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Long-term toxicity to aquatic invertebrates

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Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
Mortality tests were conducted with a flowthrough exposure system consisting of a multichannel toxicant injection system.
GLP compliance:
not specified
Specific details on test material used for the study:
sodium arsenite (Total As III )NaAsO2
Analytical monitoring:
yes
Details on sampling:
Samples were collected for analyses every 24 h during the test.
Vehicle:
no
Details on test solutions:
Stock solutions of metals were freshly prepared by dissolving the appropriate salts in deionized water in 1-L glass volumetric flasks. The sodium arsenite stock solution was prepared with deionized water (AsIII). For the mortality test, three concentrations were made using a peristaltic pump, which continuously mixed the sodium arsenite solution with river water.
Test organisms (species):
other aquatic crustacea: Asellus aquaticus
Details on test organisms:
Test organism: Asellus aquaticus
Taxonomic stage: larvae

The hypogean crustacean was collected with a manual pump (at 50 cm depth in the sediment)
Sixty organisms per species were used for each lethal toxicity experiment and thirty organisms per species per replicate for each concentration were used for bioaccumulation experiments. The organisms were transported to the laboratory in an ice-cooled container, then placed in oxygenated river water in a cooled chamber until the beginning of the experiment.
They were acclimatized to laboratory conditions for 2 days before experimentation. Organisms were collected in April 1997 for the arsenic experiments. Only last-instar larvae stages were kept for the experiments.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
10 d
Test temperature:
12°C
pH:
7.6-8.4
Dissolved oxygen:
10 mg/L
Nominal and measured concentrations:
Measured mean concentration: 20, 1230 and 4200 µg As/L in test 1 and 97.3 µg As/L in test 2.
Details on test conditions:
Mortality tests were conducted with a flowthrough exposure system consisting of a multichannel toxicant injection system, which delivered the toxicant at three concentrations and a control.
Tests were carried out with filtered river water collected with the organisms.
River water and contaminant were continuously mixed and went through compartmentalized glass tubes containing the invertebrates.

Test 1:
Each organism was separated from the others and placed in an individual compartment. Tests were performed in a room with a constant temperature of 12 ± 2°C, and light was controlled on a 14 h light/10 h dark cycle. Three replicates of five organisms per species were used for each concentration. The flow rate to each replicate was 50 ml/min for a 3-L water volume (9 L per concentration).

Test 2:
Bioaccumulation was studied in glass aquariums (5 L) for 10-day experiments at a nomial conentrationof 100 µg As/L. Arsenic solutions were continuously renewed with a peristaltic pump, and water samples were collected for analyses every 24 h during the test. The experimental design was set up also in the cooled chamber (temperature: 12 ± 2°C), with a 14/10 photoperiod. Three replicates of 30 organisms were performed for each species,
Key result
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
97.3 µg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
2 310 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Reported statistics and error estimates:
LC50 were determined by means of the Spearman-Karber methodology
Validity criteria fulfilled:
not applicable
Conclusions:
The LC10 value of the species Asellus aquaticus was reported to be 97.3 µg As/L during the larvae life stage.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
Mortality tests were conducted with a flowthrough exposure system consisting of a multichannel toxicant injection system.
GLP compliance:
not specified
Specific details on test material used for the study:
sodium arsenite (Total As III )NaAsO2
Analytical monitoring:
yes
Details on sampling:
Samples were collected for analyses every 24 h during the test.
Vehicle:
no
Details on test solutions:
Stock solutions of metals were freshly prepared by dissolving the appropriate salts in deionized water in 1-L glass volumetric flasks. The sodium arsenite stock solution was prepared with deionized water (AsIII). For the mortality test, three concentrations were made using a peristaltic pump, which continuously mixed the sodium arsenite solution with river water.
Test organisms (species):
other aquatic arthropod: Hydropsiche pellucidula
Details on test organisms:
Test organism: Hydropsiche pellucidula
Taxonomic stage: larvae

The organisms were sampled using a hand net (500 􏰍m mesh size).
Sixty organisms per species were used for each lethal toxicity experiment and thirty organisms per species per replicate for each concentration were used for bioaccumulation experiments. The organisms were transported to the laboratory in an ice-cooled container, then placed in oxygenated river water in a cooled chamber until the beginning of the experiment.
They were acclimatized to laboratory conditions for 2 days before experimentation. Organisms were collected in April 1997 for the arsenic experiments. Only last-instar larvae stages were kept for the experiments.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
10 d
Test temperature:
12°C
pH:
7.6-8.4
Dissolved oxygen:
10 mg/L
Nominal and measured concentrations:
Measured mean concentration: 20, 1230 and 4200 µg As/L in test 1 and 98 µg As/L in test 2.
Details on test conditions:
Mortality tests were conducted with a flowthrough exposure system consisting of a multichannel toxicant injection system, which delivered the toxicant at three concentrations and a control.
Tests were carried out with filtered river water collected with the organisms.
River water and contaminant were continuously mixed and went through compartmentalized glass tubes containing the invertebrates.

Test 1:
Each organism was separated from the others and placed in an individual compartment. Tests were performed in a room with a constant temperature of 12 ± 2°C, and light was controlled on a 14 h light/10 h dark cycle. Three replicates of five organisms per species were used for each concentration. The flow rate to each replicate was 50 ml/min for a 3-L water volume (9 L per concentration).

Test 2:
Bioaccumulation was studied in glass aquariums (5 L) for 10-day experiments at a nomial conentrationof 100 µg As/L. Arsenic solutions were continuously renewed with a peristaltic pump, and water samples were collected for analyses every 24 h during the test. The experimental design was set up also in the cooled chamber (temperature: 12 ± 2°C), with a 14/10 photoperiod. Three replicates of 30 organisms were performed for each species,
Key result
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
98 µg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
2 400 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Reported statistics and error estimates:
LC50 were determined by means of the Spearman-Karber methodology
Validity criteria fulfilled:
not applicable
Conclusions:
The LC10 value of the species Hydropsiche pellucidula was reported to be 98 µg As/L during the larvae life stage.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
Toxicity tests were performed with embryos of Paracentrotus lividus to investigate the toxicological effect of arsenate (AsV).
Exposures to toxicants were performed.
GLP compliance:
not specified
Specific details on test material used for the study:
Arsenic was administrated as arsenate (AsV) (Sigma-Aldrich, USA).
Analytical monitoring:
yes
Vehicle:
no
Test organisms (species):
other: Paracentrotus lividus
Details on test organisms:
TEST ORGANISM: Paracentrotus lividus
Sea urchin adults of the species P. lividus were collected in May 2012 from an intertidal rocky site along the coast of Livorno (Italy) [43°250 320 0 N, 10°230 380 0 E] and transported to the laboratory within 1 h from the collection.

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS:_
Organisms, in a minimum of 3 individuals per sex, were used to obtain gametes by injecting 1 mL of 0.5 M KCl solution into the coelom through the peristome. Sperm from each male was collected and preserved at 4°C, while oocytes from each female were shed into 50-mL beakers previously filled with 0.22 µm Filtered Sea Water (FSW). Spawned oocytes were allowed to settle and washed three times with FSW; cell solutions were then diluted to a final concentration of 1,000 oocytes/mL. Once sperm mobility had been checked, sperm density was determined by means of a hemocytometer (Thoma chamber). Sperm was then diluted into filtered seawater in order to reach a sperm: eggs ratio of 15,000:1 (Lera and Pellegrini 2006). Fertilization was executed by gently mixing the sperm and egg suspension.
Embryos were kept at 20°C in a controlled temperature chamber, a salinity of 38 ± 1 % and 9 h daylight.
Test type:
static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
72 h
Test temperature:
20 °C
Salinity:
38 ppt
Nominal and measured concentrations:
Nominal concentrations: 50, 100, and 200 µg/L
Measured arsenic in tested solutions was respectively 53±4, 98±8 and 210±6 µg As/L
Details on test conditions:
Arsenic molecules were administered to untreated plutei at the attainment of different developmental stages.
Measured arsenic in tested solutions was respectively 53±4, 98±8 and 210±6 µg/L.
Exposure ended for all the treatments after 72 h from fecundation, when pluteus stage was achieved.
Six replicates were fixed for each concentration and the final volume of the well plate was 10 mL.

Normal plutei were discriminated from incomplete developed plutei, the ones with skeletal aberrations (apex spicules cross-linked or separated) and
the ones with deformation of gastrointestinal apparatus.
The acceptability of the results was fixed at a percentage of normal plutei C80 % in negative control tests.
Reference substance (positive control):
yes
Remarks:
exposure to CuNO3*3H2O (Sigma Aldrich, USA)
Key result
Duration:
72 h
Dose descriptor:
EC10
Effect conc.:
56 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
97 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Basis for effect:
morphology
Results with reference substance (positive control):
EC50 mean value ± SD for the positive control (54.23 ± 2.13 µg/L) was consistent with value reported in literature confirming the good health of gametes
Reported statistics and error estimates:
EC50 and EC10 have been re-calculated using Trap software (log-logistic curve fitting)
Validity criteria fulfilled:
yes
Remarks:
The percentage of normal plutei in control groups ranged from 80 to 85 % in all the experiments. EC50 mean value ± SD for the positive control (54.23 ± 2.13 µg/L) was consistent with value reported in literature confirming the good health of gametes.
Conclusions:
The EC10 value of Paracentrotus lividus (marine invertebrate) was reported to be 56 µg As /L during the embryo life stage.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
28 days toxicity test of As on Cyclopoids taxa.
GLP compliance:
not specified
Specific details on test material used for the study:
NaAsO2 (Sigma-Aldrich, 98 % purity)
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
Toxicant stock solutions were made up in Milli-Q water. Arsenic was added as NaAsO2 (Sigma-Aldrich, 98 % purity).
Test organisms (species):
other: Cyclopoids taxa 1 and 2
Details on test organisms:
Sources:
Cyclopoids taxa 1: collected from a groundwater-fed, upland peat swamp in the Budderoo National Park near Robertson, NSW, Australia. Animals were
collected using a bailer with which groundwater was withdrawn from a shallow piezometer. Ten litres of water was withdrawn from the piezometer and passed through a 63-μm-mesh sieve to collect the invertebrates.

Cyclopoids taxa 2: collected from a fractured sandstone aquifer at Somersby, NSW Australia. Groundwater was pumped using a motorised inertia pump (Waterra, ON, Canada). Three hundred litres of water were removed and passed through a 63-μm-mesh sieve to collect the invertebrates.

All animals were acclimated to conditions in the environmental cabinet for at least 48 h prior to testing. Tests were commenced within 7 days of collection.
There was no evidence of organism mortality during this pretest period.
Test type:
not specified
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Hardness:
Cyclopoids taxa 1: 8-37 mg/L CaCO3
Cyclopoids taxa 2: 25-44 mg/L CaCO3
Test temperature:
18 °C
pH:
Cyclopoids taxa 1: 4.5-6
Cyclopoids taxa 2: 4.2-5.6
Dissolved oxygen:
Cyclopoids taxa 1: 18-52%
Cyclopoids taxa 2: 59-83%
Nominal and measured concentrations:
0-5000 mg As/L
Details on test conditions:
Toxicity tests were conducted in untreated 24-well tissue culture plates. Animals were placed individually into wells using a micropipette
with a known volume of diluent water. The number of individuals used in a test was dictated by the number collected.
Tests with only a small number (≤35) of individuals were repeated.
Tests were conducted using the groundwater from the site where the animals were collected as the diluent water.
Treatment concentrations were randomly allocated to each well, and the required volume of clean groundwater water was added such that the subsequent addition of toxicant solution would give a final test volume of 2.0 mL. Each test was comprised of a negative control and five or seven test concentrations, each with four or three replicate wells per plate
Key result
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
2 670 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Remarks:
Cyclopoids taxa 1
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
10 720 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Remarks:
Cyclopoids taxa 1
Key result
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
40 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Remarks:
Cyclopoids taxa 2
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
5 240 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Remarks:
Cyclopoids taxa 2
Reported statistics and error estimates:
LCx using best fitting curve from log-logistic, weibull and log-normal
Validity criteria fulfilled:
not specified
Remarks:
Control mortality was <10%; dose-response data are not provided; the organisms are groundwater organisms; Nominal As was confirmed by measurements
Conclusions:
During the 28 days toxicity test the LC10 on mortality was as follows:
Cyclopoids taxa 1 LC10: 2.670 mg As/L
Cyclopoids taxa 2 LC10: 0.040 mg As/L
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
28 days toxicity test of As on Harpactoid taxa
GLP compliance:
not specified
Specific details on test material used for the study:
NaAsO2 (Sigma-Aldrich, 98 % purity)
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
Toxicant stock solutions were made up in Milli-Q water. Arsenic was added as NaAsO2 (Sigma-Aldrich, 98 % purity).
Test organisms (species):
other: Harpactoid taxa
Details on test organisms:
collected from a fractured sandstone aquifer at Somersby, NSW Australia. Groundwater was pumped using a motorised inertia pump (Waterra, ON, Canada). Three hundred litres of water were removed and passed through a 63-μm-mesh sieve to collect the invertebrates.

All animals were acclimated to conditions in the environmental cabinet for at least 48 h prior to testing. Tests were commenced within 7 days of collection.
There was no evidence of organism mortality during this pretest period.
Test type:
not specified
Water media type:
freshwater
Limit test:
no
Total exposure duration:
14 d
Hardness:
25-44 mg/L CaCO3
Test temperature:
18 °C
pH:
4.2-5.6
Dissolved oxygen:
59-83%
Nominal and measured concentrations:
0-5000 mg As/L
Details on test conditions:
Toxicity tests were conducted in untreated 24-well tissue culture plates. Animals were placed individually into wells using a micropipette
with a known volume of diluent water. The number of individuals used in a test was dictated by the number collected.
Tests with only a small number (≤35) of individuals were repeated.
Tests were conducted using the groundwater from the site where the animals were collected as the diluent water.
Treatment concentrations were randomly allocated to each well, and the required volume of clean groundwater water was added such that the subsequent addition of toxicant solution would give a final test volume of 2.0 mL. Each test was comprised of a negative control and five or seven test concentrations, each with four or three replicate wells per plate
Key result
Duration:
14 d
Dose descriptor:
EC10
Effect conc.:
130 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
3 040 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Reported statistics and error estimates:
LCx using best fitting curve from log-logistic, weibull and log-normal
Validity criteria fulfilled:
not specified
Remarks:
Control mortality was <10%; dose-response data are not provided; the organisms are groundwater organisms; Nominal As was confirmed by measurements
Conclusions:
During the 14 days toxicity test of Harpactoid taxa on the entpoint mortality was LC10=0.130 mg As/ L.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
long-term toxicity test to Daphnia magna
GLP compliance:
not specified
Specific details on test material used for the study:
Purity: reagent grade
NaAsO2
Analytical monitoring:
yes
Details on sampling:
Samples were collected and analyzed three times weekly
Vehicle:
no
Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnids were also obtained from Environmental Research Laboratory-Duluth (ERL-D), Duluth, MN stock culture.
Life stage: < 24h neonates
They were rearedin 25°C Lake Superior water and fed a mixture of ground trout pellets and yeast(ASTM 1980).
The food mixture was prepared by homogenizing 60 g of fish food pelletswith 1.5 g of bread-makers yeast in water.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Hardness:
46.3-49.9 mg/L CaCO3
Test temperature:
20.8°C
pH:
7.2-8.1
Dissolved oxygen:
79.6 - 99.3%
Nominal and measured concentrations:
0.0728, 0.132, 0.27, 0.633, 1.32 and 2.67 mg As/L
Test concentrations were analytically confirmed.
Details on test conditions:
All tests were conductedin proportional diluters.
The diluters were calibrated to provide 0.5 dilution factors for five toxicant concentrations in duplicate. Duplicate control chambers received dilution water only. Stocksolutions were delivered to the diluter mixing cell with a metering pump. Exposure water was from the same source as culture water. Fluorescent lights provided a daily photoperiod with 16 hr of light.
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
633 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
633 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
growth
Remarks:
length
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
633 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
reproduction
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
1.32 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
other: survival, growth and offspring production
Reported statistics and error estimates:
ANOVA using Dunnett’s test
Validity criteria fulfilled:
not specified
Conclusions:
The reliable long-term NOEC value reported for freshwater invertebrates is 633 µg As/L for the survival, growth and offspring production of Daphnia magna.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
48h long toxicity study on Brachionus patulus.
GLP compliance:
not specified
Specific details on test material used for the study:
atomic absorption standards of As dissolved in 1% HNO3(As2O3), Sigma Co., Saint Louis MO, USA
Analytical monitoring:
yes
Vehicle:
no
Test organisms (species):
other: Brachionus patulus
Details on test organisms:
life stage: 72 h old; 0.2 mm length
source: The organisms were collected in Chichimeco reservoir. Aguascalientes State, Mexico, with a 120- μm-mesh-size zooplankton net.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
80-100 mg/L as CaCO3
Test temperature:
25 °C
pH:
7.5
Nominal and measured concentrations:
between 0.17 and 2.4 mg/L as As2O3
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
1 500 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
737 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Reported statistics and error estimates:
NOEC using Duncan multiple range tests
LC50 have been calculated using probit analysis
Validity criteria fulfilled:
not specified
Conclusions:
After the 48h toxicity test on Brachionus patulus, the following results were obtained: NOEC: 1500 µg As/L and LC50: 737 µg As/L, both for mortality endpoint.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
long-term saltwater toxicity test to crustacean
GLP compliance:
not specified
Specific details on test material used for the study:
NaAsO2
Analytical monitoring:
yes
Details on sampling:
Water samples from test chambers of the control and each concentration were analyzed twice weekly for the test metal (as total µg metal)
Vehicle:
no
Details on test solutions:
Primary atomic absorption standards were used as the toxicant sources in all tests. Toxicant solutions were prepared from these standards in deionized water at one hundred times the final desired concentration,The stocksolutions were metered at a constant rate with a peristaltic pump, and proportionally diluted (1:100) with 15/µm filtered sea water immediately before entering the test chambers.
Test organisms (species):
Americamysis bahia (previous name: Mysidopsis bahia)
Details on test organisms:
Animals used in these tests were from our laboratory cultures. The original culturesof M. bahiaweresuppliedby the EPA Environmental ResearchL aboratory at Gulf Breeze,Florida. The animals were acclimated from 23 ppt salinity to 30 ppt salinity by gradually adding filtered (15/µm) natural seawater over a five-day period so that salinity did not vary more than 2 ppt per 24 h. Seawater was heatedto maintain the temperatureat 23 +/- 2°C.
Mysids were cultured for several generations in flow-through 76 1(20 gal) glass aquaria according to Gentile et al. (1982b). Cultures were continuously supplied with filtered (15/µm) natural seawater at a rate of 100 ml/min, resulting in a 99% volume exchange every 24 h. The outflows were coveredby Nitex screen(250/µm) to prevent the escape of mysids or food organisms.Under gravel filters were used with a dolomite substrate one inch deepto maintain a pH of 7.8-8.2. The dolomite was covered by one inch of beachs and, the preferred substrate for these epibenthic animals. The sand was prewashed in deionized water and muffled at 500°F for 6 h to remove organic material.
Twenty-four hour post hatch nauplii of Artemia salina were added as food organisms at the rate of 7 x 104nauplii/day for each 76 1 culture (approximately 47 nauplii/mysid). This ration was adjusted according to mysid density; however, cultures were always fed ad libitum to prevent cannibalism of the young. Gentle aeration served to keep food in suspension and to provide a current conducive to feeding.
Test type:
flow-through
Water media type:
saltwater
Limit test:
no
Total exposure duration:
36 d
Test temperature:
20-25°C
pH:
7.8-8.2
Salinity:
30 ppt
Nominal and measured concentrations:
<3 (control), 144, 303, 631 and 1270 µg As/L
Details on test conditions:
The exposure system employed a siphon-flush mechanism that produced a 50% exchange of volume every 4 h. The approximate flow rates were 30 ml/min inflow, and 100 ml/min outflow. Tests were conducted with filtered (15 µm) natural seawater at 30 + / -2 mg/kg salinity and temperatures ranging from 20-25°C depending on the compound tested. Within an individual toxicity test, the temperature ranged +/ - I°C about the selected test temperature. Test systems were illuminated at 1000 lx on a 12L:12D cycle.

For the toxicity tests, 24-h-old post-larvae were randomly distributed to the exposure cups, five animals per cup. Six exposure cups were placed in each of the four toxicant concentrations and control. Mysids in eachcup were fed 24 h posthatchA. salina (7 × 102 day- 1).
The animals were exposed until sexually mature, then redistributed within each concentration to provide a consistent 2 male: 3 female ratio.
Duration:
36 d
Dose descriptor:
NOEC
Effect conc.:
631 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Duration:
36 d
Dose descriptor:
NOEC
Effect conc.:
631 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
reproduction
Reported statistics and error estimates:
ANOVA and Dunnett'stest (P <0.05)
Validity criteria fulfilled:
not applicable
Conclusions:
The 36-d NOEC for mortality and reproduction of the shrimp Americamysis bahia is 631 µg As /L for both basis of effects.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
14 days long toxicity testing on Chironomus riparius in a static renewal system in order to determine the EC10 (reproduction) value.
GLP compliance:
no
Specific details on test material used for the study:
sodium hydrogenarsenate heptahydrate, 99.998% (Sigma-Aldrich, St. Louis, MO, USA)
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
Reconstituted water was prepared using calcium chloride hexahydrate, 98%, sodium bicarbon- ate ACS reagent, 99.7–100.3%, calcium sulfate, ≥99.9% trace metals basis, magnesium sulfate heptahydrate, 98 + %, ACS reagent (Sigma- Aldrich, St. Louis, MO, USA), and potassium chloride (Fisher Scientific, Pittsburgh, PA, USA), mixed in Milli-Q HPLC grade water.
A single batch of this reconstituted water was mixed initially and 300 ml was transferred to each of the 15 beakers.
Test organisms (species):
Chironomus riparius
Details on test organisms:
Egg masses of C. riparius maintained in a colony were purchased from Environmental Consulting and Testing, Inc. (Superior, WI). After two days at 23 °C, the eggs began hatching.
Larvae of Chironomus riparius were used as test materials.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
14 d
Hardness:
1.64 mEQ Ca, 0.5 mEq Mg
Test temperature:
22.4 °C
pH:
7.1-7.8
Nominal and measured concentrations:
Nominal As(V) at 0, 0.13, 2.0, 5.3, and 13 μmol/L, all were within 15% agreement of the target concentrations.
Details on test conditions:
First instar larvae (14–20 individuals per beaker) were transferred to 600 ml glass beakers containing 300 ml of reconstituted water (described below) and factorial combinations of As(V) (at 0, 0.13, 2.0, 5.3, and 13 μmol l− 1, as sodium hydrogenarsenate heptahydrate, 99.998% (Sigma-Aldrich, St. Louis, MO, USA) and PO4 (at 0, 0.15, and 15 μmol l− 1, as potassium dihydrogen phosphate, 99.99% (Sigma-Aldrich, St. Louis, MO, USA)).
Key result
Duration:
14 d
Dose descriptor:
EC10
Effect conc.:
629.3 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
Aq
Basis for effect:
reproduction
Duration:
14 d
Dose descriptor:
EC50
Effect conc.:
1 978 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
reproduction
Reported statistics and error estimates:
EC10 have been re-calculated using Trap software (log-logistic curve fitting)
Validity criteria fulfilled:
not applicable
Remarks:
no guideline followed
Conclusions:
The EC10 Reproduction value of the species Chironomus riparius was reported to be 629.328 µg As(V) /L after a 14 days long toxicity test.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
no guideline followed
Principles of method if other than guideline:
48h toxicity test with marine molluscs
GLP compliance:
no
Specific details on test material used for the study:
sodium arsenate (Na3AsO4) (CAS no. 10048-95-0, Sigma-Aldrich, Missouri, USA)
Analytical monitoring:
yes
Details on sampling:
Each stock of seawater used for the embryotoxicity assays (at every combination of salinity and As concentration) were analysed to determine effective As concentrations for each condition.
Vehicle:
no
Details on test solutions:
Analytical grade artificial seawater (Tropic Marine Sea Salt) from the same batch was used for exposure media preparation and spawning, prepared according to the manufacturer's instructions using reverse osmosis (RO) water, 3 days before the experiments took place too achieve a salinity of 33 (i.e reference salinity). After complete salt dissolution and equilibration (24 h) seawater was filtered (0.2 μm) through cellulose acetate filters (Millipore) using a vacuum filtration unit. Seawater salinity was adjusted to obtain 3 separate batches at three different salinity levels (20, 26 and 33 ± 1) for exposure media preparation using RO water as dilution media (measured pH for all batches of seawater used ranged from 8.16 to 8.29).
Test organisms (species):
other aquatic mollusc: Crassostrea angulata
Details on test organisms:
C. gigas progenitors were obtained from Guernsey Sea Farms (UK) and C. angulata progenitors were provided by an aquaculture facility operating in the Sado estuary (Exporsado). C. angulata were collected during spawning season, from intertidal growing tables, and stimulated to spawn one day after collection. Spawning was induced by thermal stimulation, by consecutively changing oysters from seawater baths set at 18 and 28 °C in 30 min consecutive intervals.
Gamete emission and quality (oocyte shape and sperm motility) were visually inspected under a microscope. The number of male and female oysters selected for fertilization were 4 and 3 (C. gigas) and 6 and 5 (C. angulata), respectively.
Selected females were left to spawn in separate beakers, oocyte suspensions were filtered through a 100 μm nylon mesh and mixed in a final 500 ml suspension (salinity 33, 24 °C). Male gametes were collected in separate, filtered through a 45 μm nylon mesh into a mixed suspension, and left to activate for 20 min. Oocyte suspensions were fertilized by adding approximately 1 to 106 oocyte-to-sperm ratios, and fertilization success verified by microscopy.
Test type:
static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
not reported
Test temperature:
20-24-28 °C
pH:
8.16-8.29
Dissolved oxygen:
not reported
Salinity:
20-26-33 ppt
Conductivity:
not reported
Nominal and measured concentrations:
nominal: 30, 60, 120, 240, 480, 960 and 1920 μg As/L
measured:
20 ppt: 36, 68, 136, 256, 530, 1050 and 2052 µg As/L
26 ppt: 28, 72, 158, 274, 522, 1046 and 2097 µg As/L
33 ppt: 20, 74, 149, 262, 525, 1037 and 2040 µg As/L
Details on test conditions:
Exposure solutions were distributed in 24-well sterile capped polystyrene microplates (VWR), giving one microplate per salinity level, 3 wells (3 ml each) per exposure condition (As concentration) including negative controls. Each microplate corresponding to salinity and As conditions, were 3 fold replicated for incubations at different temperatures (20, 24 and 28 °C) to test two levels of 4 °C increase of temperature within projected global surface temperature rise by the end of the 21st century relative to years 1986–2005 (RCP8.5), while testing a temperature range that would allow to infer on embryo development under As exposure. All the above stated microplates were further replicated twice, to test all conditions after two different timeframes of embryonic development, to allow for the assessment of As induced delayed development to oyster embryos, while testing valid exposure time criteria of 24 h and 48 h for C. gigas. Each microplate was incubated at the desired temperature overnight in separate climatic chambers (±1 °C) before spawning induction took
place to stabilize testing media at the target temperature.
Zygotes were immediately transferred to microplates containing the exposure media to reach approximately 200 embryos per well. Microplates with the exposure media (0, 30, 60, 120, 240, 480, 960 and 1920 μg As/L) at different salinities (20, 26, 33) and fertilized oocytes were left to incubate in the dark, at different temperatures (20, 24 and 28 ± 1 °C) for 24 and 48 h post fertilization. Embryo-larval development was stopped by adding buffered formalin (4%). Analysis followed visual inspection of embryo-larval development under an inverted microscope and camera (Leica: DMIL-1; MC170 HD) by counting 100 embryos per well, and characterizing the relative frequencies of different types of development (D-shape, pre-D, protruded mantle, kidney shape, indented shell and dead larvae).
Results obtained were analysed as percentages of abnormal larvae (including pre-D and other malformations), and the validity of the experiments verified by results for negative and positive controls, considering the acceptability ranges for negative controls (> 70% D-shape larvae) and positive controls (9.47 μg. L−1 ≤ EC50≤21.72 μg/L).
Reference substance (positive control):
yes
Remarks:
Cu(NO3)2
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
23.6 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 20°C, salinity 26 ppt
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
34.6 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks:
weight
Remarks on result:
other: 24°C, salinity 26 ppt
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
44.2 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 24°C, salinity 33 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
31 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 20°C, salinity 26 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
41.8 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 24°C, salinity 26 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
18.7 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 24°C, salinity 33 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
50.5 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 28°C, salinity 26 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
15.9 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 28°C, salinity 33 ppt
Reported statistics and error estimates:
EC10 have been re-calculated using Trap software (log-logistic curve fitting)
Validity criteria fulfilled:
yes
Conclusions:
In a 48 h toxicity test, the EC10 on development of marine mollusc (Crassostrea angulata) embryos was reported to range between 23.6 and 44.2 µg As /L for different temperatures and salinities.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
no guideline followed
Principles of method if other than guideline:
48h toxicity test with marine molluscs
GLP compliance:
no
Specific details on test material used for the study:
sodium arsenate (Na3AsO4) (CAS no. 10048-95-0, Sigma-Aldrich, Missouri, USA)
Analytical monitoring:
yes
Details on sampling:
Each stock of seawater used for the embryotoxicity assays (at every combination of salinity and As concentration) were analysed to determine effective As concentrations for each condition.
Vehicle:
no
Details on test solutions:
Analytical grade artificial seawater (Tropic Marine Sea Salt) from the same batch was used for exposure media preparation and spawning, prepared according to the manufacturer's instructions using reverse osmosis (RO) water, 3 days before the experiments took place too achieve a salinity of 33 (i.e reference salinity). After complete salt dissolution and equilibration (24 h) seawater was filtered (0.2 μm) through cellulose acetate filters (Millipore) using a vacuum filtration unit. Seawater salinity was adjusted to obtain 3 separate batches at three different salinity levels (20, 26 and 33 ± 1) for exposure media preparation using RO water as dilution media (measured pH for all batches of seawater used ranged from 8.16 to 8.29).
Test organisms (species):
other aquatic mollusc: Crassostrea gigas
Details on test organisms:
C. gigas progenitors were obtained from Guernsey Sea Farms (UK) and C. angulata progenitors were provided by an aquaculture facility operating in the Sado estuary (Exporsado). C. angulata were collected during spawning season, from intertidal growing tables, and stimulated to spawn one day after collection. Spawning was induced by thermal stimulation, by consecutively changing oysters from seawater baths set at 18 and 28 °C in 30 min consecutive intervals.
Gamete emission and quality (oocyte shape and sperm motility) were visually inspected under a microscope. The number of male and female oysters selected for fertilization were 4 and 3 (C. gigas) and 6 and 5 (C. angulata), respectively.
Selected females were left to spawn in separate beakers, oocyte suspensions were filtered through a 100 μm nylon mesh and mixed in a final 500 ml suspension (salinity 33, 24 °C). Male gametes were collected in separate, filtered through a 45 μm nylon mesh into a mixed suspension, and left to activate for 20 min. Oocyte suspensions were fertilized by adding approximately 1 to 106 oocyte-to-sperm ratios, and fertilization success verified by microscopy.
Test type:
static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
not reported
Test temperature:
20-24-28 °C
pH:
8.16-8.29
Dissolved oxygen:
not reported
Salinity:
20-26-33 ppt
Conductivity:
not reported
Nominal and measured concentrations:
nominal: 30, 60, 120, 240, 480, 960 and 1920 μg As/L
measured:
20 ppt: 47, 79, 158, 256, 503, 1039 and 1932 µg As/L
26 ppt: 46, 81, 160, 258, 502, 1008 and 1997 µg As/L
33 ppt: 43, 86, 166, 254, 493, 1025 and 1966 µg As/L
Details on test conditions:
Exposure solutions were distributed in 24-well sterile capped polystyrene microplates (VWR), giving one microplate per salinity level, 3 wells (3 ml each) per exposure condition (As concentration) including negative controls. Each microplate corresponding to salinity and As conditions, were 3 fold replicated for incubations at different temperatures (20, 24 and 28 °C) to test two levels of 4 °C increase of temperature within projected global surface temperature rise by the end of the 21st century relative to years 1986–2005 (RCP8.5), while testing a temperature range that would allow to infer on embryo development under As exposure. All the above stated microplates were further replicated twice, to test all conditions after two different timeframes of embryonic development, to allow for the assessment of As induced delayed development to oyster embryos, while testing valid exposure time criteria of 24 h and 48 h for C. gigas. Each microplate was incubated at the desired temperature overnight in separate climatic chambers (±1 °C) before spawning induction took
place to stabilize testing media at the target temperature.
Zygotes were immediately transferred to microplates containing the exposure media to reach approximately 200 embryos per well. Microplates with the exposure media (0, 30, 60, 120, 240, 480, 960 and 1920 μg As/L) at different salinities (20, 26, 33) and fertilized oocytes were left to incubate in the dark, at different temperatures (20, 24 and 28 ± 1 °C) for 24 and 48 h post fertilization. Embryo-larval development was stopped by adding buffered formalin (4%). Analysis followed visual inspection of embryo-larval development under an inverted microscope and camera (Leica: DMIL-1; MC170 HD) by counting 100 embryos per well, and characterizing the relative frequencies of different types of development (D-shape, pre-D, protruded mantle, kidney shape, indented shell and dead larvae).
Results obtained were analysed as percentages of abnormal larvae (including pre-D and other malformations), and the validity of the experiments verified by results for negative and positive controls, considering the acceptability ranges for negative controls (> 70% D-shape larvae) and positive controls (9.47 μg. L−1 ≤ EC50≤21.72 μg/L).
Reference substance (positive control):
yes
Remarks:
Cu(NO3)2
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
90 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 20°C, salinity 20 ppt
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
156.4 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 20°C, salinity 26 ppt
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
306.1 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 20°C, salinity 33 ppt
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
73.1 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks:
weight
Remarks on result:
other: 24°C, salinity 20 ppt
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
171.1 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 24°C, salinity 26 ppt
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
329.9 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 24°C, salinity 33 ppt
Key result
Duration:
48 h
Dose descriptor:
EC10
Effect conc.:
82.1 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 28°C, salinity 20 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
101.1 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 20°C, salinity 20 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
161.9 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 20°C, salinity 26 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
530 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 20°C, salinity 33 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
80.71 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 24°C, salinity 20 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
214.3 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 24°C, salinity 26 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
663.5 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 24°C, salinity 33 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
103.3 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 28°C, salinity 20 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
198.9 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 28°C, salinity 26 ppt
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
493.7 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Remarks on result:
other: 28°C, salinity 33 ppt
Reported statistics and error estimates:
EC10 have been re-calculated using Trap software (log-logistic curve fitting)
Validity criteria fulfilled:
yes
Conclusions:
In a 48 h toxicity test, the EC10 on development of marine mollusc (Crassostrea gigas) embryos was reported to range between 73.1 and 329.9 µg As /L for different temperatures and salinities.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
other: EPA 600/4 89/001 Environmental Monitoring and Support Laboratory, Cincinnati, OH
GLP compliance:
not specified
Specific details on test material used for the study:
Sodium salts of arsenate (As5+)
Analytical monitoring:
yes
Details on sampling:
Water samples were collected at the beginning of the experiment on days 3 to 8 and saved for metal analysis. Samples were stored at -2O°C, then thawed and acidified with one drop of concentrated nitric acid before analysis.
Vehicle:
no
Details on test solutions:
reconstituted moderately hard water
Test organisms (species):
Ceriodaphnia dubia
Details on test organisms:
Test organism: C. dubia
Age at study initiation: less than 24 h old
Each neonate was reared in 30-ml plastic (polystyrene) cups with 15 ml of reconstituted moderately hard water (hardness 120mg/L CaCO,, alkalin- ity 34 mg/L CaCO,, pH 8.0). Culture water was a modifi- cation of the EPA moderately hard water (90 mg/L CaS0,.2H20, 96 mg/L NaHCO,, 60 mg/L MgS04, and 4 mg/L KCl).
The food source for C. dubia was a unicellular green alga, either Selenastrum capricornutum or Ankistrodesmus falca- tus. Both species were used during preliminary studies; how- ever, S. capricornutum was used during the course of the mixture experiment. The C. dubia were fed daily (4.0 x lo5 to 6.0 x lo5cells/ml in test cup) and transferred to new me- dia every other day. The organisms were cultured in a con- stant temperature room (25 k 1"C) with a 16:8-h light:dark photoperiod. The C. dubia, originally obtained from the EPA Environmental Monitoring and Support Laboratory, Newton Facility, located in Cincinnati, Ohio, have been cul- tured in our laboratory for several years.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
8 d
Hardness:
119 mg/L CaCO3
Test temperature:
25.8 "C
pH:
7.9
Dissolved oxygen:
7.5 mg/L
Nominal and measured concentrations:
measured: <0.25, 1.02, 1.14 and 1.42 mg As/L
Measured concentrations (mean f SD) differed from nominal values by 60%
Details on test conditions:
Test methods followed standard guidelines [17]; however, due to the complexity of this experiment, certain deviations were made and noted below. This toxicity assessment in- volved a three-brood static renewal test to assess the effect of metal mixtures on C. dubia reproduction (fecundity) and survival. At the beginning of each test, neonates <24 h old were pooled into a 500-ml beaker, then transferred individ- ually into a plastic cup with 15 ml of test solution. Each day during transfer of test organisms to new metal solutions, sur- vival and fecundity were recorded.
The C. dubia were fed 0.2 ml(6 x lo5celIs/ml in the test cup) of S. capricornutum daily. Food was added to the new metal solution directly be- fore or after introduction of the daphnid. Toxicity tests were conducted under the following controlled environmental conditions:
constant illumination (44-50 pE m-' s-', cool white fluorescent tubes), agitation (50 rpm using a Lab-line Junior Or- bit Shaker), and temperature (24 +- 2OC). Light intensity was measured using a LI-cor model LI-185B Quantum/Radiom- eter photometer.
Key result
Duration:
8 d
Dose descriptor:
EC10
Effect conc.:
1 360 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
reproduction
Duration:
8 d
Dose descriptor:
EC50
Effect conc.:
1 600 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
reproduction
Duration:
8 d
Dose descriptor:
NOEC
Effect conc.:
> 1 420 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Reported statistics and error estimates:
EC10 have been re-calculated using Trap software (log-logistic curve fitting)
NOEC using Fisher's test or LSD test
Validity criteria fulfilled:
not specified
Conclusions:
The EC10 Reproduction value of the species Ceriodaphnia dubia was reported to be 1.36 mg As /L.
The NOEC Mortality value was reported to be >1.42 mg As /L.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
Long term toxicity test with Hyalella azteca
GLP compliance:
not specified
Specific details on test material used for the study:
Na2HAsO4.7H2O
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
Test media consisted of de-chlorinated Burlington city tap water originating from Lake Ontario.
Stock solutions consisted of sodium arsenate (Na2HAsO4 7H2O), dissolved in de-ionized water (Milli-Q). The element was in the form As(V).
Test organisms (species):
other: Hyalella azteca
Details on test organisms:
Test organism: Hyalella azteca
Age: 0-1 week old
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Hardness:
0.87 mmol Ca; 0.351 mmol Mg
Test temperature:
25°C
pH:
8.3
Dissolved oxygen:
8.2 mg/L
Conductivity:
315 µ/cm
Nominal and measured concentrations:
20 -10000 nmol As/L (see also 'overall remarks, attachments' - Illustration)
Details on test conditions:
Twenty 0-1 week-old Hyalella were added to 250 mL of test medium with a single piece of 5x5-cm cotton gauze in 500-mL Erlenmeyer flasks
with inverted polyethylene sample cups as covers. Experiments were conducted in an incubator at 25 °C with a 16 h light/8 h dark photoperiod. Weekly, static renewal of media and contaminants were carried out during the 4-week (chronic) test. Food additions (TetraMin fish food flakes ground to 500 mm mesh size), consisted of two 2.5-mg feedings during weeks 1 and 2, three 2.5-mg feedings in week 3, and two 5.0-mg feedings in week 4. The increase in food per week was incorporated to allow for animal growth throughout the experiment. Test media consisted of de-chlorinated Burlington city tap water originating from Lake Ontario (mean ± C.I 95%: dissolved organic carbon 1.1 ± 0.36 mg/L, dissolved inorganic carbon 20 ± 0.32 mg/L, Alk 85 ± 1.06 mg/L, Cl 674 ± 0.53 µmol/L, SO4 314 ± 0.98 µmol/L, SiO2 19 ± 0.10 µmol/L, Ca 870 ± 0.41 µmol/L, Mg 351 ± 0.09 µmol/L, Na 561 ± 0.31 µmol/L, K 40 ± 0.02 µmol/L, pH 8.2 ± 0.06 and conductivity 315 ± 6.5 µ/cm.
Analyses were conducted by the National Laboratory for Environmental Testing, Environment Canada). Two replicates were run of a concentration series for each metalloid or metal per test, controls were usually run in triplicate and each test was repeated at least once.

Survival was recorded weekly and at the end of the 28-day exposure. Wet weight, dry weight and body concentrations were determined for 0 and 24 h gut-purged survivors and water samples were analyzed.
Key result
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
201 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
growth
Remarks:
weight
Duration:
28 d
Dose descriptor:
other: EC25
Effect conc.:
294 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
growth
Remarks:
weight
Duration:
28 d
Dose descriptor:
EC50
Effect conc.:
426 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
growth
Remarks:
weight
Duration:
28 d
Dose descriptor:
other: LC25
Effect conc.:
324 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Duration:
28 d
Dose descriptor:
LC50
Effect conc.:
420 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Reported statistics and error estimates:
Ecxel calculated with non-linear model
Validity criteria fulfilled:
not applicable
Conclusions:
The EC10 value for Growth (weight) of the species Hyalella azteca was reported to be 201 µg As/L.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: USEPA. 2000a. Revised Baseline Ecological Risk Assessment Hudson River PCBs Reassessment. Phase 2 Reports. Prepared by TAMS Consultants, Menzie-Cura and Associates for USEPA Region 2. Washington, DC, USA.
GLP compliance:
no
Specific details on test material used for the study:
As2O3 (analytical grade)
Analytical monitoring:
yes
Details on sampling:
not specified
Vehicle:
no
Details on test solutions:
Natural filtered seawater
Stock solutions of arsenic were freshly prepared by dissolving arsenic trioxide (As2O3) in Milli-Q water.
Test organisms (species):
other: Penaeus indicus
Details on test organisms:
Post larval stages (PL-12) of Penaeus indicus were collected from private farm in Marakkanam (Tamilnadu, India) and transported to the laboratory in air-filled plastic bags. Test organisms were acclimatized in glass aquaria with aerated natural filtered seawater for a period of 7 days with 27 PSU salinity, temperature of 28 ± 2°C, dissolved oxygen of 5.6 mg/l and pH of 8.01. After a day of acclimatization, P. indicus was then fed with pellets of mixed feed for P. indicus.
Test type:
semi-static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
28 d
Test temperature:
28 °C
pH:
8.01
Dissolved oxygen:
5.6 mg/L
Salinity:
27 ppt
Nominal and measured concentrations:
nominal: 10, 20, 40, 80 and 160 μg/L
Details on test conditions:
The experimental method includes static renewal (24-hour renewal) test by following the method of USEPA (2002a). Five concentrations (10, 20, 40, 80 and 160 μg/l) in a geometric series including control were prepared for 30 days for short-term chronic toxicity test (USEPA, 2002b). Toxicant and seawater were replaced on daily basis. Each series of test chambers consisted of triplicate with 10 animals in a 10 L glass trough. Test chambers were loosely covered to prevent loss of test animals with gentle aeration. Test animals were fed regularly three times a day.

Test organisms were subjected to physical measurements in terms of length and weight. Condition factor (K) of the experimental animal was calculated by Williams (2000) (K = 100 W / L3). The Total Length (L) of the test organism was measured from the tip of the anterior or part using ruler to the nearest centimeter. Weight was measured after blot drying with a piece of clean hand towel. Weighing was done with a tabletop digital weighing balance (Metller), to the nearest gram.
Reference substance (positive control):
no
Key result
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
13.2 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
growth
Remarks:
as condition factor
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
20 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
growth
Remarks:
weight
Reported statistics and error estimates:
EC10 have been re-calculated using Trap software (log-logistic curve fitting),
NOEC with ANOVA using Dunnett’s test
Validity criteria fulfilled:
yes
Conclusions:
During the 28 days toxicity test the LC10 on growth (condition factor ) of the species Penaeus indicus was reported to be 13.2 µg As /L
and the NOEC on growth (weight) was 20.0 µg As /L.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: Brazilian standardized protocol (ABNT, 2006). Ecotoxicologia aquática – Toxicidade crônica de curta duração – Método de ensaio com ouriço-do-mar (Echinodermata: Echinoidea), NBR15350
Version / remarks:
Embryo-larval toxicity testing (chronic short-term) with sea urchin L. variegatus
GLP compliance:
no
Specific details on test material used for the study:
Na2(AsHO4).7H2O salt (VETEC, Rio de Janeiro)
Analytical monitoring:
yes
Details on sampling:
not specified
Vehicle:
no
Details on test solutions:
Natural coastal water
Test organisms (species):
other: Lytechinus variegatus
Details on test organisms:
Embryo-larval toxicity testing (chronic short-term) with sea urchin L. variegatus
Sea urchin eggs and sperms were obtained by induced spawning of organisms collected in nature. After fertilization, embryos were exposed to seven different concentrations of As.
Test type:
static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
24 h
Hardness:
not reported
Test temperature:
not reported
pH:
not reported
Dissolved oxygen:
not reported
Salinity:
not reported
Conductivity:
not reported
Nominal and measured concentrations:
nominal: 0.06; 0.12; 0.25; 0.5; 1.0; 2.0 and 4.0 mg As/L
All measured concentrations lied close to the nominal values with a coefficient of variation ≤15 percent.
Natural concentration of As in the coastal water was 1.9 μg As/L.
Details on test conditions:
After fertilization, embryos were exposed for 24 h to seven different concentrations of As.
The exposure was terminated when the larvae of the control groups reached the larval stage known as pluteus. After fixation in formaldehyde, the number of normal or deformed pluteus larvae was determined by microscopic examination. The test was repeated two times (run in triplicates).
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Key result
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
120 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
1 330 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
morphology
Reported statistics and error estimates:
LC50 were determined by means of the Trimmed Spearman-Karber methodology; William's test (P≤0.05) was used to obtain the no-observed-effect concentration (NOEC)
Validity criteria fulfilled:
yes
Conclusions:
In a 24 h toxicity test, the EC10 on development of sea urchin (Lytechinus variegatus) embryos was reported to be 120 µg As /L.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
other: ASTM, 1984
Principles of method if other than guideline:
7 days long toxicity test on Ceriodaphnia dubia
GLP compliance:
not specified
Specific details on test material used for the study:
reported in study: regent grade sodium arsenite (Na2AsO4.7H2O)
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
Reagent grade sodium arsenite was dissolved in distilled water.
Test organisms (species):
Ceriodaphnia dubia
Details on test organisms:
The organisms were cultured in the respective water before they were tested.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
7 d
Hardness:
100 mg/L as CaCO3
Test temperature:
25°C
pH:
8.2
Nominal and measured concentrations:
0.102, 0.188, 0.404, 0.793, 1.636 and 3.250 mg As/L
Test concentrations were analytically confirmed.
Key result
Duration:
7 d
Dose descriptor:
NOEC
Effect conc.:
793 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
reproduction
Key result
Duration:
7 d
Dose descriptor:
NOEC
Effect conc.:
1 636 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Reported statistics and error estimates:
ANOVA using Dunnett’s test
Validity criteria fulfilled:
not specified
Conclusions:
7d long term toxicity on Ceriodaphnia dubia was performed for As. The obtained NOEC values are the following:
NOEC on Mortality: 1.636mg As/L,
NOEC on Reproduction: 0.793 mg As/L.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
14 days long toxicity testing on aquatic invertebrates.
GLP compliance:
no
Specific details on test material used for the study:
arsenic trioxide (As2O3)
Analytical monitoring:
yes
Details on sampling:
One- hundred-milliliter samples for water analysis were taken twice a week from the middle of each glass aquarium and were immediately analyzed.
Vehicle:
no
Details on test solutions:
Untreated Lake Superior water was heated and used in the test at 14 to 16 °C.
Stock solutions were made by dissolving reagent-grade arsenic trioxide (As203) in 1 L of deionized water. Ten grams of sodium hydroxide had to be added to sufficiently enhance the solubility of arsenic trioxide in the 1-L solution. Stock solutions were placed in syringes, and amounts were added to the test water to achieve the desired concentration.
Test organisms (species):
other aquatic crustacea: Gammarus pseudolimnaeus
Details on test organisms:
life stage: 0.004-0.02 g
They were acclimated in the flow-through test system for one week at the test temperature before the start of the test. Ten animals of each species were used per duplicatec hamber.Invertebrate species were placed in round stainless steel screen containers (6 cmd iameter x 15cm deep)that were suspended in the test aquaria.
The test organisms swere fed fall-dropped birch and aspen leaves presoaked in diluent water for at least onece a month.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
14 d
Hardness:
42-45 mg/L CaCO3
Test temperature:
14 - 16 °C
pH:
6.9 - 7.3
Dissolved oxygen:
6.2- 8.9 mg/L
Nominal and measured concentrations:
0.088 and 0.961 mg As/L
Details on test conditions:
The test was conducted with an intermittent-flow exposure system consisting of a multi-channel toxicant injection system which delivered the toxicants at two concentrations (100 and 1000 µg/L, nominal) and a control to duplicate exposure chambers.The test chambers were glass aquaria 30 x 60 x 30 cm, with a water volume of 43 L. Flow rate to each chamber was 500 ml every two min, providing a 95% replacement of the test water every hour.
Fluorescent bulbs provided a light intensity of 20 to 40 lumens at the water surface. An automatically controlled 16-hr photo period was used.
Solutions of arsenic were renewed and analyzed each week during the test.

Deaths were recorded after the fourth day and then once a week for the remainder of the test. Death was defined as complete immobilization and failure of the animal to respond to prodding.
Reference substance (positive control):
no
Key result
Duration:
14 d
Dose descriptor:
NOEC
Effect conc.:
88 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Reported statistics and error estimates:
NOEC using Duncan multiple range tests
Validity criteria fulfilled:
not applicable
Conclusions:
The 14 days long toxicity study of As2O3 on Gammarus pseudolimnaeus has yielded a NOEC of 88 µg As/L for mortality.
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
no guideline followed
Principles of method if other than guideline:
10days long chronic toxicity study of As on Gammarus pulex.
GLP compliance:
no
Specific details on test material used for the study:
HAsNa2O4·7H2O
Chemicals used were provided by SIGMA-Aldrich, UK.
Analytical monitoring:
yes
Vehicle:
no
Details on test solutions:
Stock solutions were prepared by dossolving sodium arsenate heptahydrate (AsHNa2O4; 7H2O) in milliQ water and were stabilized with 0.1 % of HNO3 (65 %, Suprapure,
Merck). Metal test solutions were prepared using mineral water (Volvic, France) as diluent of stock solutions.
Test organisms (species):
Gammarus pulex
Details on test organisms:
Only adult males of G. pulex were collected. Gammarus sampling was performed in January 2011 using a hand net in the unpolluted Meholle River [Void-Vacon, France, Lambert II E (X = 841244; Y = 2412346)]. After sampling, amphipods were quickly brought to the labora- tory in plastic bags.
Males were measured from the top of the cephalothorax to the base of the telson by image software analysis (Studio version 7; ImageJ software version 1.42q). The mean body length of selected males was 10.8 ± 1.1 mm.
Individuals were acclimatized for a week before experiments in mineral water (Volvic, France) under con- trolled conditions (10 °C in a temperature controlled room with a photoperiod of 16 h of light and 8 h of darkness). Individuals were fed with discs of conditioned leaves during acclimation up to 24 h before starting experiments.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
10 d
Test temperature:
10°C
pH:
7.52
Nominal and measured concentrations:
measured: <2, 274.7, 534.6, 1010.5 and 1502 µg As/L
Details on test conditions:
After the acclimation period, groups of 10 males of G. pulex were transferred from batches to plastic Petri dishes (100 mm width and 20 mm height; = ‘dishes’ hereafter) containing 44 mL of tested toxic solution (experimental dishes) or Volvic water (control dishes) and four glass pebbles (Ø 15 mm, convex side down). As gammarids were not fed during the whole experiment, glass pebbles served both as substrate and refuge for animals and allowed to avoid at best cannibalism.
In the acclimation temperature and photoperiod conditions, five replicates of ten gammarids were exposed during 240 h to As.
Experiments were performed under semi-static conditions with daily renewal of the solutions to maintain metal concentrations at the same level until 240 h exposure. Test containers were filled up with metal solutions 96 h before the beginning of experiments, allowing reaching equilibrium before experiments and minimizing metal adsorption during experiments.
The mortality of gammarids was recorded after 240 h exposure to each tested concentration.
Key result
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
380.3 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Remarks on result:
other: 95% confidence interval: 264.7-519.4 µg As/L
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
989.7 µg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
element (dissolved fraction)
Remarks:
As
Basis for effect:
mortality
Remarks on result:
other: 95% confidence interval: 822.2 - 1069.6 µg As/L
Reported statistics and error estimates:
LC10 and LC50 were determined using log-logistic curve fitting
Validity criteria fulfilled:
not applicable
Conclusions:
10 days long chronic toxicity test was performed on Gammarus pulex. The reported EC10 result on mortality was 380 µg As/L.

Description of key information

In total, reliable chronic toxicity data were retrieved for 12 different freshwater invertebrates. For the amphipods Gammarus pseudolimnaeus a 14 d NOEC value of 88 µg As/L (endpoint mortality) was found, for Gammarus pulex a 10 d LC10 of 380 µg As/L (endpoint mortality), while for another amphipod Hyalella azteca a 28 d EC10 of 201 µg As/L (endpoint growth) was observed.

Chronic toxicity values were also reported for different freshwater copepods, with LC10 values (endpoint mortality) between 40 and 2670 µg As/L for Cyclopoids taxa and an LC10 (endpoint mortality) of 130 µg As/L for the Harpactoid taxa.

Chronic toxicity values were also retrieved for the cladocerans Daphnia magna and Ceriodaphnia dubia. A 28 d NOEC of 633 µg As/L (endpoint growth/reproduction/mortality) was observed for D. magna, while for C. dubia the chronic toxicity values varied between 793 µg As/L (endpoint reproduction) and 1636 µg As/L (endpoint mortality).

For the insect Chironomus riparius (midge) a 14 d EC10 of 629 µg As/L (endpoint reproduction) was observed, while a lower chronic LC10 of 98 µg As/L (endpoint mortality) was found for the mayfly Hydropsiche pellucidula.

For the isopod Asellus aquaticus a chronic 10 d LC10 of 97 µg As/L (endpoint mortality) was retrieved from literature.

For the rotifer Brachionus patulus a chronic NOEC of 1500 µg As/L (endpoint mortality) was retrieved from literature.

Chronic toxicity data for different molluscs i.e. Actinonaias pectorosa, Helisoma campanulate, Physa fontinalis and Stagnicola emarginata, were found but all were unbounded and therefore not retained for PNEC derivation. Another unbounded NOEC was observed for the stonefly Pteronarcys dorsata.

Chronic toxicity data were retrieved for 6 different marine invertebrates. Chronic toxicity data to marine crustaceans were available for 2 different species, i.e. Americamysis bahia and Penaeus indicus. A chronic 36 d NOEC value of 631 µg As/L (endpoint mortality/reproduction) was observed for the mysid A. bahia, while a 28 d chronic EC10 of 13.2 µg As/L was noted for P. indicus (endpoint growth). Toxicity values for the echinoderms Paracentrotus lividus and Lytechinus variegatus revealed NOEC/EC10 values of respectively 56 µg As/L (endpoint development) and 120 µg As/L (endpoint mortality). Toxicity values for the marine oysters Crassostrea angulata and Crassostrea gigas revealed EC10 values (endpoint development) varying between 23.6 and 44.2 µg As/L for Crassostrea angulata, and between 73.1 and 329.6 µg As/L for Crassostrea gigas. All tests with both the echinoderms and molluscs are short-term toxicity tests initiated with embryonal stages.

Key value for chemical safety assessment

Additional information