Registration Dossier

Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
fertility, other
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
10 Dec 1950 - 14 Apr 1954
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented publication which meets basic scientific principles.
Cross-reference
Reason / purpose:
reference to same study

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Unnamed
Year:
1955
Reference Type:
publication
Title:
Fütterungsversuch mit Steinkohlenflugasche bei Milchkühen.
Author:
Goetze, R., und Herrmann, M.
Year:
1956
Bibliographic source:
Schriftenreihe des Vereins für Wasser-, Boden- und Lufthygiene Berlin-Dahlem 11, 81-219 (1956), Gustav Fischer Verlag, Stuttgart.
Reference Type:
publication
Title:
Die Nutzbarkeit der Steinkohlenflugasche zur Mineralstoffversorgung der Haustiere.
Author:
Goetze, R.
Year:
1956
Bibliographic source:
Schriftenreihe des Vereins für Wasser-, Boden- und Lufthygiene Berlin-Dahlem 11, 221-224 (1956), Gustav Fischer Verlag, Stuttgart.

Materials and methods

Principles of method if other than guideline:
Fertility study in grazing cows
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Fly ash from (black) coal combustion, Scholven, Germany
- Other: Further details are given in the tables below

Test animals

Species:
other: cattle
Strain:
other: Schwarzbunte Niederungsrasse
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Oldenburg, Germany (a geographical region which was assumed to be free of fly ash from coal combustion)
- Age at study initiation: 2.75 - 6.75 years
- Weight at study initiation: 535 - 710 kg
- Housing: Cattle pen; the animals were allowed to graze.
- Diet (e.g. ad libitum): fodder beed, hay and straw (ad libitum) supplemented with mash.
At their arrival the animals were carefully clinically examined. All animals were in good nutritional state and showed no symptoms of illness.
Some animals were seropositive for tuberculosis.

ENVIRONMENTAL CONDITIONS
All animals were kept on farm without contact to other cattle.
The cattle pen was always kept clean, dry and ventilated with fresh air.

IN-LIFE DATES:
Exposure period: From: 10 Dec 1950 To: 15 Dec 1953
Post exposure observation period: From: 16 Dec 1953 To: 14 Apr 1954

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test substance was mixed with the daily food.
Animals were fed twice a day in individual boxes to assure the intake of the individual portions.

DIET PREPARATION
In the high dose group, the fly ash was mixed with soaked fodder beed and mash, to improve the taste and to ascertain the daily intake of the high amounts of fly ash.
Details on mating procedure:
All animals were naturally mated.
At the beginning of the experiment, 8 out of 9 animals were already pregnant.
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
Exposure period: 3 years
Premating exposure period (females): no data
Frequency of treatment:
twice a day
Details on study schedule:
not applicable
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
300 g / animal / day
Basis:
nominal conc.
fed to the animals of group 1 and group 2
Remarks:
Doses / Concentrations:
1500 - 1800 g / animal / day
Basis:
nominal conc.
fed to the animals of group 3
Remarks:
Doses / Concentrations:
additionally approx. 300 g / animal / day
Basis:
nominal conc.
resulting from calculated amounts of coal fly ash precipitation to the grazing land in the experimental geographic area and the daily intake of the grazing test animals.
No. of animals per sex per dose:
9 cows were split into 3 groups (3 animals/group):
Group 1 (animal # 1-3): served as concurrent no treatment control animals in the first two years and received 300 g fly ash/day/animal in the 3rd year of the experiment
Group 2 (animal # 4-6): received 300 g fly ash/day/animal only in the 1st and 2nd year of the experiment, in the 3rd year concurrent no treatment.
Group 3 (animal # 7-9): received 1500 - 1800 g fly ash/day/animal during all 3 years of the experiment
Control animals:
other: yes, concurrent no treatment (for the first two years of the exposure period)
Details on study design:
To exclude possible individual differences in the sensitivity for coal fly ash, a feeding strategy conversion for animal groups 1 and 2 was performed: Animals of group 1 received fly ash only in the third year, whereas the animals of group 2 received the fly ash only in the first two years of the 3-year duration.
Positive control:
not performed

Examinations

Parental animals: Observations and examinations:
REGULAR CLINICAL OBSERVATIONS: Yes
- Time schedule:
First year: every 10 days
Second year: every 14 days
Third year: every 4 weeks

DETAILED CLINICAL OBSERVATIONS: Yes, including parasitological, bacteriological and serological analyses
- Time schedule:
1) at the arrival
2) at the end of the second year of exposure
3) at the end of the exposure period of 3 years
4) at the end of the post-exposure period

BODY WEIGHT: Yes
- Time schedule for examinations:
First year: every 10 days
Second year: every 14 days
Third year: every 4 weeks

MILK PRODUCTION:
Every day the milk production was reported by weighing.

OTHER:
Milk, blood, urine and faeces samples for chemical analyses were taken on December 10th, 1952.
Trace element analysis of liver and bone samples were performed at the end of the experiment.
Sperm parameters (parental animals):
not analysed
Litter observations:
The parturition of all calves was described as undisturbed, but the calves were not further mentioned in the report.
Postmortem examinations (parental animals):
SACRIFICE
- Maternal animals: 2 animals of each group were sacrified at the end of the exposure period, the remaining animals were euthanised at the end of the post exposure observation period.

GROSS NECROPSY
All animals were macroscopically examined.

HISTOPATHOLOGY / ORGAN WEIGHTS
Samples from : Liver, kidneys, pyloric region of the stomach, mesenterial lymph nodes, small and large intestine

OTHER:
Liver samples were chemically analysed for their trace element contents.
Postmortem examinations (offspring):
not examined
Statistics:
not applicable (n=3)
Reproductive indices:
not examined
Offspring viability indices:
not examined

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed

Details on results (P0)

see tables below

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
1 850 - 2 390 mg/kg bw/day (nominal)
Sex:
female
Basis for effect level:
other: overall effects clinical signs; mortality; body weight; gross pathology; histopathology; pregnancy rate dose level calculated on the basis of the reported body weight range (627-809 kg)

Results: F1 generation

General toxicity (F1)

Mortality / viability:
no mortality observed

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

Table 1: Details on the experimental settings

Exposure period: December 10, 1950 - December 15, 1953;

Post exposure recovery period: December 16, 1953 - April 14, 1954

Group

Animal numbers

Treatment during the 1stand 2ndyear

(until December 10, 1952)

Treatment during the 3rdyear

 

A

1-3

concurrent no treatment

300 g fly ash / day / animal

B

4-6

300 g fly ash / day / animal

concurrent no treatment

C

7-9

1500 - 1800 g fly ash /day / animal

1500 - 1800 g fly ash /day / animal

Applicant's conclusion:

Only in the first two years of the experiment, group A served as a control group. Therefore, as far as possible only results after two years of experimental duration were evaluated.

 

Table 2: Individual observations

Ani-mal

reco-very after expo-sure

Age,

body weight and

preg-nancy status

at the experi-mental

start

Clinical

signs

Preg-

nancy

ratea

 

Sexual behaviour

Body

weight

(in kg at similar

stages of preg-

nancy, measured

in 1950, 1951, 1952 and 1953)

Milk produc-tion

 (kg / d; mean values

of the

years

1951 and 1952)

1

no

5 years,

657 kg,

5thmonth

No

clinical

signs of toxico-

logical

relevance were observed.

4/6

No disturbances of conception and parturition.

657, 697, 720, 744

 

10.9

2

yes

2.75 years,

535 kg,

9thmonth

No

clinical

signs of toxico-

logical relevance were observed.

4/4

No disturbances of conception and parturition up to the 3rdbirth in 1953. During the 4thpregnancy an infection with Brucella abortus occured, followed by a preterm delivery in the 8thmonth of pregnancy.

535, n.d., 590, 682

10.7

3b

no

5 years,

585 kg,

not pregnant

No

clinical

signs of toxico-

logical relevance were observed.

3/3

No disturbances of conception and parturition up to the 2ndbirth in April, 1953. In November 1953 an abortion occured after a Brucella infection.

n.d., n.d., 554, 709

11.6

4

yes

4.75 years,

625 kg,

5thmonth

Observed clinical

signs (uterus catarrh, bronchial catarrh)

couldn´t be associated with fly ash administration, as the milk production was highest during fly ash administration

3/6

Mastitis due to a Streptococcus infection occured early after experimental start. Infection with Brucella abortus during post exposure period.

625, n.d., 695, 691

11.3

5

no

4.5 years,

645 kg,

9thmonth

No

clinical

signs of toxico-

logical relevance were observed.

4/6

No disturbances of conception and parturition up to the 3rdbirth in 1953. During the 4thpregnancy an infection with Brucella abortus occured, followed by an abortion in the 8thmonth of pregnancy.

645, 636, 686, n.d.

13.7

6

no

5 years,

710 kg,

9thmonth

No

clinical

signs of toxico-

logical relevance were observed. The occuring mastitis could not be associated with fly ash administration.

4/4

No disturbances of conception and parturition up to the 3rdbirth. During the 4thpregnancy

the animal was euthanised after an accident with fracturing of the lumbar vertebrae.

710, 727, n.d., 812

12.9

7

no

5.5 years, 627 kg,

7thmonth

No

clinical

signs of toxico-

logical relevance were observed.

4/4

Abortion in February 1951 without evidence of bacterial infection. No disturbances of conception and parturition were observed for the following 3 pregnancies.

627, 683, 685, n.d.

11.9

8

no

5.75 years,

670 kg,

7thmonth

No

clinical

signs of toxico-

logical relevance were observed.

4/7

No disturbances of conception and parturition up to the 3rdbirth. During the 4thpregnancy

the animal was euthanised.

670, 719, 732, n.d.

10.3

9

yes

6.75 years,

682 kg,

9thmonth

No

clinical

signs of toxico-

logical relevance were observed.

4/4

No disturbances of conception and parturition during all 4 pregnancies and deliveries.

682, 765, 784, 809

12.3

n.d. = not determined

anumber of pregnancies / number of natural matings

bAnimal #3 was incorporated to the experiment in March 1952 as a replacement for another animal, which was euthanised due to diseases of the lung and intestine. This was considered as not relevant, as this animal belonged to the concurrent no-treatment group.

 

Applicant's conclusion:

Various confounding factors like several infectious diseases like Brucella abortus, Tuber-culosis, liver Trematodes and others (viral infections were not mentioned at all) and complications following accidental injuries (following natural matings, for instance) exacerbate the interpretation of the individual observations. However, no dose-dependent changes of the sexual behaviour, pregnancy rate or development of conceptus and parturition were noted. The general condition of the animals seemed to be not disturbed by the test substance administration, as the milk production was even slightly higher in both treated groups compared to the control group.

Body weights of the high dose group was ranged from 627 to 809 kg. Accordingly, the applied dose was ca. 1850 -2390 mg fly ash / kg body weight / day in the high dose group.

 

Table 3:Individual macroscopic and microscopic pathological findings

Ani-

mal /

Treat-

ment

Necropsy findings

Histological findingsain kidney, liver, stomach, lymph nodes and intestine samples

1

infectious diseases: Liver-Trematodes, Tuberculosis;

changes in claws, which were considered as normal in this species under the given circumstances;

no irritation of gastro-oesophageal mucosae, no changes in bones and jointsb, which could be a consequence of a fly ash intoxication;

no histopathologic changes observed

2

infectious diseases: Liver-Trematodes, Tuberculosis, Brucella abortus;

changes in claws, which were considered as normal in this species under the given circumstances;

no irritation of gastro-oesophageal mucosae, no changes in bones and joints

liver: high glycogen deposition and fatty changes; kidney: glomerulonephritis, brownish pigmentation in tubular epithelial cells, mild nephrosis; lymph nodes: pigmentation

3

infectious diseases: Liver Trematodes, Brucella abortus;

no irritation of gastro-oesophageal mucosae;

changes in uterus and vagina were considered to be a consequence of Brucella infection;

no changes in joints;

no histopathologic changes observed

4

infectious diseases: Tuberculosis, Streptococcus, Brucella abortus;

no irritation of gastro-oesophageal mucosae;

no changes in joints;

 

kidney: glomerulonephritis, pigmentation of tubular epithelial cells; liver: fatty changes, glycogen deposits in centrolobular cells.

5

infectious diseases: Liver Trematodes, Tuberculosis, Brucella abortus;

petechial changes of the gall bladder mucosa, which were considered as not toxicologically relevant;

no irritation of gastro-oesophageal mucosae;

no changes in joints;

 

no histopathologic changes observed

6

infectious diseases: Tuberculosis;

fracture of the lumbar vertebra;

a necropsy of this animal was not considered, as the animal suffered for 6 weeks from the downer cow syndrome due to the lumbar vertebra fracture.

kidneys: lymphocytic interstitial infiltration; liver: mild perivascular lymphocytic infiltration;

7

infectious diseases: Liver Trematodes;

mild erythemacin the pyloric mucosa of the abomasum, the amounts of sand in the gastro-oesophageal mucosa was considered as normal, no defects, bleeding or necrosis in the mucosa of the gastro-oesophageal tract, no other findings of toxicological relevance; no changes in bones and joints;

no histopathologic changes observed with exception for a focal fatty change (fine drops in only one place in the liver)

8

infectious diseases: Liver Trematodes;

the amounts of sand in the gastro-oesophageal mucosa was considered as normal;

no irritation of gastro-oesophageal mucosae;

no changes in bones and joints;

no histopathologic changes observed

9

infectious diseases: Liver Trematodes;

the amounts of sand in the gastro-oesophageal mucosa was considered as normal;

mild erythema in the abomasal mucosa, no defects, bleeding or necrosis in the mucosa of the gastro-oesophageal tract,

no changes in bones and joints;

kidney: chronic glomerulonephritis, focal nephritis chronica interstitialis, protein deposition in tubular epithelial cells;

liver: hydropic degeneration with cell lysis, mild fatty change and glycogen deposits in the centrolobular region; lymph nodes: pigmentation

aThe histologic findings detected in animals #2, #4 and #9 (groups A, B and C) were considered to be without toxicological relevance, as these finding were without a dose-response relation.

bThe joints which were previously described to be affected in animals chronically intoxicated with fly ash

cThe mild erythemas in some gastrooesophageal tracts were considered to be of no toxicological relevance, as this finding was also observed in 25 out of 30 animals analysed at the butchery of Hannover, Germany.

 

Applicant's conclusion:

The pathological findings observed after 3 years of experimental duration are of low toxicological relevance, as too many known and unknown confounding factors like infectious diseases were present and due to the fact, that the treatment regime of animal groups A and B were exchanged after 2 years. However, no major pathological defects of the gastro-intestinal mucosa or other target organs like the joints occurred. Therefore it can be summarized that oral administration of the fly ash up to 1850 mg/kg/day to female cattle for three years did not result in major pathological changes.

 

Table4: Analysis of milk samples collected on December 10, 1952:

(Average values in % determined from two different laboratories in parallel)

Animal

Treatment up to the sampling date

Dry residue

Ash

CaO

MgO

P2O5

1

concurrent no treatment

12.9

0.74

0.172

0.0144

0.212

2

concurrent no treatment

15.8

0.89

0.267

0.0222

0.276

3

concurrent no treatment

10.3

0.71

0.136

0.0155

0.185

4

300 g fly ash per day

11.2

0.78

0.173

0.0170

0.235

5

300 g fly ash per day

14.7

0.82

0.197

0.0194

0.299

6

300 g fly ash per day

14.1

0.71

0.152

0.0204

0.159

7

1500 - 1800 g fly ash per day

15.3

0.69

0.161

0.0161

0.220

8

1500 - 1800 g fly ash per day

14.1

0.73

0.155

0.0144

0.213

9

1500 - 1800 g fly ash per day

10.3

0.73

0.151

0.0153

0.224

Applicant's conclusion:

Two years of oral administration of two different doses of fly ash did not influence the composition of the milk regarding the analysed parameters.

 

Table5: Analysis of blood samples collected on December 10, 1952:

(Average values in % determined from two different laboratories in parallel)

Animal

Treatment up to the sampling date

Dry residue

Ash

CaO

MgO

P2O5

Fe2O3

1

concurrent no treatment

19.2

1.37

0.0127

0.0058

0.0374

0.0534

2

concurrent no treatment

19.9

1.03

0.0110

0.0060

0.0380

0.0544

3

concurrent no treatment

18.7

1.15

0.0116

0.0061

0.0371

0.0435

4

300 g fly ash per day

19.5

1.22

0.0133

0.0062

0.0396

0.0502

5

300 g fly ash per day

19.2

1.36

0.0094

0.0077

0.0333

0.0572

6

300 g fly ash per day

19.6

1.13

0.0101

0.0068

0.0350

0.0430

7

1500 - 1800 g fly ash per day

20.3

1.16

0.0105

0.0071

0.0359

0.0480

8

1500 - 1800 g fly ash per day

18.3

0.98

0.0112

0.0062

0.0362

0.0489

9

1500 - 1800 g fly ash per day

19.3

1.12

0.0116

0.0077

0.0345

0.0547

Applicant's conclusion:

Two years of oral administration of two different doses of fly ash did not influence the composition of the blood regarding the analysed parameters.

 

Table 6: Analysis of urine samples collected on December 10, 1952

(Average values in % determined from two different laboratories in parallel)

Animal

Treatment up to the sampling date

pH

Glucose

Protein

Dry residue

CaO

MgOa

P2O5

1

concurrent no treatment

8.14

neg.

neg.

6.84

0.0055

0.0170

0.0027

2

concurrent no treatment

8.20

neg.

neg.

5.18

0.0018

0.0221

0.0024

3

concurrent no treatment

8.18

neg.

neg.

5.41

0.0011

0.0261

0.0016

4

300 g fly ash per day

8.36

neg.

neg.

5.33

0.0066

0.0090

0.0013

5

300 g fly ash per day

8.26

neg.

neg.

4.53

0.00097

0.0080

0.0014

6

300 g fly ash per day

8.34

neg.

neg.

5.11

0.0043

0.0302

0.0031

7

1500 - 1800 g fly ash per day

8.08

neg.

neg.

3.85

0.0084

0.0100

0.0013

8

1500 - 1800 g fly ash per day

8.40

neg.

neg.

6.33

0.0041

0.0357

0.0022

9

1500 - 1800 g fly ash per day

8.38

neg.

neg.

5.80

0.0233

0.0411

0.0025

aMgO was only analysed from one laboratorium

Applicant's conclusion:

Two years of oral administration of two different doses of fly ash did not influence the composition of the urine regarding the analysed parameters.

 

Table8: Analysis of faeces sampled on December 10, 1952

(Values in % of fresh faeces)

Animal

Treatment up to the sampling date

Ash

SiO2

Al2O3

Fe2O3

MgO

CaO

SO3

P2O5

1

concurrent no treatment

2.77

1.51

0.119

0.066

0.118

0.270

0.057

0.183

2

concurrent notreatment

2.23

1.11

0.086

0.054

0.150

0.250

0.044

0.164

3

concurrent no treatment

2.28

1.13

0.091

0.062

0.111

0.259

0.052

0.167

4

300 g fly ash per day

2.68

1.32

0.296

0.142

0.131

0.255

0.054

0.190

5

300 g fly ash per day

2.36

1.17

0.240

0.148

0.111

0.240

0.054

0.170

6

300 g fly ash per day

2.98

1.55

0.431

0.201

0.131

0.350

0.077

0.270

7

1500 - 1800 g fly ash per day

4.81

2.49

0.716

0.615

0.286

0.372

0.107

0.182

8

1500 - 1800 g fly ash per day

6.21

2.84

0.827

0.643

0.352

0.477

0.135

0.246

9

1500 - 1800 g fly ash per day

4.76

2.31

0.644

0.506

0.288

0.492

0.105

0.208

aMgO was only analysed from one laboratorium

Applicant's conclusion:

Two years of oral administration of two different doses of fly ash influenced the ash and mineral contents of the faeces in a dose-dependent manner.

 

Analysis of trace elements in liver and bone samples:

Liver and bone samples of all animals were analysed for their contents of Arsenic, Manganese, Lead, Iron, Copper, Zinc and Cobalt. Bones were additionally analysed for their CaO and P2O5content.

In summary, there was no significant influence of the fly ash administration on the trace element contents in liver and bone samples.

Applicant's summary and conclusion

Conclusions:
A NOAEL for female cattle of ca. 1850-2390 mg/kg bw/day could be identified. Male reproduction parameters were not examined. Litters were not examined.