Registration Dossier
Registration Dossier
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EC number: - | CAS number: -
Endpoint summary
Administrative data
Description of key information
There is no data available for the registered substance on repeated-dose toxicity. However, there is data available for the source substances FeNaEDDHA and manganese salts.
The values for the manganese salts were converted to the registered substance under consideration of the maximum percentage of manganese included in the substance. This data is used within a frame of a weight-of-evidence approach to assess the toxicity of the target substance. The whole dataset included in the weight-of evidence approach is shown under 'Additional information'. Based on this the key value for manganese was chosen from the the two year repeated dose toxicity study (NTP 1993, 2a). The NOAEL was determined to be 987.6 mg/kg bw/day for the manganese moiety of the target substance, based on the effects on the thyroid gland (Follicular dilatation) in female mice.
For Fe(Na)EDDHA a NOAEL of 10 mg/kg bw/day is calculated from the subchronic oral toxicity study in rats (Novartis Crop Protection AG, 1998). Further information can be found under 'Additional information'.
Key value for chemical safety assessment
- Toxic effect type:
- dose-dependent
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- short-term repeated dose toxicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Please refer to Read Across Statement attached in Section 13
- Reason / purpose for cross-reference:
- read-across source
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- There were no test item related clinical findings.
- Mortality:
- no mortality observed
- Description (incidence):
- No deaths occurred.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Mean body weight was reduced and there was a mean body weight gain depression at 200 and 1000 mg/kg bw/day.
Compared to the control group, the body weight development of animals in groups 3 and 4 (200 and 1000 mg/kg) was impaired. At week 4, the mean weights of group 4 (1000 mg/kg bw) were 25% (males) and 22% (females), and the mean body weight gains (week -1 to week 4) 44% (males) and 54% (females) below that of the control group, respectively. The mean body weights of group 3 (200 mg/kg bw) were 6% (males) and 11% (females), and the mean body weight gains 11% (males) and 27% (females) below that of the control group, respectively. Body weight development in group 2 (50 mg/kg bw) was not affected by treatment. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Overall food consumption (week 1 to 4) was reduced by 29% in males and 26% in females of group 4 (1000 mg/kg bw), and by 10% in males and 8% in females of group 3 (200 mg/kg bw).
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- effects observed, treatment-related
- Description (incidence and severity):
- The mean water consumption was markedly increased at 1000 mg/kg bw/day.
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Anaemia was observed at 200 and 1000 mg/kg bw/day: "It was accompanied by a slight hyperchromasia of red blood cells. The release of reticulocytes from erythropoietic organs was not increased, suggesting a lack of an adaptive response. Furthermore, males of group 4 (1000 mg/kg bw) had slightly lower values of white blood cells, predominantly of lymphocytes and basophils. In addition, males of groups 3 and 4 (200 mg/kg bw and 1000 mg/kg bw) had minimally higher platelet counts. Slightly lower values of prothrombin time, as recorded for males of group 2 (50 mg/kg), and for animals of groups 3 and 4 (200 and 1000 mg/kg bw) are considered without toxicological relevance".
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Minimal or slight changes in several clinical chemistry parameters were noted at 1000 mg/kg bw/day. Minimally increased creatinine concentrations were observed in males at 200 mg/kg bw/day:
"Plasma creatinine concentrations were minimally increased in males of group 3 (200 mg/kg) and slightly increased in males and females of group 4 (1000 mg/kg). In group 4 (1000 mg/kg bw), plasma cholesterol levels were slightly increased in males and females and plasma potassium concentrations were decreased in females. Furthermore, in the high dose group (1000 mg/kg) plasma bilirubin levels were increased in males, plasma albumin concentrations were minimally elevated and plasma globulins were minimally decreased in females, resulting in an increased A/G ratio". - Urinalysis findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Excretion of a larger amount of reddish discoloured urine and mild bilirubinuria were observed in males at 1000 mg/kg bw/day. There was no evidence that treatment with the test article had affected other urine parameters investigated.
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- The mean body weight relative weights of the heart, kidneys, adrenals and spleen were changed at 200 and/or 1000 mg/kg bw/day as compared to the concurrent control group:
"At necropsy the mean carcass weight was decreased in group 3 (200 mg/kg; males -8%, females -10%) and in group 4 (1000 mg/kg; males -27%, females -23%), compared to that of the control group.
Heart to body weight ratio was significantly increased in males of group 3 (+12%) and group 4 (+30%), and in females of group 4 (+18%). Kidney to body weight ratio was increased in males of group 3 (+12%) and group 4 (+51%), and in females of group 4 (+31%). Adrenal to body weight ratio was increased in males of group 2 (+11%), group 3 (+13%) and group 4 (+28%). Spleen to body weight ratio was increased in females of group 4 (+24%).
Changes in absolute organ weights of liver, thymus, adrenals, testis and spleen, which attained statistical significances in group 4, are considered consequent to the body weight decrease in this group, and are therefore of no toxicological relevance. Other differences which attained a level of statistical significance were dose-independent and, therefore, not considered of experimental relevance". - Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- There were no test item related macroscopical findings.
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Microscopical examination revealed cytoplasmic vacuolisation of cortical tubules of the kidneys at 1000 mg/kg bw/day:
"On morphological grounds this lesion was compatible with osmotic nephrosis (C. Gopinath et al, 1987) which is known to be induced in rats by hypertonic sugar solutions including dextran and some chelating agents (P. Greaves, 1990)". - Histopathological findings: neoplastic:
- not specified
- Dose descriptor:
- NOAEL
- Basis for effect level:
- other: This study was a range finding study for a 90-day repeated dose oral toxicity study (Novartis Crop Protection AG, 1998). The latter study was used for NOEL derivation.
- Remarks on result:
- not measured/tested
- Remarks:
- no NOAEL identified
- Critical effects observed:
- not specified
- Conclusions:
- No NOAEL is identified. This study was used as scientific basis for dose level selection for a subsequent 90 -day repeated dose oral toxicity study in the rat. The same result is expected for the organic constituents of target substance since it has the same composition as the source substance.
- Executive summary:
In a dose range-finding subacute oral toxicity study (CIBA-GEIGY Limited, 1996), FeNaEDDHA in 0.5 % CMC and 0.1 % Tween 80 in distilled water was administered to 5 Sprague-Dawley derived rats/sex/dose level by oral gavage (10 mL/kg bw) at dose levels of 50, 200 or 1000 mg/kg bw/day for a period of 28 days. Male and female animals of the concurrent control group were treated with the vehicle only. Treatment with the test item resulted in impaired body weight development of rats treated at 200 and 1000 mg/kg bw/day and correspondent lower food intake. An anaemia without erythropoietic response was noted at 200 and 1000 mg/kg bw/day. At the same dose levels, the kidney was revealed as target organ by microscopical examination, by blood chemistry data and by organ weight evaluation. In addition, body weight relative organ weight changes were noted in the heart, adrenals and spleen. However, the relevance of these findings was considered as equivocal. This study was used as scientific basis for dose level selection for a subsequent 90 -day repeated dose oral toxicity study in the rat.
This subacute oral toxicity study in the rat is acceptable and satisfies the requirement for test guideline OECD 407.
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Please refer to Read Across Statement attached in Section 13
- Reason / purpose for cross-reference:
- read-across source
- Species:
- rat
- Clinical signs:
- no effects observed
- Description (incidence and severity):
- During this study neither changes to the behavior nor relevant clinical signs were observed.
- Mortality:
- no mortality observed
- Description (incidence):
- There was no mortality during this study which was attributed to the treatment with the test article. In the third treatment week one female animal (no. 86) died. However, there were no indications of a test article-related occurrence.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Decreased mean body weights were noted from weeks 5 and 8 onwards at 200 mg/kg bw/day in male and female rats, respectively. The mean body weight gains were decreased by the end of the treatment period in animals treated with 200 mg/kg bw/day. During the recovery period, body weight gain in animals previously treated with 200 mg/kg bw/day was higher as compared to controls.
The body weight development in the other treated groups was not influenced by treatment. - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Food consumption and food consumption ratios were decreased at 200 mg/kg bw/day in both sexes (decreased food consumption of 13% in males and of 8% in females, compared to the controls). During the recovery period food intake improved.
The mean food consumption of the other treated groups was not influenced by treatment. - Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- no effects observed
- Description (incidence and severity):
- The mean water consumption was not influenced by treatment.
- Ophthalmological findings:
- no effects observed
- Description (incidence and severity):
- The examinations of the eyes (lids and surrounds, conjunctiva, pupillary reflex, cornea, sclera, anterior chamber, lens, vitreous, and fundus) towards treatment end (day 87) and towards recovery end (day 115) did not reveal any treatment-related findings.
- Haematological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- At the end of the treatment period, a normochromic anaemia with lower erythrocyte count, haemoglobin concentration and haematocrit was observed in males and females at 200 mg/kg bw/day and males at 50 mg/kg bw/day. A higher reticulocyte count associated with higher MCV and MCH values and reduced white blood cell, basophil, lymphocyte and monocyte counts were confined to males at 200 mg/kg bw/day. A higher platelet count was recorded for males at 50 and 200 mg/kg bw/day and a higher prothrombin activity was recorded for males and females at 200 mg/kg bw/day. Evidence of reversibility for all the above parameters was apparent after the recovery period.
- Clinical biochemistry findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Several clinical chemistry parameters including creatinine, urea, protein, globulin, cholesterol and sodium concentration were increased at 50 and/or 200 mg/kg bw/day. Lower potassium levels were noted in males at 200 mg/kg bw/day. All values were similar to the control group values after the 4-week recovery period.
- Urinalysis findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Excretion of more acidic red-brown (males) or yellow-brown (females) discoloured urine was observed at 200 mg/kg bw/day. More acidic urine was also excreted by males at 50 mg/kg bw/day. By the end of the recovery period, the colour and pH of the urine excreted by male and females previously treated at 200 mg/kg bw/day was similar to that of control group animals.
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- The mean carcass weights were decreased at the end of the treatment period at 200 mg/kg bw/day. Males at 200 mg/kg bw/day showed an elevated mean heart to body weight ratio which still was higher than the concurrent control value at the end of the recovery period.
- Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- There were no test item related findings.
- Histopathological findings: non-neoplastic:
- no effects observed
- Description (incidence and severity):
- There were no test item related findings.
- Histopathological findings: neoplastic:
- not specified
- Dose descriptor:
- NOAEL
- Effect level:
- 10 mg/kg bw/day (actual dose received)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- other: haematology parameters (anaemia) The NOAEL was estimated based on the LOAEL of 50 mg/kg bw/day applying an assessment factor of 5.
- Critical effects observed:
- not specified
- Conclusions:
- NOEL of 5 mg/kg bw was established in this study (for males and females). NOAEL of 10 mg/kg bw was derived based on LOAEL of 50 mg/kg bw. The same result is expected for the organic constituents of target substance since it has the same composition as the source substance.
- Executive summary:
In a subchronic toxicity study (Novartis Crop Protection AG, 1998), FeNaEDDHA in 0.5 % CMC and 0.1 % Tween 80 in distilled water was administered to 10 Sprague-Dawley derived rats/sex/dose level by oral gavage (10 mL/kg bw) at dose levels of 5, 50 or 200 mg/kg bw/day for a period of 90 days. Male and female animals of the concurrent control group were treated with the vehicle only. Additional 10 animals/sex of the high dose and control group were kept on control diet for a 4 -week recovery period before sacrifice. Treatment with the test item resulted in lower food intake and impaired body weight development of rats treated at 200 mg/kg bw/day. Reversible effects on the red blood cell (normochromic anaemia) and white blood cell parameters, and higher values of platelets and prothrombin activity were noted at 50 and/or 200 mg/kg bw/day. In addition, there were changes of blood chemistry and urine parameters concerning the liver and kidneys. The body weight relative heart weight was increased in males at 200 mg/kg bw/day. Under the conditions of this study, the NOAEL for FeNaEDDHA when administered by daily oral gavage for three months was 10 mg/kg bw/day (estimated from the LOAEL).
This subchronic toxicity study in the rat is acceptable and satisfies the guideline requirement for a subchronic oral study (OECD 408).
Estimation of NOAEL:
A more realistic NOAEL was estimated from the LOAEL of 50 mg/kg bw/day applying an assessment factor of 5. This method is applicable and scientifically justified for this test, as only slight adverse effects were observed at 50 mg/kg bw/day (haematology parameters - anaemia). To take the relatively large concentration gap between 5 (clear NOEL) and 50 mg/kg bw/day into account, 5 mg/kg bw/day was not taken as NOAEL, but extrapolated form the LOAEL. The guidance on information requirements and CSA, R.8 (ECHA, 2008 -2010) recommends a factor between 3 and 10 for extrapolation from LOAEL to NOAEL. An assessment factor of 5 seems to be approprate and conservative enough for the current study as only slight effects were observed at the LOAEL of 50 mg/kg bw/day. Consequently a NOAEL of 10 mg/kg bw/day is calculated for this study.
- Endpoint:
- sub-chronic toxicity: oral
- Remarks:
- other: combined subacute, subchronic, carcinogenicity
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- Please refer to Read Across Statement attached in Section 13
- Reason / purpose for cross-reference:
- read-across source
- Clinical signs:
- not specified
- Mortality:
- mortality observed, treatment-related
- Description (incidence):
- Mice: One control male mouse and one female mouse receiving 3,130 ppm died of unknown causes.
- Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: The mean body weight gain in males receiving 3,130 ppm was marginally lower than that of the controls and was significantly lower in the three highest female dose groups than the controls. Final mean body weights of all exposed animals were within 5% of those of the controls.
Mice: Mean body weight gains of all exposed males were significantly lower than that of the control group, and the final mean body weight of the 50,000 ppm group was 13% lower than that of the controls. The mean body weight gain of 50,000 ppm females was significantly lower than that of the controls. - Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- Rats: Feed consumption by exposed rats was similar to that by the controls.
Mice: Feed consumption by exposed male and female mice was similar to that by the controls. - Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: Although the total leukocyte counts were similar in exposed and control males, neutrophil counts were significantly higher in all exposed male groups, whereas lymphocyte counts were significantly lower in the 6,250, 12,500, and 25,000 ppm groups. In contrast, the total leukocyte counts of 6,250, 12,500, and 25,000 ppm females were significantly lower, primarily because of lower lymphocyte counts. A marginal but significant increase in percent hematocrit and erythrocyte counts occurred in males exposed to 6,250, 12,500, or 25,000 ppm.
Mice: The percent hematocrit, hemoglobin concentrations, and mean erythrocyte volumes of 50,000 ppm male and female mice were significantly lower than those of the controls. - Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: Absolute and relative liver weights of all exposed males and of the female 25,000 ppm group were significantly lower than those of the controls. The absolute and relative lung weights of all exposed females were also significantly lower than those of controls.
Mice: The absolute and relative liver weights of 50,000 ppm male mice were significantly lower than those of the controls; absolute and relative liver weights of females were similar to those of the controls. - Gross pathological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Mice: A few mice in the male and female exposure groups exhibited fight wounds. Three 50,000 ppm males had mild epithelial hyperplasia and hyperkeratosis of the forestomach.
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Key result
- Dose descriptor:
- NOAEL
- Remarks:
- rats
- Effect level:
- ca. 1 005 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted target substance
- Sex:
- female
- Basis for effect level:
- other: body weight gain; leucocyte count
- Remarks on result:
- other: Effect levels calculated from feed intake
- Dose descriptor:
- LOAEL
- Remarks:
- rats
- Effect level:
- ca. 2 443 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- female
- Basis for effect level:
- other: body weight gain; leucocyte count
- Remarks on result:
- other: Effect levels calculated from feed intake
- Dose descriptor:
- NOAEL
- Remarks:
- rats
- Effect level:
- ca. 7 363 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male
- Basis for effect level:
- other: mortality; body weight; organ weights except lung, liver; leukocyte count dose calculated from feed ingestion 1700 mg/kg bw/d was the highest dose tested, so the actual NOAEL could be even much higher.
- Dose descriptor:
- NOAEL
- Remarks:
- mice
- Effect level:
- ca. 14 640 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- female
- Basis for effect level:
- other: body weight, hematocrit, hemoglobin concentrations, mean erythrocyte volumes
- Remarks on result:
- other: Effect levels calculated from feed intake
- Dose descriptor:
- LOAEL
- Remarks:
- mice
- Effect level:
- ca. 29 280 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- female
- Basis for effect level:
- other: body weight, hematocrit, hemoglobin concentrations, mean erythrocyte volumes
- Remarks on result:
- other: Effect levels calculated from feed intake
- Dose descriptor:
- NOAEL
- Remarks:
- mice
- Effect level:
- ca. 15 883 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male
- Basis for effect level:
- other: body weight, hematocrit, hemoglobin concentrations, mean erythrocyte volumes; Three 50000 ppm males had mild epithelial hyperplasia and hyperkeratosis of the forestomach.
- Remarks on result:
- other: Effect levels calculated from feed intake
- Dose descriptor:
- LOAEL
- Remarks:
- mice
- Effect level:
- ca. 31 762 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male
- Basis for effect level:
- other: body weight, hematocrit, hemoglobin concentrations, mean erythrocyte volumes; Three 50000 ppm males had mild epithelial hyperplasia and hyperkeratosis of the forestomach.
- Remarks on result:
- other: Effect levels calculated from feed intake
- Critical effects observed:
- no
- Conclusions:
- In 90-day repeated dose oral toxicity test (feed study), the ingestion of diets containing manganese (II) sulfate monohydrate had no effects on feed consumption, mortality and most organ weights (except liver) in both rats and mice.
In rats, no clinical or histopathologic findings were attributed to the administration of MnSO4, whereas the application of 6250 ppm MnSO4 had impact on the body weight gain, lymphocyte and leukocyte count in females.
Some mice show pathologic results, e.g. mild epithelial hyperplasia and hyperkeratosis of the forestomach, in high dose level, i.e. 50000 ppm in diet. These are not regarded as very relevant due to the mildness of the effects and low incidence.
So, female rats are considered to be the more sensitive sex of the most sensitive species in this testing scheme and should consequently be taken into account during risk assessment. The NOAEL was determined to be 3130 ppm in diet, which corresponds to 232 mg MnSO4/kg bw/day in female rats. Hence, converted to the manganese moiety in target substance the NOAEL of the most sensitive species (female rat) is 1005 mg/kg bw/day. - Executive summary:
In a subchronic oral toxicity study equivalent to OECD guideline 408, Manganese sulphate monohydrate, a read-across substance for the manganese moiety of the target substance, was administered over 13 weeks to each five F344/N rats and B6C3F1 mice per sex and per dose at dose levels of 0, 3130, 6250, 12500, 25000 and 50000 ppm (mice) and 0, 1600, 3130, 6250, 12500, 25000 ppm in diet (rats).
There were no substance-related effects on mortality, body weight, organ weights, except for lung and liver, and leukocyte count in male rats and on feed consumption in female rats, whereas 6250 ppm had effect on body weight gain and leucocyte count in female rats.
50000 ppm had no effects on mortality in mice, whereas 25000 ppm MnSO4 had influence on body weight, hematocrit, hemoglobin concentrations and mean erythrocyte volumes in both sexes. The effect concentrations were converted to the corresponding moiety of manganese in the target substance (see below in brackets).
The LOAEL in male mice is 6760 (= 29280 mg/kg bw/day) and in female mice 7333 mg/kg bw/d (= 31762 mg/kg bw/day), based on body weight, hematocrit, hemoglobin concentrations and mean erythrocyte volumes with a subsequent NOAEL for female mice of 3380 (= 14640 mg/kg bw/day) and for male mice of 3667 mg/kg bw/d (= 15883 mg/kg bw/day).The LOAEL in rats is 564 mg/kg bw/d (= 2443 mg/kg bw/day), based on body weight gain and leucocyte count in females, the NOAEL is 232 mg/kg bw/d (= 1005 mg/kg bw/day).
So, female rats are considered to be the more sensitive sex of the most sensitive species in this testing scheme and should consequently be taken into account during risk assessment. The NOAEL was determined to be 3130 ppm in diet, which corresponds to 232 mg MnSO4/kg bw/day. Hence, converted to the manganese moiety of the target substance the NOAEL of the most sensitive species (female rat) is 1005 mg/kg bw/day.
This subchronic toxicity study in rats and mice is acceptable and satisfies the guideline requirement for a subchronic oral study equivalent to OECD study 408.
- Endpoint:
- short-term repeated dose toxicity: oral
- Remarks:
- other: combined subacute, subchronic, carcinogenicity
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- Please refer to Read Across Statement attached in Section 13
- Reason / purpose for cross-reference:
- read-across source
- Clinical signs:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: Males exposed to 50,000 ppm and all exposed groups of females exhibited diarrhea during the second week.
- Mortality:
- mortality observed, treatment-related
- Description (incidence):
- Mice: One female mouse in the 25,000 ppm group died of unknown causes on day 1.
Please refer to table 1 and 2 - Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: The mean body weight gain of the male 50,000 ppm group at the end of the 14-day study was 57% less than that of the control group, and the final mean body weight of this group was 13% lower than that of the controls. The mean body weight gain of 50,000 ppm females was 20% less than that of the controls and the final mean body weight was 7% lower than that of the controls.
Mice: No significant evidence of toxicity was observed except possible body weight effects in both sexes. However, no conclusions can be made regarding the body weight data because of poor randomization of animals at study initiation.
Please refer to table 1 and 2 - Food consumption and compound intake (if feeding study):
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: During the first week, feed consumption by 50,000 ppm males was 19% lower than controls, whereas that by 50,000 ppm females was 15% lower. During the second week, however, feed consumption by both male and female 50,000 ppm groups was similar to that by controls.
Please refer to table 1 and 2 - Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: The total leukocyte and neutrophil counts were significantly increased in 50,000 ppm groups, particularly males.
Mice: No chemical-related differences in hematology parameters were observed.
Please refer to table 5 and 6 - Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Description (incidence and severity):
- Rat: the absolute and relative liver weights of 50,000 ppm male rats were significantly lower than those of the controls. Absolute or relative organ weight differences in some of the exposure groups were probably related to body weight differences between exposed and control groups. Mice: No organ weight differences were attributed to manganese (II) sulfate monohydrate exposure.
Please refer to table 3 and 4 - Gross pathological findings:
- not examined
- Histopathological findings: non-neoplastic:
- not examined
- Histopathological findings: neoplastic:
- not examined
- Other effects:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: Manganese concentrations in the livers of 50,000 ppm males and females were more than twice those of controls.
Mice: Manganese concentrations in the livers of 50,000 ppm mice were 8 to 15 times higher than those found in controls. - Dose descriptor:
- LOAEL
- Remarks:
- rats
- Effect level:
- ca. 16 892 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male
- Basis for effect level:
- other: body weight gain
- Remarks on result:
- other: Value calculated from food consumption
- Dose descriptor:
- NOAEL
- Remarks:
- rats
- Effect level:
- ca. 8 416 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male
- Basis for effect level:
- other: body weight gain
- Remarks on result:
- other: Value calculated from food consumption
- Dose descriptor:
- NOAEL
- Remarks:
- mice
- Effect level:
- ca. 44 873 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male/female
- Basis for effect level:
- other: mortality, body weight, hematology parameters
- Remarks on result:
- other: Value calculated from food consumption. Since mg/kg bw/d was the highest dose tested, the actual NOAEL might be even much higher.
- Critical effects observed:
- no
- Conclusions:
- In a 14 day repeated dose toxicity test (feed study) in rats and mice with the read-across substance MnSO4, no substance-related effects were observed regarding mortality in both species up to the highest dose tested, 50000 ppm MnSO4 in diet. In mice, also no substance-related effects were observed regarding body and organ weight and hematology parameters. Consequently, the actual NOAEL could be even much higher since no effects were observed in the highest dose tested. In rats, only slight adverse effects such as diarrhea, elevated leukocyte and neutrophile count and liver weight were observed at the highest dose tested, whereas the effects on hematology could be also a consequence of the observed diarrhea. Only the male 50000 ppm group showed a reduced body weight gain. So, the dose of 50000 ppm in diet should be considered as NOAEL for mice and LOAEL in rats. In mice this is approx. 10360 mg MnSO4/kg bw/day (=44873 mg/kg bw/day for the manganese moiety in the target substance), and in rats it is 3900 mg/kg bw/day (=16892 mg/kg bw/day for the manganese moiety in the target substance). The NOAEL in rats is 1943 mg/kg bw/day (= 8416 mg/kg bw/day for the manganese moiety in the target substance).
- Executive summary:
A subacute oral toxicity study equivalent to OECD guideline 407 conducted with the source substance Manganese sulphate monohydrate is available. The test item was administered over 14 days to each five F344/N rats and B6C3F1 mice per sex and per dose at dose levels of 0, 3130, 6250, 12500, 25000 and 50000 ppm in diet. There were no substance-related effects on mortality, food consumption or hematology parameters in both rats and mice, as well as no effects on body weight in rats (except the male 50000 ppm group); no conclusion can be drawn for the body weight of mice due to the poor randomization of the animals.
The effect concentrations were converted to the corresponding manganese moiety of the the target substance (in brackets).
The LOAEL is 3900 mg/kg bw/day (=18906 mg/kg bw/day for the manganese moiety in the target substance) in rats, based on body weight gain in males, with a consequent NOAEL of 1943 mg/kg bw/day (= 8416 mg/kg bw/day for the manganese moiety in the target substance). The NOAEL in mice is 10360 mg/kg bw/day (=44873 mg/kg bw/day for the manganese moiety in the target substance) (highest tested dose), based on mortality, body weight and hematology parameters.
This subacute toxicity study in rats and mice is acceptable and satisfies the guideline requirement for a subacute oral study equivalent to OECD study 407.
- Endpoint:
- chronic toxicity: oral
- Remarks:
- combined repeated dose and carcinogenicity
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- Please refer to Read Across Statement attached in Section 13
- Reason / purpose for cross-reference:
- read-across source
- Clinical signs:
- no effects observed
- Mortality:
- mortality observed, treatment-related
- Description (incidence):
- Rats: Survival of 15,000 ppm male rats was significantly lower than that of the controls; survival of 1,500 and 5,000 ppm males and all exposed groups of females was similar to that of controls. The significant reduction in survival of 15,000 ppm males was attributed to increased severity of nephropathy and renal failure. The decreased survival did not occur until approximately week 93 of the study.
Mice: Survival of exposed males and females was similar to that of the control groups. No clinical findings were attributed to the administration of manganese (II) sulfate monohydrate. - Body weight and weight changes:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: The mean body weights of 1,500 and 5,000 ppm male rats were similar to those of controls throughout the 2-year study. The mean body weights of 15,000 ppm male rats were within 5% of that of controls until week 89. From week 89, the mean body weights ranged from 8% to 13% lower than that of controls; at the end of the 2-year study, the final mean body weight of 15,000 ppm males was 10% lower than that of controls. Mean body weights of exposed females were similar to that of controls throughout the study.
Mice: After week 37, mean body weights of all exposed groups of females were lower than that of the controls; the final mean body weights for the 1,500, 5,000, and 15,000 ppm groups were 6%, 9%, and 13% lower than that of the control group. - Food consumption and compound intake (if feeding study):
- no effects observed
- Description (incidence and severity):
- Rats: Feed consumption by exposed groups was similar to that by control groups.
Mice: Feed consumption by exposed male and female mice was similar to that of the control groups. - Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not examined
- Haematological findings:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: Slight differences in some parameters between exposed and control groups were not considered related to the ingestion of manganese (II) sulfate monohydrate.
Mice: Slight differences in some parameters between exposed and control groups are found and not consistent with the findings in the 13-week study and their significance is uncertain. - Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Rats: At the 9- and 15-month interim evaluations, the absolute kidney weights of exposed rats were similar to those of the controls.
Mice: At the 9-month interim evaluation, absolute liver weights of 15,000 ppm males and of 5,000 and 15,000 ppm females were significantly lower than those of controls. Since these groups also had lower mean body weights, and relative liver weights were similar to controls, the lower absolute liver weights are not considered chemical related. At the 15-month interim evaluation, absolute and relative liver weights of exposed mice were similar to controls. - Gross pathological findings:
- no effects observed
- Description (incidence and severity):
- no chemical related significant incidences over control in pancreas or kidney
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats:
Pancreas: Hyperplasia or adenoma of the pancreatic islets occurred in a few males in each of the exposure groups but not in the control group. However, neither the trend test nor pairwise comparisons were significant, and the incidences in each of the dose groups were within the range of NTP historical control groups.
Kidney: Chronic nephropathy occurred in all male rats examined at both interim evaluations and most of the control and exposed males at the end of the study. The average severity of nephropathy was slightly greater in the high-dose group, but the difference was not statistically significant. Because of the subjective nature of the severity grading, an additional evaluation of the kidney of high-dose and control male rats was performed without knowledge of the previous diagnoses. The result of the additional evaluation confirmed the presence of a marginally increased severity of nephropathy in the high-dose group, and the difference was significant (P=0.04) by a two-sided MannWhitney U test. The severity of nephropathy varied from minimal to marked. Minimal nephropathy was characterized by a few sparsely scattered cortical foci of regenerating tubules with increased epithelial cytoplasmic basophilia and slightly thickened glomerular basement membranes. Nephropathy of mild severity had similar morphologic features, but these features occurred with greater frequency. Also present were occasional dilated tubules filled with homogenous hyaline material and lined by flattened epithelial cells. Nephropathy of moderate to marked severity had similar but more severe and extensive tubule lesions. In addition, variable interstitial fibrosis and mineralization with mononuclear leukocyte infiltration, variable tubule loss and atrophy, and degenerative glomerular changes occurred. In the most severe cases, cystic tubules lined by cuboidal or attenuated epithelial cells were present.
The incidences of several lesions commonly associated with advanced nephropathy and renal failure were significantly increased in 15,000 ppm male rats. These lesions included mineralization of blood vessels, mineralization of the glandular stomach, fibrous osteodystrophy of the femur, and parathyroid gland hyperplasia.
Mice:
Thyroid Gland: At the 9- and 15-month interim evaluations, thyroid follicle dilatation was present in 15,000 ppm males and females but not in the controls. At the end of the 2-year study, the incidence of follicular dilatation increased significantly in 15,000 ppm males and 5,000 and 15,000 ppm females. A significantly increased incidence of focal hyperplasia of follicular epithelium also occurred in 15,000 ppm males and in all exposed females. Follicular dilatation at the 9-month evaluation was characterized by a uniform increase in the follicular diameter throughout the gland. Follicular dilatation in mice at the end of the study differed from that observed in mice at the 9-month interim evaluation in that the dilated follicles were limited to the periphery of the glands. The affected follicles contained pale eosinophilic colloid and were lined by a single layer of flat to slightly cuboidal follicular epithelial cells. Follicular cell hyperplasia and adenoma constitute a morphological continuum. Follicular cell hyperplasia consisted of single or multiple collections of variably sized follicles with irregular hypertrophy and increased cellularity of the follicular epithelium. Minimal to mild follicular cell hyperplasia consisted of one or several follicles lined by columnar epithelium with small and infrequent papillary infoldings. Moderate to marked hyperplasia involved clusters of variably sized follicles with more prominent papillary formations.
Forestomach: A statistically significant increased incidence of focal squamous hyperplasia of the forestomach occurred in the 15,000 ppm males and females, accompanied by ulceration/erosion and inflammation. Hyperplasia of the squamous epithelium occurred focally at various sites of the forestomach mucosa. The lesion was characterized by broad-based areas of either proliferative epithelial thickening and hyperkeratosis or by polypoid projections of thickened epithelium protruding directly from the mucosa into the lumen of the stomach. Inflammation of the lamina propria and submucosa subjacent to the ulcerative lesions consisted of a mixture of infiltrating neutrophils and mononuclear leukocytes. - Histopathological findings: neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats:
Pancreas: Hyperplasia or adenoma of the pancreatic islets occurred in a few males in each of the exposure groups but not in the control group. In addition, a carcinoma of the pancreatic islets was found in one 15,000 ppm male. However, neither the trend test nor pairwise comparisons were significant, and the incidences in each of the dose groups were within the range of NTP historical control groups.
Adrenal Gland: In females, medullary hyperplasia occurred with a significant negative trend and a significantly decreased incidence in the 15,000 ppm group. Benign pheochromocytomas of the adrenal medulla in males occurred with a significant negative trend, but the decreases were not significant by pairwise comparison the incidence of medullary hyperplasia in exposed males was similar to that of the controls
Mice:
Thyroid Gland: Follicular cell adenomas were found in three (6%) 15,000 ppm males. This rate is marginally higher than the average rate of 2% and just within the range of 0%-6% for historical control male mice. The incidence of this neoplasm was 10% in 15,000 ppm females, which is slightly above the average of 3% and range of 0%-9% for historical control female mice. The incidences of adenoma in 15,000 ppm males and females were not significantly greater than those of the controls. Follicular cell adenomas were generally more discrete collections of altered follicles compressing the surrounding parenchyma at the 9-month evaluation.
Liver: One male in the 15,000 ppm group and two females in the 5,000 ppm group had hepatocellular adenomas at the 15-month interim evaluation. At the end of the 2-year study, hepatocellular adenomas occurred with a statistically significant negative trend in males that was also significant by pairwise comparison in the 5,000 and 15,000 ppm groups. Hepatocellular foci did not occur in an exposure-related pattern. The incidences of adenoma or foci in exposed females were similar to those of the controls. - Other effects:
- effects observed, treatment-related
- Description (incidence and severity):
- Rats: At both the 9- and 15-month interim evaluations, the manganese levels in the liver of 5,000 and 15,000 ppm males and females were significantly greater than those in controls. The hepatic iron concentrations for these exposure groups were lower than for controls. The concentrations of manganese in the brain, kidney, and pancreas of exposed and control rats were variable; 15,000 ppm males had a significantly higher concentration of manganese in the brain and kidney at the 9-month interim evaluation and in the brain, kidney, and pancreas at the 15-month interim evaluation. Copper levels in the kidney of 15,000 ppm males at 9 months and in 15,000 ppm females at 9 and 15 months were significantly greater than those of the controls.
Mice: At the 9- and 15-month interim evaluations, tissue concentrations of manganese were significantly elevated in the livers of the 5,000 and 15,000 ppm groups. Hepatic iron levels were significantly lower in exposed females at the 9- and 15-month interim evaluations and in 5,000 and 15,000 males at the 15-month interim evaluation. Tissue concentrations of manganese in the brain (except 1,500 and 5,000 ppm females at 15 months), kidney, and pancreas (except 1,500 males at 9 months and 1,500 ppm females at 15 months) of exposed groups were significantly greater those of controls (Tables H3 and H4). - Dose descriptor:
- NOAEL
- Remarks:
- rat
- Effect level:
- ca. 3 032 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male
- Basis for effect level:
- other: nephropathy, mean survival
- Remarks on result:
- other: systemic effects
- Dose descriptor:
- NOAEL
- Remarks:
- rat
- Effect level:
- ca. 3 421.8 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- female
- Basis for effect level:
- other: nephropathy, mean survival
- Remarks on result:
- other: systemic effects
- Dose descriptor:
- NOAEL
- Remarks:
- mouse
- Effect level:
- ca. 2 598.8 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male
- Basis for effect level:
- other: Thyroid gland, Follicular dilatation and focal hyperplasia in follicular cells
- Remarks on result:
- other: systemic effects
- Dose descriptor:
- NOAEL
- Remarks:
- mouse
- Effect level:
- ca. 987.6 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- female
- Basis for effect level:
- other: Thyroid gland, Follicular dilatation
- Remarks on result:
- other: systemic effects
- Dose descriptor:
- NOAEL
- Remarks:
- mouse
- Effect level:
- ca. 2 598.8 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- male
- Basis for effect level:
- other: forestomach, focal squamous hyperplasia
- Remarks on result:
- other: local effects
- Dose descriptor:
- NOAEL
- Remarks:
- mouse
- Effect level:
- ca. 3 430.4 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- female
- Basis for effect level:
- other: Forestomach, focal squamous hyperplasia
- Remarks on result:
- other: local effects
- Critical effects observed:
- no
- Conclusions:
- This chronic / carcinogenicity study was performed according to OECD guideline study 451 on MnSO4 and is well documented. Therefore, this study can be considered to be reliable. Additionally, MnSO4 can serve as a read-across substance for the target substance and a chronic / carcinogenicity study as surrogate for a subchronic repeated dose study. Consequently, the study fulfils the requirements for this endpoint under REACH.
Under the conditions of these 2-year feed studies, there was no evidence of carcinogenic activity of manganese (II) sulfate monohydrate in male or female F344/N rats receiving 1,500, 5,000, or 15,000 ppm. There was equivocal evidence of carcinogenic activity of manganese (II) sulfate monohydrate in male and female B6C3F1 mice, based on the marginally increased incidences of thyroid gland follicular cell adenoma and the significantly increased incidences of follicular cell hyperplasia. The ingestion of a diet containing manganese (II) sulfate monohydrate was associated with an increased severity of nephropathy in male rats, focal squamous hyperplasia of the forestomach in male and female mice, and ulcers and inflammation of the forestomach in male mice.
The lowest NOAEL derived from this chronic study is 228 mg/kg bw/day (female mice, Thyroid gland, Follicular dilatation), which corresponds to 987.6 mg/kg bw/day for the manganese moiety of the target substance.
In conclusion, based on the outcome of this study, the target substance can be considered as non-carcinogenic and relatively non-toxic. - Executive summary:
In a combined chronic / carcinogenicity study according to OECD Guideline 451, Manganese (II) sulfate monohydrate was administered to each 70 F344/N rats and B6C3F1 mice in feed at dose levels of 0, 1500, 5000 and 15000 ppm in feed (average food consumption was determined and absolute amount were calculated accordingly) over 2 years. No evidence for carcinogenicity of MnSO4 was found in rats, and only equivocal evidence of carcinogenic activity was found in mice. No clinical findings such as body weights and feed consumption or incidence of neoplasms and non-neoplasms were chemical related, effects on pancreas, kidneys or adrenal gland were not significantly over control in rats, and only slight effects compared to control were noted in mice. The NOAELs are based on nephropathy, effects on thyroid gland, forestomach and relative liver weight. The lowest NOAEL derived from this chronic study is 228 mg/kg bw/day (female mice, Thyroid gland, Follicular dilatation), which corresponds to 987.6 mg/kg bw/day for the manganese moiety of the target substance.
This chronic / carcinogenicity study in rats and mice is acceptable to fulfil the requirements under this endpoint, satisfies the guideline requirement for a chronic / carcinogenicity study in rats or mice and meets scientific principles.
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- weight of evidence
- Justification for type of information:
- Please refer to Read Across Statement attached in Section 13
- Reason / purpose for cross-reference:
- read-across source
- Clinical signs:
- not specified
- Description (incidence and severity):
- no data on clinical signs
- Mortality:
- no mortality observed
- Description (incidence):
- no mortality occurred
- Body weight and weight changes:
- not examined
- Food consumption and compound intake (if feeding study):
- not examined
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- not examined
- Haematological findings:
- not examined
- Clinical biochemistry findings:
- not examined
- Urinalysis findings:
- effects observed, treatment-related
- Description (incidence and severity):
- visual exermination, male animals were noted to have viscous, gritty urine in the urinary bladder at necropsy
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- not specified
- Description (incidence and severity):
- kidneys were weighted but no data is reported
- Gross pathological findings:
- not specified
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Description (incidence and severity):
- incidences of urinary tract lesions: Tubular protein nephropathy, tubulointerstitial nephritis, glomerulosclerosis/glomerulonephritis, Urolithiasis and Papillitis (females) (see tables 1-4)
- Histopathological findings: neoplastic:
- not specified
- Dose descriptor:
- NOAEL
- Effect level:
- > 7 776.97 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Remarks:
- converted to target substance
- Sex:
- female
- Basis for effect level:
- other: Tubular protein nephropathy, tubulointerstitial nephritis, glomerulpsclerosis/glomerulonephritis, urolithiasis, papillitis
- Remarks on result:
- other: Highest dose tested; therefore, the actual NOAEL can be also much higher
- Critical effects observed:
- no
- Conclusions:
- Since the given data indicate, that the study in its original design is well-performed, the obtained results can be considered to be reliable. The incidence of tubular protein nephropathy in male rats in study A does not increase time-dependently, whereas in study B a dose-dependent increase was obtained. In both studies, the incidence of tubulointerstitial nephritis, the ratio glomerulosclerosis / glomerulonephritis and urolithiasis increases dose-dependently. On the other hand, in both studies in female rats, no increase over background of above mentioned incidents was observed. On the contrary, the incidence of these adverse effects even decreases in study A; in general, no incidence of urolithiasis or papillitis (except 33% in study A, 1838 mg/kg bw) was observed. So, the most likely reason for the difference in the incidence of adverse effects between male and female rats is the induction of a α2u-globulin related nephropathy. This could be observed in this experiment in a dose-dependent (study B) manner or not directly dose-correlated (study A), which matches, as mean values, the expected supralinear increase of the incidence of a α2u-globulin related nephropathy with given doses. The other observed adverse effects are expected to be related to the phenomenon of this nephropathy. The α2u-globulin related nephropathy is an adverse effect which only occurs in male rats and has no relevance for female rats, which was also demonstrated by the lack of dose-related adverse effects. This special type of nephropathy has also no relevance for humans.
Consequently, it can be concluded that the observed effect in female rats are the only relevant ones for human risk assessment. Since no adverse effects were observed over background in this experiment, it can be concluded that the NOAEL must be ≥ 1838 mg/kg bw/day. This corresponds to ≥ 7776.97 mg/kg bw/day for the manganese moiety of the target substance. - Executive summary:
In an subchronic, 63 d repeated dose toxicity study, Manganese acetate was administered to 12 Sprague-Dawley rats per sex and dose by oral gavage at dose levels of 306, 612, 1225, 1838 mg/kg manganese per day.
There was a dose-dependent increase in the incidence of lesions in the urinary tract of male rats, which is not considered relevant for female rats or humans. In the more relevant female rats, no compound related toxicity was observed; even a decrease of incidences of adverse effects in the female urinary tract was observed. Consequently, no LOAEL could be derived. The NOAEL was determined to be ≥ 1838 mg/kg bw/day. This corresponds to ≥ 7776.97 mg/kg bw/day for the manganese moiety of the target substance.
Referenceopen allclose all
CHEMICAL ANALYSIS OF DOSE FORMULATIONS
The calculated mean contents of the test item in the dose formulations used for the low, mid and high dose groups were 105, 112 and 107 % of the nominal concentrations, respectively.
CHEMICAL ANALYSIS OF DOSE FORMULATIONS
The calculated overall mean contents of the test item in the dose formulations used for the low, mid and high dose groups were 96.9, 104.2 and 101.0 % of the nominal concentrations, respectively (RCC project no. 651688). In a previous 28 -day dose range finding study with the same test item and vehicle, the stability and homogeneity of dose formulations were within the limits of acceptance (RCC project no. 393322).
Estimation of NOAEL:
A more realistic NOAEL was estimated from the LOAEL of 50 mg/kg bw/day applying an assessment factor of 5. This method is applicable and scientifically justified for this test, as only slight adverse effects were observed at 50 mg/kg bw/day (haematology parameters - anaemia). To take the relatively large concentration gap between 5 (clear NOEL) and 50 mg/kg bw/day into account, 5 mg/kg bw/day was not taken as NOAEL, but extrapolated form the LOAEL. The guidance on information requirements and CSA, R.8 (ECHA, 2008 -2010) recommends a factor between 3 and 10 for extrapolation from LOAEL to NOAEL. An assessment factor of 5 seems to be approprate and conservative enough for the current study as only slight effects were observed at the LOAEL of 50 mg/kg bw/day.
Table 3: Survival, Body Weights, and Feed Consumption of Rats in the 13-Week Feed Study of Manganese (II) Sulfate Monohydrate
| Concentration (ppm) | Survivala | Mean Body Weight and Weight Changesb(g) Relative | Final Weight Feed to Controls (%) | Consumptionc | |||
| Initial | Final | Change | Week 1 | Week 13 | |||
| Male | |||||||
| 0 | 10/10 | 136 ± 5 | 291 ± 4 | 155 ± 4 | 14.9 | 13.1 | |
| 1,600 | 10/10 | 142 ± 4 | 294 ± 5 | 152 ± 4 | 101 | 14.5 | 13.5 |
| 3,130 | 10/10 | 149 ± 3 | 291 ± 4 | 141 ± 4 | 100 | 14.8 | 13.6 |
| 6,250 | 10/10 | 148 ± 2 | 294 ± 3 | 146 ± 3 | 101 | 15.0 | 9.6 |
| 12,500 | 10/10 | 150 ± 11 | 290 ± 6 | 140 ± 11 | 99 | 14.9 | 14.9 |
| 25,000 | 10/10 | 140 ± 4 | 284 ± 6 | 144 ± 4 | 97 | 14.1 | 14.4 |
| Female | |||||||
| 0 | 10/10 | 99 ± 1 | 184 ± 2 | 84 ± 2 | 10.7 | 9.2 | |
| 1,600 | 10/10 | 103 ± 1 | 181 ± 2 | 79 ± 2 | 99 | 10.8 | 9.3 |
| 3,130 | 10/10 | 96 ± 1 | 175 ± 2* | 80 ± 3 | 95 | 10.9 | 9.2 |
| 6,250 | 10/10 | 101 ± 1 | 176 ± 2* | 75 ± 1** | 96 | 10.7 | 14.3 |
| 12,500 | 10/10 | 106±1** | 178 ± 1* | 73 ± 2** | 97 | 10.7 | 10.5 |
| 25,000 | 10/10 | 104±1** | 174 ± 3** | 70 ± 2** | 95 | 12.1 | 10.3 |
* Significantly different (P≤0.05) from the control group by Williams' or Dunnett's test.
** P≤0.01
a Number of animals surviving at 13 weeks/number initially in group
b Weights given as mean ± standard error.
c Feed consumption is expressed as grams per animal per day.
Table 4: Survival, Body Weights, and Feed Consumption of Mice in the 13-Week Feed Study of Manganese (II) Sulfate Monohydrate
| Concentration (ppm) | Survivala | Mean Body Weight and Weight Changesb(g) Relative | Final Weight Feed to Controls (%) | Consumptionc | |||
| Initial | Final | Change | Week 1 | Week 13 | |||
| Male | |||||||
| 0 | 9/10d | 25.0 ± 0.5 | 31.4 ± 0.6 | 6.6 ± 0.5 | 3.4 | 3.0 | |
| 3,130 | 10/10 | 25.7 ± 0.3 | 30.5 ± 0.5 | 4.8 ± 0.4** | 97 | 3.0 | 3.0 |
| 6,250 | 10/10 | 26.3 ± 0.2* | 31.0 ± 0.3 | 4.7 ± 0.4** | 99 | 3.4 | 3.3 |
| 12,500 | 10/10 | 26.0 ± 0.3 | 30.9 ± 0.4 | 4.9 ± 0.4** | 98 | 3.4 | 3.8 |
| 25,000 | 10/10 | 25.9 ± 0.2 | 30.6 ± 0.5 | 4.7 ± 0.5** | 97 | 2.6 | 3.3 |
| 50,000 | 10/10 | 25.1 ± 0.4 | 27.4 ± 0.3** | 2.3 ± 0.4** | 87 | 3.0 | 4.7 |
| Female | |||||||
| 0 | 10/10 | 20.0 ± 0.2 | 24.2 ± 0.3 | 4.2 ± 0.3 | 2.4 | 2.3 | |
| 3,130 | 9/10e | 20.0 ± 0.3 | 24.2 ± 0.5 | 4.1 ± 0.3 | 100 | 3.3 | 2.4 |
| 6,250 | 10/10 | 20.5 ± 0.2 | 24.3 ± 0.3 | 3.8 ± 0.3 | 100 | 2.8 | 2.2 |
| 12,500 | 10/10 | 21.0 ± 0.3 | 24.5 ± 0.3 | 3.5 ± 0.3 | 101 | 2.7 | 3.0 |
| 25,000 | 10/10 | 20.3 ± 0.3 | 24.2 ± 0.4 | 3.9 ± 0.4 | 100 | 3.1 | 3.4 |
| 50,000 | 10/10 | 20.1 ± 0.2 | 22.8 ± 0.3** | 2.7 ± 0.2** | 94 | 2.8 | 3.0 |
* Significantly different (P≤0.05) from the control group by Williams' or Dunnett's test.
** P≤0.01
a Number of animals surviving at 13 weeks/number initially in group
b Weights given as mean ± standard error.
c Feed consumption is expressed as grams per animal per day.
d Week of death: 11
e Week of death: 6
Table 5: Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats in the 13-Week Feed Study of Manganese (II) Sulfate Monohydratea
| 0 ppm | 1,600 ppm | 3,130 ppm | 6,250 ppm | 12,500 ppm | 25,000 ppm | |
| Male | ||||||
| n | 10 | 10 | 10 | 10 | 10 | 10 |
| Necropsy body wt | 301 ± 4 | 300 ± 5 | 296 ± 4 | 299 ± 4 | 296 ± 6 | 298 ± 11 |
| Brain | ||||||
| Absolute | 1.723 ± 0.015 | 1.708 ± 0.013 | 1.731 ± 0.017 | 1.707 ± 0.045 | 1.721 ± 0.010 | 1.715 ± 0.029 |
| Relative | 5.73 ± 0.06 | 5.70 ± 0.08 | 5.86 ± 0.07 | 5.72 ± 0.16 | 5.84 ± 0.11 | 5.82 ± 0.21 |
| Heart | ||||||
| Absolute | 0.720 ± 0.016 | 0.733 ± 0.019 | 0.744 ± 0.009 | 0.739 ± 0.021 | 0.742 ± 0.023 | 0.710 ± 0.014 |
| Relative | 2.39 ± 0.05 | 2.44 ± 0.05 | 2.52 ± 0.02 | 2.47 ± 0.08 | 2.51 ± 0.04 | 2.40 ± 0.08 |
| R. Kidney | ||||||
| Absolute | 0.899 ± 0.016 | 0.858 ± 0.014 | 0.852 ± 0.015 | 0.904 ± 0.028 | 0.861 ± 0.019 | 0.892 ± 0.025 |
| Relative | 2.99 ± 0.05 | 2.86 ± 0.03 | 2.88 ± 0.04 | 3.02 ± 0.08 | 2.91 ± 0.04 | 3.01 ± 0.09 |
| Liver | ||||||
| Absolute | 10.688 ± 0.282 | 9.535 ± 0.205** | 9.129 ± 0.165** | 9.441 ± 0.211** | 8.950 ± 0.274** | 9.014 ± 0.274** |
| Relative | 35.49 ± 0.71 | 31.78 ± 0.50** | 30.85 ± 0.25** | 31.57 ± 0.55** | 30.25 ± 0.49** | 30.43 ± 1.00** |
| Lungs | ||||||
| Absolute | 1.164 ± 0.037 | 1.073 ± 0.031 | 1.080 ± 0.019 | 1.111 ± 0.042 | 1.325 ± 0.060* | 1.011 ± 0.032* |
| Relative | 3.86 ± 0.10 | 3.57 ± 0.08 | 3.65 ± 0.06 | 3.72 ± 0.15 | 4.49 ± 0.21** | 3.42 ± 0.13 |
| L. Testis | ||||||
| Absolute | 1.292 ± 0.033 | 1.414 ± 0.103 | 1.294 ± 0.021 | 1.315 ± 0.011 | 1.245 ± 0.031 | 1.290 ± 0.019 |
| Relative | 4.29 ± 0.10 | 4.74 ± 0.39 | 4.38 ± 0.08 | 4.40 ± 0.06 | 4.22 ± 0.11 | 4.36 ± 0.11 |
| Thymus | ||||||
| Absolute | 0.146 ± 0.017 | 0.144 ± 0.016 | 0.109 ± 0.013 | 0.170 ± 0.021 | 0.174 ± 0.018 | 0.118 ± 0.018 |
| Relative | 0.48 ± 0.05 | 0.48 ± 0.05 | 0.37 ± 0.04 | 0.56 ± 0.07 | 0.58 ± 0.06 | 0.39 ± 0.05 |
| Female | ||||||
| n | 10 | 10 | 10 | 10 | 10 | 10 |
| Necropsy body wt | 187 ± 2 | 182 ± 2 | 178 ± 3* | 178 ± 2* | 181 ± 1* | 178 ± 3** |
| Brain | ||||||
| Absolute | 1.638 ± 0.012 | 1.596 ± 0.021 | 1.608 ± 0.017 | 1.613 ± 0.025 | 1.638 ± 0.016 | 1.608 ± 0.007 |
| Relative | 8.78 ± 0.10 | 8.76 ± 0.12 | 9.05 ± 0.13 | 9.08 ± 0.14 | 9.05 ± 0.10 | 9.05 ± 0.12 |
| Heart | ||||||
| Absolute | 0.485 ± 0.015 | 0.478 ± 0.008 | 0.443 ± 0.016 | 0.477 ± 0.013 | 0.472 ± 0.011 | 0.465 ± 0.016 |
| Relative | 2.60 ± 0.07 | 2.62 ± 0.04 | 2.49 ± 0.07 | 2.68 ± 0.06 | 2.61 ± 0.06 | 2.61 ± 0.08 |
| R. Kidney | ||||||
| Absolute | 0.509 ± 0.011 | 0.505 ± 0.014 | 0.498 ± 0.008 | 0.493 ± 0.013 | 0.511 ± 0.011 | 0.517 ± 0.015b |
| Relative | 2.73 ± 0.06 | 2.77 ± 0.08 | 2.80 ± 0.04 | 2.77 ± 0.07 | 2.82 ± 0.06 | 2.89 ± 0.08b |
| Liver | ||||||
| Absolute | 5.754 ± 0.225 | 5.689 ± 0.119 | 5.343 ± 0.146 | 5.363 ± 0.125 | 5.281 ± 0.084* | 5.008 ± 0.143** |
| Relative | 30.80 ± 1.08 | 31.20 ± 0.53 | 30.04 ± 0.70 | 30.16 ± 0.59 | 29.17 ± 0.37 | 28.10 ± 0.52** |
| Lungs | ||||||
| Absolute | 1.006 ± 0.061 | 0.847 ± 0.031** | 0.878 ± 0.030** | 0.761 ± 0.021** | 0.834 ± 0.025** | 0.712 ± 0.036** |
| Relative | 5.40 ± 0.34 | 4.65 ± 0.17* | 4.94 ± 0.16* | 4.28 ± 0.11** | 4.62 ± 0.16** | 4.00 ± 0.20** |
| Thymus | ||||||
| Absolute | 0.130 ± 0.011 | 0.117 ± 0.008 | 0.123 ± 0.019 | 0.113 ± 0.011 | 0.126 ± 0.013 | 0.121 ± 0.008 |
| Relative | 0.70 ± 0.06 | 0.64 ± 0.04 | 0.69 ± 0.11 | 0.63 ± 0.06 | 0.70 ± 0.07 | 0.68 ± 0.04 |
* Significantly different (P≤0.05) from the control group by Williams' or Dunnett's test
** P≤0.01
a Organ weights and body weights are given in grams; organ-weight-to-body-weight ratios are given as mg organ weight/g body weight (mean ± standard error)
b n=9
Table 6: Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice in the 13-Week Feed Study of Manganese (II) Sulfate Monohydratea
| 0 ppm | 3,130 ppm | 6,250 ppm | 12,500 ppm | 25,000 ppm | 50,000 ppm | |
| Male | ||||||
| n | 9 | 10 | 10 | 10 | 10 | 10 |
| Necropsy body wt | 32.8 ± 0.5 | 32.3 ± 0.5 | 29.9 ± 0.5 | 31.6 ± 0.8 | 32.1 ± 0.4 | 29.8 ± 0.7** |
| Brain | ||||||
| Absolute | 0.453 ± 0.004 | 0.434 ± 0.009 | 0.440 ± 0.009 | 0.427 ± 0.020 | 0.453 ± 0.004 | 0.424 ± 0.006 |
| Relative | 13.85 ± 0.22 | 13.48 ± 0.42 | 14.76 ± 0.42 | 13.64 ± 0.82 | 14.13 ± 0.17 | 14.31 ± 0.41 |
| Heart | ||||||
| Absolute | 0.146 ± 0.004 | 0.134 ± 0.003 | 0.139 ± 0.003 | 0.132 ± 0.004* | 0.135 ± 0.004* | 0.115 ± 0.004** |
| Relative | 4.44 ± 0.10 | 4.15 ± 0.10 | 4.66 ± 0.13 | 4.23 ± 0.25 | 4.20 ± 0.08 | 3.89 ± 0.20* |
| R. Kidney | ||||||
| Absolute | 0.268 ± 0.008 | 0.267 ± 0.006 | 0.263 ± 0.008 | 0.255 ± 0.008 | 0.272 ± 0.008 | 0.222 ± 0.006** |
| Relative | 8.16 ± 0.16 | 8.28 ± 0.22 | 8.80 ± 0.21 | 8.17 ± 0.49 | 8.47 ± 0.20 | 7.46 ± 0.17 |
| Liver | ||||||
| Absolute | 1.528 ± 0.097 | 1.452 ± 0.082 | 1.202 ± 0.032* | 1.338 ± 0.076* | 1.401 ± 0.076* | 1.063 ± 0.045** |
| Relative | 46.51 ± 2.69 | 44.78 ± 2.06 | 40.27 ± 1.18 | 42.76 ± 3.10 | 43.55 ± 2.10 | 35.54 ± 0.72** |
| Lungs | ||||||
| Absolute | 0.160 ± 0.006 | 0.169 ± 0.005 | 0.188 ± 0.006** | 0.162 ± 0.005b | 0.169 ± 0.007 | 0.147 ± 0.00b |
| Relative | 4.88 ± 0.17 | 5.26 ± 0.23 | 6.29 ± 0.18** | 5.22 ± 0.32b | 5.28 ± 0.24 | 4.90 ± 0.19b |
| L. Testis | ||||||
| Absolute | 0.109 ± 0.002 | 0.107 ± 0.003b | 0.111 ± 0.002 | 0.107 ± 0.001 | 0.117 ± 0.003 | 0.094 ± 0.003** |
| Relative | 3.32 ± 0.07 | 3.32 ± 0.15b | 3.74 ± 0.11 | 3.41 ± 0.14 | 3.67 ± 0.14 | 3.19 ± 0.15 |
| Thymus | ||||||
| Absolute | 0.034 ± 0.004 | 0.035 ± 0.002 | 0.039 ± 0.003 | 0.034 ± 0.003 | 0.039 ± 0.002 | 0.031 ± 0.003 |
| Relative | 1.04 ± 0.11 | 1.09 ± 0.07 | 1.29 ± 0.10 | 1.08 ± 0.10 | 1.22 ± 0.06 | 1.04 ± 0.11 |
| Female | ||||||
| n | 10 | 9 | 10 | 10 | 10 | 10 |
| Necropsy body wt | 24.8 ± 0.4 | 25.8 ± 0.7 | 24.9 ± 0.3 | 26.4 ± 0.7 | 25.3 ± 0.5 | 24.7 ± 0.6 |
| Brain | ||||||
| Absolute | 0.452 ± 0.010 | 0.456 ± 0.005 | 0.454 ± 0.014 | 0.473 ± 0.006 | 0.451 ± 0.006 | 0.438 ± 0.005 |
| Relative | 18.29 ± 0.55 | 17.61 ± 0.43 | 18.28 ± 0.65 | 18.03 ± 0.56 | 17.88 ± 0.40 | 17.80 ± 0.34 |
| Heart | ||||||
| Absolute | 0.110 ± 0.003 | 0.109 ± 0.005 | 0.108 ± 0.002b | 0.120 ± 0.003 | 0.115 ± 0.003 | 0.106 ± 0.006 |
| Relative | 4.44 ± 0.13 | 4.18 ± 0.11 | 4.31 ± 0.06b | 4.56 ± 0.11 | 4.55 ± 0.10 | 4.31 ± 0.24 |
| R. Kidney | ||||||
| Absolute | 0.169 ± 0.003 | 0.169 ± 0.005 | 0.172 ± 0.003 | 0.188 ± 0.004 | 0.182 ± 0.006 | 0.173 ± 0.003 |
| Relative | 6.82 ± 0.07 | 6.50 ± 0.13 | 6.91 ± 0.11 | 7.17 ± 0.26 | 7.20 ± 0.21 | 7.03 ± 0.16 |
| Liver | ||||||
| Absolute | 1.041 ± 0.036 | 1.153 ± 0.082c | 0.992 ± 0.025 | 1.220 ± 0.075 | 1.056 ± 0.038 | 0.959 ± 0.051 |
| Relative | 41.95 ± 1.16 | 44.21 ± 1.84c | 39.82 ± 0.80 | 45.89 ± 1.81 | 41.65 ± 0.94 | 38.64 ± 1.27 |
| Lungs | ||||||
| Absolute | 0.157 ± 0.005 | 0.167 ± 0.017 | 0.157 ± 0.004 | 0.159 ± 0.008 | 0.153 ± 0.007 | 0.143 ± 0.003 |
| Relative | 6.34 ± 0.21 | 6.34 ± 0.45 | 6.31 ± 0.19 | 6.07 ± 0.37 | 6.05 ± 0.28 | 5.81 ± 0.16 |
| Thymus | ||||||
| Absolute | 0.033 ± 0.003 | 0.040 ± 0.002 | 0.043 ± 0.004* | 0.047 ± 0.001b**b | 0.044 ± 0.003b**b | 0.047 ± 0.002** |
| Relative | 1.34 ± 0.12 | 1.55 ± 0.05 | 1.74 ± 0.18* | 1.80 ± 0.05*b | 1.76 ± 0.13*b | 1.90 ± 0.07** |
* Significantly different (P≤0.05) from the control group by Williams' or Dunnett's test
** P≤0.01
a Organ weights and body weights are given in grams; organ-weight-to-body-weight ratios are given as mg organ weight/g body weight (mean ± standard error)
b n=9
c n=8
Table 7: Hematology Data for Rats in the 13 -Week Feed Study of Manganese (II) Sulfate Monohydratea
| 0 ppm | 1,600 ppm | 3,130 ppm | 6,250 ppm | 12,500 ppm | 25,000 ppm | |
| Male | ||||||
| n | 10 | 10 | 10 | 10 | 10 | 10 |
| Hematocrit (%) | 40.8 ± 1.2 | 42.3 ± 0.9 | 42.2 ± 1.0 | 44.1 ± 0.6* | 45.2 ± 1.4* | 45.8 ± 1.5* |
| Hemoglobin (g/dL) | 15.4 ± 0.3 | 15.4 ± 0.2 | 15.8 ± 0.3 | 16.0 ± 0.2 | 16.0 ± 0.3 | 15.7 ± 0.4 |
| Erythrocytes (106/µL) | 7.87 ± 0.15 | 8.20 ± 0.18 | 8.13 ± 0.17 | 8.60 ± 0.08** | 8.33 ± 0.13** | 8.82 ± 0.19** |
| Mean cell volume (fL) | 52.7 ± 0.6 | 51.9 ± 0.5 | 52.1 ± 0.4 | 51.4 ± 0.4 | 54.2 ± 0.9 | 52.2 ± 0.8 |
| Leukocytes (103/µL) | 3.17 ± 0.17 | 3.32 ± 0.14 | 3.27 ± 0.17 | 3.18 ± 0.09 | 3.94 ± 0.19* | 3.07 ± 0.18 |
| Segmented neutrophils (103/µL) | 0.61 ± 0.06 | 1.19 ± 0.06** | 1.15 ± 0.12** | 1.29 ± 0.10** | 1.73 ± 0.12** | 1.30 ± 0.08** |
| Lymphocytes (103/µL) | 2.51 ± 0.19 | 2.08 ± 0.12 | 2.05 ± 0.12 | 1.80 ± 0.10** | 2.17 ± 0.15* | 1.73 ± 0.17** |
| Monocytes (103/µL) | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.01 ± 0.01 | 0.02 ± 0.02 | 0.01 ± 0.00 |
| Eosinophils (103/µL) | 0.04 ± 0.02 | 0.05 ± 0.01 | 0.06 ± 0.01 | 0.05 ± 0.01 | 0.05 ± 0.02 | 0.04 ± 0.01 |
| Female | ||||||
| n | 10 | 10 | 10 | 10 | 10 | 10 |
| Hematocrit (%) | 42.0 ± 0.8 | 40.4 ± 0.7 | 41.3 ± 0.8 | 41.1 ± 0.8 | 42.3 ± 0.8 | 43.1 ± 1.1 |
| Hemoglobin (g/dL) | 15.7 ± 0.2 | 15.5 ± 0.2 | 15.1 ± 0.2 | 15.5 ± 0.3 | 15.6 ± 0.3 | 16.1 ± 0.3 |
| Erythrocytes (106/µL) | 7.34 ± 0.13 | 7.09 ± 0.15 | 7.39 ± 0.16 | 7.16 ± 0.16 | 7.59 ± 0.15 | 7.91 ± 0.16* |
| Mean cell volume (fL) | 57.3 ± 0.7 | 57.0 ± 0.7 | 56.0 ± 0.5 | 56.5 ± 0.8 | 56.0 ± 0.6 | 54.6 ± 0.5** |
| Leukocytes (103/µL) | 3.79 ± 0.33 | 2.98 ± 0.09* | 3.09 ± 0.16 | 2.78 ± 0.09** | 2.75 ± 0.13** | 2.78 ± 0.21** |
| Segmented neutrophils (103/µL) | 1.05 ± 0.17 | 0.72 ± 0.08 | 0.76 ± 0.09 | 0.58 ± 0.06** | 0.73 ± 0.08 | 0.87 ± 0.11 |
| Lymphocytes (103/µL) | 2.70 ± 0.23 | 2.23 ± 0.08 | 2.26 ± 0.13 | 2.12 ± 0.09* | 1.95 ± 0.12** | 1.82 ± 0.12** |
| Monocytes (103/µL) | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 |
| Eosinophils (103/µL) | 0.04 ± 0.01 | 0.03 ± 0.01 | 0.07 ± 0.02 | 0.06 ± 0.02 | 0.07 ± 0.01 | 0.09 ± 0.02* |
* Significantly different (P≤0.05) from the control group by Dunn's or Shirley's test
** P≤0.01
a Mean ± standard error
Table 8:
Hematology Data for Mice in the 13-Week Feed Study of Manganese (II) Sulfate Monohydratea
| 0 ppm | 3,130 ppm | 6,250 ppm | 12,500 ppm | 25,000 ppm | 50,000 ppm | |
| Male | ||||||
| n | 9 | 10 | 10 | 10 | 10 | 10 |
| Hematocrit (%) | 46.1 ± 0.9 | 47.6 ± 1.1 | 52.6 ± 2.1 | 45.2 ± 1.0 | 45.6 ± 1.3 | 38.3 ± 1.5** |
| Hemoglobin (g/dL) | 14.4 ± 0.2 | 15.2 ± 0.4 | 16.4 ± 0.5 | 14.5 ± 0.3 | 14.2 ± 0.4 | 12.1 ± 0.5* |
| Erythrocytes (106/µL) | 8.77 ± 0.24 | 9.28 ± 0.18 | 10.06 ± 0.33* | 9.02 ± 0.17 | 9.17 ± 0.24 | 8.71 ± 0.36 |
| Mean cell volume (fL) | 51.4 ± 0.4 | 51.3 ± 0.6 | 52.0 ± 0.6 | 50.3 ± 0.3 | 50.0 ± 0.6 | 43.6 ± 1.2** |
| Leukocytes (103/µL) | 3.48 ± 0.13 | 2.52 ± 0.25* | 3.28 ± 0.12 | 3.24 ± 0.43 | 2.40 ± 0.38** | 2.85 ± 0.54** |
| Segmented neutrophils (103/µL) | 1.52 ± 0.27 | 0.92 ± 0.16 | 1.12 ± 0.10 | 1.60 ± 0.27 | 0.81 ± 0.10 | 1.04 ± 0.22b |
| Lymphocytes (103/µL) | 1.87 ± 0.21 | 1.49 ± 0.15 | 2.04 ± 0.13 | 1.55 ± 0.28 | 1.54 ± 0.32 | 1.87 ± 0.39b |
| Monocytes (103/µL) | 0.01 ± 0.01 | 0.00 ± 0.00 | 0.01 ± 0.01 | 0.01 ± 0.01 | 0.00 ± 0.00 | 0.01 ± 0.01b |
| Eosinophils (103/µL) | 0.08 ± 0.02 | 0.10 ± 0.03 | 0.10 ± 0.03 | 0.05 ± 0.02 | 0.05 ± 0.01 | 0.03 ± 0.01b |
| Female | ||||||
| n | 10 | 6 | 8 | 9 | 10 | 10 |
| Hematocrit (%) | 45.4 ± 1.7 | 50.1 ± 1.7 | 49.4 ± 1.6 | 47.9 ± 1.5 | 46.4 ± 1.2 | 40.7 ± 1.5 |
| Hemoglobin (g/dL) | 14.4 ± 0.5 | 15.3 ± 0.4 | 14.9 ± 0.4 | 14.4 ± 0.3 | 14.8 ± 0.4 | 12.8 ± 0.5* |
| Erythrocytes (106/µL) | 8.54 ± 0.33 | 9.57 ± 0.22 | 8.87 ± 0.24 | 8.95 ± 0.19 | 8.89 ± 0.21 | 8.80 ± 0.35 |
| Mean cell volume (fL) | 52.4 ± 1.4 | 49.7 ± 3.2c | 54.6 ± 1.9 | 52.6 ± 0.7 | 52.3 ± 0.2 | 46.5 ± 1.1** |
| Leukocytes (103/µL) | 2.90 ± 0.44 | 2.78 ± 0.34 | 2.54 ± 0.39 | 2.19 ± 0.17 | 2.11 ± 0.21 | 2.46 ± 0.32 |
| Segmented neutrophils (103/µL) | 0.62 ± 0.13 | 1.07 ± 0.14 | 1.09 ± 0.22 | 0.78 ± 0.13 | 0.72 ± 0.14 | 0.88 ± 0.18 |
| Lymphocytes (103/µL) | 1.89 ± 0.44 | 1.65 ± 0.23 | 1.37 ± 0.17 | 1.37 ± 0.13 | 1.33 ± 0.18 | 1.55 ± 0.17 |
| Monocytes (103/µL) | 0.01 ± 0.01 | 0.03 ± 0.01 | 0.02 ± 0.02 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 |
| Eosinophils (103/µL) | 0.08 ± 0.02 | 0.03 ± 0.02 | 0.05 ± 0.03 | 0.04 ± 0.01 | 0.06 ± 0.01 | 0.03 ± 0.01 |
* Significantly different (P≤0.05) from the control group by Dunn's or Shirley's test
** P≤0.01
a Mean ± standard error
b n=9
c n=7
Table 1: Survival, Body Weights, and Feed Consumption of Rats in the 14-Day Feed Study of Manganese (II) Sulfate Monohydrate
| Concentration (ppm) | Survivala | Mean Body Weight and Weight Changesb(g) Relative | Final Weight Feed to Controls (%) | Consumptionc | |||
| Initial | Final | Change | Week 1 | Week 2 | |||
| Male | |||||||
| 0 | 5/5 | 183 ± 14 | 241 ± 12 | 58 ± 2 | 17.2 | 17.2 | |
| 3,130 | 5/5 | 176 ± 6 | 235 ± 5 | 59 ± 3 | 98 | 16.6 | 17.2 |
| 6,250 | 5/5 | 182 ± 15 | 242 ± 13 | 60 ± 3 | 101 | 16.7 | 17.7 |
| 12,500 | 5/5 | 176 ± 7 | 235 ± 6 | 58 ± 3 | 98 | 16.7 | 17.7 |
| 25,000 | 5/5 | 186 ± 7 | 243 ± 6 | 57 ± 1 | 101 | 17.0 | 18.1 |
| 50,000 | 5/5 | 185 ± 5 | 210 ± 6* | 25 ± 2** | 87 | 13.9 | 16.8 |
| Female | |||||||
| 0 | 5/5 | 140 ± 3 | 165 ± 4 | 25 ± 2 | 12.2 | 11.4 | |
| 3,130 | 5/5 | 144 ± 5 | 171 ± 5 | 27 ± 3 | 104 | 14.2 | 11.8 |
| 6,250 | 5/5 | 134 ± 4 | 157 ± 3 | 23 ± 2 | 95 | 11.8 | 11.0 |
| 12,500 | 5/5 | 136 ± 5 | 163 ± 6 | 27 ± 1 | 99 | 13.2 | 11.9 |
| 25,000 | 5/5 | 139 ± 5 | 166 ± 4 | 27 ± 1 | 101 | 13.3 | 12.0 |
| 50,000 | 5/5 | 134 ± 5 | 153 ± 5 | 20 ± 1 | 93 | 10.4 | 12.1 |
* Significantly different (P≤0.05) from the control group by Williams' or Dunnett's test.
** P≤0.01
a Number of animals surviving at 14 days/number initially in group
b Weights given as mean ± standard error.
c Feed consumption is expressed as grams per animal per day.
Table 2: Survival, Body Weights, and Feed Consumption of Mice in the 14-Day Feed Study of Manganese (II) Sulfate Monohydrate
| Concentration (ppm) | Survivala | Mean Body Weight and Weight Changesb(g) Relative | Final Weight Feed to Controls (%) | Consumptionc | |||
| Initial | Final | Change | Week 1 | Week 2 | |||
| Male | |||||||
| 0 | 5/5 | 21.4 ± 0.6 | 25.6 ± 1.2 | 4.2 ± 0.8 | 4.2 | 4.2 | |
| 3,130 | 5/5 | 23.8 ± 0.4* | 26.8 ± 0.7 | 3.0 ± 0.5 | 105 | 2.8 | 3.2 |
| 6,250 | 5/5 | 24.4 ± 0.5** | 26.0 ± 1.0 | 1.6 ± 0.5** | 102 | 3.0 | 3.5 |
| 12,500 | 5/5 | 24.6 ± 0.7** | 24.0 ± 0.7 | -0.6 ± 0.5** | 94 | 3.7 | 4.3 |
| 25,000 | 5/5 | 24.8 ± 0.4** | 24.4 ± 0.2 | -0.4 ± 0.2** | 95 | 5.1 | 4.6 |
| 50,000 | 5/5 | 19.2 ± 0.4* | 21.8 ± 0.7* | 2.6 ± 0.5** | 85 | 3.2 | 4.9 |
| Female | |||||||
| 0 | 5/5 | 15.6 ± 0.6 | 21.0 ± 1.0 | 5.4 ± 0.8 | 3.3 | 4.2 | |
| 3,130 | 5/5 | 18.4 ± 0.2** | 18.0 ± 0.3** | -0.4 ± 0.2** | 86 | 3.8 | 4.8 |
| 6,250 | 5/5 | 17.8 ± 0.2** | 17.2 ± 0.4** | -0.6 ± 0.4** | 82 | 4.1 | 4.3 |
| 12,500 | 5/5 | 18.6 ± 0.7** | 16.8 ± 0.6** | -1.8 ± 0.4** | 80 | 4.1 | 5.2 |
| 25,000 | 4/5d | 18.2 ± 0.4** | 17.0 ± 0.4** | -1.3 ± 0.3** | 81 | 4.8 | |
| 50,000 | 5/5 | 18.6 ± 0.4** | 15.2 ± 0.5** | -3.4 ± 0.2** | 72 | 3.5 | 3.9 |
* Significantly different (P≤0.05) from the control group by Williams' or Dunnett's test.
** P≤0.01
a Number of animals surviving at 14 days/number initially in group
b Weights given as mean ± standard error.
c Feed consumption is expressed as grams per animal per day.
d Day of death: 1
Table 3: Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats in the 14-Day Feed Study of Manganese (II) Sulfate Monohydratea
| 0 ppm | 3,130 ppm | 6,250 ppm | 12,500 ppm | 25,000 ppm | 50,000 ppm | |
| Male | ||||||
| n | 5 | 5 | 5 | 5 | 5 | 5 |
| Necropsy body wt | 241 ± 12 | 235 ± 5 | 242 ± 13 | 235 ± 6 | 243 ± 6 | 210 ± 6* |
| Brain | ||||||
| Absolute | 1.654 ± 0.089 | 1.718 ± 0.025 | 1.640 ± 0.101 | 1.674 ± 0.054 | 1.732 ± 0.077 | 1.748 ± 0.034 |
| Relative | 6.95 ± 0.51 | 7.32 ± 0.08 | 6.85 ± 0.57 | 7.14 ± 0.23 | 7.15 ± 0.37 | 8.35 ± 0.21* |
| Heart | ||||||
| Absolute | 0.692 ± 0.038 | 0.668 ± 0.031 | 0.728 ± 0.049 | 0.678 ± 0.022 | 0.700 ± 0.016 | 0.602 ± 0.019 |
| Relative | 2.88 ± 0.05 | 2.84 ± 0.07 | 3.00 ± 0.09 | 2.89 ± 0.05 | 2.88 ± 0.03 | 2.87 ± 0.04 |
| R. Kidney | ||||||
| Absolute | 0.880 ± 0.048 | 0.844 ± 0.037 | 0.870 ± 0.057 | 0.838 ± 0.019 | 0.884 ± 0.023 | 0.802 ± 0.018 |
| Relative | 3.66 ± 0.04 | 3.59 ± 0.08 | 3.58 ± 0.07 | 3.57 ± 0.03 | 3.64 ± 0.02 | 3.83 ± 0.07 |
| Liver | ||||||
| Absolute | 8.988 ± 0.553 | 9.028 ± 0.324 | 9.092 ± 0.402 | 8.114 ± 0.256 | 8.598 ± 0.141 | 7.528 ± 0.264** |
| Relative | 37.27 ± 0.48 | 38.37 ± 0.58 | 37.62 ± 0.57 | 34.57 ± 0.82* | 35.41 ± 0.40* | 35.87 ± 0.57* |
| Lungs | ||||||
| Absolute | 1.004 ± 0.058 | 1.082 ± 0.039 | 1.176 ± 0.095 | 1.030 ± 0.035 | 1.030 ± 0.046 | 1.024 ± 0.045 |
| Relative | 4.17 ± 0.11 | 4.61 ± 0.14 | 4.83 ± 0.16* | 4.39 ± 0.13 | 4.24 ± 0.15 | 4.89 ± 0.20* |
| L. Testis | ||||||
| Absolute | 1.256 ± 0.063 | 1.225 ± 0.026b | 1.256 ± 0.043 | 1.236 ± 0.025 | 1.272 ± 0.029 | 1.234 ± 0.044 |
| Relative | 5.22 ± 0.03 | 5.17 ± 0.04 | 5.21 ± 0.14 | 5.27 ± 0.08 | 5.24 ± 0.07 | 5.89 ± 0.17** |
| Thymus | ||||||
| Absolute | 0.261 ± 0.014 | 0.266 ± 0.033 | 0.272 ± 0.013 | 0.241 ± 0.013 | 0.252 ± 0.015 | 0.241 ± 0.011 |
| Relative | 1.10 ± 0.08 | 1.13 ± 0.14 | 1.13 ± 0.08 | 1.03 ± 0.05 | 1.04 ± 0.07 | 1.16 ± 0.08 |
| Female | ||||||
| n | 5 | 5 | 5 | 5 | 5 | 5 |
| Necropsy body wt | 165 ± 4 | 169 ± 6 | 157 ± 3 | 163 ± 6 | 166 ± 4 | 153 ± 5 |
| Brain | ||||||
| Absolute | 1.660 ± 0.024 | 1.684 ± 0.038 | 1.662 ± 0.036 | 1.590 ± 0.069 | 1.660 ± 0.048 | 1.672 ± 0.020 |
| Relative | 10.09 ± 0.22 | 9.99 ± 0.35 | 10.61 ± 0.36 | 9.77 ± 0.42 | 10.03 ± 0.39 | 10.94 ± 0.28 |
| Heart | ||||||
| Absolute | 0.522 ± 0.019 | 0.556 ± 0.018 | 0.502 ± 0.015 | 0.492 ± 0.021 | 0.530 ± 0.013 | 0.482 ± 0.024 |
| Relative | 3.17 ± 0.08 | 3.29 ± 0.08 | 3.20 ± 0.09 | 3.01 ± 0.05 | 3.19 ± 0.04 | 3.14 ± 0.06 |
| R. Kidney | ||||||
| Absolute | 0.604 ± 0.017 | 0.620 ± 0.030 | 0.556 ± 0.019 | 0.568 ± 0.018 | 0.610 ± 0.034 | 0.566 ± 0.022 |
| Relative | 3.67 ± 0.09 | 3.66 ± 0.06 | 3.54 ± 0.06 | 3.48 ± 0.08 | 3.67 ± 0.13 | 3.69 ± 0.04 |
| Liver | ||||||
| Absolute | 6.018 ± 0.212 | 5.634 ± 0.278 | 5.274 ± 0.260 | 5.228 ± 0.297 | 5.586 ± 0.122 | 5.132 ± 0.197* |
| Relative | 36.49 ± 0.70 | 33.26 ± 0.83 | 33.58 ± 1.43 | 31.96 ± 0.99* | 33.76 ± 1.22 | 33.49 ± 0.95 |
| Lungs | ||||||
| Absolute | 0.824 ± 0.044 | 0.908 ± 0.065 | 0.890 ± 0.044 | 0.816 ± 0.070 | 0.922 ± 0.076 | 0.746 ± 0.052 |
| Relative | 4.99 ± 0.21 | 5.35 ± 0.25 | 5.68 ± 0.29 | 4.99 ± 0.37 | 5.53 ± 0.38 | 4.85 ± 0.21 |
| Thymus | ||||||
| Absolute | 0.290 ± 0.016 | 0.284 ± 0.019 | 0.241 ± 0.011 | 0.242 ± 0.025 | 0.287 ± 0.012 | 0.260 ± 0.020 |
| Relative | 1.76 ± 0.09 | 1.67 ± 0.08 | 1.54 ± 0.06 | 1.49 ± 0.18 | 1.73 ± 0.07 | 1.69 ± 0.10 |
* Significantly different (P≤0.05) from the control group by Williams' or Dunnett's test
** P≤0.01
a Organ weights and body weights are given in grams; organ-weight-to-body-weight ratios are given as mg organ weight/g body weight (mean ± standard error)
b n=4
Table 4: Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice in the 14-Day Feed Study of Manganese (II) Sulfate Monohydratea
| 0 ppm | 3,130 ppm | 6,250 ppm | 12,500 ppm | 25,000 ppm | 50,000 ppm | |
| Male | ||||||
| n | 5 | 5 | 5 | 5 | 5 | 5 |
| Necropsy body wt | 25.6 ± 1.2 | 26.8 ± 0.7 | 26.0 ± 1.0 | 24.0 ± 0.7 | 24.4 ± 0.2 | 21.8 ± 0.7** |
| Brain | ||||||
| Absolute | 0.452 ± 0.005 | 0.460 ± 0.009 | 0.442 ± 0.014 | 0.416 ± 0.013 | 0.450 ± 0.004 | 0.436 ± 0.006 |
| Relative | 17.80 ± 0.77 | 17.23 ± 0.67 | 17.16 ± 1.12 | 17.43 ± 0.94 | 18.45 ± 0.26 | 20.11 ± 0.84 |
| Heart | ||||||
| Absolute | 0.132 ± 0.014 | 0.120 ± 0.004 | 0.125 ± 0.006b | 0.120 ± 0.005 | 0.124 ± 0.004 | 0.102 ± 0.004** |
| Relative | 5.13 ± 0.39 | 4.49 ± 0.19 | 4.95 ± 0.22b | 4.99 ± 0.11 | 5.08 ± 0.17 | 4.68 ± 0.06 |
| R. Kidney | ||||||
| Absolute | 0.208 ± 0.012 | 0.216 ± 0.002 | 0.224 ± 0.014 | 0.220 ± 0.007 | 0.218 ± 0.004 | 0.194 ± 0.009 |
| Relative | 8.11 ± 0.17 | 8.08 ± 0.22 | 8.61 ± 0.40 | 9.17 ± 0.14* | 8.93 ± 0.10* | 8.90 ± 0.27* |
| Liver | ||||||
| Absolute | 1.154 ± 0.107 | 1.222 ± 0.082 | 1.346 ± 0.081 | 1.226 ± 0.086 | 1.206 ± 0.022 | 0.920 ± 0.038* |
| Relative | 44.75 ± 2.21 | 45.55 ± 2.62 | 51.61 ± 1.60 | 50.96 ± 2.64 | 49.43 ± 0.82 | 42.17 ± 0.47 |
| Lungs | ||||||
| Absolute | 0.152 ± 0.011 | 0.160 ± 0.003 | 0.176 ± 0.009 | 0.192 ± 0.009* | 0.204 ± 0.014* | 0.158 ± 0.007* |
| Relative | 5.98 ± 0.47 | 5.98 ± 0.15 | 6.81 ± 0.45 | 8.01 ± 0.33** | 8.35 ± 0.52** | 7.27 ± 0.36** |
| L. Testis | ||||||
| Absolute | 0.100 ± 0.003 | 0.101 ± 0.005 | 0.099 ± 0.003 | 0.095 ± 0.010 | 0.100 ± 0.011 | 0.101 ± 0.004 |
| Relative | 3.92 ± 0.17 | 3.77 ± 0.11 | 3.81 ± 0.16 | 3.94 ± 0.36 | 4.12 ± 0.45 | 4.64 ± 0.25 |
| Thymus | ||||||
| Absolute | 0.041 ± 0.002 | 0.040 ± 0.003 | 0.043 ± 0.006 | 0.030 ± 0.003 | 0.059 ± 0.004* | 0.032 ± 0.007 |
| Relative | 1.59 ± 0.04 | 1.50 ± 0.10 | 1.66 ± 0.27 | 1.28 ± 0.17 | 2.43 ± 0.17* | 1.48 ± 0.32 |
| Female | ||||||
| n | 5 | 5 | 5 | 5 | 4 | 5 |
| Necropsy body wt | 21.0 ± 1.0 | 18.0 ± 0.3* | 18.8 ± 1.3* | 16.8 ± 0.6** | 17.0 ± 0.4** | 15.2 ± 0.5** |
| Brain | ||||||
| Absolute | 0.440 ± 0.013 | 0.454 ± 0.005 | 0.430 ± 0.008 | 0.440 ± 0.015 | 0.453 ± 0.010 | 0.416 ± 0.017 |
| Relative | 21.03 ± 0.52 | 25.25 ± 0.52** | 23.23 ± 1.35** | 26.20 ± 0.40** | 26.62 ± 0.10** | 27.36 ± 0.71** |
| Heart | ||||||
| Absolute | 0.094 ± 0.004 | 0.096 ± 0.005 | 0.090 ± 0.000 | 0.100 ± 0.006 | 0.098 ± 0.005 | 0.082 ± 0.004 |
| Relative | 4.48 ± 0.08 | 5.33 ± 0.27 | 4.87 ± 0.29 | 5.94 ± 0.23** | 5.73 ± 0.18** | 5.39 ± 0.16** |
| R. Kidney | ||||||
| Absolute | 0.148 ± 0.008 | 0.148 ± 0.011 | 0.136 ± 0.006 | 0.152 ± 0.010 | 0.150 ± 0.006 | 0.126 ± 0.005 |
| Relative | 7.04 ± 0.16 | 8.24 ± 0.65 | 7.36 ± 0.54 | 9.01 ± 0.32** | 8.82 ± 0.24* | 8.29 ± 0.17** |
| Liver | ||||||
| Absolute | 0.964 ± 0.052 | 0.866 ± 0.033 | 0.784 ± 0.032 | 0.916 ± 0.039 | 0.870 ± 0.033 | 0.716 ± 0.089** |
| Relative | 45.84 ± 0.57 | 48.05 ± 1.13 | 42.62 ± 3.53 | 54.59 ± 1.76 | 51.16 ± 1.22 | 47.24 ± 6.01 |
| Lungs | ||||||
| Absolute | 0.150 ± 0.015 | 0.160 ± 0.007 | 0.144 ± 0.009 | 0.152 ± 0.006 | 0.175 ± 0.013 | 0.132 ± 0.006 |
| Relative | 7.12 ± 0.60 | 8.88 ± 0.31 | 7.78 ± 0.66 | 9.05 ± 0.14* | 10.30 ± 0.77** | 8.69 ± 0.29** |
| Thymus | ||||||
| Absolute | 0.056 ± 0.002 | 0.058 ± 0.003 | 0.057 ± 0.008 | 0.049 ± 0.003 | 0.059 ± 0.005 | 0.042 ± 0.003* |
| Relative | 2.69 ± 0.15 | 3.19 ± 0.10 | 3.10 ± 0.52 | 2.94 ± 0.13 | 3.50 ± 0.35 | 2.82 ± 0.24 |
* Significantly different (P≤0.05) from the control group by Williams' or Dunnett's test
** P≤0.01
a Organ weights and body weights are given in grams; organ-weight-to-body-weight ratios are given as mg organ weight/g body weight (mean ± standard error)
b n=4
Table 5: Hematology Data for Rats in the 14-Day Feed Study of Manganese (II) Sulfate Monohydratea
| 0 ppm | 3,130 ppm | 6,250 ppm | 12,500 ppm | 25,000 ppm | 50,000 ppm | |
| Male | ||||||
| n | 5 | 5 | 5 | 5 | 5 | 5 |
| Hematocrit (%) | 43.8 ± 1.0 | 42.6 ± 1.2 | 42.9 ± 0.6 | 44.3 ± 1.0 | 40.3 ± 1.4 | 41.6 ± 1.0 |
| Hemoglobin (g/dL) | 15.3 ± 0.3 | 15.9 ± 0.2 | 15.7 ± 0.3 | 15.9 ± 0.3 | 14.9 ± 0.3 | 14.8 ± 0.3 |
| Erythrocytes (106/µL) | 8.10 ± 0.21 | 7.80 ± 0.25 | 7.96 ± 0.15 | 8.04 ± 0.08 | 7.54 ± 0.26 | 7.97 ± 0.19 |
| Mean cell volume (fL) | 53.0 ± 0.3 | 53.6 ± 0.6 | 52.8 ± 0.4 | 53.8 ± 0.9 | 52.4 ± 0.4 | 51.2 ± 0.2* |
| Leukocytes (103/µL) | 2.58 ± 0.36 | 3.38 ± 0.24 | 3.84 ± 0.37* | 3.60 ± 0.26* | 3.58 ± 0.22* | 6.32 ± 0.82** |
| Segmented neutrophils (103/µL) | 0.76 ± 0.11 | 1.03 ± 0.11 | 0.90 ± 0.19 | 0.93 ± 0.10 | 0.92 ± 0.12 | 3.52 ± 0.28** |
| Lymphocytes (103/µL) | 1.70 ± 0.36 | 2.24 ± 0.22 | 2.77 ± 0.24 | 2.53 ± 0.18 | 2.26 ± 0.16 | 2.46 ± 0.62 |
| Monocytes (103/µL) | 0.10 ± 0.03 | 0.07 ± 0.03 | 0.12 ± 0.03 | 0.10 ± 0.03 | 0.38 ± 0.11 | 0.33 ± 0.09* |
| Eosinophils (103/µL) | 0.02 ± 0.01 | 0.04 ± 0.02 | 0.05 ± 0.03 | 0.04 ± 0.02 | 0.02 ± 0.01 | 0.01 ± 0.01 |
| Female | ||||||
| n | 5 | 5 | 5 | 5 | 5 | 5 |
| Hematocrit (%) | 42.9 ± 1.4 | 43.6 ± 0.5 | 45.1 ± 0.5 | 43.4 ± 0.7 | 43.9 ± 0.8 | 39.3 ± 1.1 |
| Hemoglobin (g/dL) | 15.3 ± 0.3 | 16.0 ± 0.2 | 16.3 ± 0.1 | 16.0 ± 0.3 | 15.7 ± 0.2 | 14.0 ± 0.4 |
| Erythrocytes (106/µL) | 7.87 ± 0.16 | 8.10 ± 0.09 | 8.28 ± 0.08 | 8.03 ± 0.11 | 7.88 ± 0.02 | 7.42 ± 0.22 |
| Mean cell volume (fL) | 53.4 ± 0.7 | 52.6 ± 0.4 | 53.6 ± 0.2 | 52.8 ± 0.4 | 54.6 ± 0.9 | 51.8 ± 0.4 |
| Leukocytes (103/µL) | 2.72 ± 0.26 | 3.38 ± 0.09 | 3.22 ± 0.41 | 3.00 ± 0.64 | 2.72 ± 0.25 | 4.26 ± 0.32* |
| Segmented neutrophils (103/µL) | 0.55 ± 0.07 | 0.75 ± 0.06 | 0.86 ± 0.23 | 0.68 ± 0.25 | 0.55 ± 0.12 | 1.62 ± 0.23* |
| Lymphocytes (103/µL) | 2.02 ± 0.18 | 2.35 ± 0.10 | 2.08 ± 0.23 | 2.05 ± 0.37 | 1.95 ± 0.25 | 2.23 ± 0.23 |
| Monocytes (103/µL) | 0.07 ± 0.02 | 0.25 ± 0.04** | 0.25 ± 0.13* | 0.20 ± 0.04* | 0.18 ± 0.05* | 0.34 ± 0.14** |
| Eosinophils (103/µL) | 0.08 ± 0.04 | 0.04 ± 0.01 | 0.02 ± 0.01 | 0.07 ± 0.04 | 0.04 ± 0.02 | 0.07 ± 0.03 |
* Significantly different (P≤0.05) from the control group by Dunn's or Shirley's test
** P≤0.01
a Mean ± standard error
Table 6: Hematology Data for Mice in the 14-Day Feed Study of Manganese (II) Sulfate Monohydratea
| 0 ppm | 3,130 ppm | 6,250 ppm | 12,500 ppm | 25,000 ppm | 50,000 ppm | |
| Male | ||||||
| n | 3 | 5 | 5 | 5 | 5 | 5 |
| Hematocrit (%) | 45.5 ± 3.9 | 49.3 ± 3.1 | 46.3 ± 3.0 | 46.9 ± 1.1 | 46.9 ± 0.9 | 46.6 ± 1.0 |
| Hemoglobin (g/dL) | 15.0 ± 0.4 | 14.9 ± 0.5 | 14.5 ± 0.9 | 15.1 ± 0.5 | 14.9 ± 0.3 | 14.8 ± 0.5 |
| Erythrocytes (106/µL) | 9.08 ± 0.39 | 9.18 ± 0.37 | 8.82 ± 0.58 | 8.98 ± 0.15 | 9.18 ± 0.18 | 9.04 ± 0.25 |
| Mean cell volume (fL) | 50.7 ± 2.3 | 53.8 ± 1.1 | 52.8 ± 0.2 | 52.4 ± 0.5 | 51.6 ± 0.5 | 51.8 ± 1.1 |
| Leukocytes (103/µL) | 2.30 ± 0.23 | 3.96 ± 0.33 | 3.94 ± 0.50 | 5.18 ± 2.23 | 2.58 ± 0.44 | 3.54 ± 1.20 |
| Segmented neutrophils (103/µL) | 0.84 ± 0.23 | 1.61 ± 0.25 | 1.53 ± 0.60 | 2.65 ± 1.47 | 0.74 ± 0.17 | 0.83 ± 0.11 |
| Lymphocytes (103/µL) | 1.35 ± 0.08 | 2.10 ± 0.30 | 2.18 ± 0.29 | 2.26 ± 0.70 | 1.59 ± 0.26 | 2.49 ± 1.11 |
| Monocytes (103/µL) | 0.08 ± 0.02 | 0.22 ± 0.07 | 0.22 ± 0.07 | 0.26 ± 0.10 | 0.21 ± 0.04 | 0.21 ± 0.07 |
| Eosinophils (103/µL) | 0.03 ± 0.02 | 0.03 ± 0.01 | 0.02 ± 0.01 | 0.02 ± 0.01 | 0.04 ± 0.01 | 0.01 ± 0.01 |
| Female | ||||||
| n | 4 | 5 | 4 | 5 | 3 | 5 |
| Hematocrit (%) | 43.6 ± 1.3 | 47.6 ± 0.8 | 51.2 ± 1.1** | 47.8 ± 0.4 | 44.5 ± 2.5 | 42.5 ± 2.9 |
| Hemoglobin (g/dL) | 14.0 ± 0.3 | 15.2 ± 0.2 | 16.2 ± 0.3** | 15.1 ± 0.1 | 14.7 ± 0.6 | 13.7 ± 1.0 |
| Erythrocytes (106/µL) | 8.41 ± 0.19 | 9.15 ± 0.16 | 9.75 ± 0.20** | 9.11 ± 0.06 | 8.97 ± 0.43 | 8.46 ± 0.52 |
| Mean cell volume (fL) | 52.0 ± 1.2 | 52.2 ± 0.2 | 53.0 ± 0.7 | 52.6 ± 0.2 | 50.0 ± 1.5 | 50.4 ± 0.5 |
| Leukocytes (103/µL) | 1.833 ± 0.24 | 1.76 ± 0.09 | 3.20 ± 0.60 | 2.66 ± 0.18 | 2.67 ± 0.29 | 5.74 ± 4.14 |
| Segmented neutrophils (103/µL) | 0.35 ± 0.14b | 0.59 ± 0.11 | 0.87 ± 0.18* | 0.99 ± 0.14* | 0.84 ± 0.14 | 3.84 ± 3.23 |
| Lymphocytes (103/µL) | 1.25 ± 0.14b | 1.03 ± 0.10 | 2.23 ± 0.47 | 1.53 ± 0.12 | 1.67 ± 0.29 | 1.65 ± 0.70 |
| Monocytes (103/µL) | 0.06 ± 0.01b | 0.11 ± 0.04 | 0.07 ± 0.03 | 0.12 ± 0.04 | 0.09 ± 0.03 | 0.21 ± 0.17 |
| Eosinophils (103/µL) | 0.03 ± 0.02b | 0.02 ± 0.01 | 0.03 ± 0.03 | 0.01 ± 0.01 | 0.07 ± 0.02 | 0.22c |
* Significantly different (P≤0.05) from the control group by Dunn's or Shirley's test
** P≤0.01
a Mean ± standard error
b n=3
c n=1; no standard error calculated
TABLE 4 Survival of Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Control | 1,500 ppm | 5,000 ppm | 15,000 ppm | |
| Male | ||||
| Animals initially in study | 70 | 70 | 70 | 70 |
| 9-Month interim evaluationa | 8 | 10 | 10 | 10 |
| 15-Month interim evaluationa | 10 | 9 | 9 | 8 |
| Moribund | 21 | 24 | 24 | 38 |
| Natural deaths | 6 | 10 | 5 | 7 |
| Animals surviving to study termination | 25 | 17 | 22b | 7 |
| Percent probability of survival at end of studycMean survival (days)d | 49 | 34 | 43 | 14 |
| 581 | 573 | 579 | 571 | |
| Survival analysese | P=0.004 | P=0.381 | P=0.872 | P=0.006 |
| Female | ||||
| Animals initially in study | 70 | 70 | 70 | 70 |
| 9-Month interim evaluationa | 10 | 10 | 10 | 10 |
| 15-Month interim evaluationa | 10 | 10 | 9 | 10 |
| Accidental deathsa | 0 | 0 | 1 | 0 |
| Moribund | 6 | 11 | 6 | 11 |
| Natural deaths | 7 | 2 | 2 | 1 |
| Missexeda | 0 | 0 | 0 | 2 |
| Animals surviving to study termination | 37 | 37 | 42 | 36 |
| Percent probability of survival at end of study | 74 | 74 | 85 | 75 |
| Mean survival (days) | 608 | 594 | 596 | 607 |
| Survival analyses | P=0.914N | P=0.986 | P=0.378N | P=0.984N |
a Censored from survival analyses
b Includes one animal that died the last week of study
c Kaplan-Meier determinations
d Mean of all deaths (uncensored, censored, and terminal sacrifice)
e The result of the life table trend test (Tarone, 1975) is in the control column, and the results of the life table pairwise comparisons (Cox, 1972) with the controls are in the exposed columns. A negative trend or lower mortality in an exposure group is indicated by N.
TABLE 5 Mean Body Weights and Survival of Male Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Weeks on Study |
0 ppm | 1,500 ppm | 5,000 ppm | 15,000 ppm | |||||||
| Av. Wt. (g) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
|
| 1 | 126 | 70 | 124 | 98 | 70 | 126 | 100 | 70 | 123 | 98 | 70 |
| 2 | 167 | 70 | 166 | 100 | 70 | 167 | 101 | 70 | 164 | 98 | 70 |
| 3 | 201 | 70 | 197 | 98 | 70 | 202 | 101 | 70 | 193 | 96 | 70 |
| 4 | 227 | 70 | 220 | 97 | 70 | 228 | 100 | 70 | 220 | 97 | 70 |
| 5 | 247 | 70 | 244 | 98 | 70 | 251 | 101 | 70 | 244 | 99 | 70 |
| 6 | 272 | 70 | 268 | 98 | 70 | 274 | 101 | 70 | 268 | 98 | 70 |
| 7 | 283 | 70 | 280 | 99 | 70 | 287 | 102 | 70 | 280 | 99 | 70 |
| 8 | 297 | 70 | 295 | 99 | 70 | 303 | 102 | 70 | 293 | 99 | 70 |
| 9 | 314 | 70 | 307 | 98 | 70 | 313 | 100 | 70 | 306 | 98 | 70 |
| 10 | 327 | 70 | 318 | 97 | 70 | 328 | 100 | 70 | 321 | 98 | 70 |
| 11 | 334 | 70 | 327 | 98 | 70 | 334 | 100 | 70 | 324 | 97 | 70 |
| 12 | 343 | 70 | 335 | 98 | 70 | 343 | 100 | 70 | 335 | 98 | 70 |
| 13 | 353 | 70 | 347 | 98 | 70 | 351 | 99 | 70 | 341 | 97 | 70 |
| 18 | 386 | 69 | 375 | 97 | 70 | 383 | 99 | 70 | 376 | 97 | 70 |
| 21 | 399 | 69 | 394 | 99 | 69 | 394 | 99 | 70 | 386 | 97 | 70 |
| 25 | 415 | 69 | 408 | 98 | 69 | 411 | 99 | 70 | 399 | 96 | 70 |
| 28 | 431 | 69 | 423 | 98 | 69 | 425 | 98 | 70 | 416 | 96 | 70 |
| 32 | 447 | 69 | 440 | 98 | 69 | 441 | 99 | 70 | 429 | 96 | 70 |
| 36 | 462 | 69 | 455 | 99 | 69 | 453 | 98 | 70 | 442 | 96 | 70 |
| 40a | 460 | 60 | 453 | 98 | 60 | 452 | 98 | 60 | 441 | 96 | 58 |
| 44 | 472 | 60 | 450 | 95 | 60 | 451 | 96 | 60 | 441 | 93 | 58 |
| 48 | 480 | 60 | 474 | 99 | 58 | 476 | 99 | 60 | 466 | 97 | 58 |
| 53 | 484 | 60 | 479 | 99 | 57 | 479 | 99 | 60 | 471 | 97 | 57 |
| 57 | 496 | 60 | 487 | 98 | 57 | 487 | 98 | 59 | 482 | 97 | 57 |
| 61 | 497 | 60 | 485 | 98 | 57 | 490 | 99 | 58 | 481 | 97 | 57 |
| 65a | 505 | 50 | 488 | 97 | 48 | 493 | 98 | 49 | 488 | 97 | 49 |
| 69 | 509 | 49 | 487 | 96 | 46 | 491 | 97 | 49 | 486 | 95 | 49 |
| 73 | 510 | 48 | 493 | 97 | 46 | 497 | 97 | 47 | 493 | 97 | 47 |
| 77 | 509 | 47 | 490 | 96 | 45 | 496 | 98 | 43 | 490 | 96 | 46 |
| 81 | 498 | 46 | 479 | 96 | 45 | 484 | 97 | 42 | 477 | 96 | 45 |
| 85 | 492 | 43 | 479 | 97 | 44 | 484 | 99 | 41 | 478 | 97 | 44 |
| 89 | 486 | 38 | 469 | 97 | 40 | 469 | 97 | 39 | 449 | 92 | 43 |
| 93 | 472 | 35 | 451 | 96 | 38 | 460 | 97 | 35 | 432 | 91 | 35 |
| 97 | 449 | 31 | 426 | 95 | 32 | 443 | 99 | 32 | 409 | 91 | 26 |
| 101 | 433 | 26 | 431 | 100 | 23 | 423 | 98 | 28 | 378 | 87 | 18 |
| 104 | 402 | 26 | 402 | 100 | 20 | 393 | 98 | 24 | 360 | 90 | 10 |
| Mean for weeks | |||||||||||
| 1-13 | 269 | 264 | 98 | 270 | 100 | 262 | 97 | ||||
| 14-52 | 439 | 430 | 98 | 432 | 98 | 422 | 96 | ||||
| 53-104 | 482 | 468 | 97 | 471 | 98 | 455 | 94 | ||||
a Interim evaluations occurred during weeks 39 and 65.
TABLE 6 Mean Body Weights and Survival of Female Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Weeks on Study |
0 ppm | 1,500 ppm | 5,000 ppm | 15,000 ppm | |||||||
| Av. Wt. (g) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
|
| 1 | 104 | 70 | 103 | 100 | 70 | 104 | 100 | 70 | 103 | 99 | 70 |
| 2 | 127 | 70 | 128 | 101 | 70 | 128 | 101 | 70 | 126 | 99 | 69 |
| 3 | 143 | 70 | 143 | 100 | 70 | 143 | 100 | 70 | 142 | 99 | 69 |
| 4 | 154 | 70 | 154 | 100 | 70 | 153 | 99 | 70 | 151 | 98 | 68 |
| 5 | 161 | 70 | 164 | 102 | 70 | 160 | 99 | 70 | 160 | 99 | 68 |
| 6 | 170 | 70 | 173 | 102 | 70 | 170 | 100 | 70 | 168 | 99 | 68 |
| 7 | 174 | 70 | 179 | 103 | 70 | 176 | 101 | 70 | 175 | 101 | 68 |
| 8 | 181 | 70 | 184 | 102 | 70 | 181 | 100 | 69 | 181 | 100 | 68 |
| 9 | 186 | 70 | 190 | 102 | 70 | 186 | 100 | 69 | 186 | 100 | 68 |
| 10 | 190 | 70 | 194 | 102 | 70 | 190 | 100 | 69 | 190 | 100 | 68 |
| 11 | 191 | 70 | 195 | 102 | 70 | 192 | 100 | 69 | 191 | 100 | 68 |
| 12 | 196 | 70 | 198 | 101 | 70 | 196 | 100 | 68 | 195 | 100 | 68 |
| 13 | 199 | 70 | 201 | 101 | 70 | 199 | 100 | 68 | 198 | 100 | 68 |
| 18 | 205 | 70 | 208 | 101 | 70 | 206 | 100 | 68 | 205 | 100 | 68 |
| 21 | 210 | 70 | 213 | 101 | 70 | 211 | 100 | 68 | 211 | 100 | 68 |
| 25 | 217 | 70 | 221 | 102 | 70 | 218 | 100 | 68 | 217 | 100 | 68 |
| 28 | 225 | 70 | 225 | 100 | 70 | 222 | 99 | 68 | 223 | 99 | 68 |
| 32 | 232 | 70 | 236 | 102 | 70 | 231 | 100 | 68 | 231 | 100 | 68 |
| 36 | 237 | 70 | 244 | 103 | 70 | 240 | 101 | 68 | 237 | 100 | 68 |
| 40a | 241 | 60 | 245 | 102 | 60 | 240 | 101 | 58 | 243 | 101 | 58 |
| 44 | 249 | 60 | 257 | 103 | 60 | 251 | 101 | 58 | 251 | 101 | 58 |
| 48 | 258 | 60 | 260 | 101 | 60 | 261 | 101 | 58 | 266 | 103 | 58 |
| 53 | 272 | 60 | 277 | 102 | 60 | 270 | 99 | 58 | 277 | 102 | 58 |
| 57 | 277 | 60 | 287 | 104 | 60 | 279 | 101 | 58 | 287 | 104 | 58 |
| 61 | 286 | 60 | 294 | 103 | 59 | 287 | 101 | 58 | 295 | 103 | 58 |
| 65a | 296 | 50 | 305 | 103 | 48 | 298 | 101 | 49 | 303 | 103 | 48 |
| 69 | 305 | 50 | 313 | 103 | 48 | 306 | 101 | 49 | 312 | 102 | 48 |
| 73 | 311 | 50 | 320 | 103 | 47 | 316 | 101 | 49 | 318 | 102 | 47 |
| 77 | 321 | 50 | 327 | 102 | 47 | 328 | 102 | 48 | 329 | 103 | 46 |
| 81 | 327 | 49 | 332 | 102 | 43 | 333 | 102 | 48 | 335 | 103 | 46 |
| 85 | 335 | 48 | 339 | 101 | 42 | 340 | 102 | 47 | 342 | 102 | 46 |
| 89 | 336 | 47 | 346 | 103 | 41 | 346 | 103 | 45 | 346 | 103 | 46 |
| 93 | 340 | 43 | 342 | 101 | 41 | 345 | 102 | 45 | 345 | 102 | 45 |
| 97 | 334 | 43 | 335 | 100 | 39 | 341 | 102 | 43 | 337 | 101 | 44 |
| 101 | 336 | 38 | 330 | 98 | 39 | 337 | 100 | 42 | 333 | 99 | 38 |
| 104 | 327 | 38 | 326 | 100 | 37 | 336 | 103 | 42 | 336 | 103 | 36 |
| Mean for weeks | |||||||||||
| 1-13 | 167 | 170 | 102 | 168 | 101 | 167 | 100 | ||||
| 14-52 | 230 | 234 | 102 | 231 | 100 | 232 | 101 | ||||
| 53-104 | 315 | 320 | 102 | 319 | 101 | 321 | 102 | ||||
a Interim evaluations occurred during weeks 39 and 65.
TABLE 7 Incidence and Severity of Nephropathy of Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Dose (ppm) | 0 | 1,500 | 5,000 | 15,000 |
| Males | ||||
| 9-Month Interim Evaluation | ||||
| Kidneya | 8 | 10 | 10 | 10 |
| Nephropathyb | 8 (1.3)c | 10 (1.1) | 10 (1.1) | 10 (1.3) |
| 15-Month Interim Evaluation | ||||
| Kidney | 10 | 9 | 9 | 8 |
| Nephropathy | 10 (1.6) | 9 (1.9) | 9 (1.9) | 8 (2.0) |
| 2-Year Study | ||||
| Kidney | 52 | 50 | 51 | 52 |
| Nephropathy (initial evaluation) | 50 (2.9) | 49 (3.0) | 51 (3.0) | 50 (3.2) |
| Nephropathy (additional evaluation) | 52 (2.8) | - | - | 51 (3.1)* |
| Females | ||||
| 2-Year Study | ||||
| Kidney | 50 | 50 | 51 | 48 |
| Nephropathy | 48 (1.8) | 50 (1.5) | 49 (1.7) | 48 (1.9) |
* Significantly different (P0.05) from the control group by two-sided Mann-Whitney U test
a Number of animals with organ examined microscopically
b Number of animals with lesion
c Average severity grade of lesions in all animals (0=normal; 1=minimal; 2=mild; 3=moderate; 4=marked)
TABLE 10 Survival of Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Control | 1,500 ppm | 5,000 ppm | 15,000 ppm | |
| Male | ||||
| Animals initially in study | 70 | 70 | 70 | 70 |
| 9-Month interim evaluationa | 10 | 10 | 10 | 9 |
| 15-Month interim evaluationa | 10 | 10 | 9 | 10 |
| Accidental deathsa | 0 | 0 | 0 | 1 |
| Moribund | 2 | 3 | 2 | 1 |
| Natural deaths | 2 | 3 | 3 | 3 |
| Animals surviving to study termination | 46b | 44b | 46 | 46 |
| Percent probability of survival at end of studycMean survival (days)d | 92 | 88 | 91 | 93 |
| 620 | 619 | 615 | 615 | |
| Survival analysese | P=0.920N | P=0.708 | P=0.992 | P=0.748 |
| Female | ||||
| Animals initially in study | 70 | 70 | 70 | 70 |
| 9-Month interim evaluationa | 10 | 10 | 10 | 10 |
| 15-Month interim evaluationa | 9 | 10 | 9 | 9 |
| Accidental deathsa | 1 | 0 | 0 | 0 |
| Moribund | 6 | 4 | 6 | 4 |
| Natural deaths | 2 | 0 | 6 | 5 |
| Missinga | 0 | 0 | 1 | 0 |
| Animals surviving to study termination | 42 | 46 | 38 | 42 |
| Percent probability of survival at end of study | 85 | 92 | 77 | 83 |
| Mean survival (days) | 605 | 623 | 594 | 614 |
| Survival analyses | P=0.563 | P=0.318N | P=0.456 | P=0.961 |
a Censored from survival analyses
b Includes one animal that died during the last week of the study.
c Kaplan-Meier determinations
d Mean of all deaths (uncensored, censored, and terminal sacrifice)
e The result of the life table trend test (Tarone, 1975) is in the control column, and the results of the life table pairwise comparisons (Cox, 1972) with the controls are in the exposed columns. A negative trend or lower mortality in an exposure group is indicated by N.
TABLE 11 Mean Body Weights and Survival of Male Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Weeks on Study |
0 ppm | 1,500 ppm | 5,000 ppm | 15,000 ppm | |||||||
| Av. Wt. (g) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
|
| 1 | 21.1 | 70 | 21.1 | 100 | 70 | 21.1 | 100 | 70 | 20.6 | 98 | 70 |
| 2 | 22.9 | 70 | 22.7 | 99 | 70 | 22.8 | 100 | 70 | 22.1 | 97 | 70 |
| 3 | 23.8 | 70 | 23.9 | 100 | 70 | 23.6 | 99 | 70 | 23.1 | 97 | 70 |
| 4 | 24.7 | 70 | 25.1 | 102 | 70 | 24.8 | 100 | 70 | 24.2 | 98 | 70 |
| 5 | 25.6 | 70 | 26.1 | 102 | 70 | 25.7 | 100 | 70 | 25.0 | 98 | 70 |
| 6 | 26.2 | 70 | 26.9 | 103 | 70 | 26.4 | 101 | 70 | 25.8 | 99 | 70 |
| 7 | 27.4 | 70 | 28.1 | 103 | 70 | 27.6 | 101 | 70 | 26.9 | 98 | 70 |
| 8 | 27.7 | 70 | 28.4 | 103 | 70 | 27.6 | 100 | 70 | 26.9 | 97 | 70 |
| 9 | 28.7 | 70 | 29.2 | 102 | 70 | 28.8 | 100 | 70 | 27.8 | 97 | 70 |
| 10 | 29.1 | 70 | 29.4 | 101 | 70 | 29.1 | 100 | 70 | 28.1 | 97 | 70 |
| 11 | 29.9 | 70 | 30.3 | 101 | 70 | 30.1 | 101 | 70 | 29.0 | 97 | 70 |
| 12 | 30.5 | 70 | 31.3 | 103 | 70 | 30.9 | 101 | 70 | 29.5 | 97 | 70 |
| 13 | 31.6 | 70 | 32.5 | 103 | 70 | 31.8 | 101 | 70 | 30.6 | 97 | 70 |
| 17 | 34.1 | 70 | 35.2 | 103 | 70 | 34.6 | 102 | 70 | 32.8 | 96 | 70 |
| 21 | 36.9 | 70 | 37.9 | 103 | 70 | 37.1 | 101 | 70 | 35.4 | 96 | 69 |
| 25 | 39.0 | 70 | 40.1 | 103 | 70 | 39.3 | 101 | 70 | 37.6 | 96 | 69 |
| 29 | 41.4 | 70 | 42.1 | 102 | 70 | 41.4 | 100 | 70 | 39.3 | 95 | 69 |
| 33 | 43.3 | 70 | 43.7 | 101 | 70 | 43.1 | 100 | 70 | 41.1 | 95 | 69 |
| 37 | 43.8 | 70 | 44.5 | 102 | 70 | 43.9 | 100 | 69 | 42.1 | 96 | 69 |
| 41a | 44.5 | 60 | 45.2 | 102 | 60 | 44.8 | 101 | 59 | 43.2 | 97 | 60 |
| 45 | 45.4 | 60 | 46.5 | 102 | 60 | 46.0 | 101 | 59 | 44.4 | 98 | 60 |
| 49 | 46.8 | 60 | 48.2 | 103 | 60 | 47.4 | 101 | 59 | 45.8 | 98 | 60 |
| 54 | 46.7 | 60 | 48.0 | 103 | 60 | 47.3 | 101 | 59 | 46.0 | 99 | 60 |
| 57 | 46.0 | 60 | 47.6 | 103 | 60 | 46.5 | 101 | 59 | 45.7 | 99 | 60 |
| 61 | 46.6 | 60 | 48.0 | 103 | 60 | 47.1 | 101 | 58 | 46.4 | 100 | 60 |
| 65a | 47.0 | 50 | 48.4 | 103 | 50 | 47.3 | 100 | 49 | 46.4 | 99 | 49 |
| 69 | 46.9 | 49 | 48.2 | 103 | 50 | 47.1 | 100 | 49 | 45.7 | 97 | 49 |
| 73 | 46.8 | 49 | 48.1 | 103 | 50 | 46.5 | 99 | 49 | 45.3 | 97 | 49 |
| 78 | 46.4 | 49 | 48.3 | 104 | 50 | 47.1 | 102 | 49 | 46.0 | 99 | 49 |
| 82 | 47.7 | 49 | 49.3 | 103 | 50 | 48.2 | 101 | 49 | 46.6 | 98 | 49 |
| 86 | 47.8 | 49 | 48.9 | 102 | 49 | 48.2 | 101 | 48 | 46.8 | 98 | 49 |
| 90 | 48.4 | 49 | 48.9 | 101 | 48 | 48.7 | 101 | 48 | 47.4 | 98 | 49 |
| 94 | 47.0 | 49 | 47.6 | 101 | 47 | 47.0 | 100 | 48 | 46.2 | 98 | 46 |
| 98 | 46.6 | 49 | 48.1 | 103 | 46 | 47.0 | 101 | 48 | 45.7 | 98 | 46 |
| 101 | 45.9 | 48 | 46.9 | 102 | 44 | 44.9 | 98 | 47 | 44.0 | 96 | 46 |
| 104 | 45.2 | 47 | 46.3 | 102 | 44 | 44.9 | 99 | 47 | 44.1 | 98 | 46 |
| Mean for weeks | |||||||||||
| 1-13 | 26.9 | 27.3 | 102 | 26.9 | 100 | 26.1 | 97 | ||||
| 14-52 | 41.7 | 42.6 | 102 | 42.0 | 101 | 40.2 | 96 | ||||
| 53-104 | 46.8 | 48.0 | 103 | 47.0 | 100 | 45.9 | 98 | ||||
a Interim evaluations occurred during weeks 39 and 65.
TABLE 12 Mean Body Weights and Survival of Female Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Weeks on Study |
0 ppm | 1,500 ppm | 5,000 ppm | 15,000 ppm | |||||||
| Av. Wt. (g) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
Av. Wt. (g) |
Wt. (% of controls) |
No. of Survivors |
|
| 1 | 17.2 | 70 | 17.0 | 99 | 70 | 17.1 | 99 | 70 | 17.0 | 99 | 70 |
| 2 | 18.4 | 70 | 18.4 | 100 | 70 | 18.5 | 101 | 70 | 18.0 | 98 | 70 |
| 3 | 19.5 | 70 | 19.5 | 100 | 70 | 19.5 | 100 | 70 | 19.2 | 99 | 70 |
| 4 | 20.6 | 70 | 20.7 | 101 | 70 | 20.7 | 101 | 69 | 20.0 | 97 | 70 |
| 5 | 21.6 | 70 | 21.7 | 101 | 70 | 21.5 | 100 | 69 | 20.7 | 96 | 70 |
| 6 | 21.9 | 70 | 22.0 | 101 | 70 | 21.9 | 100 | 69 | 21.3 | 97 | 70 |
| 7 | 23.0 | 70 | 22.8 | 99 | 70 | 22.9 | 100 | 69 | 22.3 | 97 | 70 |
| 8 | 23.4 | 70 | 23.8 | 102 | 70 | 23.6 | 101 | 69 | 22.9 | 98 | 70 |
| 9 | 24.3 | 70 | 24.3 | 100 | 70 | 24.3 | 100 | 69 | 23.9 | 98 | 70 |
| 10 | 24.5 | 70 | 24.4 | 100 | 70 | 24.4 | 100 | 69 | 23.8 | 97 | 70 |
| 11 | 25.1 | 70 | 25.1 | 100 | 70 | 25.1 | 100 | 69 | 24.6 | 98 | 70 |
| 12 | 25.6 | 70 | 25.6 | 100 | 70 | 25.7 | 100 | 69 | 25.2 | 98 | 70 |
| 13 | 25.6 | 70 | 26.2 | 102 | 70 | 26.3 | 103 | 69 | 25.5 | 100 | 70 |
| 17 | 28.1 | 70 | 28.8 | 103 | 70 | 28.6 | 102 | 68 | 27.7 | 99 | 70 |
| 21 | 30.4 | 70 | 31.4 | 103 | 70 | 31.5 | 104 | 68 | 30.5 | 100 | 70 |
| 25 | 33.0 | 70 | 33.7 | 102 | 70 | 33.9 | 103 | 68 | 32.8 | 99 | 70 |
| 29 | 36.0 | 70 | 35.7 | 99 | 70 | 36.0 | 100 | 68 | 34.5 | 96 | 70 |
| 33 | 38.5 | 70 | 38.3 | 100 | 70 | 38.2 | 99 | 68 | 37.0 | 96 | 70 |
| 37 | 40.3 | 70 | 39.7 | 99 | 70 | 39.3 | 98 | 68 | 37.9 | 94 | 70 |
| 41a | 41.9 | 59 | 41.6 | 99 | 60 | 40.9 | 98 | 58 | 39.0 | 93 | 60 |
| 45 | 43.8 | 59 | 43.6 | 100 | 60 | 42.7 | 98 | 58 | 41.6 | 95 | 60 |
| 49 | 46.3 | 58 | 46.6 | 101 | 60 | 45.4 | 98 | 58 | 43.6 | 94 | 60 |
| 54 | 47.6 | 58 | 47.0 | 99 | 60 | 45.8 | 96 | 58 | 43.8 | 92 | 60 |
| 57 | 47.6 | 57 | 47.4 | 100 | 60 | 46.0 | 97 | 58 | 43.9 | 92 | 60 |
| 61 | 48.6 | 57 | 48.4 | 100 | 60 | 46.6 | 96 | 57 | 45.0 | 93 | 60 |
| 65a | 49.7 | 48 | 48.6 | 98 | 50 | 47.0 | 95 | 48 | 45.3 | 91 | 50 |
| 69 | 49.8 | 48 | 49.5 | 99 | 50 | 47.8 | 96 | 48 | 45.6 | 92 | 49 |
| 73 | 51.0 | 48 | 50.7 | 99 | 50 | 48.1 | 94 | 47 | 46.3 | 91 | 48 |
| 78 | 50.6 | 47 | 50.5 | 100 | 50 | 48.4 | 96 | 47 | 46.3 | 92 | 48 |
| 82 | 52.7 | 47 | 51.6 | 98 | 50 | 50.1 | 95 | 46 | 47.9 | 91 | 48 |
| 86 | 53.1 | 47 | 52.0 | 98 | 50 | 50.0 | 94 | 46 | 47.8 | 90 | 48 |
| 90 | 53.5 | 45 | 51.8 | 97 | 50 | 50.8 | 95 | 44 | 48.3 | 90 | 48 |
| 94 | 53.8 | 45 | 51.0 | 95 | 50 | 50.5 | 94 | 44 | 47.7 | 89 | 47 |
| 98 | 52.4 | 45 | 49.7 | 95 | 49 | 48.6 | 93 | 44 | 46.2 | 88 | 46 |
| 102 | 51.6 | 43 | 48.4 | 94 | 48 | 47.2 | 92 | 40 | 44.4 | 86 | 44 |
| 104 | 50.8 | 42 | 47.8 | 94 | 46 | 46.2 | 91 | 40 | 44.3 | 87 | 43 |
| Mean for weeks | |||||||||||
| 1-13 | 22.4 | 22.4 | 100 | 22.4 | 100 | 21.9 | 98 | ||||
| 14-52 | 37.6 | 37.7 | 100 | 37.4 | 99 | 36.1 | 96 | ||||
| 53-104 | 50.9 | 49.6 | 97 | 48.1 | 94 | 45.9 | 90 | ||||
a Interim evaluations occurred during weeks 39 and 65.
TABLE 13 Incidences of Selected Lesions of the Thyroid Gland of Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Dose (ppm) | 0 | 1,500 | 5,000 | 15,000 |
| Males | ||||
| 9-Month Interim Evaluation | ||||
| Thyroid Glanda | 10 | 0 | 0 | 9 |
| Follicular Dilatationb | 0 | _c | - | 6**(1.0)d |
| 15-Month Interim Evaluation | ||||
| Thyroid Gland | 10 | 2 | 1 | 10 |
| Follicular Dilatation | 0 | 0 | 0 | 9**(1.0) |
| Follicular Cell Adenoma | 0 | 0 | 0 | 1 |
| 2-Year Study | ||||
| Thyroid Gland | 50 | 49 | 51 | 50 |
| Follicular Dilatation | 2 (1.0) | 2 (1.5) | 5 (1.0) | 23** (1.2) |
| Follicular Cell, Hyperplasia, Focal | 5 (1.0) | 2 (1.5) | 8 (1.5) | 27** (1.9) |
| Follicular Cell Adenomae | ||||
| Overall ratef | 0/50 (0%) | 0/49 (0%) | 0/51 (0%) | 3/50 (6%) |
| Adjusted rateg | 0.0% | 0.0% | 0.0% | 6.5% |
| Terminal rateh | 0/46 (0%) | 0/44 (0%) | 0/46 (0%) | 3/46 (7%) |
| First incidence (days) | - | - | - | 729 (T) |
| Logistic regression testi | P=0.015 | - | - | P=0.121 |
| Females | ||||
| 9-Month Interim Evaluation | ||||
| Thyroid Gland | 10 | 0 | 1 | 10 |
| Follicular Dilatation | 0 | - | 1 (1.0) | 7** (1.0) |
| 15-Month Interim Evaluation | ||||
| Thyroid Gland | 9 | 0 | 2 | 9 |
| Follicular Dilatation | 0 | 0 | 5* (1.0) | |
| 2-Year Study | ||||
| Thyroid Gland | 50 | 50 | 49 | 51 |
| Follicular Dilatation | 1 (1.0) | 5 (1.0) | 11** (1.4) | 24** (1.2) |
| Follicular Cell, Hyperplasia, Diffuse | 1 (1.0) | 1 (1.0) | 0 | 0 |
| Follicular Cell, Hyperplasia, Focal | 3 (2.3) | 15** (1.5) | 27** (1.5) | 43** (2.1) |
| Follicular Cell Adenomaj | ||||
| Overall rates | 2/50 (4%) | 1/50 (2%) | 0/49 (0%) | 5/51 (10%) |
| Adjusted rates | 4.8% | 2.2% | 0.0% | 11.9% |
| Terminal rates | 2/42 (5%) | 1/46 (2%) | 0/37 (0%) | 5/42 (12%) |
| First incidence (days) | 729 (T) | 729 (T) | 729 (T) | |
| Logistic regression tests | P=0.037 | P=0.468N | P=0.267N | P=0.216 |
(T)Terminal Sacrifice
* Significantly different (P0.05) from the control group by the Fisher exact test (interim evaluations) or by the logistic regression test (2-year study)
** P≤0.01
a Number of animals with organ examined microscopically
b Number of animals with lesion
c Not applicable; tissue not examined microscopically in this group
d Average severity grade of lesions in affected animals (1=minimal; 2=mild; 3=moderate; 4=marked)
e Historical incidence for 2-year feed studies with untreated control groups (mean ± standard deviation): 19/1,105 (1.7% ± 1.7%); range 0%-4%
f Number of neoplasm-bearing animals/number of animals microscopically examined
g Kaplan-Meier estimated neoplasm incidence at the end of the study after adjustment for intercurrent mortality
h Observed incidence at terminal kill
i Beneath the control incidence are the P values associated with the trend test. Beneath the exposed group incidence are the P values corresponding to pairwise comparisons between the controls and that exposed group. The logistic regression test regards these neoplasms as nonfatal. A lower incidence in an exposure group is indicated by N.
j Historical incidence: 27/1,099 (2.5% ± 2.9%); range 0%-9%
TABLE 14 Incidences of Selected Lesions of the Forestomach of Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
| Dose (ppm) | 0 | 1,500 | 5,000 | 15,000 |
| Males | ||||
| 2-Year Study | ||||
| Forestomacha | 50 | 49 | 51 | 50 |
| Erosion, Focalb | 0 | 0 | 0 | 2 (3.0)c |
| Squamous Hyperplasia, Focal | 2 (2.0) | 1 (2.0) | 5 (1.2) | 14** (2.3) |
| Inflammation, Chronic Active | 0 | 0 | 0 | 5* (2.0) |
| Ulcer | 0 | 0 | 0 | 6* (2.5) |
| Squamous Cell Papilloma | 1 | 1 | 0 | 0 |
| Females | ||||
| 9-Month Interim Evaluation | ||||
| Forestomach | 10 | 0 | 0 | 9 |
| Squamous Hyperplasia, Focal | 0 | 0 | 0 | 1 (2.0) |
| 15-Month Interim Evaluation | ||||
| Forestomach | 9 | 0 | 1 | 9 |
| Squamous Hyperplasia, Focal | 0 | 0 | 1 (2.0) | 1 (2.0) |
| Inflammation, Chronic Active | 0 | 0 | 0 | 1 (1.0) |
| 2-Year Study | ||||
| Forestomach | 51 | 50 | 49 | 50 |
| Squamous Hyperplasia, Focal | 1 (2.0) | 3 (1.7) | 3 (2.0) | 9**(2.4) |
| Ulcer | 2 (2.0) | 0 | 0 | |
| 3 (2.7) | ||||
| Inflammation, Chronic Active | 0 | 1 (2.0) | 1 (2.0) | 3 (1.7) |
| Squamous Cell Papilloma | 1 | 0 | 0 | 0 |
* Significantly different (P0.05) from the control group by the logistic regression test
** P0.01
a Number of animals with organ examined microscopically
b Number of animals with lesion
c Average severity grade of lesions in affected animals (1=minimal; 2=mild; 3=moderate; 4=marked)
TABLE A1 Summary of the Incidence of Neoplasms in Male Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydratea
See attached backgroud material
TABLE A2 Individual Animal Tumor Pathology of Male Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE A3 Statistical Analysis of Primary Neoplasms in Male Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE A4a Historical Incidence of Pancreatic Islets Neoplasms in Untreated Male F344/N Rats
See attached backgroud material
TABLE A4b Historical Incidence of Adrenal Medulla Pheochromocytomas in Untreated Male F344/N Rats
See attached backgroud material
TABLE A5 Summary of the Incidence of Nonneoplastic Lesions in Male Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE B1 Summary of the Incidence of Neoplasms in Female Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE B2 Individual Animal Tumor Pathology of Female Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE B3 Statistical Analysis of Primary Neoplasms in Female Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE B4 Summary of the Incidence of Nonneoplastic Lesions in Female Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE C1 Summary of the Incidence of Neoplasms in Male Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE C2 Individual Animal Tumor Pathology of Male Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE C3 Statistical Analysis of Primary Neoplasms in Male Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE C4 Historical Incidence of Thyroid Gland Follicular Cell Neoplasms in Untreated Male B6C3F1 Mice
See attached backgroud material
TABLE C5 Summary of the Incidence of Nonneoplastic Lesions in Male Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE D1 Summary of the Incidence of Neoplasms in Female Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE D2 Individual Animal Tumor Pathology of Female Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE D3 Statistical Analysis of Primary Neoplasms in Female Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE D4 Historical Incidence of Thyroid Gland Follicular Cell Neoplasms in Untreated Female B6C3F1 Mice
See attached backgroud material
TABLE D5 Summary of the Incidence of Nonneoplastic Lesions in Female Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE F3 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats at the 9-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE F4 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Rats at the 15-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE F7 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice at the 9-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE F8 Organ Weights and Organ-Weight-to-Body-Weight Ratios for Mice at the 15-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE G3 Hematology and Clinical Chemistry Data for Rats at the 9-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE G4 Hematology and Clinical Chemistry Data for Rats at the 15-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE G7 Hematology and Clinical Chemistry Data for Mice at the 9-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE G8 Hematology and Clinical Chemistry Data for Mice at the 15-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE H1 Tissue Metal Concentration Analyses for Rats at the 9-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE H2 Tissue Metal Concentration Analyses for Rats at the 15-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE H3 Tissue Metal Concentration Analyses for Mice at the 9-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE H4 Tissue Metal Concentration Analyses for Mice at the 15-Month Interim Evaluation in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE J1 Feed and Compound Consumption by Male Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE J2 Feed and Compound Consumption by Female Rats in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE J3 Feed and Compound Consumption by Male Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
TABLE J4 Feed and Compound Consumption by Female Mice in the 2-Year Feed Study of Manganese (II) Sulfate Monohydrate
See attached backgroud material
Table 1: Percentage of incidence of male urinary tract lesions – Study A
Dose (mg/kg)
|
Tubular protein nephropathy |
Tubulointerstitial nephritis |
Glomerulosclerosis/ glomerulonephritis |
Urolithiasis |
0 |
100 |
66 |
0 |
0 |
306 |
66 |
66 |
66 |
33 |
612 |
100 |
100 |
33 |
66 |
1225 |
50 |
100 |
100 |
50 |
1838 |
100 |
100 |
100 |
33 |
Table 2: Percentage of incidence of male urinary tract lesions – Study B
Dose (mg/kg)
|
Tubular protein nephropathy |
Tubulointerstitial nephritis |
Glomerulosclerosis/ glomerulonephritis |
Urolithiasis |
0 |
33 |
33 |
0 |
0 |
306 |
33 |
33 |
33 |
0 |
612 |
66 |
66 |
33 |
33 |
1225 |
100 |
100 |
50 |
50 |
1838 |
100 |
100 |
100 |
33 |
Table 3: Percentage of incidence of female urinary tract lesions – Study A
Dose (mg/kg)
|
Tubular protein nephropathy |
Tubulointerstitial nephritis |
Glomerulosclerosis/ glomerulonephritis |
Urolithiasis |
Papillitis |
0 |
100 |
66 |
66 |
0 |
0 |
306 |
100 |
33 |
33 |
0 |
0 |
612 |
100 |
33 |
33 |
0 |
0 |
1225 |
0 |
33 |
33 |
0 |
0 |
1838 |
0 |
0 |
33 |
0 |
33 |
Table 4: Percentage of incidence of female urinary tract lesions – Study B
Dose (mg/kg)
|
Tubular protein nephropathy |
Tubulointerstitial nephritis |
Glomerulosclerosis/ glomerulonephritis |
Urolithiasis |
Papillitis |
0 |
66 |
0 |
0 |
0 |
0 |
306 |
33 |
33 |
0 |
0 |
0 |
612 |
66 |
33 |
66 |
0 |
0 |
1225 |
33 |
33 |
0 |
0 |
0 |
1838 |
66 |
33 |
0 |
0 |
0 |
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- NOAEL
- 10 mg/kg bw/day
- Study duration:
- subchronic
- Species:
- rat
Repeated dose toxicity: inhalation - systemic effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: inhalation - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - systemic effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Repeated dose toxicity: dermal - local effects
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
EDDHA
The subchronic toxicity of Fe(Na)EDDHA (CAS 84539 -55 -9) by oral route was investigated in rats (Novartis Crop Protection AG, 1998). The test item was administered to 10 Sprague-Dawley derived rats/sex/dose level by oral gavage at 5, 50 or 200 mg/kg bw/day for 90 days. A concurrent control group was treated with the vehicle only. Additional 10 animals/sex of the high dose and control group were kept on control diet for a 4 -week recovery period before sacrifice. Treatment with the test item resulted in lower food intake and impaired body weight development at 200 mg/kg bw/day. Reversible effects on red blood cell (normochromic anaemia) and white blood cell parameters, and higher values of platelets and prothrombin activity were noted at 50 and/or 200 mg/kg bw/day. In addition, changes of blood chemistry and urine parameters concerning the liver and kidneys were noted. The body weight relative heart weight was increased in males at 200 mg/kg bw/day. Under the conditions of this study, the NOAEL for Fe(Na)EDDHA when administered daily by oral gavage for three months was 10 mg/kg bw/day (estimated from the LOAEL).
Estimation of NOAEL:
A more realistic NOAEL was estimated from the LOAEL of 50 mg/kg bw/day applying an assessment factor of 5. This method is applicable and scientifically justified for this test, as only slight adverse effects were observed at 50 mg/kg bw/day (haematology parameters - anaemia). To take the relatively large concentration gap between 5 (clear NOEL) and 50 mg/kg bw/day into account, 5 mg/kg bw/day was not taken as NOAEL, but extrapolated from the LOAEL. The guidance on information requirements and CSA, R.8 (ECHA, 2008 -2010) recommends a factor between 3 and 10 for extrapolation from LOAEL to NOAEL. An assessment factor of 5 seems to be appropriate and conservative enough for the current study as only slight effects were observed at the LOAEL of 50 mg/kg bw/day. Consequently a NOAEL of 10 mg/kg bw/day is calculated for this study.
In a dose range-finding subacute oral toxicity study (CIBA-GEIGY Limited, 1996a), Fe(Na)EDDHA was administered to 5 Sprague-Dawley derived rats/sex/dose level by oral gavage at 50, 200 or 1000 mg/kg bw/day for 28 days. A concurrent control group was treated with the vehicle only. Treatment with the test item resulted in impaired body weight development at 200 and 1000 mg/kg bw/day and correspondend lower food intake. Anaemia without erythropoietic response was noted at 200 and 1000 mg/kg bw/day. At the same dose levels, the kidney was revealed as target organ by microscopical examination, by blood chemistry data evaluation and by organ weight evaluation. In addition, body weight relative organ weight changes were noted in the heart, adrenals and spleen. However, the relevance of these findings was considered as equivocal. This study was used as scientific basis for dose level selection for the above-mentioned 90 -day repeated dose oral toxicity study in the rat.
Manganese salts
The effect concentrations of manganese compounds were converted to the manganese moiety of the target substance under consideration of the molecular weight and the purity.
In a 2 year repeated-dose study with MnSO4 on rats and mice, no carcinogenic effects were detected (NTP, 1993, MnSO4, 2a, rats, mice). The lowest NOAEL was determined to be approx. 228 mg MnSO4/kg bw/day (= 987.6 mg/kg bw/day for the manganese moiety of the target substance) based on the effects of MnSO4 on the thyroid gland (Follicular dilatation) in female mice. These effects were also detected at a very high dose of MnSO4 over the whole lifetime of the animals, which does neither lead to the conclusion of a specific organ toxicity of the test item nor to a high toxicity in general.
The NOAELs determined in two other studies on MnSO4 at shorter test durations (NTP 1993, 14 days and 13 weeks), as well as on Manganese (II) acetate (Ponnapakkam 2003, 63 d) are mainly of the magnitude of > 1 g/kg bw/day. The observed lesions in the male urinary tract in the latter study can be neglected because these are most likely caused by a sex- and species-specific α2u-globulin related nephropathy which is of no relevance for humans. Furthermore, the effects on body weight gain and leucocyte count in female rats after 13 weeks can be considered as transient or irrelevant because a reduced weight gain or any signs of infection were not detected in the 2 year study as well as in male rats. As MnSO4 serves as a source substance for the manganese moiety of the target substance, these conclusions can also be drawn for the latter.
Conclusion
The two year repeated dose toxicity study (NTP 1993, 2a) was chosen to derive the hazard value, because this study covers the lifetimes of the test species and was performed on two different species, rats and mice, and therefore all possible substance-related effects including cancer due to long-term exposure to the target substance can be detected.
The NOAEL derived for MnSO4 in this study was converted to the registered substance under consideration of the molecular weight and the purity of the target substance. The NOAEL was determined to be 987.6 mg/kg bw/day for the manganese moiety of the target substance, based on the effects on the thyroid gland (Follicular dilatation) in female mice.
Justification for classification or non-classification
Based on the available data the registered substance is not subject to classification and labelling according to the Regulation (EC) No 1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
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