Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
Growth inhibition by soil components
for degradation of dioxins using white rot fungus
11
Shinya SuzukiShinya Suzuki, , Ayako Tachifuji, Yasushi MatsufujiAyako Tachifuji, Yasushi Matsufuji
Department of Civil Engineering, Fukuoka University8-19-1 Nanakuma, Johnan-ku, Fukuoka, 814-0180 JapanE-mail: [email protected]@fukuoka-u.ac.jp
O
Cl
Cl
O
OCl
O
Cl
Cl
Cl
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
1-1 Background in Japan1) Dioxin compound
- risk to cause long-term contamination in soil- chemically stable structure- accumulate in environment
2) There are still many contaminated soils- not only around MSW incineration facilities - but also around rice field
- included in pesticide as impurities about 40 years ago
3) Physicochemical treatment methods- already developed, but- disadvantage from economic and energetic point of view
1) Dioxin compound- risk to cause long-term contamination in soil- chemically stable structure- accumulate in environment
2) There are still many contaminated soils- not only around MSW incineration facilities - but also around rice field
- included in pesticide as impurities about 40 years ago
3) Physicochemical treatment methods- already developed, but- disadvantage from economic and energetic point of view
4) So, for remediation of contaminated soil - which has low concentration of dioxins- more effective to use bioremediation method
4) So, for remediation of contaminated soil - which has low concentration of dioxins- more effective to use bioremediation method
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
1-2 Objective
Objective of this study- to accomplish effective bioremediation method
- for low levels of dioxins contaminated soil- by using “Phlebia brevispora”
- to clarify inhibitory factor of soil components - to evaluate relationship between growth of fungi and degradation of dioxins
Objective of this study- to accomplish effective bioremediation method
- for low levels of dioxins contaminated soil- by using “Phlebia brevispora”
- to clarify inhibitory factor of soil components - to evaluate relationship between growth of fungi and degradation of dioxins
In particular, - method using white rot fungi attracts attention- but living spheres of such fungi are usually in dead trees, fallen trees, etc.
It is still unclear whether - such fungi can survive and degrade dioxins in soil environment.
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
Soils: composed from various materials such as clay minerals, organic substances
- to compare difference of degradability according to organic substances
Soils: composed from various materials such as clay minerals, organic substances
- to compare difference of degradability according to organic substances
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
2-1 Materials
Organic-poor soil Organic-rich soil
2,7-DCDD- degraded easily, shorter incubation period- simplified extraction
O
Cl
Cl
O
1,3,6,8-TCDD-main components of CNP/PCP originating dioxins- longer incubation period
OCl
O
Cl
Cl
Cl
Phlebia brevispora (TMIC 33929)- can degrade 1,3,6,8-TCDD- maintained on potato dextrose agar (PDA) medium
Phlebia brevispora (TMIC 33929)- can degrade 1,3,6,8-TCDD- maintained on potato dextrose agar (PDA) medium
unitorganic-poor
soilorganic-rich
soil
ｐＨ(CaCl2) - 4.3 5.4
Ignition Loss (%) 4.0 12.8
TOC mg/g 0.4 31.7
TN mg/g 0.1 3.1
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
2-2 Experimental condition2,7-DCDD 1,3,6,8-TCDD
no soil ○ ○- -
organic-poor soil ○ -(L/S=6) -
organic-rich soil ○ ○(L/S=6) (L/S=6)
○(L/S=9)○
(L/S=12)○
(L/S=15)
Influence of soil property- L/S=6
Influence of Liquid-Solid (L/S) ratio- 1,3,6,8-TCDD- organic-rich soil
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
2-3 Analytical procedurePre Incubation
-Kirks HCLN medium: 30mL(autoclaved)
-Fungus body- Incubation: 14 or 30days-Temp.: 25℃-shaking every time
Growth of fungi“Weight analysis”
Growth of fungi“Weight analysis”
Degradation of dioxins- “2,7-DCDD”- “1,3,6,8-TCDD”
Degradation of dioxins- “2,7-DCDD”- “1,3,6,8-TCDD”
-Incubation: 5days-Temp.: 25℃
Incubation
Analysis (n=3)Mixing
or-Distilled water: 15mL(L/S = 9)
30mL(L/S = 12)45mL(L/S = 15)
no addition0mL(L/S = 6)
Simulation of contaminated soil
Organic-poor soil: 5g (autoclaved)
-2,7-DCDD
Organic-rich soil: 5g (autoclaved)
or
-1,3,6,8-TCDD
or
2,7-DCDD or 1,3,6,8-TCDD
Liquid-state condition
Slurry-state condition
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
0
20
40
60
80
100
120
no soil2,7-DCDD
organic-poorsoil
2,7-DCDD
organic-richsoil
2,7-DCDD
reco
very
rat
e of
2,7
-DC
DD
(%)
control
treatment
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
3-1-1) Influence of soil property L/S = 6
O
O Cl
Cl O
O ClCl
ClCl
O
OCl
Cl
OH
O
OClCl
ClCl
OHOH
O
OCl
Cl
OCH3
O
OClCl
ClCl
OCH3OCH3
2,7-DCDD
- monohydroxy-DCDD, monomethoxy-DCDD were obtained as metabolite
- monohydroxy-DCDD, monomethoxy-DCDD were obtained as metabolite
Degradation rate of 1,3,6,8-TCDD: only 1.1% in slurry-state with organic-rich soil
Degradation rate of 1,3,6,8-TCDD: only 1.1% in slurry-state with organic-rich soil
76.3% 66.1% 12.7%
0
20
40
60
80
100
120
no soil1,3,6,8-TCDD
organic-richsoil
1,3,6,8-TCDD
reco
very
rat
e of
1,3
,6,8
-TC
DD
(%)
control
treatment
35.0%1.1%
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
3-1-2) Liquid-solid ratio 1,3,6,8-TCDD in organic-rich soil
Dissolved matters from organic-rich soil had inhibition activity?- this improvement probably caused by dilution of inhibition material in the soil ?
Dissolved matters from organic-rich soil had inhibition activity?- this improvement probably caused by dilution of inhibition material in the soil ?
0
20
40
60
80
100
120
L/S=6 L/S=9 L/S=12 L/S=15
reco
very
rat
e of
1,3
,6,8
-TC
DD
(%)
control
treatment
1.1% 18.9% 32.2% 27.0%
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
3-2-1) Soil extractionSo, in order to examine inhibition activity of soluble matters only
- organic-rich soil was washed in advance to fractionate into…,So, in order to examine inhibition activity of soluble matters only
- organic-rich soil was washed in advance to fractionate into…,
organic-rich soilorganic-rich soil dissolved matterdissolved matter
washed organic-rich soilwashed organic-rich soil
itemsoil growth amount degradation rate
no soil ○ ○- -
organic-rich soil ○ ○(L/S=6) (L/S=6)
2,7-DCDD
dissolved matter ○ ○- -
washed organic-rich soil ○(L/S=6)
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
3-2-2) Analytical procedurePre Incubation
-Kirks HCLN medium: 30mL(autoclaved)
-Fungus body-Incubation: 14days-Temp.: 25℃-shaking every time
Growth of fungi“Weight analysis”
Growth of fungi“Weight analysis”
Degradation of dioxins- “2,7-DCDD”
Degradation of dioxins- “2,7-DCDD”
-Incubation: 5days-Temp.: 25℃
Incubation
Analysis (n=3)
2,7-DCDD Extraction
-24hours-shaking
Solvent: distilled water
Only soluble matters from organic-rich soil were added into flask
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
0.45
0.50
0
10
20
30
40
50
60
70
80
90
100
pre incubationDay 0
incubationDay 0
incubationDay 10
incubationDay 14
grow
th a
mou
nt o
f fun
gi (
g)
degr
adat
ion
rate
of 2
,7-D
CD
D (%
)
incubation time
no soil
dissolved matter
organic-rich soil
no soil
dissolved matter
organic-rich soil
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
3-2-3) Growth and degradation rate
In case of dissolved matters, growth amount of fungus: much smaller- Growth inhibition was clearly confirmed during pre-incubation period
In case of dissolved matters, growth amount of fungus: much smaller- Growth inhibition was clearly confirmed during pre-incubation period
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
3-2-4) Relationship growth amount v.s. degradation rate
Degradation rate of dioxins in liquid-state only depended on growth amount- soluble matters from organic-rich soil had growth inhibition activity,- it was a part of whole inhibitory effect from organic-rich soil.
Degradation rate of dioxins in liquid-state only depended on growth amount- soluble matters from organic-rich soil had growth inhibition activity,- it was a part of whole inhibitory effect from organic-rich soil.
y = 293.01xR² = 0.917
0
10
20
30
40
50
60
70
80
90
100
0.00 0.05 0.10 0.15 0.20 0.25 0.30
degr
adat
ion
rate
of 2
,7-D
CD
D (%
)
growth amount of fungi (g)
no soil
dissoleved matter
organic-rich soil
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
0
20
40
60
80
100
120
organic-rich soil2,7-DCDD
washed organic-richsoil
2,7-DCDD
no soil2,7-DCDD
reco
very
rat
e of
2,7
-DC
DD
(%)
control
treatment
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
3-2-5) Influence of growth inhibition
Difference of degradation rate of 2,7-DCDD- between “organic-rich soil” and “no soil” was about 60%, - growth inhibition: about 35%, other inhibition: about 25%
Difference of degradation rate of 2,7-DCDD- between “organic-rich soil” and “no soil” was about 60%, - growth inhibition: about 35%, other inhibition: about 25%
25%
35%
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
4-1 Conclusions Growth inhibition by soil components
For degradation of dioxins using white rot fungusGrowth inhibition by soil components
For degradation of dioxins using white rot fungus
2) Organic-rich soil has large inhibition activity- soluble matters has growth inhibition effect
2) Organic-rich soil has large inhibition activity- soluble matters has growth inhibition effect
1) Degradation rate of 1,3,6,8-TCDD: - only 1.1% in case of organic-rich soil- increased according to change liquid-solid ratio
1) Degradation rate of 1,3,6,8-TCDD: - only 1.1% in case of organic-rich soil- increased according to change liquid-solid ratio
3) Difference of degradation rate of 2,7-DCDD between with and without “organic-rich soil”: about 60%- of this, growth inhibition accounted for about 35%
3) Difference of degradation rate of 2,7-DCDD between with and without “organic-rich soil”: about 60%- of this, growth inhibition accounted for about 35%
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
1. Introduction 2. Materials and Methods 3. Results and Discussion 4. Conclusions
4-2 Conclusions
In order to establish remediation method for soil- to important to consider soil components- to control water content
In order to establish remediation method for soil- to important to consider soil components- to control water content
- Water content: most important factor in remediation of contaminated soil- This improvement in degradation of dioxins
- probably caused by dilution of growth inhibition material in soil- In fact, degradation of dioxins increased by soil washing in advance
- Water content: most important factor in remediation of contaminated soil- This improvement in degradation of dioxins
- probably caused by dilution of growth inhibition material in soil- In fact, degradation of dioxins increased by soil washing in advance
Shinya Suzuki, Fukuoka University, Japan. Sardinia 2011
Thank you very much for your attention !Thank you very much for your attention !
