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Environmental fate & pathways

Biodegradation in soil

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Endpoint:
biodegradation in soil: simulation testing
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because the substance is readily biodegradable
Endpoint:
biodegradation in soil, other
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Test conditions and methods not described, no quantitative results.
Principles of method if other than guideline:
no data
GLP compliance:
no
Test type:
laboratory
Radiolabelling:
no
Oxygen conditions:
not specified
Soil classification:
not specified
Details on soil characteristics:
no data
Initial conc.:
0.25 mg/kg soil d.w.
Parameter followed for biodegradation estimation:
test mat. analysis
Details on experimental conditions:
Vanillin degradation was tested for 4 different soils: garden soil, rendzina forest soil, raw humus sampled under Rhododendron ferrugineum, and
river deposits.
0.25 g/l of solutions of vanillin were inoculated with each soil type at the dilution rate of 1x10-3.

During this study, the authors had isolated various microorganism genera or species able to degrade vanillin: Fusarium sp., Trichoderma virile,
Penicillium sp. and a gram negative coccoid bacterium. These microorganisms are supposed to be able to carry out the three main metabolic
mechanisms allowing to an ultimate degradation of vanillin: oxidation, demethylation, cycle opening.
Remarks on result:
other: no degradation rate available
Transformation products:
yes
No.:
#1
Details on transformation products:
No degradation rate was presented by the authors. Two degradation products were observed, vanillic acid and a compound that reacts with phenylhydrazine.
The vanillin degradation efficiency differs between each soil type. In an decreasing order, the soil's degradation potency is: rendzina forest soil >
garden soil >>> raw humus sampled = river deposits.
Evaporation of parent compound:
not specified
Volatile metabolites:
not specified
Residues:
not specified

No more data on results.

Endpoint:
biodegradation in soil, other
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test conditions and methods well described.
Guideline:
other: no data
GLP compliance:
not specified
Test type:
laboratory
Radiolabelling:
no
Oxygen conditions:
aerobic
Soil classification:
other: natural soil amended with 9 % montmorillonite
Year:
1984
% Clay:
9
% Silt:
34
% Sand:
57
% Org. C:
5.8
pH:
5.6
CEC:
8.2 meq/100 g soil d.w.
Details on soil characteristics:
SOIL COLLECTION AND STORAGE
- Geographic location: Brooklyn Botanic Garden, Ossining, New York, US
- Pesticide use history at the collection site: No data
- Collection procedures: No data
- Sampling depth (cm): No data
- Storage conditions: Room temperature
- Storage length: 8 years
- Soil preparation (e.g., 2 mm sieved; air dried etc.): Air dried, 2 mm sieved and remaining soil were moistened (6 to 7% water) and mixed daily for 1 week.

PROPERTIES OF THE SOILS (in addition to defined fields)
Mineral soil composition: Vermiculite, kaolinite, illite, mica but not montmorillonite, N 0.1%, P 28 µg/g, K 62 µg/g, Ca 360 µg/g
3 different types of soil were used: amended with 9% (vol/vol) of kaolinite or montmorillonite or not amended.
Initial conc.:
1 190 mg/kg soil d.w.
Parameter followed for biodegradation estimation:
CO2 evolution
Temp.:
25 °C
Humidity:
7 g water/100g soil dry weight
Details on experimental conditions:
EXPERIMENTAL DESIGN
- Soil preincubation conditions (duration, temperature if applicable): 16h at 4 deg.C. After 1 week of incubation, no significant amounts of CO2 were detected. A suspension of active "garden soil" was prepared (10 % wt/vol) and diluted 10-fold. 2ml aliquots were inoculated into each soil in an
attempt to increase the number of organisms capable of utilising vanillin. The soils were then incubated for another 3 weeks.
- Soil (g/replicate): 100g
- No. of replication control and treatments: 3 replicates
- Test apparatus (Type/material/volume): Soil samples were placed in incubation vessel, which were connected to a manifold and continuously
aerated with water saturated CO2-free air at 25+/-3 deg.C
Other details: 0, 0.1, 0.3, 0.6, 1.2, or 2ml concentrated H2SO4 mixed in sufficient water to bring the soil to their 1/3 atm tension water content
(20.7% for soil without added clays, 21.8 and 23.4% for the soils amended with kaolinite and montmorillonite, respectively. Then the soils were stored for 16h at 4 deg.C and passed through a 2mm sieve.

OXYGEN CONDITIONS
- Methods used to create the an/aerobic conditions: Test vessels were continuously aerated with water saturated CO2-free air at 25+/-3 deg.C
% Degr.:
41
Parameter:
CO2 evolution
Sampling time:
21 d
Transformation products:
not measured
Evaporation of parent compound:
not specified
Volatile metabolites:
not specified
Residues:
not specified
Details on results:
%DEGRADATION OF TEST SUBSTANCE:
After 3 weeks of incubation, pH = 5-5.6, based on CO2 evolution
- Montmorillonite-amended soil: 41 %
- Kaolinite-amended soil: 11%
- Unamended soil: 9%

MAIN RESULTS
In the montmorillonite-amended soils, there was a progressive decrease in the total amount of carbon mineralized as the soil acidity increased, with complete inhibition in the soil pH of 1.6. pH was not alone responsible for the low utilization of vanillin in the kaolinite-amended and clay-unamended soils.
At the end of the experiment, the pH of the montmorillonite-amended soils increased from 0.3 to 0.9 pH units.

Table 1: Total carbon evolves as CO2 (mgC/100g soil) from soil-clay  mixtures treated with H2SO4 and amended with vanillin 

(yield 75mg C/100 soil) and the % of vanillin mineralisation after 3 weeks of incubation  (mean +/- standard error).

Ml H2SO4/      No clay added                9% kaolinite          9%  montmorillonite
  100g soil       pH        C        %           pH         C      %        pH           C          %
        0             5.1    7+/-2.3    9            5     8+/-1.1   11       5.6      31+/-2.2    41
       0.1           3.8    4+/-0.9    5          3.8    3+/-0.4     4       4.2      30+/-1.9    40
       0.3           3.2    2+/-0.6    3          3.2    4+/-1.1     5       3.4      26+/-4       35
       0.6           2.8    2+/-0.5    3          2.9    4+/-1.1     5       2.8      22+/-0.2    29
       1.2           2.3    3+/-0.7    4          2.2    3+/-0.6     4       2.2      10+/-2.1    13
        2             1.8    3+/-0.3    4          1.8    3+/-0.6     4       1.6      2+/-0.4        3

Endpoint:
biodegradation in soil, other
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Test conditions and methods poorly described, results obtained by graphical interpretation.
Principles of method if other than guideline:
Measurement of O2 consumption in aqueous soil suspension
GLP compliance:
no
Test type:
laboratory
Radiolabelling:
no
Oxygen conditions:
not specified
Soil classification:
not specified
% Org. C:
1.9
pH:
8
Details on soil characteristics:
SOIL COLLECTION AND STORAGE
- Geographic location: Chernozem soil (Libeznice, Czech Republic)
- Sampling depth (cm): Superficial top soil (0-15 cm)
- Storage conditions: no data
- Storage length: no data
- Soil preparation (e.g., 2 mm sieved; air dried etc.): 2-5 mm aggregates

SOIL PROPERTIES
- Soil type: Chernozem
- % total organic carbon: 1.92%
- % nitrogen: 0.22%
- % phosphorus: 0.009%
- pH: 8 in aqueous suspension (1 : 2.5)
- Water holding capacity: 48.7%
Initial conc.:
2 360 mg/kg soil d.w.
Parameter followed for biodegradation estimation:
O2 consumption
Details on experimental conditions:
EXPERIMENTAL DESIGN
A. First experiment:
- Soil preincubation conditions: no data
- Soil condition: air dried
- Soil (g/replicate):0.5
- No. of replication controls, if used: 2
- No. of replication treatments: 2
- Test apparatus (Type/material/volume): Warburg manometric method
Test material application
- Volume of test solution used/treatment: In addition to 2 ml soil suspension containing 0.58 g of dry soil, 1 ml of water (control) or vanillin was added into Warburg respiration vessel. Vanillin concentration was equal to 0.49 mg C in the added ml, that is 2.360 g vanillin/kg dry soil.
% Degr.:
ca. 69
Parameter:
O2 consumption
Sampling time:
72 h
DT50:
35 h
Type:
(pseudo-)first order (= half-life)
Remarks on result:
other: no data available on T°C"
Transformation products:
not measured
Details on results:
The vanillin dissipation half-life was estimated by graphical  interpretation; the value is around 35 hours. Vanillin degradation kinetic reaches a 
plateau tending to 75% beyond 80 hours of experiments.  The degradation rate increases of 10 % during the last 40th hours.
Endpoint:
biodegradation in soil, other
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Test conditions and methods not described, results obtained by graphical interpretation.
Principles of method if other than guideline:
not detailed
GLP compliance:
no
Test type:
laboratory
Radiolabelling:
no
Oxygen conditions:
not specified
Soil classification:
not specified
Details on soil characteristics:
SOIL PROPERTIES
The soil contains vermiculite, kaolinite, illite and mica, but not montmorillonite.
It was amended with approximately 5, 10 or 20% (v/v) with montmorillonite just before experimentation (fresh) or 3 years previously (old).
Samples of soil had also been amended 3 years previously with 5, 10 or 20% (v/v) kaolinite or 5% (v/v) mica-vermiculite).
- Soil preparation (e.g., 2 mm sieved; air dried etc.): The soils were air dried and sieved. Aggregates between 0.5 and 2 mm were used.
Initial conc.:
1 205 mg/kg soil d.w.
Details on experimental conditions:
CONCENTRATION OF PARENT COMPOUNDS
The soils were amended with 50 mg C/100 g of air dried soil, which corresponds to a vanillin of 1.205 g/kg of dried soil.

No more details are provided on test conditions, soil preparation and physicochemical properties, these have been previously described in Kunc and Stotzky (1974).

Kunc F, Statzky G (1974).Effect of glucose on vanillic acid oxidation in Cellulomonas sp. Fol. Microbiol. 19, pp 29-37.
DT50:
ca. 2.3 - ca. 5 d
Type:
(pseudo-)first order (= half-life)
Remarks on result:
other: no data available on T°C
Transformation products:
not measured
Evaporation of parent compound:
not specified
Volatile metabolites:
not specified
Residues:
not specified
Details on results:
The results presented in the table below are obtained by graphic interpretation. The vanillin degradation rate increases with the montmorillonite load in the soil. The dissipation half-life time of vanillin is 5 days in the control soil and less than 2.5 days in a soil amended with 20% (v/v)
montmorillonite.

Table: Vanillin dissipation half-life time related to the montmorillonite  load in the soil.
Montmorillonite load (%)        0     5      10      20
DT50 (days)                        5.0     4.1     3.9     2.3

Endpoint:
biodegradation in soil, other
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Test conditions and methods poorly described, results obtained by graphical interpretation.
Principles of method if other than guideline:
no data
GLP compliance:
no
Test type:
laboratory
Radiolabelling:
no
Oxygen conditions:
not specified
Soil classification:
not specified
Soil type:
other: arable land on sandy soil
% Org. C:
1.2
pH:
6.2
Details on soil characteristics:
SOIL COLLECTION AND STORAGE
- Geographic location: Holland
- Sampling depth (cm): 0-5 cm
- Storage conditions: no data
- Storage length: no data
- Soil preparation: the soil was air dried to 30% of water holding capacity and passed through a 2 mm sieve.

SOIL PROPERTIES
No other properties mentionned.
Duration:
150 - 300 h
Initial conc.:
4.8 mg/kg soil d.w.
Parameter followed for biodegradation estimation:
CO2 evolution
Temp.:
20 °C
Humidity:
60 other: % of water-holding capacity
Details on experimental conditions:
EXPERIMENTAL DESIGN
- Soil condition: Soil previously dried to 30% of the water holding capacity and the moisture was adjusted to 60% just before the experiment and
periodically humidified to keep the water content constant.
- Soil (g/replicate): for the experiment performed at 20 deg. C, the soil amount was 10 g, and for the one performed at 13 deg. C the sample amount was 50g.
- Test apparatus (Type/material/volume): Not specified
- CO2 concentration was measured as previously described in Huntjens (1979).
ref: Huntjens JLM (1979). A sensitive method for continous measurement of the carbon dioxide evolution rate of soil samples. Plant and Soil. Vol 53, pp 529-534.

INITIAL TEST SUBSTANCE CONCENTRATION:
Two experiments with two different temperatures and vanillin concentrations were performed:
T=20 deg. C: [C] ca. 1000 µgC/g dry soil = 4.8 g vanillin/kg dry soil
T=13 deg. C: [C] ca. 540 µgC/g dry soil = 2.6 g vanillin/kg dry soil
% Degr.:
ca. 55
Parameter:
CO2 evolution
Sampling time:
300 h
DT50:
ca. 250 h
Type:
(pseudo-)first order (= half-life)
Temp.:
20 °C
Transformation products:
not measured
Evaporation of parent compound:
not specified
Volatile metabolites:
not specified
Residues:
not specified
Details on results:
All the results presented are obtained by graphical interpretation. At 20 deg. C, vanillin was decomposed very slowly during the first 100 hours.
After 150 hours of incubation less than 10% of the compound was degraded. The dissipation half life is around 250 hours and at the end of the
experiment, after 300 hours of incubation, the degradation rate was near 55%. At 13 deg C, the only degradation value available is for an incubation time of 150 hours. After this delay, vanillin was decomposed at about 20%.

Description of key information

None of the publications dealing with the degradation of vanillin in soil have been selected as key study. However, they can be used in a weight of evidence approach to report that vanillin can be degraded in different soils, in different conditions of temperature (13 and 20°C) and that vanillic acid, which is itself degraded in soil, is one of the degradation product.

Key value for chemical safety assessment

Additional information

It is to be noted that since vanillin is readily biodegradable, further degradation in soil would not be necessary to investigate.