84.62%, and percent

14

moisture, ranging from 7.87% to 30.65%. Five calibration standards at concentrations of

50 µg/L, 150 µg/L, 250 µg/L, 350 µg/L, and 500 µg/L were prepared for both chlordane

and dieldrin. Also, six calibration standards at concentrations of 200 µg/L, 250 µg/L, 300

µg/L, 350 µg/L, 400 µg/L, and 450 µg/L were prepared for the surrogate,

decachlorobiphenyl. The retention time windows were established for each compound.

Chlordane had four peaks and had a retention time of approximately 26.4 minutes for

peak 1, 26.6 minutes for peak 2, 26.8 minutes for peak 3, and 27.0 minutes for peak 4.

For calculations, the sum of the area of the chlordane peaks was used. Dieldrin had one

peak and had a retention time of approximately 28.0 minutes. Decachlorobiphenyl had

one peak and had retention time of approximately 32.8 minutes. The calibration factor

(CF), mean calibration factor (mean CF), standard deviation (SD), and relative standard

deviation (RSD) were calculated for each compound using the equations presented in

equation (1) to (4).

If the RSD was greater than 20%, a new set of calibration standards were prepared. The

concentration of each compound was calculated using the equation presented in equation

(1)

(2)

(3)

(4)

15

(5), where Ax is the area of the peak, Vt is the total volume of the concentrated extract

(µL), D is the dilution factor, is the mean calibration factor (area/ng), Vi is the volume

of the extract injected (µL), and Ws is the weight of sample extracted (g).

The surrogate recovery was calculated using the equation presented in equation (6).

The percent moisture was calculated using the equation presented in equation (7), where

Ws is the weight of sample (g) and Wds is the weight of dry sample (g).

The concentration of chlordane and dieldrin was corrected using the surrogate recovery

and percent moisture. The corrected concentration was calculated using the equation

presented in equation (8).

(5)

(6)

(7)

(8)

16

2.3 Results

2.3.1 Concentration Profiles of Sediment Cores

The sediment cores were analyzed to determine the concentration of chlordane

and dieldrin. The concentration profiles for each sediment collection site are shown

below (Figure 2.4 – 2.8).

Figure 2.4: Site 1 Concentration Profile

Figure 2.5: Site 2 Concentration Profile

0

10

20

30

40

50

60

0 50 100 150 200

Sed

ime

nt

De

pth

(cm

)

Concentration (µg/kg dw)

Site 1 Chlordane Site 1 Dieldrin

0

10

20

30

40

50

60

0 50 100 150 200

Sed

ime

nt

De

pth

(cm

)

Concentration (µg/kg dw)

Site 2 Chlordane Site 2 Dieldrin

17

Figure 2.6: Site 3 Concentration Profile

Figure 2.7: Site 4 Concentration Profile

Figure 2.8: Site 5 Concentration Profile

0

10

20

30

40

50

60

0 50 100 150 200

Sed

ime

nt

De

pth

(cm

)

Concentration (µg/kg dw)

Site 3 Chlordane Site 3 Dieldrin

0

10

20

30

40

50

60

0 50 100 150 200

Sed

ime

nt

De

pth

(cm

)

Concentration (µg/kg dw)

Site 4 Chlordane Site 4 Dieldrin

0

10

20

30

40

50

60

0 50 100 150 200

Sed

ime

nt

De

pth

(cm

)

Concentration (µg/kg dw)

Site 5 Chlordane Site 5 Dieldrin

18

The concentration profiles showed two different trends. For sediment cores collected

along Mānoa-Pālolo Drainage Canal and between Mānoa-Pālolo Drainage Canal and the

Drainage Canal near Isenberg Street, site 1, 2, and 3, the concentration decreases with

depth with fluctuations. For sediment cores collected between the Drainage Canal near

Isenberg Street and the McCully Street Bridge, site 4 and 5, the concentration increases

with depth with fluctuations.

The concentration profiles are different due to the location and how sediment is

transported to these locations. For site 1, 2, and 3, the concentration decreases with

depth, so this could mean that chlordane and dieldrin is depleting from the environment

but is still entering the environment through soil erosion and runoff. For site 4 and 5, the

concentration increases with depth, so this could mean that degradation of chlordane and

dieldrin by microbes might be occurring on the surface or that sediment containing

chlordane and dieldrin have not been settling in that area recently.

2.3.2 Comparison of the Concentration of Surface Sediments to Aquatic Life Protection

Guidelines

The concentrations of chlordane and dieldrin were compared with the Canadian

Sediment Quality Guidelines (CSQG) for aquatic life to determine if chlordane and

dieldrin in Ala Wai Canal sediment are affecting aquatic life in the Ala Wai Canal. The

probable effect level (PEL) was 8.87 µg/kg dry weight for chlordane and 6.67 µg/kg dry

weight for dieldrin [4, 31].

The concentration of the sediments at the surface ranged from 19.06 µg/kg dry

weight to 169.26 µg/kg dry weight for chlordane and 2.13 µg/kg dry weight to 35.63

µg/kg dry weight for dieldrin. Plots comparing the concentration of chlordane and

19

dieldrin in surface sediment to the probable effect level are shown below (Figure 2.9 and

2.10). At all the sites, the concentration of chlordane and dieldrin in surface sediment

exceeded the probable effect level, except for dieldrin at site 5.

Figure 2.9: Chlordane Concentration at the Surface for Each Site and the Probable

Effect Level (PEL) of Chlordane

Figure 2.10: Dieldrin Concentration at the Surface for Each Site and the Probable

Effect Level (PEL) of Dieldrin

0

20

40

60

80

100

120

140

160

180

200

Site 1 Site 2 Site 3 Site 4 Site 5

Co

nce

ntr

atio

n (

µg/

kg d

w)

Chlordane Conc.

Chlordane PEL

0

5

10

15

20

25

30

35

40

Site 1 Site 2 Site 3 Site 4 Site 5

Co

nce

ntr

atio

n (

µg/

kg d

w)

Dieldrin Conc.

Dieldrin PEL

20

2.4 Discussion

The Ala Wai Canal is a basin where sediment is deposited. The sediment is

transported from streams and drainage canals that discharge into the Ala Wai Canal. The

Ala Wai Canal reduces the amount of sediment transported into the nearshore coastal

waters of Waikīkī, which serves as a recreation area for tourists and residents. Large

quantities of sediment are transported to the Ala Wai Canal through storm water runoff

from surrounding areas, particularly during periods of high intensity rainfall [3].

The concentration profiles showed two different trends. The concentration of

chlordane and dieldrin either decreases or increases with depth. When the concentration

decreases with depth, this could mean that chlordane and dieldrin that was introduced in

the environment before the 1970s has been depleting but is still entering the environment

through soil erosion and runoff after the 1970s. When the concentration increases with

depth, this could mean that degradation of chlordane and dieldrin by microbes might be

occurring on the surface or that sediment containing chlordane and dieldrin have not been

settling in that area recently. Where the sediment is transported from, how the sediment

is transported, and disturbance near or at the sampling site can contribute to this

difference.

Higher concentrations of chlordane and dieldrin at the surface where found at site

1, 2, and 3, along the Mānoa-Pālolo Drainage Canal and near the outlet of the Mānoa-

Pālolo Drainage Canal. Chlordane and dieldrin were both used as a termiticide before

being banned in 1988 for chlordane and in 1985 for dieldrin. In Hawaii, 50,000 pounds

of chlordane and approximately 9000 pounds of aldrin (degrades to dieldrin) was used in

1977 for pest control [4]. Chlordane and dieldrin are transported through soil erosion and

21

runoff, so the areas that were treated for termites contain chlordane and dieldrin that can

be transported to water sources. Higher concentrations of chlordane and dieldrin were

found at these sites since the soil and sediment from the Pālolo and Mānoa areas contains

more chlordane and dieldrin than the McCully/Mōʻiliʻili and Makiki areas. The areas of

Pālolo and Mānoa contain many older homes that were treated for termites while the

areas of McCully/Mōʻiliʻili and Makiki contain apartments and businesses that were not

treated as often for termites. Based on a census conducted in 2000 on selected housing

characteristics, around 80% to 84% of the structures in the Pālolo, Mānoa,

McCully/Mōʻiliʻili, and Makiki areas were built in 1979 or earlier and 80% of the

structures in the McCully/Mōʻiliʻili area and 84% of the structures in the Makiki area

contained 5 or more units while 19% of the structures in the Pālolo area and 23% of the

structures in the Mānoa area contained 5 or more units [36].

Most of the sediment deposited in the Ala Wai Canal comes from the Mānoa-

Pālolo Drainage Canal. Sediment is transported to the Ala Wai Canal through water

containing suspended sediment particles that flow to Ala Wai Canal and then eventually

settle to the bottom of the Ala Wai Canal depending on the flow rate and the size of the

sediment particles. More sediment particles settle to the bottom if the flow rate of the

water is slow and the water contains large suspended particles [3]. At the outlet of the

Mānoa-Pālolo Drainage Canal the flow rate decreases since the water coming from the

Mānoa-Pālolo Drainage Canal is entering a wider and deeper area [3]. This reduces the

ability to carry more suspended sediment particles, which eventually settles out overtime

and results in an area accumulated with sediment [3]. Sediment particles, around the size

of a sand particle, were found at areas near the outlet of the Mānoa-Pālolo Drainage

22

Canal [3]. The proportion of slit to sand also increased with distance away from the

outlet of the Mānoa-Pālolo Drainage Canal [3]. Also, a lot of slit was found in the

unlined section of the Mānoa-Pālolo Drainage Canal [3]. The Mānoa-Pālolo Drainage

Canal, site 2, and the areas near the outlet of the Mānoa-Pālolo Drainage Canal, site 1 and

3, contained higher concentrations of chlordane and dieldrin than the areas further away

from the outlet of the Mānoa-Pālolo Drainage Canal, site 4 and 5. This can be due to the

amount of sediment transported and how the sediment is transported to the Ala Wai

Canal. The most amount of sediment was collected at the Mānoa-Pālolo Drainage Canal,

site 2, and the least amount of sediment was collected at the area furthest away from the

outlet of the Mānoa-Pālolo Drainage Canal, site 5. The sediment at the Mānoa-Pālolo

Drainage Canal, site 2, was composed mainly of slit, while the sediment near the outlet of

the Mānoa-Pālolo Drainage Canal, site 1 and 3, was composed of more sand than slit

when compared to the areas further away from the outlet of the Mānoa-Pālolo Drainage

Canal, site 4 and 5.

Construction can contribute to the contamination of chlordane and dieldrin

through soil erosion or runoff since construction areas may be dug up to construction a

new structure, building, or house. Also, the sediment layers may be disturbed due to

construction being done at or near the waterway. Site 4 and 5 is near the McCully Street

Bridge. There was a construction zone located at site 5. The construction occurring at

site 5 could have disturbed the sediment layers since the least amount of sediment was

collected at this site. Site 4 was near site 5 and had the same concentration profile trend.

The concentration profile at these sites was different from site 1, 2, and 3 and could have

been due the construction near these sites.

23

Sediment was analyzed to determine the concentration of chlordane and dieldrin.

Chlordane and dieldrin are persistent, toxic, and can bioaccumulate in the food chain [27,

30]. There is concern about the harmful effects chlordane and dieldrin has on humans

since chlordane and dieldrin effects the nervous, immune, and reproductive system and

may cause cancer [32, 33]. The concentrations of chlordane and dieldrin for the

sediments at the surface exceeded the Canadian Sediment Quality Guidelines (CSQG) for

aquatic life at all sites, except for dieldrin at site 5. Therefore, residents and state and city

officials should be warned about the chlordane and dieldrin levels in the Ala Wai Canal

and especially in the input streams and drainage canals since there are no warnings about

the potentially contaminated fish or shellfish that resident might catch and consume in

those sites.

Through this investigation of the horizontal and vertical distribution of chlordane

and dieldrin in Ala Wai Canal sediment, the contaminations levels for chlordane and

dieldrin were determined for the sampling sites. The highest levels of contamination

come from the Pālolo and Mānoa areas, which contain older homes that were treated

before the 1980s with chlordane or dieldrin to control termites. The soil from these older

homes still contains chlordane and dieldrin that enter water sources through soil erosion

and runoff. Also, the concentration of chlordane and dieldrin in Ala Wai Canal sediment

are affecting aquatic life in the Ala Wai Canal. Plans need to be developed and

implemented to restore stream and channel banks which can help control erosion and to

educate residents about these contaminants that can be harmful to their health if the

contaminated fish and shellfish are consumed. Also, periodic testing is needed to

determine the presence of chlordane and dieldrin and the contamination levels at sites

24

contributing to the transportation of chlordane and dieldrin down to the Ala Wai Canal

and at sites at the Ala Wai Canal and continuing monitoring of the contaminations levels

for chlordane and dieldrin is needed to identify trends.

25

CHAPTER 3 DETERMINE WHETHER OR NOT CHLORDANE CAN BE

BIODEGRADED IN ALA WAI CANAL SEDIMENT

3.1 Introduction

Chlordane and dieldrin were both detected in Ala Wai Canal sediment. However,

higher concentrations were detected for chlordane. In this study, chlordane was used to

determine whether or not chlordane can be biodegraded in Ala Wai Canal sediment since

high concentrations of chlordane was detected in Ala Wai Canal sediment.

Chlordane was produced in 1947 and was sold as an insecticide for agricultural

applications and termiticide for protecting structures from termites [30, 33]. The United

States of America (U.S.A) produced 9.5 million kg of chlordane in 1974 [33]. In 1978,

the EPA banned the use of chlordane on food crops and phased out other applications of

chlordane used above the ground over the following 5 years [30]. From 1983 and 1988,

chlordane was only approved to be used as a termiticide for underground treatment

around home foundations [30]. In 1988, all uses of chlordane were banned [30].

Chlordane is no longer used today, but is still being detected in environmental

samples worldwide [4, 5, 7, 10, 11, 13, 14, 16, 20, 23, 27, 30, 32, 33, 39]. Chlordane has

been listed by the EPA’s Great Waters Program and the Stockholm Convention as a

pollutant of concern [8, 10, 20, 30, 35]. Chlordane can remain intact in the environment

for long periods, be widely distributed throughout the environment as a result of natural

processes through soil and water, accumulate in the fatty tissue of living organisms and

therefore becoming more concentrated higher up the food chain, and is toxic to aquatic

life, wildlife, and humans [10, 30, 39]. Therefore, data of contamination levels are

needed for regulation purposes, to pin point sources, to determine the risks involved with

26

chlordane remaining in the environment, and to discover trends on how chlordane is

being degraded naturally. This information can be useful in determining how chlordane

can be degraded.

Microbial degradation can be used to remove pollutants from sediments.

However, not much research has been done showing chlordane degradation from

sediments and about the degradation pathways or degradation products of chlordane [8,

10, 20]. Chlordane refers to a mixture of chlordane isomers (cis-chlordane and trans-

chlordane), other chlorinated hydrocarbons, and by-products such as heptachlor and

nonachlor [8, 10, 20, 33]. Since chlordane contains a mixture of various compounds and

each compound degrades independently, this makes chlordane hard to degrade and in

determining the degradation pathways or degradation products of chlordane [10, 20, 33].

Anaerobic microbial degradation has been reported by Hirano et al. [10].

Degradation of chlordane occurred after 4 weeks for sediment collected at one sampling

site [10]. At another sampling site degradation of chlordane occurred immediately after

the experiment started while no degradation of chlordane was also observed at another

sampling site [10]. This was due to the different contamination levels and amount of

organic carbon found at the various sampling sites where sediment was collected [10]. If

the sediment is highly contaminated with chlordane and contains a lot of organic carbon,

the microbial population is sufficient enough to induce degradation of chlordane, while

for sediment less contaminated with chlordane and containing little organic carbon, the

microbial population needs time to grow before degradation of chlordane will occur [10].

This study is focused on determining whether or not chlordane in Ala Wai Canal

sediment can be biodegraded. This was accomplished by conducting laboratory

27

microcosm experiments and analyzing the samples taken from the microcosms

approximately once a month to determine the percentage of chlordane remaining in the

microcosms and observing the chromatographs for any dechlorinating peaks.

3.2 Methods

3.2.1 Sample Site and Collection

Two sediment cores were collected at site 1 (Figure 3.1) on November 24, 2011

using an AMS multi-stage sludge and sediment core sampler (American Falls, Idaho,

U.S.A). The sediment below the surface (anaerobic layer) was kept and placed in a glass

jar. The sediment sample was stored in an ice cooler with ice packs and was immediately

taken to the laboratory and stored in a COY Laboratories anaerobic chamber (Grass Lake,

Michigan, U.S.A).

Figure 3.1: Sediment Core Collection Site [34]

ALA WAI CANAL

28

3.2.2 Culture Preparation and Sampling

Microcosms were prepared (in triplicate) using 125 mL serum bottles (Figure

3.2). Each bottle contained approximately 20 g of wet sediment, 100 mL of culture

medium, 39.3 µL of chlordane at 3.05 M, and 1 mL of sodium propionate at 20 mM

(electron donor). The culture medium contained (per liter of deionized (DI) water) 0.001

g of resazurin, 0.1 g of MgCl2·6H2O, 0.1 g of CaCl2·6H2O, 3.0 g of Na2CO3, 0.5 g of

NH4Cl, 0.6 g of Na2HPO4, 8.7 g of K2HPO4, 10 mL of vitamin solution, and 10 mL of

mineral solution [6, 19, 21]. The bottles were capped with Teflon stoppers and aluminum

crimps. The bottles then were vigorously shaken and kept in a COY Laboratories

anaerobic chamber (Grass Lake, Michigan, U.S.A). Microcosms were also prepared (in

triplicate) as described above, except sodium sulfate was also added to the bottles (Figure

3.2). Sterilized control microcosms were also prepared (in triplicate) as described above

and then the bottles were wrapped in aluminum foil and autoclaved overnight at 103 °C

to 105 °C (Figure 3.2).

Table 3.1: Set Up of the Laboratory Microcosm Experiments

Set Components

A Ala Wai Canal Sediment, Chlordane, and Culture Medium

B Components from Set A and Sulfate

C Components from Set A and Autoclaved

Samples were taken from each microcosm approximately every 1 month for

chlordane analysis. The bottles were vigorously hand-shaken for 2 minutes and

approximately 2 mL of completely mixed sample was removed with a glass pipette and

placed in a 22.2 mL glass vial for extraction. An additional 2 mL of completely mixed

sample was removed and placed in a 2 mL centrifuge tube for measuring pH. The

29

sample in the vial was measured to determine the weight of the completely mixed

sample. Each sample that will be extracted was mixed with the surrogate, 40 µL of

decachlorobiphenyl at 250 µg/L. The pH was determined by centrifuging the additional

sample taken for 10 minutes at 13,000 rpm. After centrifuging the sample, 1 mL of

supernatant was removed and placed in a glass tube with 2 mL of deionized (DI) water.

The pH was then measured using a Denver Instruments pH meter (Bohemia, New York,

U.S.A). The remaining supernatant was removed and discarded and the cells remaining

in the centrifuge tube was kept and stored in the freezer at -20 °C.

3.2.3 Extraction and Cleanup

The samples were extracted in a similar manner to the methods described by

Hirano et al. (2007) [10]. The samples were extracted twice with 6.25 mL of acetone.

The vials were shaken for 2.5 minutes on a shaker and placed in an ultrasonic cleaner and

then the acetone layer was transferred to a 125 mL serum bottle. The samples were then

extracted with 31.25 mL of deionized (DI) water containing 5% (by volume) NaCl and

12.5 mL of hexane. The bottles were shaken for 2.5 minutes on a shaker and the hexane

layer was transferred to a 22.2 mL glass vial and stored in the freezer at -20 °C.

The extracts were cleaned using SW-846 Method 3665A and 3660B [37]. Only 2

mL of extract was used for cleanup. First, 2 mL of the hexane layer from the extraction

was transferred to a 15 mL centrifuge tube containing 5 mL of deionized (DI) water

containing 50% (by volume) sulfuric acid. The extracts were then vortex for a minute

and set aside for a minute to allow the phases to separate. The hexane layer was

transferred to a glass tube and 1 mL of hexane was added to the centrifuge tube and then

hand-shaken. The hexane layer was removed and combined with the hexane layer in the

30

glass tube. The hexane layer from the sulfuric acid cleanup was then cleaned using

copper powder to remove sulfur. Copper powder was filled up approximately to the 0.5

mL mark (2 g of copper powder) and 1 mL of hexane layer from the sulfuric acid cleanup

was transferred to a 2 mL centrifuge tube containing copper powder. The tubes were

shaken for a minute on a shaker and set aside to allow the phases to separate. The hexane

layer was transferred to a 2 mL gas chromatography (GC) vial. The remaining hexane

layer from the sulfuric acid cleanup was cleaned with copper powder and combined with

the hexane layer in the GC vial as stated above. The GC vials were stored in the freezer

at -20 °C until analysis.

3.2.4 Calibration and Analysis

The concentration of chlordane in the samples was determined using SW-846

Method 8081B [37]. Chlordane in the samples were analyzed using a Hewlett Packard

5890 Series II Gas Chromatograph equipped with an electron capture detector (GC-ECD)

(Palo Alto, California, U.S.A) with the following operating conditions. The oven

temperature was initially held at 100 °C for 2 minutes, ramped from 100 °C to 160 °C at

15 °C/min, and then ramped from 160 °C to 270 °C at 5 °C/min. The carrier gas was

helium (ultra high purity) and the pressure was set to 60 psi. The makeup gas for the

detector was nitrogen and the pressure was set to 25 psi. The detector temperature was

set to 300 °C. The GC-ECD also had an Agilent GC AutoSampler Controller and 6890

Series Injector (Santa Clara, California, U.S.A) to inject 2 µL of sample. The injector

temperature was set to 225 °C. Also, a J&W DB-XLB column of 30 m × 0.533 mm ×

150 µm size was used (Folsom, California, U.S.A).

31

Chlordane concentrations were quantified using calibration standards and were

corrected by surrogate recoveries that were greater than 100%. Five calibration standards

at concentrations of 50 µg/L, 150 µg/L, 250 µg/L, 350 µg/L, and 500 µg/L were prepared

for chlordane. Also, five calibration standards at concentrations of 250 µg/L, 300 µg/L,

350 µg/L, 400 µg/L, and 450 µg/L were prepared for the surrogate, decachlorobiphenyl.

The retention time windows were established during the analysis of chlordane and

dieldrin in the sediment samples. Standard curves (Figure 3.2 and 3.3) were used to

determine the concentration of chlordane and decachlorobiphenyl from peak areas and

then adjusted based on the amount of hexane used for extraction and the amount of

completely mixed sample removed from the microcosms. The concentration of

chlordane was also corrected using surrogate recoveries.

Figure 3.2: Decachlorobipheyl Calibration Curve

y = 75685x R² = 0.9629

0

5000000

10000000

15000000

20000000

25000000

30000000

35000000

40000000

0 100 200 300 400 500

Are

a

Concentration (µg/L)

32

Figure 3.3: Chlordane Calibration Curve

The concentration of decachlorobiphenyl was calculated using the equation presented in

equation (9).

The surrogate recovery was calculated using the equation presented in equation (10).

The concentration of chlordane was calculated using the equation presented in equation

(11).

y = 81767x R² = 0.9976

0

5000000

10000000

15000000

20000000

25000000

30000000

35000000

40000000

45000000

0 100 200 300 400 500 600

Are

a

Concentration (µg/L)

(9)

(10)

(11)

33

3.3 Results

Laboratory microcosm experiments were performed to determine whether or not

chlordane in Ala Wai Canal sediment can be depleted. For a period of 81 days

(approximately 11 weeks), the microcosm experiments did not show any indications of

degradation of chlordane (Figure 3.4). After the initial sampling, each set of microcosm

experiments still had 97% or more chlordane remaining, except for the samples taken on

day 54 (approximately 8 weeks). The samples taken on day 54 had more chlordane than

the initial samples taken. This could have probably been due to the culture not being

mixed well and then a sample with a high spike of chlordane was removed from the

culture.

Figure 3.4: Chlordane Concentration of Samples Taken from Microcosms

(each point represents the average plus and minus one standard deviation of the mean)

-50

0

50

100

150

200

250

300

350

400

0 20 40 60 80 100

Ch

lord

ane

Co

nce

ntr

atio

n (

µg/

kg w

w)

Time (day)

Set A

Set B

Set C

34

The chromatographs for the microcosm experiments also did not show any dechlorinating

peaks since the chromatographs for the chlordane containing microcosms (set A) and the

chlordane and sulfate containing microcosms (set B) contained the same peaks as the

sterilized control microcosms (set C).

3.4 Discussion

Degradation of chlordane did not occur during a period of 81 days (approximately

11 weeks). This means that degradation of chlordane did not start or that chlordane

cannot be depleted from Ala Wai Canal sediment. For degradation of chlordane to occur,

the microbial population needs time to grow to a sufficient size, known as the acclimation

period, before microbes could start degrading chlordane [10, 12]. During the acclimation

period, slight or no degradation occurs and is shown as a lag period on degradation

profiles. Suitable conditions are needed for degradation to occur. Sediment

contaminated with high levels of chlordane and contain a high percentage of organic

carbon can provide suitable conditions for the microbial population to grow [10]. Hirano

et al. (2007) reported that degradation of chlordane occurred immediately for sediment

contaminated with chlordane and containing the highest percentage of organic carbon

while no degradation occurred for sediment not contaminated with chlordane and

containing the least percentage of organic carbon [10]. A lag period was noticed for all

microcosm experiments containing Ala Wai Canal sediment. The Ala Wai Canal

sediment used for the microcosm experiments were contaminated with chlordane so the

sediment probably did not have enough microbes and needs more time for the microbial

population to increase before degradation will occur or the Ala Wai Canal sediment did

not have the right conditions to entice microbial degradation.

35

CHAPTER 4 CONCLUSIONS AND RECOMMENDATIONS

4.1 Conclusions

Through this study about the horizontal and vertical distribution of chlordane and

dieldrin in Ala Wai Canal sediment and whether or not chlordane in Ala Wai Canal

sediment can be biodegraded, a chlordane and dieldrin concentration survey was obtained

and it was determined that no degradation of chlordane in Ala Wai Canal sediment

occurred for a period of 81 days (approximately 11 week).

The concentration of chlordane and dieldrin in Ala Wai Canal sediment are

affecting aquatic life in the Ala Wai Canal. This is concerning since chlordane and

dieldrin are persistent, toxic, and can bioaccumulate in the food chain [27, 30]. The

highest levels of contamination come from the Pālolo and Mānoa areas, which contain

older homes that were treated before the 1980s with chlordane or dieldrin to control

termites. The soil from these older homes still contains chlordane and dieldrin that enter

water sources through soil erosion and runoff. Therefore, plans need to be developed and

implemented to restore stream and channel banks which can help control erosion.

Periodic testing and continuing monitoring is also needed to determine the presence of

chlordane and dieldrin and the contamination levels at sites contributing to the

transportation of chlordane and dieldrin down to the Ala Wai Canal and at sites at the Ala

Wai Canal. Also, residents and immigrants that speak no or little English need be

educated about pollution and the harmful effects of consuming contaminated fish and

shellfish and drinking contaminated water.

Degradation of chlordane in Ala Wai Canal sediment was not observed for a

period of 81 days (approximately 11 weeks). The microbial population in the sediment

36

needs to be large enough before degradation will occur, which depends on the

contamination levels, temperature, and carbon and energy source [2, 7, 8, 10, 13, 14, 20].

The Ala Wai Canal sediment used for the microcosm experiments were contaminated

with chlordane so the sediment probably did not have enough microbes and needs more

time for the microbial population to increase before degradation will occur or the right

conditions to entice microbial degradation.

4.2 Recommendations

The Ala Wai Canal is contaminated and the City and County of Honolulu and the

State of Hawaiʻi are seeking ways to clean up the Ala Wai Canal [18]. First, the sources

contributing to the contamination in the Ala Wai Canal needs to be determined. By

determining the source of contamination, these areas can be continuously monitored to

discover trends in the contamination levels, which can be used to determine how the

contamination is transported and what is contributing to the high levels of contamination.

Since the Ala Wai Canal contains contaminated fish and shellfish, periodic testing of the

fish and shellfish is also needed to determine the contamination levels in the fish tissue

and in the shellfish. Secondly, plans need to be developed and implemented to control

soil erosion, which contributes to contamination entering water sources, such as streams

and drainage canals. This can be done by restoring stream and channel banks and

preventing soil from entering nearby water sources when land is cleared or during

construction. By having these plans and monitoring the contamination levels, then state

and city officials can determine whether or not the soil erosion reduction practices are

effective. Lastly, residents and immigrants that speak no or little English need to be

educated about the contamination found in water sources and harmful effects of

37

consuming contaminated fish and shellfish and drinking contaminated water. The Ala

Wai Canal has signs warning people not to swim and catch and consume the fish and

shellfish in the Ala Wai Canal. However, the contamination found in Ala Wai Canal is

also found at streams and drainage canals that discharge water and sediment to the Ala

Wai Canal [4, 5, 9, 17]. Therefore, signs are also needed at those contaminated sites to

prevent consumption of contaminated fish and shellfish and awareness and ways

residents can help out with the cleaning efforts are promoted through education.

Microbial degradation is one method that can be used to deplete contaminants

from the environment. The laboratory microcosm experiments performed in anaerobic

conditions did not show any signs of chlordane being depleted from Ala Wai Canal

sediment, however this can be due to the sediment probably not having enough microbes

and more time is needed for the microbial population to increase before degradation will

occur or the right conditions to entice microbial degradation. Therefore, future research

is needed to determine the conditions needed to promote microbial degradation. Other

techniques can be used to determine whether or not contaminants can be degraded from

the environment such as microbial degradation under aerobic conditions and

photodegradation [8, 13, 14, 20]. Future research, using these techniques could be used

to determine if these techniques are more suitable for degrading the contaminants found

in the Ala Wai Canal. Data pertaining to the characteristics of the sediment found in the

Ala Wai Canal was not gather for this study, however future research involving

degradation should include sediment characteristics, which can help determine what is

inhibiting and stimulating degradation of contaminants. If the containments in the Ala

38

Wai Canal can be degraded, this can lead to more information that can be used to develop

and find ways to detect and eventually deplete the containments from the Ala Wai Canal.

39

APPENDIX A

Table A.1: Dry Weights of Sediment Collected for Set A

Site Foil (g) Hood Dry (g) 1st Oven Dry + Foil (g) 2nd Oven Dry + Foil (g) 1st Oven Dry (g) 2nd Oven Dry (g) Dry Weight - 1st Dry Weight - 2nd RPD (< 4%)

1.1.1 1.7910 4.7169 5.6939 5.5978 3.9029 3.8068 82.74% 80.71% 2.49%

1.1.2 1.5527 5.1261 5.2874 5.2092 3.7347 3.6565 72.86% 71.33% 2.12%

1.1.3 1.4610 4.9233 4.9746 4.9024 3.5136 3.4414 71.37% 69.90% 2.08%

1.1.4 1.6546 5.1270 5.6772 5.5910 4.0226 3.9364 78.46% 76.78% 2.17%

1.1.5 1.3982 5.2896 5.4369 5.3418 4.0387 3.9436 76.35% 74.55% 2.38%

1.2.1 1.6762 5.1079 5.896 5.8139 4.2198 4.1377 82.61% 81.01% 1.96%

1.2.2 1.6100 4.6539 5.4731 5.4166 3.8631 3.8066 83.01% 81.79% 1.47%

2.1.1 1.3466 4.8947 5.2369 5.1233 3.8903 3.7767 79.48% 77.16% 2.96%

2.1.2 1.5912 4.7586 5.7939 5.6841 4.2027 4.0929 88.32% 86.01% 2.65%

2.1.3 1.3348 4.5704 5.2826 5.1612 3.9478 3.8264 86.38% 83.72% 3.12%

2.1.4 1.3742 4.7137 5.2663 5.1395 3.8921 3.7653 82.57% 79.88% 3.31%

2.1.5 1.3190 4.6444 4.9124 4.7903 3.5934 3.4713 77.37% 74.74% 3.46%

3.1.1 1.3664 4.7321 5.1247 5.0067 3.7583 3.6403 79.42% 76.93% 3.19%

3.1.2 1.3749 4.5855 5.1028 4.9896 3.7279 3.6147 81.30% 78.83% 3.08%

3.1.3 1.4703 5.3477 5.4254 5.3365 3.9551 3.8662 73.96% 72.30% 2.27%

3.1.4 1.4926 5.5514 5.5727 5.4639 4.0801 3.9713 73.50% 71.54% 2.70%

3.1.5 1.3229 4.8586 4.9578 4.8666 3.6349 3.5437 74.81% 72.94% 2.54%

4.1.1 1.3978 4.7994 5.1542 5.0373 3.7564 3.6395 78.27% 75.83% 3.16%

4.1.2 1.3602 4.9525 5.2626 5.1450 3.9024 3.7848 78.80% 76.42% 3.06%

4.1.3 1.3733 4.8288 5.1419 5.0302 3.7686 3.6569 78.04% 75.73% 3.01%

4.1.4 1.3848 4.8338 4.9446 4.8295 3.5598 3.4447 73.64% 71.26% 3.29%

4.1.5 1.3296 4.6529 5.2768 5.1546 3.9472 3.825 84.83% 82.21% 3.14%

4.2.1 1.4029 5.0559 5.5284 5.4215 4.1255 4.0186 81.60% 79.48% 2.63%

4.2.2 1.3451 4.9167 4.9153 4.8126 3.5702 3.4675 72.61% 70.52% 2.92%

5.1.1 1.4144 4.7803 5.2850 5.1801 3.8706 3.7657 80.97% 78.78% 2.75%

5.1.3 1.7077 4.8922 5.6334 5.6270 3.9257 3.9193 80.24% 80.11% 0.16%

5.1.4 1.7134 5.2300 5.6029 5.5004 3.8895 3.787 74.37% 72.41% 2.67%

5.2.2 1.5198 5.0714 5.5345 5.4134 4.0147 3.8936 79.16% 76.78% 3.06%

40

Table A.2: Dry Weights of Sediment Collected for Set B

Site Foil (g) Hood Dry (g) 1st Oven Dry + Foil (g) 2nd Oven Dry + Foil (g) 1st Oven Dry (g) 2nd Oven Dry (g) Dry Weight - 1st Dry Weight - 2nd RPD (< 4%)

1.1.1 1.4243 5.4325 6.2115 6.1348 4.7872 4.7105 88.12% 86.71% 1.62%

1.1.2 1.5141 6.6205 7.6137 7.5286 6.0996 6.0145 92.13% 90.85% 1.40%

1.1.3 1.4799 5.6688 6.5145 6.4380 5.0346 4.9581 88.81% 87.46% 1.53%

1.1.4 1.4682 6.0164 6.6566 6.5673 5.1884 5.0991 86.24% 84.75% 1.74%

1.1.5 1.7829 5.0461 6.2452 6.1791 4.4623 4.3962 88.43% 87.12% 1.49%

2.1.1 1.4359 4.5471 5.1231 5.0473 3.6872 3.6114 81.09% 79.42% 2.08%

2.1.2 1.4707 4.6910 4.8753 4.7747 3.4046 3.3040 72.58% 70.43% 3.00%

2.1.3 1.7621 4.7167 5.2983 5.1861 3.5362 3.4240 74.97% 72.59% 3.22%

2.1.4 1.4622 5.5374 5.7530 5.6512 4.2908 4.1890 77.49% 75.65% 2.40%

2.1.5 1.5107 4.8700 5.1758 5.0934 3.6651 3.5827 75.26% 73.57% 2.27%

2.2.1 1.5496 5.9345 6.1425 6.0435 4.5929 4.4939 77.39% 75.72% 2.18%

2.2.2 1.7018 5.0640 5.3456 5.2502 3.6438 3.5484 71.95% 70.07% 2.65%

2.2.3 1.4302 6.4875 6.0840 5.9971 4.6538 4.5669 71.73% 70.40% 1.88%

2.2.4 1.6243 5.9217 5.8238 5.7222 4.1995 4.0979 70.92% 69.20% 2.45%

2.2.5 1.4525 4.7742 4.7634 4.6761 3.3109 3.2236 69.35% 67.52% 2.67%

3.1.1 1.5947 5.8309 5.8785 5.7950 4.2838 4.2003 73.47% 72.04% 1.97%

3.1.2 1.5821 5.8743 6.3072 6.2148 4.7251 4.6327 80.44% 78.86% 1.97%

3.1.3 1.6388 7.1634 6.9458 6.8397 5.3070 5.2009 74.08% 72.60% 2.02%

3.1.4 1.5298 5.0132 5.2234 5.1459 3.6936 3.6161 73.68% 72.13% 2.12%

3.1.5 1.4116 5.9844 6.1571 6.0720 4.7455 4.6604 79.30% 77.88% 1.81%

3.2.1 1.4978 6.1198 6.6975 6.6043 5.1997 5.1065 84.97% 83.44% 1.81%

3.2.2 1.3995 5.3476 5.5567 5.5481 4.1572 4.1486 77.74% 77.58% 0.21%

4.1.1 1.4500 5.6815 5.7455 5.6526 4.2955 4.2026 75.61% 73.97% 2.19%

4.1.2 1.8342 5.6015 6.3167 6.2238 4.4825 4.3896 80.02% 78.36% 2.09%

4.1.3 1.6638 6.8588 6.7731 6.6828 5.1093 5.0190 74.49% 73.18% 1.78%

4.1.4 1.6542 5.4424 5.7558 5.6678 4.1016 4.0136 75.36% 73.75% 2.17%

5.1.1 1.7177 6.4005 6.4712 6.3661 4.7535 4.6484 74.27% 72.63% 2.24%

5.1.2 1.4650 5.9795 6.3039 6.2152 4.8389 4.7502 80.92% 79.44% 1.85%

41

Table A.3: Concentration, % Moisture, and % Recovery of Sediment Collected for Set A

Site % Moisture Decachlorobiphenyl Chlordane Dieldrin

Concentration (µg/kg) % Recovery Corrected Concentration (µg/kg) Corrected Concentration (µg/kg)

1.1.1 17.2571 38.6005 38.6005 4.6335 ND

1.1.2 27.1434 13.5235 13.5235 55.6066 12.8850

1.1.3 28.6332 24.6492 24.6492 29.5375 21.5654

1.1.4 21.5409 32.9427 32.9427 13.2811 8.0632

1.1.5 23.6483 28.7887 28.7887 50.0547 26.4352

1.2.1 17.3868 36.4656 36.4656 46.4633 103.1289

1.2.2 16.9922 7.0042 7.0042 149.7152 26.7666

2.1.1 20.5202 15.1120 15.1120 166.8800 36.9672

2.1.2 11.6820 19.7568 19.7568 94.8871 ND

2.1.3 13.6224 6.0449 6.0449 239.7678 37.8866

2.1.4 17.4300 15.2658 15.2658 248.0344 43.2297

2.1.5 22.6294 19.4735 19.4735 198.3966 29.1487

3.1.1 20.5786 9.3815 9.3815 98.2920 34.2479

3.1.2 18.7024 18.6311 18.6311 85.2549 26.8242

3.1.3 26.0411 32.6484 32.6484 12.3294 ND

3.1.4 26.5032 5.9299 5.9299 97.2823 6.9805

3.1.5 25.1863 13.1496 13.1496 329.7045 67.9782

4.1.1 21.7319 22.6872 22.6872 117.2141 ND

4.1.2 21.2034 15.8907 15.8907 186.8065 ND

4.1.3 21.9558 14.8456 14.8456 95.2276 ND

4.1.4 26.3561 15.6488 15.6488 174.1233 ND

4.1.5 15.1669 19.5134 19.5134 224.5136 11.5480

4.2.1 18.4023 84.6204 84.6204 5.9259 3.2471

4.2.2 27.3863 41.0242 41.0242 38.4477 16.6358

5.1.1 19.0302 20.1114 20.1114 42.4497 68.0089

5.1.3 19.7559 4.4452 4.4452 79.5071 23.7331

5.1.4 25.6310 3.7704 3.7704 80.0446 51.4833

5.2.2 20.8365 4.5594 4.5594 38.1209 3.4332

42

Table A.4: Concentration, % Moisture, and % Recovery of Sediment Collected for Set B

Site % Moisture Decachlorobiphenyl Chlordane Dieldrin

Concentration (µg/kg) % Recovery Corrected Concentration (µg/kg) Corrected Concentration (µg/kg)

1.1.1 11.8785 1.6058 1.6058 272.6451 215.9787

1.1.2 7.8680 18.7285 18.7285 20.9477 4.7021

1.1.3 11.1876 5.4508 5.4508 2.8008 18.4174

1.1.4 13.7624 2.0673 2.0673 26.5557 20.9703

1.1.5 11.5693 2.0546 2.0546 61.0219 25.9825

2.1.1 18.9110 2.3487 2.3487 18.2532 ND

2.1.2 27.4227 2.2599 2.2599 29.5816 ND

2.1.3 25.0281 1.6988 1.6988 17.3368 ND

2.1.4 22.5124 25.4988 25.4988 2.8129 ND

2.1.5 24.7413 12.1535 12.1535 8.1374 ND

2.2.1 22.6068 17.9397 17.9397 9.2735 1.7002

2.2.2 28.0450 23.3906 23.3906 16.2134 3.4918

2.2.3 28.2651 4.1196 4.1196 33.6085 1.6746

2.2.4 29.0829 15.9097 15.9097 35.3632 4.2993

2.2.5 30.6502 10.2810 10.2810 46.9967 4.8068

3.1.1 26.5328 17.1221 17.1221 3.7832 2.6617

3.1.2 19.5632 0.5296 0.5296 5.3565 7.6469

3.1.3 25.9151 12.0620 12.0620 3.4210 0.7448

3.1.4 26.3225 0.4502 0.4502 133.5747 38.5814

3.1.5 20.7022 20.2479 20.2479 21.0320 2.8413

3.2.1 15.0348 15.9929 15.9929 11.2492 2.0478

3.2.2 22.2605 33.4727 33.4727 8.8245 3.2870

4.1.1 24.3950 15.2549 15.2549 5.5754 1.4339

4.1.2 19.9768 23.2928 23.2928 17.0707 3.9741

4.1.3 25.5074 1.4729 1.4729 110.5043 ND

4.1.4 24.6362 18.7748 18.7748 7.9908 3.9870

5.1.1 25.7324 22.1639 22.1639 10.2112 2.9091

5.1.2 19.0752 3.4479 3.4479 ND 0.8294

43

APPENDIX B

Table B.1: Years Structures Built in the Pālolo, Mānoa, McCully/Mōʻiliʻili, and Makiki Areas [36]

Year Structure Built Pālolo Mānoa McCully/Mōʻiliʻili Makiki

1999 - March 2000 30 59 9 215

1995 - 1998 126 260 149 854

1990 - 1994 143 377 353 431

1980 - 1989 475 568 1698 1746

1970 - 1979 587 1216 5021 5497

1960 - 1969 823 1692 3416 4193

1940 - 1959 2054 2275 2864 2481

1939 or earlier 486 1106 588 859

Table B.2: Number of Units in Structures in the Pālolo, Mānoa, McCully/Mōʻiliʻili, and Makiki Areas [36]

Units in Structure Pālolo Mānoa McCully/Mōʻiliʻili Makiki

1 unit, detached 2870 4788 831 1383

1 unit, attached 587 587 370 321

2 units 176 175 447 300

3 or 4 units 157 247 1187 552

5 - 9 units 311 379 2212 1262

10 - 19 units 216 429 1786 2214

20 or more units 402 941 7241 10235

other 5 7 24 9

44

APPENDIX C

Table C.1: Chlordane Concentration of Samples Taken from Microcosms

Date 1/10/2012 2/4/2012 3/10/2012 3/31/2012

Microcosm

#

Chlordane Conc.

(μg/kg)

Chlordane Conc.

(μg/kg)

Chlordane Conc.

(μg/kg)

Chlordane Conc.

(μg/kg)

Set A

C1 4.7538

44.8350

1.8411

14.9072

5.9514

47.9261

0.5149

4.3744 C2 15.4045 3.3444 8.9018 0.8492

C3 114.3467 39.5362 128.9250 11.7591

Set B

C4 113.6751

86.3150

48.4190

32.1544

144.3607

91.9541

13.1589

8.6294 C5 59.1320 23.5465 62.1205 5.0001

C6 86.1378 24.4977 69.3813 7.7293

Set C

C7 256.2968

231.3326

88.5691

85.0907

378.8256

320.7271

29.6408

24.2284 C8 211.8748 43.1056 319.4936 25.2333

C9 225.8261 123.5973 263.8621 17.8111

45

Table C.2: pH of Samples Taken from Microcosms

Date 1/10/2012 2/4/2012 3/10/2012 3/31/2012

Microcosm # pH pH pH pH

Set A

C1 7.09

6.98

7.46

7.36

7.71

7.62

7.84

7.74 C2 6.96 7.35 7.61 7.73

C3 6.89 7.28 7.54 7.65

Set B

C4 6.97

6.98

7.34

7.34

7.91

7.90

8.03

8.03 C5 6.98 7.34 7.83 7.99

C6 6.99 7.35 7.97 8.07

Set C

C7 6.82

6.82

7.39

7.32

7.46

7.44

7.53

7.52 C8 6.83 7.31 7.47 7.55

C9 6.81 7.26 7.40 7.48

46

Figure C.1: pH of Samples Taken from Microcosms

6.6

6.8

7

7.2

7.4

7.6

7.8

8

8.2

0 20 40 60 80 100

pH

Time (day)

Set A

Set B

Set C

47

Figure C.2: Chromatographs of Samples Taken on January 14, 2012

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 1:04:50 PM Page 1 of 1

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.11

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.18

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.53

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Figure C.3: Chromatographs of Samples Taken on February 4, 2012

Chrom Perfect Chromatogram Report

Printed on 2/5/2012 12:13:00 PM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 12:16:33 PM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 12:21:02 PM Page 1 of 1

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.81

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 12:45:57 PM Page 1 of 1

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.12

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 12:49:20 PM Page 1 of 1

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.10

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.93

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.15

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.67

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 12:50:08 PM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 1:09:23 PM Page 1 of 1

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.44

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.51

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 1:10:14 PM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 2/5/2012 1:11:02 PM Page 1 of 1

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Figure C.4: Chromatographs of Samples Taken on March 10, 2012

Chrom Perfect Chromatogram Report

Printed on 3/11/2012 11:00:20 AM Page 1 of 1

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.17

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.95

21

.17

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.68

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.04

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.32

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Chrom Perfect Chromatogram Report

Printed on 3/11/2012 11:01:48 AM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 3/11/2012 11:04:56 AM Page 1 of 1

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Printed on 3/11/2012 11:07:31 AM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 3/11/2012 11:09:29 AM Page 1 of 1

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Printed on 3/11/2012 11:08:32 AM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 3/11/2012 11:12:31 AM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 3/11/2012 11:13:17 AM Page 1 of 1

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Chrom Perfect Chromatogram Report

Printed on 3/11/2012 11:14:10 AM Page 1 of 1

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Figure C.5: Chromatographs of Samples Taken on March 31, 2012

Chrom Perfect Chromatogram Report

Printed on 4/8/2012 11:47:16 AM Page 1 of 1

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.43

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.92

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.16

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