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Long-term toxicity to fish

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Endpoint:
fish early-life stage toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
unknown
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study generally meets accepted scientific standards, well documented and acceptable for assessment. Some testing information is not provided, such as some water quality parameters.
Qualifier:
equivalent or similar to guideline
Guideline:
other: OECD 229
Deviations:
yes
Remarks:
test not performed with recommended test species
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
All test chambers were tested every week
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: NP was diluted in acetone and mixed with distilled water
- Controls: dilution water control and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.01%
Test organisms (species):
other: Gobiocypris rarus
Details on test organisms:
TEST ORGANISM
- Common name: Chinese Rare Minnow
- Source: Institute of Hydrobiology, Chinese Academy of Sciences

ACCLIMATION
- Brood stock kept in dechlorinated tap water at 25 °C and photoperiod 16L:8D

FEEDING
- Type/source of feed:generally fed with commercial granular food and dry brine shrimp
- Amount given: 0.1% body weight/day

Test organisms were adults about 9 months old, with weights of 2.3 +/-0.5 g and lenths of 48.3 +/-5.6 mm.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
21 d
Post exposure observation period:
none
Hardness:
not provided
Test temperature:
25 °C
pH:
not provided
Dissolved oxygen:
not provided
Salinity:
NA
Nominal and measured concentrations:
nominal: control, solvent control, 5, 10, 20 µg/L
average measured: 4.52, 9.13 and 18.53 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 18 L tank
- Type of flow-through (e.g. peristaltic or proportional diluter): info not given
- Renewal rate of test solution (frequency/flow rate): 10 L/h
- No. of organisms per vessel: 2 (1 breeding pair)
- No. of vessels per concentration (replicates): 8
- No. of vessels per control (replicates): 8
- No. of vessels per vehicle control (replicates): 8

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): survival, reproductive behavior and spawning assessed daily

Eggs were removed, counted and checked for fertility daily. Embryos from each spawn were put in groups of 50 and maintained in respective test treatment concentration to determine hatching success. Newly hatched fry (n=50) were maintained in respective test treatments and assessed for growth and mortality. Experiment was terminated when offspring were 15 days-post hatch when offspring were measured for body lengths and weight.
Reference substance (positive control):
no
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
> 0.02 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
adult mortality
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
> 0.02 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
fertility
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
> 0.02 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
reproduction
Details on results:
Adult survival was 100% in all treatments/controls. Growth was not significantly different from controls. Fecundity and fertility were not significantly different from controls in all treatments. Hatching rates of offspring were not significantly different from controls. Larvae offspring lengths and weights and survival were also not affected by treatments.
Other effects (estrogenic) assessed were VTG production and target organ effects. Male fish in NP treatments showed testis-ova phenomenon, with those in highest test concentration showing significant effects. Lesions on liver tissue were significantly different than controls in highest test concentration.
Reported statistics and error estimates:
Statistical analysis was performed by using Statistica. Homogeneity of variance across treatments was assessed using Levene's test. ANOVA and Dunnett's multiple comparison tests were used. If nonhomogeneous, Kruskal-Wallis test was used followed by Mann-Whitney U-test with Bonferroni's adjustment.

Hatchability, Survival and Growth of Rare Minnow Larvae Exposed to Nonylphenol to 15 -days Post Hatch

  Control  Sovent Control  5 (ug/L)  10 (ug/L)   20 (ug/L)
Hatch Rate (%)  95.3  93.6  88.2  80.7   79.4
Body Length (mm) 9.5  9.6  9.1 9.5   8.8
Body Weight (mg) 6.1  6.2  4.7  5.0   4.1
Accumulative Death Rates (%)  11.2  13.1  9.8  14.3   12.4
Validity criteria fulfilled:
not specified
Remarks:
missing DO and temperature of each test solution information
Conclusions:
Study authors conclude that the rare minnow can be used in short-term reproductive bioassay. Of the two chemicals tested, EE2 was more toxic than nonylphenol showing increased plasma VTG, liver lesions and spawning intervals. The rare minnow is more sensitive to EE2 than medaka.
Executive summary:

The 21– day short-term chronic toxicity of nonylphenol to reproduction stage of the Chinese Rare Minnow (Gobiocypris rarus) was studied under flow through conditions. Paired adult fish, 8 pairs per test concentration of Gobiocypris rarus were exposed to control, solvent control and measured concentrations of 5.0, 10, and 20 µg/L nonylphenol.  The test system was maintained at 25 ºC.  The 21– day NOEC value, based on sublethal effects (survival of adults, adult fecundity, offspring hatchability, survival, growth), was >0.02 mg/L.   Mean percent hatch of any test concentration was not significantly different from controls. Growth was not significantly different from controls. Fecundity and fertility were not significantly different from controls in all treatments. Hatching rates of offspring were not significantly different from controls. Larvae offspring lengths and weights and survival were also not affected by treatments.

This toxicity study is classified as acceptable supporting study and satisfies the guideline requirement for toxicity study with fish.

 

Results Synopsis

 

Test Organism Size/Age (mean wet weight or length): paired sexually mature adults

Test Type (Flow through, Static, Static Renewal): flow through

 

NOEC: >0.02 mg/L nonylphenol

Endpoint(s) Effected: survival, fertility, fecundity, larval hatchability, growth

Zha et al. (2008) was chosen as a supporting study because it was a well documented study, performed on a sensitive organism, Gobiocypris rarus.

Endpoint:
fish life cycle toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
unknown
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study meets generally accepted scientific standards, is well documented and acceptable for assessment. Some testing information is missing such as water quality characteristics during testing.
Qualifier:
equivalent or similar to guideline
Guideline:
EPA OPPTS 850.1500 (Fish Life Cycle Toxicity)
Principles of method if other than guideline:
Generally follows fish life cycle test procedures, but extends length of test to cover two generations. Embryological development, hatching success, posthatch survival, growth, sexual differentiation, and reproduction of the first generation and progeny are assessed.
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
Test solutions were sampled every two weeks through out exposure period.
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: dilution water and solvent
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.1ml/L
Test organisms (species):
Oryzias latipes
Details on test organisms:
TEST ORGANISM
- Common name: Medaka
- Source: original stock from local fish farm in Kumamoto, Japan. Used in house breeding stock maintained for several years.

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Numbers of parental fish (i.e. of females used to provide required number of eggs): about 100 breeding pairs kept for at least 2 weeks in breeding conditions.
- Method of collection of fertilised eggs: collected a few hours after fertilization, pooled in a petri dish with 5 ml of dechlorinated tap water
- Subsequent handling of eggs: eggs were checked for fertilization and development under a microscope and then placed in test chambers.

POST-HATCH FEEDING
- Start date:< 24 h after hatching
- Type/source of feed: Artemia nauplii
- Frequency of feeding: twice a day
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
104 d
Remarks on exposure duration:
F1 generation was exposed 60days post-hatch
Post exposure observation period:
none
Hardness:
44-73.5 mg/L CaCO3
Test temperature:
24 °C for reproductive phase
28 °C for remaining of test
pH:
7.4-7.5 s.u.
Dissolved oxygen:
not given
Salinity:
NA
Nominal and measured concentrations:
nominal: control, solvent control, 1.85, 5.56, 16.7, 50, and 150 µg/L nonylphenol
measured: ND, ND, 4.2, 8.2, 17.7, 51.5, 183 µg/L nonylphenol
Details on test conditions:
TEST SYSTEM
- Test vessel: glass chamber holding 1.8 L of test solution
- Type of flow-through (e.g. peristaltic or proportional diluter): dilutor
- Renewal rate of test solution (frequency/flow rate): 14 times a day
- No. of fertilized eggs/embryos per vessel: 15
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D

EFFECT PARAMETERS MEASURED (with observation intervals if applicable)
Daily observation, Embryological development, hatching success, posthatch survival, growth, sexual differentiation, and reproduction of the first generation and progeny are assessed.

VEHICLE CONTROL PERFORMED: yes

RANGE-FINDING STUDY
7-day range-finder performed
- Results used to determine the conditions for the definitive study: yes

POST-HATCH DETAILS
60 days post-hatch, 5 fish from each of four test chambers were randomly removed and observed for sex characteristics and weighed.
70 days post-hatch, sex of surviving fish assessed, water temp increased to promote spawning, eggs collected and viability assessed. Eggs spawned at day 103-104 post hatch of parents.
F1 phase - semistatic exposure until hatching.
F1 hatching larvae were transferred to test chambers (60 larvae per treatment) and exposed to NP until 60 days post hatch.

Duration:
104 d
Dose descriptor:
NOEC
Effect conc.:
0.008 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
other: F0 generation post swim-up mortality
Duration:
104 d
Dose descriptor:
LOEC
Effect conc.:
0.018 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
other: F0 generation post swim-up mortality
Details on results:
Study states that results above reflect overall toxic effect of nonylphenol throughout complete life-cycle of the F0 generation.
For the progeny generation, there was no effect on hatching success, mortality, behaviour, appearance, or growth of hatched larvae. An effect on sexual differentiation was seen at 60 d post-hatch.
Reported statistics and error estimates:
Chi-square or Student's t-test or Bonferroni's U tests used to determine differences between solvent and control groups. Groups were pooled if no differences found. Homogeneity of variances were checked by Levene's test and normality was checked by Shapiro-Wilk's test. Data further assessed using analysis of variance and Dunnett's multiple comparison test. Significant difference at p<0.05 level.

Testing Results of F0 Generation. Body wt and Sex Ratio at 60 -day Post Hatch

NP Concentration (µg/L) Hatchability (%)  Time to Hatch (d)  Body wt (mg)  Sex Ratio (M:F) 
Control  90  9.4  178  9:11 
Solvent control  93.9  9.9  170  8:12 
4.2  95  9.5  167  12:8 
8.2  90  9.6  197  13:7 
17.7  88.3  10.2  179  9:11 
51.5  93.3  10.2  167  0:20 
183  46.7*  9.6  -  - 

*Significantly different from pooled controls

Post swim-up mortality had a positive correlation with NP concentration with significant effects seen at 51.5 and 17.7 µg/L concentrations.

Fecundity of F0 generation was not affected by NP treatments, but fertility was decreased in the 17.7 µg/L concentration. Hatchability, time to hatch and juvenile mortality and growth of progeny were not significantly affected by NP exposures. However, assessment of sex characteristics shows affects at the 17.7 µg/L concentration.

Validity criteria fulfilled:
not specified
Remarks:
water quality parameters during testing are not provided
Conclusions:
Study results indicate that reproductive effects of nonylphenol to Medaka can be exerted by nonylphenol at concentrations as low as 0.0177 µg/L.
Executive summary:

The 104– day chronic toxicity of nonylphenol to a full-life cycle of Medaka (Oryzias latipes) was studied under flow through conditions through two generations.  Embryos, 60 eggs per test concentration of Medaka were exposed to control, solvent control and measured concentrations of 4.2, 8.2, 17.7, 51.5, 183 µg/L nonylphenol.  The test system was maintained near 25 ºC and elevated to 28 ºC during spawning phase.  The 104– day NOEC value, based on complete life-cycle effects was 8.2 µg/L nonylphenol for the F0 generation. The most sensitive effect was on mortality of post swim-up larvae of the F0 generation.   Post swim-up mortality had a positive coorelation with NP concentration with significant effects seen at 51.5 and 17.7 µg/L concentrations. Fecundity of F0 generation was not affected by NP treatments, but fertility was decreased in the 17.7 µg/L concentration. Hatchability, time to hatch and juvenile mortality and growth of progeny were not significantly affected by NP exposures. However, assessment of sex characteristics shows affects at the 17.7 µg/L concentration.  

This toxicity study is classified as acceptable supporting study.

Results Synopsis

 

Test Organism Size/Age: full life cycle

Test Type (Flowthrough, Static, Static Renewal): flow through

 

NOEC survival:  0.0082 mg/L nonylphenol

Endpoint(s) Effected: reproduction

Yakota et al (2001) was chosen as a supporting study because it was a well documented study, performed on a common sensitive organism, Oryzias latipes.

Results of this study are show that this species is one of the more sensitive species to nonylphenol. Results are in general agreement with other studies with the same species Nimrod and Benson (1998).
Endpoint:
fish life cycle toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 Aug 99 - 25 Feb 00
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study meets generally accepted scientific standards and is described in sufficient detail
Qualifier:
according to guideline
Guideline:
OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)
Deviations:
yes
Remarks:
reduction in surveillance dates
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
Details not provided, but known that sampling occurred multiple times during each phase of the test.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: not given
- Controls: dilution water control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): none
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: zebra fish
- Source: cultured at Fraunhofer Laboratories
- Strain: from West Aquarium GmbH in 37431 Bad Lauterberg, Germany

Parental fish held in same water as used in test at 26 °C 12L:12D and fed daily with TetraMin and Artemia nauplii. Fish were without diseases and abnormalities.

POST-HATCH FEEDING DURING TEST
- Type/source of feed: Artemia nauplii
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
185 d
Post exposure observation period:
none
Hardness:
not provided
Test temperature:
26 +/- 0.3 °C
pH:
7.5-8.8 s.u.
Dissolved oxygen:
mean values ranged from 84-86% saturation
Salinity:
NA
Nominal and measured concentrations:
Nominal: 0, 1.2, 3.7, 11.9, 38 µg/L
mean measured for Period 1: 1.0, 2.8, 10.7, 35.4 µg/L
mean measured for Period 2: 1.3, 3.2, 11.4, 34.7 µg/L
mean measured for Period 3: 1.2, 2.9, 12.6, 35.8 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Days 1-14, cages placed in 13.4 L glass aquaria, Days 14-38, 13.4 L glass aquaria, Day 38-185, 29 L glass aquaria
- Material, size, headspace, fill volume: Days 14-38= 12 L, Days 38-185 = 25 L
- Type of flow-through (e.g. peristaltic or proportional diluter): membrane pumps
- Renewal rate of test solution (frequency/flow rate): 2 fold exchange of test medium/day
- No. of fertilized eggs/embryos per vessel: 100
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: purified drinking water according to OECD 210
- Culture medium different from test medium: not different from holding water
- Intervals of water quality measurement: all working days

OTHER TEST CONDITIONS
- Photoperiod: 12L12D


EFFECT PARAMETERS MEASURED (with observation intervals if applicable): hatch rate, survival, F1 growth, fecundity, hatch of F2, survival of F2, growth of F2

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations: not provided
- Results used to determine the conditions for the definitive study: yes 96h LC50 = 0.38 mg/L

POST-HATCH DETAILS
- Begin of post-hatch period: Day 6
- No. of hatched eggs (alevins)/treatment released to the test chamber: 50
- Release of alevins from incubation cups to test chamber on day no.: F1 on day 35, F2 on day 155

FERTILIZATION SUCCESS STUDY
- Number of eggs used: 100 per vessel
Duration:
78 d
Dose descriptor:
NOEC
Effect conc.:
0.012 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks:
F1 generation
Duration:
38 d
Dose descriptor:
NOEC
Effect conc.:
> 0.035 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
F1 generation
Duration:
151 d
Dose descriptor:
NOEC
Effect conc.:
0.012 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
reproduction
Details on results:
no significant effect in % hatch in F1 generation at highest concentration
significant effect in time to reach sexual maturity of F1 generation at 0.035 mg/L concentration
significant effect in number of eggs produced at 0.035 mg/L concentration
no significant effect in F2 survival at 0.035 mg/L concentration
significant effect in F2 growth at 0.035 mg/L

Therefore, most sensitive endpoint = growth of both F1 and F2 generation and fecundity NOEC = 0.012 mg/L
EC10 infertilization = 0.0135 mg/L
Reported statistics and error estimates:
ANOVA followed by Dunnett's Test or Williams Tests. EC/LC10 calculated by Probit.
Validity criteria fulfilled:
yes
Conclusions:
The aim of this project is to contribute to the development of testing strategies for endocrine disrupters.
Executive summary:

The 185 – day chronic toxicity of octylphenol to full life cycle of zebra fish, Danio rerio was studied under flow through conditions.  Fertilized eggs/embryos (100/embryos per chamber of Danio rerio were exposed to control and test chemical average measured concentrations of 1.0, 2.8, 10.7, 35.4 ug /L octylphenol.  The test system was maintained at 26 +/- 0.3 ºC and a pH of 7.5 to 8.8 s.u.  The 78 day and 151 day NOEC values, based on sublethal effects (growth of F1 and F2 generation and reproduction of F1 generation), 0.012 mg/L octylphenol for all effects.   The most sensitive end point was growth and fecundity.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for early life toxicity study with fish.

 

Results Synopsis

 

Test Organism Size/Age (mean wet weight or length): eggs

Test Type (Flow through, Static, Static Renewal): flowthrough

 

LOEC:  0.035 mg octylphenol /L}

NOEC:  0.012 mg octylphenol./L}

Endpoint(s) Effected:  growth of F1 and F2 generation, fecundity

 

Wenzel et al. (2001) was chosen as a Key study because results from this study provided sensitivity information to octylphenol for the test organism, Danio rerio, which is a common aquatic toxicity test species. No other reliable freshwater chronic toxicity data was available for inclusion in this assessment.

Endpoint:
fish life cycle toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Jun 22, 1995 - Sept 12, 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study was conducted to generally accepted scientific standards, was well documented and accpetable for assessment. Some testing water quality characteristics were not provided.
Qualifier:
equivalent or similar to guideline
Guideline:
EPA OPPTS 850.1500 (Fish Life Cycle Toxicity)
Deviations:
not specified
Principles of method if other than guideline:
Test was performed in three phases, exposure to juvenile fish for 28 days, growout phase and reproduction phase until Day 83. Fish not exposed to toxicant in growout or reproduction phases.
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
Exposures were sampled twice a week and on the last day of exposure. Samples were refrigerated or frozen for storage before analysis.
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: dilution water and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
Test organisms (species):
Oryzias latipes
Details on test organisms:
TEST ORGANISM
- Common name: Japanese medaka
- Source: eggs collected from in-house breeding cultures

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Method of collection of fertilised eggs: eggs collected within 5 hours of spawning
- Subsequent handling of eggs: eggs were pooled and separated by rubbing egg mass on mesh screen. Eggs maintained in hatching solution with elevated calcium to promote hatching in 25 °C water.

POST-HATCH FEEDING
Tests were initiated with 5-8 day old fish. Fish were fed live brine shrimp in the morning and Tetramin flake food in afternoon
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Post exposure observation period:
Yes, fish were observed for survival, growth and reproduction after 28 days of exposure. Post exposure observation was recorded until 83 Days from test initiation.
Hardness:
not given
Test temperature:
25 °C
pH:
not given
Dissolved oxygen:
not given but aerated
Salinity:
NA
Nominal and measured concentrations:
nominal: control, solvent control, 0.5, 2.0, 8.0 µg/L
average measured: ND, 0.44, 0.54, 0.77, 1.93 µg/L
Details on test conditions:
TEST SYSTEM
- Emybro cups (if used, type/material, size, fill volume):
- Test vessel: 40 L aquaria
- Type of flow-through (e.g. peristaltic or proportional diluter): peristaltic pump
- Renewal rate of test solution (frequency/flow rate): 40 ml/min
- No. of larvae per vessel: 240
- No. of vessels per concentration (replicates): 1 for initial exposure, 3 for growout phase
- No. of vessels per control (replicates): 1 for initial exposure, 3 for growout phase
- No. of vessels per vehicle control (replicates): 1 for initial exposure, 3 for growout phase

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap
- Culture medium different from test medium: yes in that the eggs were cultured in hatching solution high in calcium to facilitat hatching

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D


EFFECT PARAMETERS MEASURED (with observation intervals if applicable): daily assessed for survival, growth at day 31, hatchability

VEHICLE CONTROL PERFORMED: yes

RANGE-FINDING STUDY
- Test concentrations: unknown
- Results used to determine the conditions for the definitive study: yes


Experiment was conducted in three phases; exposure, growout and reproduction. On Day 28 of test, toxicant stock was no longer delivered to test chambers. Day 31, fish were removed from test aquaria and placed in clean water in 40 L aquaria, three replicates/ treatment group. 60 fish per aquaria for growout phase in static conditions. Remaining fish measured for length/weight. On day 56 of test, some fish were removed and assessed for secondary sex characteristics. Fish from each treatment were randomly assigned to 20L aquaria until there were 10 breeding pairs. These fish were maintained as in growout phase and egg production assessed. Eggs were collected and cultured in hatching solution. On day 83, remaining fish were assessed for length and weight.
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
> 0.002 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
> 0.002 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Details on results:
The authors found no significant difference in mortality or growth at Day 28 from treatments and controls.
Reported statistics and error estimates:
ANOVA or Kruskal-Wallis ANOVA was used.

Alive:Dead Ratios for Each Test Phase and Growth Results at Day 31

 Phase Blank  Acetone  0.5 mg/L  0.77 ug/L  1.93 ug/L 
Exposure  221:19  223:17  232:8  220:20  228:12 
Growout  179:1  179:1  177:3  177:3  180:0 
 Reproduction 78:2  80:0  80:0  79:0  80:0 
 Length (mm) 18.2 (pooled)  -  18.6  18.4  18.9 

Among treatment groups, no differences were found in percent viable eggs. Significant differences in fecundity were found between the 0.5 mg/L treatment group and the solvent control, but not the blank control or 1.93 mg/L treatment.

Validity criteria fulfilled:
not specified
Conclusions:
The study authors conclude that, at the concentrations tested, nonylphenol did not result in observable developmental or reproductive toxicity.
Executive summary:

The 83– day chronic toxicity of nonylphenol to a semi-full-life cycle of Medaka (Oryzias latipes) was studied under flow through conditions.  Larvae, 240 fish (5 -8 days old) per test concentration of Medaka were exposed to control, solvent control and measured concentrations of 0.5, 0.77, 1.93 ug/L nonylphenol.  The test system was maintained near 25 ºC.  The 28– day NOEC value, based on survival and growth was >0.00193 ug/L nonylphenol. At Day 28, fish were no longer exposed to nonylphenol and were transferred to clean water for growout and reproduction phases.

The authors found no significant difference in mortality or growth at Day 28 from treatments and controls.

 

This toxicity study is classified as acceptable supporting study.

 

Results Synopsis

 

Test Organism Size/Age: 5 -8 day old larvae

Test Type (Flowthrough, Static, Static Renewal): flow through

 

NOEC survival:  >0.00193 mg/L nonylphenol

Endpoint(s) Effected: mortality, growth

Nimrod and Benson (1998) was chosen as a supporting study because it was a somewhat well documented study and performed on a common sensitive organism, Oryzias latipes.

Although this study does not provide a definitive endpoint result, results show that this species may be one of the more sensitive species to nonylphenol. Results are in general agreement with Yakota (2001) and show that the sensitivity of

Oryzias latipes is not more sensitive than Yakota (2001).

.
Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
February 1999 and 2000
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study meets generally accepted scientific standards and acceptable for assessment. Some testing information is missing.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 215 (Fish, Juvenile Growth Test)
Deviations:
yes
Remarks:
did not measure growth
Principles of method if other than guideline:
Generally followed fish juvenile test except purpose of test was to determine effects on gill ATPase activity, plasma vitellogenin levels and hypoosmoregulatory performance. However, mortality was also measured.
GLP compliance:
not specified
Analytical monitoring:
not specified
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: solvent
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not provided
Test organisms (species):
Salmo salar
Details on test organisms:
TEST ORGANISM
- Common name: Atlantic salmon
- Source: Marine Harvest-MacConnell, Loch Shiel hatchery, Scotland
- Age at study initiation (mean and range, SD): smolt
- Length at study initiation (mean and range, SD): 148.3 +/-2.6 mm


FEEDING DURING TEST
- Food type: not provided
- Amount: not provided
- Frequency: not provided

Fish were fed to satiation twice a day with commercial salmon pellets during acclimation before test.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
30 d
Post exposure observation period:
3 months in seawater
Hardness:
not provided
Test temperature:
12 +/- 1 °C
pH:
not provided
Dissolved oxygen:
not provided
Salinity:
NA
Nominal and measured concentrations:
nominal: solvent control, 5, 10, 15, 20 µg/L nonylphenol
Details on test conditions:
TEST SYSTEM
- Test vessel:glass tanks with 63 L volume
- Type of flow-through (e.g. peristaltic or proportional diluter): peristaltic
- Renewal rate of test solution (frequency/flow rate): 1L/min
- No. of organisms per vessel: 12
- No. of vessels per concentration (replicates): unknown
- No. of vessels per vehicle control (replicates): unknown


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap
-Culture medium differentfrom test medium: fish were acclimated in dechlorinated tap

OTHER TEST CONDITIONS
- Photoperiod: natural

VEHICLE CONTROL PERFORMED: yes



Reference substance (positive control):
no
Duration:
30 d
Dose descriptor:
NOEC
Effect conc.:
> 0.02 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
Study states no mortality of fish up to 3 months after exposure.
No significant differences in levels of gill Na, K ATPase activity between treatment and controls.
Reported statistics and error estimates:
Statistics not performed on mortality assessment since no mortality occurred. Differences in ATPase activity were determined by one-way ANOVA
Validity criteria fulfilled:
not specified
Remarks:
water quality parameters during testing are not provided
Conclusions:
Study results indicate that low levels of nonylphenol did not induce an increase in plasma VTG or have significant effects on gill ATPase activity in Atlantic salmon smolts.
Executive summary:

The 30– day chronic toxicity of nonylphenol to Atlantic salmon (Salmo salar) smolts was studied under flow through conditions.  Juveniles, 12 fish per test concentration

were exposed to solvent control and nominal concentrations of 5, 10, 15, and 20 µg/L nonylphenol.  The test system was maintained near 12 ºC.  The 30– day NOEC value, based on mortality was >20 µg/L nonylphenol.

 

This toxicity study is classified as acceptable supporting study.

 

Results Synopsis

 

Test Organism Size/Age: juvenile

Test Type (Flowthrough, Static, Static Renewal): flow through

 

NOEC survival:  >0.020 mg/L nonylphenol

Endpoint(s) Effected: mortality

Moore et al (2003) was chosen as a supporting study because it was performed on a sensitive organism, Salmo salar.

There were no other reliable toxicity studies with nonylphenol for this species. Results are in general agreement with other studies with the similar species (Afonso et al 2002).
Endpoint:
long-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
unknown
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study generally meets accepted scientific standards, well documented and acceptable for assessment. Some test water quality information is not provided.
Qualifier:
equivalent or similar to guideline
Guideline:
other: OECD 229
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
All test chambers were tested once a week.
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: NP was dissolved in solvent
- Controls: dilution water control and solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dimethylsulfoxide
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.0001%
Test organisms (species):
Oryzias latipes
Details on test organisms:
TEST ORGANISM
- Common name: Japanese medaka
- Source: broodstock maintained at Chemicals Evaluation and Research Institute laboratory

FEEDING
- Type/source of feed: artemia nauplii
- Frequency: twice a day

Test organisms were adults about 4 months post hatch, with weights of 396 +/-54 mg and lengths of 35 +/-1.5 mm.

60 mating pairs were placed in 1 L chambers with test dilution water at 24 °C to assess fertility for three weeks. Spawned eggs were assessed for fertility. Of the 60 acclimated pairs, 48 pairs with mean fertility > 95% were chosen to use in the toxicity tests.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
21 d
Post exposure observation period:
none
Hardness:
not provided
Test temperature:
24.3 +/-0.5 °C
pH:
7.3 +/- 0.1 s.u.
Dissolved oxygen:
7.6 +/- 0.2 mg/L
Salinity:
NA
Nominal and measured concentrations:
nominal: control, solvent control, 25, 50, 100, 200 µg/L
average measured: ND, ND, 24.8, 50.9, 101, 184 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass chamber 15.0 cm x 17.5 cm holding 1.8 L test solution
- Type of flow-through (e.g. peristaltic or proportional diluter): mini-diluter
- Renewal rate of test solution (frequency/flow rate): 12 chamber volumes/24 h
- No. of organisms per vessel: 2 (1 breeding pair) per compartment, 4 compartments per test vessel
- No. of vessels per concentration (replicates): 4 (each vessel with 4 compartments)
- No. of vessels per control (replicates): 4 (each vessel with 4 compartments)
- No. of vessels per vehicle control (replicates): 4 (each vessel with 4 compartments)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D
- Fish fed artemia nauplii twice a day
- pH, DO, temperature measured once a week

RANGE FINDING TESTS
- Tested concentrations: 0, 50, 100, 200 µg/L
- Results used to determine the conditions for the definitive study: yes

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): survival, fecundity, fertility and spawning assessed daily. At test termination, morphological changes in anal fin were assessed. Gonads were removed and weighted to determine GSI. Hepatic VTG was also measured.

Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
0.051 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
fertility
Remarks:
and fecundity
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
0.101 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
fertility
Remarks:
and fecundity
Details on results:
No fish in the control or solvent controls died. No females fish in the treatments died. However three male fish in the 184 µg/L treatment died. Also, there was no significant difference between treatments and controls in the numbers of papillary processes in anal fins.
A significant decrease in fecundity occurred in the 101 µg/L concentration during days 15 to 21. A significant decrease in mean fertility was also seen in the 184 µg/L concentration.
Reported statistics and error estimates:
Arc-sine/square root transformation was used for fertility data assessment. Student's t-test was used to compare control groups. Homogeneity of variance was tested by using Leven's test. ANOVA followed by Dunnett's multiple comparison tests was used for data analysis.
Validity criteria fulfilled:
yes
Conclusions:
Study authors conclude that exposure to 4-nonylphenol, at the concentrations tested, affects reproduction of mating pairs of medaka.
Executive summary:

The 21– day short-term chronic toxicity of nonylphenol to reproduction stage of the Japanese medaka (Oryzias latipes) was studied under flow through conditions.  Paired adult fish, 16 pairs per test concentration of Oryzias latipes were exposed to control, solvent control and average measured concentrations of 24.8, 50.9, 101, 184 µg/L nonylphenol.  The test system was maintained at 24.3 +/-0.5 ºC.  The 21– day NOEC value, based on sublethal effects (fecundity, fertility), was 0.0509 mg/L.   There was no mortality in the control treatments.

This toxicity study is classified as acceptable Supporting study and satisfies the guideline requirement for toxicity study with fish.

 

Results Synopsis

 

Test Organism Size/Age (mean wet weight or length): paired sexually mature adults

Test Type (Flowthrough, Static, Static Renewal): flow through

 

NOEC:  0.0509 mg/L nonylphenol

Endpoint(s) Effected: fertility, fucundity

Kang et al. (2003) was chosen as a supporting study because tests were performed on a common sensitive fish species. Results of this study are in general agreement with other studies with medaka (Yakota et al. (2001) and Nimrod and Benson (1998)).

Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Aug-Sept 1993
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study is comparable to a guidelines study with acceptable restrictions which do not impair the overall conclusion from the data. No information is given regarding statistical procedures used. No information is given regarding GLP.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: ASTM 1993 Standard Practice for Conducting Bioconcentration Tests with Fishes and Saltwater Bivalve Molluscs. E1022-84
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
other: USEPA
Deviations:
not specified
Remarks:
no method number or title given
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 215 (Fish, Juvenile Growth Test)
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
All test chambers were tested at 4 h after test initiation and three times a week thereafter. Duplicate samples were collected.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: no solvents used to generate stock solution. Solution was stirred with magnetic stirrer. Test concentrations were prepared by withdrawling stock solution by pump. Solution was withdrawn near the bottom of the flask.
- Controls: dilution water control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): None
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): NA
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none
Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Common name: fathead minnow
- Source: brood cultures maintained at Univ. of WI - Superior


Cultures were hatched and reared for 30 days in laboratory water. Fry were fed live brine shrimp nauplii three times a day.
Fish were 4 weeks old (juvenile) at test initiation.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Post exposure observation period:
none
Hardness:
49.1 +/-0.3 mg/L CaCO3
Test temperature:
25.1+/-0.02 °C
pH:
7.6 +/- 0.02 s.u.
Dissolved oxygen:
6.5 +/- 0.6 mg/L
Salinity:
NA
Nominal and measured concentrations:
measured: < MDL, 9.3, 19.2, 38.1, 77.5, and 193 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 2.5 L glass exposure chambers
- Type of flow-through (e.g. peristaltic or proportional diluter): diluter
- Flow rate: 16 ml/min
- No. of fertilized fish per vessel: 20
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 0

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap
- Alkalinity: 48.6 mg/L
- TOC: assumed <5.0 mg/L as used dechlorinated tap for dilution water
- Intervals of water quality measurement: pH, temp, DO measured daily, hardness, alkalinity, and conductivity measured weekly for duration of test

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality assessed daily, growth

VEHICLE CONTROL PERFORMED: no




Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
0.038 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks:
wet weight
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
0.077 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
0.077 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
0.193 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
no mortality occurred in the controls
Reported statistics and error estimates:
no information

Wet Weight and Mortality of Fathead Minnows at Day 28

  Control  9.3 (ug/L)  19.2 (ug/L)  38.1 (ug/L)  77.5 (ug/L)  193 (ug/L) 
wet weight (g) 0.448  0.390  0.327  0.374  0.266* 0.206*
 # dead 0  0  0  1   0 14* 

*Significantly different from control

Validity criteria fulfilled:
yes
Conclusions:
Study author just presents study results and makes no formal overall conclusions.
Executive summary:

The 28– day chronic toxicity of nonylphenol to juvenile life stage of Fathead minnow (Pimephales promelas) was studied under flow through conditions.  Juvenile fish, 40 fish per test concentration of Fathead minnow were exposed to control, and measured concentrations of 9.3, 19.2, 38.1, 77.5, and 193 µg/L nonylphenol.  The test system was maintained near 25 ºC and a pH of 7.6 s.u.  The 28– day NOEC value, based on sublethal effects (growth), was 38.1 µg/L. The 28 -day NOEC value, based on mortality was 77.5 µg/L. The most sensitive end point was growth.

 

This toxicity study is classified as acceptable supporting study.

 

Results Synopsis

 

Test Organism Size/Age: juvenile

Test Type (Flow through, Static, Static Renewal): flow through

 

NOEC growth:  0.038 mg/L nonylphenol

NOEC survival:  0.0775 mg/L nonylphenol

Endpoint(s) Effected: mortality, growth

Brooke (1993b) was chosen as a supporting study because it was a well documented study, performed equivalent to guideline study on a preferred organism, Pimephales promelas. Results of this study are in agreement with another chronic study, but is slightly less sensitive.

Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Aug-Sept 1993
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study is comparable to a guidelines study with acceptable restrictions which do not impair the overall conclusion from the data. No information is given regarding statistical procedures used. No information is given regarding GLP.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: ASTM 1993 Standard Practice for Conducting Bioconcentration Tests with Fishes and Saltwater Bivalve Molluscs. E1022-84
Deviations:
not specified
Qualifier:
according to guideline
Guideline:
other: USEPA
Deviations:
not specified
Remarks:
no method number or title given
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 215 (Fish, Juvenile Growth Test)
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
All test chambers were tested at 4 h after test initiation and three times a week thereafter. Duplicate samples were collected.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: no solvents used to generate stock solution. Solution was stirred with magnetic stirrer. Test concentrations were prepared by withdrawling stock solution by pump. Solution was withdrawn near the bottom of the flask.
- Controls: dilution water control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): None
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): NA
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
TEST ORGANISM
- Common name: bluegill
- Source: certified disease free, acquired from Klaubec Fish Farms, Amana, IA

Juvenile (10-12 weeks old) were acclimated in laboratory water for at least 7 days prior to test initiation.
Fish were fed live brine shrimp twice a day during acclimation and during the test.

Fish were likely 11-13 weeks old at test initiation. Study states that "Juvenile bluegills (4 wk old) were exposed..." However, this is likely a typo.
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Post exposure observation period:
none
Hardness:
49.0 +/-0.3 mg/L CaCO3
Test temperature:
25.0+/-0.02 °C
pH:
7.55 +/- 0.01 s.u.
Dissolved oxygen:
6.1 +/- 0.5 mg/L
Salinity:
NA
Nominal and measured concentrations:
measured: < MDL, 5.6, 12.4, 27.6, 59.5, 126 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 2.5 L glass exposure chambers
- Type of flow-through (e.g. peristaltic or proportional diluter): diluter
- Flow rate: 16 ml/min
- No. of fertilized fish per vessel: 20
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 0

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap
- Alkalinity: 48.9 mg/L
-TOC: assumed <5.0 mg/L as used dechlorinated tap for dilution water
- Intervals of water quality measurement: pH, temp, DO measured daily, hardness, alkalinity, and conductivity measured weekly for duration of test

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D


EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality assessed daily, growth

VEHICLE CONTROL PERFORMED: no




Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
0.059 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
0.126 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
No mortality occurred in the controls. Although growth was measured, no significant effect from the control was seen.
Reported statistics and error estimates:
no information

Wet Weight and Mortality of Fathead Minnows at Day 28

  Control  5.6(ug/L)  12.4 (ug/L)  27.6 (ug/L)  59.5 (ug/L)  126 (ug/L) 
wet weight (g) 0.400  0.406  0.361  0.455  0.345 0.402
 # dead 0  0  0  10  1 3* 

*Significantly different from control

Validity criteria fulfilled:
yes
Conclusions:
Study author just presents study results and makes no formal overall conclusions.
Executive summary:

The 28– day chronic toxicity of nonylphenol to juvenile life stage of Bluegill (Lepomis macrochirus) was studied under flow through conditions.  Juvenile fish, 40 fish per test concentration of Bluegill were exposed to control, and measured concentrations of 5.6, 12.4, 27.6, 59.5, 126 µg/L nonylphenol.  The test system was maintained near 25 ºC and a pH of 7.6 s.u.  The 28– day NOEC value, based on mortality was 59.5 µg/L.   Although growth was measured, no significant effect from the control was seen.  

This toxicity study is classified as acceptable supporting study.

 

Results Synopsis

 

Test Organism Size/Age: juvenile

Test Type (Flow through, Static, Static Renewal): flow through

 

NOEC survival:  0.0595 mg/L nonylphenol

Endpoint(s) Effected: mortality

Brooke (1993b) was chosen as a supporting study because it was a well documented study, performed equivalent to guideline study on a sensitive organism, Lepomis macrochirus. Results of this study are the only acceptable data for this species. Results are in general agreement with other species such as Fathead minnow (Brooke (1993b).

Endpoint:
fish early-life stage toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
unknown
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study generally meets accepted scientific standards and is described in sufficient detail.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: ASTM
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
yes
Details on sampling:
All test chambers were tested twice a week for the duration of the test.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: no solvents used to generate stock solution. Solution was stirred with magnetic stirrer. Test concentrations were prepared by withdrawling stock solution by pump. Solution was withdrawn near the bottom of the flask.
- Controls: dilution water control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): None
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): NA
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: rainbow trout
- Source: certified disease-free embryos supplied as ova and sperm from Mount Lassen Trout Farms, Inc., Red Bluff, CA

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Method of collection of fertilised eggs: 5,000 ova from three females and sperm from four males were combined in 1 L water.
- Subsequent handling of eggs: Embryos were immediately drawn into a glass tube after fertilization, counted and placed randomly into incubation cups for testing

POST-HATCH FEEDING
- Start date: day 45 of test
- Type/source of feed: trout chow
- Amount given: enough so there was excess at bottom of test chamber
- Frequency of feeding: 3 times a day
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
91 d
Post exposure observation period:
none
Hardness:
54.0 +/-6.4 mg/L CaCO3
Test temperature:
10.7+/-0.7 °C
pH:
6.97 +/- 0.39 s.u.
Dissolved oxygen:
9.4 +/- 0.7 mg/L
Salinity:
NA
Nominal and measured concentrations:

measured: < MDL, 6.0, 10.3, 23.1, 53.0, 114 ug/L
Details on test conditions:
TEST SYSTEM
- Embryo cups (if used, type/material, size, fill volume): oscillating incubation chambers of 100 ml beakers with bottoms removed and replaced with nylon mesh
- Test vessel: 2 L exposure chambers
- Type of flow-through (e.g. peristaltic or proportional diluter): diluter
- No. of fertilized eggs/embryos per vessel: 100
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 0

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- Alkalinity: 41.0+/- 3 mg/L
- Intervals of water quality measurement: temp measured daily, DO measured ever 48 h, hardness, alkalinity, pH, and conductivity measured weekly for duration of test

OTHER TEST CONDITIONS
- Photoperiod: 16L:8D


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

VEHICLE CONTROL PERFORMED: no

POST-HATCH DETAILS
- Begin of post-hatch period: day 29-39 days
- No. of hatched eggs (alevins)/treatment released to the test chamber: 15 fry per test chamber
- Release of alevins from incubation cups to test chamber on day no.: approx day 45



Duration:
91 d
Dose descriptor:
NOEC
Effect conc.:
0.006 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
91 d
Dose descriptor:
LOEC
Effect conc.:
0.01 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Details on results:
NOEC and LOEC are based on total length, and wet and dry weights
Reported statistics and error estimates:
NOEC and LOECs determined by ANOVA test. Dunnett's test was used to determine significant effects from controls.

Hatchability, Survival and Growth of Rainbow Trout Exposed to Nonylphenol for 91 Days

  Control  6.0 (ug/L)  10.3 (ug/L)  23.1 (ug/L)  53.0 (ug/L)  114 (ug/L) 
Mean % hatch  83.5  89  88.5  92  92.5  88.5 
Mean % survival at Day 51d post-hatch  93.3  93.3  90  33.3  0  0 
Mean wet weight (g) at 51d post-hatch  0.734  0.716  0.516  0.375  0  0 
Mean dry weight (g) 51d post-hatch  0.135  0.133  0.095  0.066  0  0 
Mean standard length (mm) 51d post-hatch  35.8  38.0  33.1  29.8  0  0 
Mean biomass/chamber  1.89  1.87  1.28  0.33  0  0 
Conclusions:
Study author just presents study results and makes no formal overall conclusions.
Executive summary:

The 91– day chronic toxicity of nonylphenol to early life stage of Rainbow Trout (Oncorhynchus mykiss) was studied under flow through conditions.  Fertilized eggs/embryos 100 fertilized eggs per test concentration of Oncorhynchus mykiss

were exposed to {control, and measured concentrations of 6.0, 10.3, 23.1, 53.0, 114 ug/L nonylphenol.  The test system was maintained at 10.7 +/- 0.7 ºC and a pH of 6.97 +/- 0.39 s.u.  The 91– day NOEC value, based on sublethal effects (growth), was 6.0 mg/L.   Mean percent hatch of any test concentration was not significantly different from controls. Time to hatch was 39 +/- 5 days with swim-up at approximately at day 45. The most sensitive end point was growth.

 

This toxicity study is classified as acceptable Key study and satisfies the guideline requirement for early life toxicity study with fish.

 

Results Synopsis

 

Test Organism Size/Age (mean wet weight or length): freshly fertilized eggs

Test Type (Flowthrough, Static, Static Renewal): flow through

 

LOEC:  0.0103 mg/L nonylphenol

NOEC:  0.006 mg/L nonylphenol

Endpoint(s) Effected: growth

Endpoint:
fish short-term toxicity test on embryo and sac-fry stages
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2001
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study meets generally accepted scientific standards and is acceptable for assessment. Some testing information is missing such as water quality characteristics during testing. Concentrations are nominal
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 212 (Fish, Short-term Toxicity Test on Embryo and Sac-Fry Stages)
Deviations:
yes
Principles of method if other than guideline:
Study authors state no specific guidelines were followed. Salmon alevins (sac-fry) were exposed for 29 days post hatch in static-renewal test to different test concentrations. Mortality as well as genetic sex was determined using Y-chromosomal DNA markers at 103 and 179 days post hatch.
GLP compliance:
not specified
Analytical monitoring:
no
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Controls: dilution water and solvent
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): ethanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.1%
Test organisms (species):
Oncorhynchus tschawytscha
Details on test organisms:
TEST ORGANISM
- Common name: Chinook salmon
- Source: Big Qualicom River Hatchery
- Age at study initiation (mean and range, SD): post hatch
- Feeding during test: none as alevins were sac-fry

Exposure started at hatching

ACCLIMATION
- Eggs were obtained from hatchery, placed in incubation trays with well water at 10 °C.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
29 d
Post exposure observation period:
Fish were sampled at 103 and 179 days post hatch to determine sex by DNA markers.
Hardness:
not provided
Test temperature:
10 °C
pH:
not provided
Dissolved oxygen:
not provided
Salinity:
NA
Nominal and measured concentrations:
nominal: control, solvent control, 0.001, 1, 10 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: plastic perforated chambers within glass aquaria holding 4 L of test solution
- Renewal rate of test solution (frequency/flow rate): every second day
- No. of fertilized eggs/embryos per vessel: 100 alevins per replicate
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: well water
- Culture medium different from test medium: unknown

OTHER TEST CONDITIONS
- Photoperiod: darkness. Study authors state Salmon alevin are grown in darkness to mimic their natal incubation in gravel redds in rivers

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality assessed before renewal

VEHICLE CONTROL PERFORMED: yes

Reference substance (positive control):
yes
Remarks:
17B-estradiol, testosterone
Duration:
29 d
Dose descriptor:
NOEC
Effect conc.:
> 0.01 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
Survival ranged from 97-100% in all treatments.
Reported statistics and error estimates:
none provided
Validity criteria fulfilled:
not specified
Remarks:
water quality parameters during testing are not provided
Conclusions:
Study concluded that DNA markers can provide a useful tool for investigating influences of environmental factors on sex determination and differentiation.
Executive summary:

The 29– day chronic toxicity of nonylphenol to a alevins of Chinook Salmon (Oncorhynchus tshawytscha) was studied under static renewal conditions.  Alevins, 200 per test concentration were exposed to control, solvent control and nominal concentrations of 0.001, 1.0, 10 ug/L nonylphenol.  The test system was maintained near 10 ºC.  The 29– day NOEC value, based on mortality was >10 ug/L nonylphenol.

 

This toxicity study is classified as acceptable supporting study.

 

Results Synopsis

 

Test Organism Size/Age: alevins

Test Type (Flowthrough, Static, Static Renewal): static renewal

 

NOEC survival:  >0.01 mg/L nonylphenol

Endpoint(s) Effected: mortality

Afonso et al. (2002) was chosen as a supporting study because it was a performed on a commercially important organism, Oncorhynchus tshawytscha. There is no other reliable aquatic toxicity of nonylphenol to this species.

Results are in general agreement with other studies with the similar species.
Endpoint:
fish life cycle toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
unknown
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study meets generally accepted scientific standards, is adequately documented and acceptable for assessment. Some testing procedure specifics are not provided. Study was not performed according to GLP.
Qualifier:
equivalent or similar to guideline
Guideline:
EPA OPP 72-5 (Fish Life Cycle Toxicity)
Principles of method if other than guideline:
Study author refers to OECD Guideline 204 for analytical requirements. However, this test was a full life cycle test on rainbow trout, starting with eyed-eggs through one year of life. Experiment tested only two different nonylphenol concentrations.
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
NA
Analytical monitoring:
yes
Details on sampling:
Test concentrations were sampled every three weeks throughout the 1 year experiment.
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 1 g/L of NP was mixed with solvent and spring water
- Controls: yes 0.01 % solvent control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dimethylsulfoxide 0.1%
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: rainbow trout
- Source: Swiss Federal Institute for Environmental Science and Technology brood stock

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
trout eggs were obtained by artificial fertilization of in house brood stock

POST-HATCH FEEDING
- Start date: "when fish started feeding"
- Type/source of feed: BioMar A/S Brande, Denmark
- Amount given: 1.5 % of body weight
- Frequency of feeding: once per day
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
365 d
Post exposure observation period:
no
Hardness:
21.3 dGH
Test temperature:
10 +/- 1 °C
pH:
7.7-7.8 s.u.
Dissolved oxygen:
80 % saturation
Salinity:
NA
Nominal and measured concentrations:
nominal: control, solvent control, 1, and 10 µg/L
measured: ND, ND, 1.05, and 10.17 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 100 L aquaria
- Renewal rate of test solution (frequency/flow rate): 9 L/h
- No. of fertilized eggs/embryos per vessel: 1400 fertilized eggs per group
- No. of vessels per concentration (replicates): not given


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: spring water
- Culture medium different from test medium: no
- Intervals of water quality measurement: regularly checked

OTHER TEST CONDITIONS
- Photoperiod: 12L:12D


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality checked continuously until hatching, hatch time also noted

VEHICLE CONTROL PERFORMED: yes



Duration:
365 d
Dose descriptor:
NOEC
Effect conc.:
> 10 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
365 d
Dose descriptor:
NOEC
Effect conc.:
> 10 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
growth rate
Duration:
365 d
Dose descriptor:
NOEC
Effect conc.:
> 10 µg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
number hatched
Details on results:
There were no significant effects in either test concentration from the dilution water control or the solvent control.
Mortality rate in the eyed-egg stage in the test treatments and solvent control were 13-15% and 10% in the blank control. Total mortalities did not result in significant reduced hatch rates in any treatment compared to the control.
No significant affects on body weight were seen in the experiment from exposure to NP for one year at test concentrations.
Reported statistics and error estimates:
Body weight data were log-transformed. Normality of data was determined by probability plots. Student's t-test was used to determine differences between blank and solvent control. ANOVA was used to determine statistical significance.
Validity criteria fulfilled:
yes
Conclusions:
Study authors' conclusions are that sex-reversal or testis-ova in rainbow trout are not induced at the NP concentrations tested. Vitellogenin, VG mRNA and zona radiata protein (ZRP) expression were also measured. There was increased VG expression in the trout liver, but VG mRNA levels were not elevated. Although typical concentrations found in sewage treatment wastewater can induce VG and ZRP expression, these concentrations do not affect sexual differentiation in rainbow trout.
Executive summary:

The 365 – day chronic toxicity of nonylphenol to 1 year life cycle of Rainbow trout (Oncorhynchus mykiss) was studied under flow through conditions.  Fertilized eggs/embryos {14000 eggs/test concentration} of Oncoryhnchus mykiss

were exposed to control, solvent control, measured concentrations of 0.001 and 0.01mg/L NP.  The test system was maintained at 10 +/- 1.0 ºC and a pH of 7.7 to 7.8  The 365 – day NOEC values, based on mortality/growth and hatchability, were >0.01 mg/L.   There were no significant effects in mortality, growth or hatchability in either test concentration from the dilution water control or the solvent control..

 

 

Results Synopsis

 

Test Organism Size/Age (mean wet weight or length): started with eyed-eggs

Test Type (Flowthrough, Static, Static Renewal): flowthrough

 

LOEC:  >0.01 mg/L nonylphenol for survival, growth, and hatchability

 

 

Description of key information

Wenzel et al. (2001), performed according to OECD 212, provided a NOEC of 0.012 mg/L octylphenol based on growth and fecundity for the zebrafish, Danio rerio.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Dose descriptor:
NOEC
Effect concentration:
0.012 mg/L

Additional information

Review of toxicity test results for long-term exposure of octylphenol to fish resulted in a key study of reliable data by Wenzel et al.,(2001) with the freshwater fish Danio rerio.  The key study byWenzel et al.,(2001) provided two endpoint NOEC values at a 78 day exposure that included 0.012 mg octylphenol/L for inhibition of reproduction and >0.035 mg octylphenol/L for survival of Danio rerio.