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Environmental fate & pathways

Bioaccumulation: aquatic / sediment

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Reference
Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from peer reviewed journal
Justification for type of information:
Data is from peer reviewed journal
Qualifier:
according to guideline
Guideline:
other: as mentioned below
Principles of method if other than guideline:
Bioaccumulation study was conducted on test organism Bluegill sunfish (Leponis Macrochirus) for evaluating the bioconcentration factor (BCF value) of test chemical.
GLP compliance:
not specified
Specific details on test material used for the study:
- Other: Pure chemicals were purchased from Monsanto Co. and radioactive materials, with respective activities and radiochemical purity were synthesized,
14C-SPME, 0.54 mCi/g., > 99%
Radiolabelling:
yes
Details on sampling:
- Sampling intervals/frequency for test medium samples: On each sample day triplicate 5-mL water samples were collected for analysis.
- Sampling method: Radioactive test chemical stock solutions were prepared once at the beginning of the studies by the addition of a sufficient amount of radiotracer and unlabeled chemical in deionized water to achieve desired concentrations with an acceptable specific activity for each study.
- Sample storage conditions before analysis: The stock solutions were refrigerated in the dark to minimize thermal and photodegradation during storage
Vehicle:
yes
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
TEST ORGANISM
- Common name: Bluegill sunfish (Leponis Macrochirus)
- Source: Test organism Bluegill sunfish was obtained from Nebraska fish farm.
- Length at study initiation (length definition, mean, range and SD): 67±12 mm
- Weight at study initiation (mean and range, SD): 4.9±1.7 g
- Food type: fish was fed a dry pelleted ration ad libitum each day.


ACCLIMATION
- Acclimation period: Test organism was acclimatized in holding tanks at the laboratory for at least 30 days prior to radiotracer exposure
Route of exposure:
aqueous
Test type:
not specified
Water / sediment media type:
natural water: freshwater
Total exposure / uptake duration:
28 d
Total depuration duration:
7 d
Hardness:
35 mg/l CaCo3
Test temperature:
18-20°C
pH:
7.1
Dissolved oxygen:
> 5 mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel: aquaria was used as a test vessel for the study.
- Material, size, headspace, fill volume: Aquaria of 30 l volume was used for the study.
- Aeration: Yes
- Type of flow-through (e.g. peristaltic or proportional diluter): proportional dilution
- No. of organisms per vessel: 30 to 50 fish

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: well water
- Intervals of water quality measurement: Water temperatures were recorded daily while dissolved oxygen was measured at least weekly throughout the study.

Nominal and measured concentrations:
Measured test concentration 0, 6.3 ± 0.3 mg/l and 60.3 ± 2.5 mg/l.
Nominal test concentrations: 0.5 and 50 mg/l
Reference substance (positive control):
not specified
Details on estimation of bioconcentration:
Results from residue study:
Radioactivity present in aqueous samples were determined by addition of the 5-mL samples to scintillation vials containing cocktail and counted by a liquid scintillation counter. Portions of fish carcass were either combusted (14C samples) or digested (35S samples) prior to radioanalysis of the trapped 14C02 or dissolved tissue, respectively. All measurements of radioactivity were made using a Model 2112 Packard Tri-Carb Liquid Scintillation Spectrometer respectively. All data were corrected for background, dilution, quenching and counting efficiency. Minimum detection limits were established for each sample based on the size and radiotracer counting error associated with that sample. Higher detection limits generally were applicable to the visceral tissue data, which was much smaller in sample size than the muscle tissue.
Key result
Temp.:
>= 18 - <= 20 °C
pH:
7.1
Type:
BCF
Value:
< 6.3 dimensionless
Basis:
not specified
Time of plateau:
28 d
Remarks on result:
other: Other details not known
Key result
Temp.:
>= 18 - <= 20 °C
pH:
7.1
Type:
BCF
Value:
< 60.3 dimensionless
Basis:
not specified
Time of plateau:
28 d
Remarks on result:
other: Other details not known
Key result
Elimination:
no
Parameter:
other: No data available
Depuration time (DT):
7 d
Details on results:
Other biological observations:
The mean measured concentrations of 14C-residues present in the muscle of bluegill exposed to 6.3 or 60.3 mg/L 14C-SPME exhibited little, if any, significant accumulation above minimum detectable limits during the entire exposure period. Thus, the maximum bio-concentration factor in the muscle portion of bluegill exposed to either concentration of 14C-SPME was
Incidents in the course of the test which might have influenced the results: During study days 25-28 feeding activity decreased, some fish became dark and lethargic. Death occurred in 28% of this test group. Necropsies conducted on moribund or dead fish failed to find internal or external parasites as a causative agent. Within 2 days of transfer of surviving bluegill to flowing, uncontaminated water these fish resumed a normal behavior and feeding activity increased. These observations suggest that the stress and mortality present in the bluegill population exposed to 60.3 mg/L 14C-SPME may be related to treatment. However, these effects were not substantiated by the toxicity tests conducted.

Radiometric analysis indicated a small but consistent accumulation of radioactivity in visceral portions of fish during the 28-day exposure period. This level never exceeded 4X the minimum detectable limits. No detectable residues were found during depuration.
Validity criteria fulfilled:
not specified
Conclusions:
The bioconcentration factor (BCF value) of test chemical on Bluegill sunfish (Leponis Macrochirus) was determined to be < 6.3 and < 60.3, respectively.
Executive summary:

Bioaccumulation study was conducted on test organism Bluegill sunfish (Leponis Macrochirus)for evaluating the bioconcentration factor (BCF value) of test chemical.Test fish used for the study wasBluegill sunfish (Leponis Macrochirus) obtained from Nebraska fish farmwith a mean wet weight of 4.9±1.7 g and a mean length of 67±12 mm, respectively. All fish wereacclimatized in holding tanks for atleast 30 days prior to radiotracer exposure. Fish were fed a dry pelleted ration ad libitum each day. A modified continuous-flow proportional dilution apparatus was used to establish and maintain the desired chemical concentrations in the test chambers throughout the studies. Thirty-liter aquaria with 30 to 50 fish each were used to expose the test organisms. Aerated well water (pH 7.1, hardness 35 mg/L CaCo3, dissolved oxygen >5 mg/L, temperature 18-20°C was provided to each unit at a flow rate of 5 L/hr. Water temperatures were recorded daily while dissolved oxygen was measured at least weekly throughout the study. Radioactive test chemical stock solutions were prepared once at the beginning of the studies by the addition of a sufficientamount of radiotracer and unlabeled chemical in deionized water toachieve desired concentrations. The stock solutions were refrigerated in the dark to minimize thermal and photodegradation during storage. Test chemical conc. used for the study was 0, 6.3 ± 0.3 and 60.3 ± 2.5 mg/l (Measured test concentration) and 0.5 and 50 mg/l (Nominal test concentrations), respectively. Aquaria of 30 l volume was used as a test vessel for the study.Water and fish were sampled from each aquarium after 1, 7, 10, 21 and 28 days of exposure. Fish remaining after 28 days were transferred to clean flowing water and samples taken on the following days post exposure: 1, 3, 7. Control fish and water were sampled on days 1 and 28 of exposure. On each sample day triplicate 5-mL water samples and 5 fish were collected for analysis. The fish were eviscerated and filleted to provide samples of edible tissues (muscle) and a composite of the remaining portions of the fish. Duplicate portions of muscle tissue (fillets) and individual visceral tissue (viscera and internal organs) of fish from each test group were analyzed by standard radiometric methods.Radioactivity present in aqueous samples were determined by addition of the 5-mL samples to scintillation vials containing cocktail and counted by a liquid scintillation counter. Portions of fish carcass were either combusted (14C samples) or digested (35S samples) prior to radioanalysis of the trapped 14C02 or dissolved tissue, respectively. All measurements of radioactivity were made using a Model 2112 Packard Tri-Carb Liquid Scintillation Spectrometer respectively. All data were corrected for background, dilution, quenching and counting efficiency. Minimum detection limits were established for each sample based on the size and radiotracer counting error associated with that sample. Higher detection limits generally were applicable to the visceral tissue data, which was much smaller in sample size than the muscle tissue. The mean measured concentration of 14C-residues present in the muscle tissue remained below minimum detectable limits throughout the 7 day depuration period.The bioconcentration factor (BCF value) of test chemical on Bluegill sunfish (Leponis Macrochirus) was determined to be <6.3 and <60.3, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the chemical is not expected to bioaccumulate in the food chain.

Description of key information

Bioaccumulation study was conducted on test organism Bluegill sunfish (Leponis Macrochirus)for evaluating the bioconcentration factor (BCF value) of test chemical F. R. Johannsen et. al., 1988). Test fish used for the study wasBluegill sunfish (Leponis Macrochirus) obtained from Nebraska fish farmwith a mean wet weight of 4.9±1.7 g and a mean length of 67±12 mm, respectively. All fish wereacclimatized in holding tanks for atleast 30 days prior to radiotracer exposure. Fish were fed a dry pelleted ration ad libitum each day. A modified continuous-flow proportional dilution apparatus was used to establish and maintain the desired chemical concentrations in the test chambers throughout the studies. Thirty-liter aquaria with 30 to 50 fish each were used to expose the test organisms. Aerated well water (pH 7.1, hardness 35 mg/L CaCo3, dissolved oxygen >5 mg/L, temperature 18-20°C was provided to each unit at a flow rate of 5 L/hr. Water temperatures were recorded daily while dissolved oxygen was measured at least weekly throughout the study. Radioactive test chemical stock solutions were prepared once at the beginning of the studies by the addition of a sufficientamount of radiotracer and unlabeled chemical in deionized water toachieve desired concentrations. The stock solutions were refrigerated in the dark to minimize thermal and photodegradation during storage. Test chemical conc. used for the study was 0, 6.3 ± 0.3 and 60.3 ± 2.5 mg/l (Measured test concentration) and 0.5 and 50 mg/l (Nominal test concentrations), respectively. Aquaria of 30 l volume was used as a test vessel for the study.Water and fish were sampled from each aquarium after 1, 7, 10, 21 and 28 days of exposure. Fish remaining after 28 days were transferred to clean flowing water and samples taken on the following days post exposure: 1, 3, 7. Control fish and water were sampled on days 1 and 28 of exposure. On each sample day triplicate 5-mL water samples and 5 fish were collected for analysis. The fish were eviscerated and filleted to provide samples of edible tissues (muscle) and a composite of the remaining portions of the fish. Duplicate portions of muscle tissue (fillets) and individual visceral tissue (viscera and internal organs) of fish from each test group were analyzed by standard radiometric methods.Radioactivity present in aqueous samples were determined by addition of the 5-mL samples to scintillation vials containing cocktail and counted by a liquid scintillation counter. Portions of fish carcass were either combusted (14C samples) or digested (35S samples) prior to radioanalysis of the trapped 14C02 or dissolved tissue, respectively. All measurements of radioactivity were made using a Model 2112 Packard Tri-Carb Liquid Scintillation Spectrometer respectively. All data were corrected for background, dilution, quenching and counting efficiency. Minimum detection limits were established for each sample based on the size and radiotracer counting error associated with that sample. Higher detection limits generally were applicable to the visceral tissue data, which was much smaller in sample size than the muscle tissue. The mean measured concentration of 14C-residues present in the muscle tissue remained below minimum detectable limits throughout the 7 day depuration period.The bioconcentration factor (BCF value) of test chemical on Bluegill sunfish (Leponis Macrochirus) was determined to be <6.3 and <60.3, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the chemical is not expected to bioaccumulate in the food chain.

Key value for chemical safety assessment

BCF (aquatic species):
60.3 dimensionless

Additional information

Experimental key study & predicted data of the test chemical and various supporting studies for its structurally similar read across substance were reviewed were reviewed for the bioaccumulation end point which are summarized as below:

 

In an experimental key study from peer reviewed journal (F. R. Johannsen et. al., 1988),bioaccumulation experiment was conducted on test organism Bluegill sunfish (Leponis Macrochirus)for evaluating the bioconcentration factor (BCF value) of test chemical. Test fish used for the study was Bluegill sunfish (Leponis Macrochirus) obtained from Nebraska fish farm with a mean wet weight of 4.9±1.7 g and a mean length of 67±12 mm, respectively. All fish were acclimatized in holding tanks for atleast 30 days prior to radiotracer exposure. Fish were fed a dry pelleted ration ad libitum each day. A modified continuous-flow proportional dilution apparatus was used to establish and maintain the desired chemical concentrations in the test chambers throughout the studies. Thirty-liter aquaria with 30 to 50 fish each were used to expose the test organisms. Aerated well water (pH 7.1, hardness 35 mg/L CaCo3, dissolved oxygen >5 mg/L, temperature 18-20°C was provided to each unit at a flow rate of 5 L/hr. Water temperatures were recorded daily while dissolved oxygen was measured at least weekly throughout the study. Radioactive test chemical stock solutions were prepared once at the beginning of the studies by the addition of a sufficient amount of radiotracer and unlabeled chemical in deionized water to achieve desired concentrations. The stock solutions were refrigerated in the dark to minimize thermal and photodegradation during storage. Test chemical conc. used for the study was 0, 6.3 ± 0.3 and 60.3 ± 2.5 mg/l (Measured test concentration) and 0.5 and 50 mg/l (Nominal test concentrations), respectively. Aquaria of 30 l volume was used as a test vessel for the study. Water and fish were sampled from each aquarium after 1, 7, 10, 21 and 28 days of exposure. Fish remaining after 28 days were transferred to clean flowing water and samples taken on the following days post exposure: 1, 3, 7. Control fish and water were sampled on days 1 and 28 of exposure. On each sample day triplicate 5-mL water samples and 5 fish were collected for analysis. The fish were eviscerated and filleted to provide samples of edible tissues (muscle) and a composite of the remaining portions of the fish. Duplicate portions of muscle tissue (fillets) and individual visceral tissue (viscera and internal organs) of fish from each test group were analyzed by standard radiometric methods. Radioactivity present in aqueous samples were determined by addition of the 5-mL samples to scintillation vials containing cocktail and counted by a liquid scintillation counter. Portions of fish carcass were either combusted (14C samples) or digested (35S samples) prior to radioanalysis of the trapped 14C02 or dissolved tissue, respectively. All measurements of radioactivity were made using a Model 2112 Packard Tri-Carb Liquid Scintillation Spectrometer respectively. All data were corrected for background, dilution, quenching and counting efficiency. Minimum detection limits were established for each sample based on the size and radiotracer counting error associated with that sample. Higher detection limits generally were applicable to the visceral tissue data, which was much smaller in sample size than the muscle tissue. The mean measured concentration of 14C-residues present in the muscle tissue remained below minimum detectable limits throughout the 7 day depuration period. The bioconcentration factor (BCF value) of test chemical on Bluegill sunfish (Leponis Macrochirus) was determined to be <6.3 and <60.3, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the chemical is not expected to bioaccumulate in the food chain.

 

In a prediction done using the BCFBAF Program (v3.01) of Estimation Programs Interface, the bioconcentration factor (BCF) of test chemical was estimated. The bioconcentration factor (BCF) of test chemical was estimated to be 3.162 L/kg whole body w.w (at 25 deg C) which does not exceed the bio concentration threshold of 2000, indicating that the test chemical is not expected to bioaccumulate in the food chain.

 

From CompTox Chemistry Dashboard using OPERA (OPEn (quantitative) structure-activity Relationship Application)  V1.02 model in which calculation based on PaDEL descriptors (calculate molecular descriptors and fingerprints of chemical), the bioaccumulation i.e BCF for test chemical was estimated to be 6.46 dimensionless . The predicted BCF result based on the 5 OECD principles. Thus based on the result it is concluded that the test chemical is non-bioaccumulative in nature.

 

In a supporting study from authoritative database (J-CHECK, 2018) for the test chemical,bioaccumulation experiment was conducted on test organism Cyprinus carpio for 6 weeks for evaluating the bioconcentration factor (BCF value) of test chemical. The study was performed according to “OECD Guideline 305 C (Bioaccumulation: Test for the Degree of Bioconcentration in Fish)” and other guideline "Bioaccumulation test of a chemical substance in fish or shellfish" provided in "the Notice on the Test Method Concerning New Chemical Substances", respectively. Cyprinus carpio was used as a test organism for the study. Test chemical nominal conc. used for the study was 0.5 mg/land 0.05 mg/l, respectively. Analytical method involve therecovery ratio: Fish : 84.8 %, - Limit of quantitation : Test water : 1st concentration area : 17 microg/L, 2nd concentration area :1.7 microg/L, Fish : 200 ng/g. Range finding study involves the LC50 (48 hr) ≥ 500 mg/l (w/v) on Rice fish (Oryzias latipes). Lipid content of the test organism Cyprinus carpio was determined to be 3.7% at the start of exposure. The bioconcentration factor (BCF value) of test chemical on Cyprinus carpio was determined to be<= 0.4L/Kg at a conc. of 0.5 mg/l and <= 4.1 L/Kg at a conc. of 0.05 mg/l, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the test chemical is not expected to bioaccumulate in the food chain.

 

For the test chemical, bioaccumulation experiment was conducted on test organism Cyprinus carpio for 6 weeks for evaluating the bioconcentration factor (BCF value) of test chemical (authoritative database, 2018). The study was performed according to other guideline "Bioaccumulation test of a chemical substance in fish or shellfish" provided in "the Notice on the Test Method Concerning New Chemical Substances", respectively. Cyprinus carpio was used as a test organism for the study. Test chemical nominal conc. used for the study was 1 mg/l and 0.1 mg/l, respectively. Analytical method involve the recovery ratio: Test water: 1st concentration area: 83.9 %, 2nd concentration area: 82.4 %, Fish : 88.1 %, - Limit of detection : Fish : 0.38 ppm. Range finding study involves the TLm (48 hr) ≥ 1000 mg/l (w/v) on Rice fish (Oryzias latipes). Lipid content of the test organism Cyprinus carpio was determined to be 2.8%. The bioconcentration factor (BCF value) of test chemical on Cyprinus carpio was determined to be<= 0.43L/Kg at a conc. of 1 mg/l and<= 3.8 L/Kg at a conc. of 0.1 mg/l, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the test chemical is not expected to bioaccumulate in the food chain.

 

On the basis of above results for test chemical, it can be concluded that the BCF value of test chemical was evaluated to be upto 60.3, respectively, which does not exceed the bioconcentration threshold of 2000, indicating that the test chemical is not expected to bioaccumulate in the food chain.