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Administrative data

Key value for chemical safety assessment

Toxic effect type:
concentration-driven

Effects on fertility

Description of key information

The endpoint reproductive toxicity of the vanadium category substance is not addressed by substance-specific information but rather by read-across of data available for soluble tri-, tetra- and pentavalent vanadium substances. Therefore, the remaining text in this chapter is generic for all vanadium substances and has not been adapted on a substance-specific basis.

All vanadium substances within the read-across concept represent inorganic substances, including salts or oxides, and form the vanadium category. For the substances of the inorganic vanadium substances category, it is assumed that the in vivo bioavailability of the vanadium varies in a predictable manner, which is dependent on the in vitro bioaccessibility of the respective vanadium substance, i.e. all members of the category liberate vanadium ions in aqueous media at different rates, inter alia depending on the chemical structure. Thus, this concept is based on the chemistry / composition of all substances and on experimental studies for (i) water solubility and (ii) in-vitro bioaccessibility: assessment of the solubility and speciation of vanadium substances in five different artificial physiological fluids. It is hypothesised that the systemic toxicity is dependent on the ratio at which vanadium ions are liberated during e.g. gastric passage – the toxicity correlated with the in vitro bioaccessibility. Thus, the vanadium substances, showing similar in vitro bioaccessibility are placed in the same read-across group. Further details on the read-across concept are presented in the report attached to IUCLID section 13.

Link to relevant study records

Referenceopen allclose all

Endpoint:
fertility, other
Remarks:
90- day toxicity study with evaluation of reproductive organs
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-09-20 (first exposure) to 1990-12-21/22 (necropsy date)
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Well reported study. Test procedure in accordance with generally accepted scientific standards and described in sufficient detail.
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
no guideline followed
Principles of method if other than guideline:
Groups of 10 male and 10 female mice were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 1, 2, 4, 8, or 16 mg/m3 (equivalent to 0, 0.56, 1.12, 2.25, 4.50, and 8.99 mg V/m³) by inhalation, 6 hours per day, 5 days per week for 3 months. Effects on fertility and sexual function of male and female mice were evaluated after 90 days of inhalation exposure.
GLP compliance:
yes
Limit test:
no
Justification for study design:
not applicable
Specific details on test material used for the study:
not specified
Species:
mouse
Strain:
B6C3F1
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Taconic Farms, Inc. (Germantown, NY)
- Age at study initiation: average age: 6 or 7 weeks old on the first day of the study (4 weeks old on receipt)
- Weight at study initiation: range of mean body weights in the exposure groups: 25-26 g (males) and 20-21 g (females)
- Housing: housed individually; stainless steel wire mesh (Lab Products, Inc., Maywood, NJ), changed weekly
- Diet: ad libitum, except during exposure periods; NIH-07 open formula pelleted diet (Zeigler Brothers, Inc., Gardners, PA)
- Water: ad libitum; tap water (City of Chicago municipal supply) via automatic watering system
- Acclimation period: quarantined for 10 or 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 23.9± ca.2°C (75° ± 3° F)
- Humidity (%): 55% ± 15%
- Air changes (per hr): 15/hour
- Photoperiod (hrs dark / hrs light): 12 hours/day
Route of administration:
inhalation: aerosol
Type of inhalation exposure (if applicable):
whole body
Mass median aerodynamic diameter (MMAD):
>= 1 - <= 1.3 µm
Remarks on MMAD:
GSD: 2.1 - 3.0
Vehicle:
air
Details on exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- For the 3-month studies, vanadium pentoxide aerosol generation was based on the principle of pneumatic dispersion and consisted of two major components: a screw feeder (Model 310, Accurate, White Water, WI) that metered vanadium pentoxide powder at a constant rate and a Jet-O-Mizer jetmill (Fluid Energy Corp., Harfield, PA) that used compressed air to disperse the metered powder and form the aerosol.
- Aerosol leaving the jetmill passed through a one-stage impactor and a vertical elutriator to eliminate or deagglomerate the large particles before entering a plenum and manifold distribution system. The aerosol delivery system consisted of three holding chambers that diluted the aerosol in three stages. A metered amount of diluted aerosol was removed and mixed with conditioned air at the inlet to each exposure chamber to achieve the appropriate exposure concentration. The electrical charge buildup on the aerosol particles was neutralized by mixing the aerosol with high concentrations of bipolar ions, which were generated using a Pulse Gun (Static Control Services, Palm Springs, CA) air nozzle. For the 3-month study, a transvector air pump was installed at the aerosol inlet to each exposure chamber to provide additional control of the aerosol flow rate and improve stability of the chamber concentration.
- The stainless-steel inhalation exposure chambers (Lab Products, Inc., Maywood NJ), were designed so that uniform aerosol concentrations could be maintained throughout the chambers when catch pans were in place. The total active mixing volume of each chamber was 1.7 m³.

CHAMBER ATMOSPHERE CHARACTERIZATION
- The particle size distribution in each chamber was determined prior to the start, during the first week of the 3-month study, and monthly during the 3-month study.
- For the 3-month study, a 10-stage Quartz Crystal Microbalance-based cascade impactor was used to separate the aerosol particles into sequential size ranges; the mass median aerodynamic diameter was calculated from the corresponding mass fraction of particles at each stage.

OTHER
- The uniformity of aerosol concentration in the inhalation exposure chambers without animals was evaluated before each of the studies began; concentration uniformity with animals present in the chambers was also measured. During the 3-month study, minor excursions in chamber uniformity values were observed in one or more exposure chambers, but these excursions had no impact on the studies.
- The stability of vanadium pentoxide in the exposure system was tested with XRD analysis. XRD analyses indicated no detectable build-up of degradation products at a detection limit of approximately 1%.
Details on mating procedure:
not specified
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
- Chamber aerosol concentrations were monitored with real-time aerosol monitors (RAMs) that used a pulsed-light-emitting diode in combination with a silicon detector to sense light scattered over a forward angular range of 45° to 95° by particles traversing the sensing volume. The instruments respond to particles 0.1 to 20 μm in diameter.
- During the 3-month study, an individual monitor was used for each exposure chamber. The voltage output of the online monitors was read and recorded, and the calibration curve was applied to the voltages measured by the RAM to convert the measured voltages to exposure chamber concentrations.
- Each RAM was calibrated daily during the 3-month study by correlating the measured voltage with vanadium pentoxide concentrations determined by gravimetric analysis of glass fiber filters.
Duration of treatment / exposure:
3 months (91-92 days)
Frequency of treatment:
6 hours per day, 5 days per week
Details on study schedule:
not specified
Dose / conc.:
1 mg/m³ air (analytical)
Remarks:
SD. 0.0; target concentration: 1 mg/m³; equivalent to 0.56 mg V/m³
Dose / conc.:
2 mg/m³ air (analytical)
Remarks:
SD. 0.0; target concentration: 2 mg/m³; equivalent to 1.12 mg V/m³
Dose / conc.:
4 mg/m³ air (analytical)
Remarks:
SD. 0.1; target concentration: 4 mg/m³; equivalent to 2.25 mg V/m³
Dose / conc.:
7.9 mg/m³ air (analytical)
Remarks:
SD. 0.2; target concentration: 8 mg/m³; equivalent to 4.50 mg V/m³
Dose / conc.:
15.9 mg/m³ air (analytical)
Remarks:
SD. 0.4; target concentration: 16 mg/m³; equivalent to 8.99 mg V/m³
No. of animals per sex per dose:
10 male and 10 female animals
Control animals:
yes
Details on study design:
- Dose selection rationale: based on decreased survival in the 32 mg/m3 males and body weight decreases in 32 mg/m3 males and females in a 16-day study, an exposure concentration of 32 mg/m3 was considered too high for use in a 3-month study. Therefore, the exposure concentrations selected for the 3-month inhalation study in rats were 0, 1, 2, 4, 8, and 16 mg/m3.
- Rationale for animal assignment (if not random): randomly into groups of approximately equal initial mean body weights.
Positive control:
not specified
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
Clinical findings were recorded weekly.

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: initially, weekly, and at the end of the studies.

FOOD CONSUMPTION AND COMPOUND INTAKE: No data
WATER CONSUMPTION AND COMPOUND INTAKE: No data
OPHTHALMOSCOPIC EXAMINATION: No data
HAEMATOLOGY: No data
CLINICAL CHEMISTRY: No data
URINALYSIS: No data
NEUROBEHAVIOURAL EXAMINATION: No data
IMMUNOLOGY: No data



Oestrous cyclicity (parental animals):
- At the end of the 3-month study, samples were collected for vaginal cytology evaluations from female mice exposed to 4, 8, or 16 mg/m3.
- For 12 consecutive days prior to scheduled terminal sacrifice, the vaginal vaults of the females were moistened with saline, if necessary, and samples of vaginal fluid and cells were stained. Relative numbers of leukocytes, nucleated epithelial cells, and large squamous epithelial cells were determined and used to ascertain oestrous cycle stage (i.e., diestrus, proestrus, estrus, and metestrus).
Sperm parameters (parental animals):
- At the end of the 3-month study, samples were collected for sperm motility evaluations from male mice exposed to 4, 8, or 16 mg/m3.
- Male animals were evaluated for sperm count and motility. The left testis and left epididymis were isolated and weighed. The tail of the epididymis (cauda epididymis) was then removed from the epididymal body (corpus epididymis) and weighed. Modified Tyrode’s buffer was applied to slides and a small incision was made at the distal border of the cauda epididymis. The sperm effluxing from the incision were dispersed in the buffer on the slides, and the numbers of motile and nonmotile spermatozoa were counted for five fields per slide by two observers.
- Following completion of sperm motility estimates, each left cauda epididymis was placed in buffered saline solution. Caudae were finely minced, and the tissue was incubated in the saline solution and then heat fixed at 65° C. Sperm density was then determined microscopically with the aid of a hemacytometer.
- To quantify spermatogenesis, the testicular spermatid head count was determined by removing the tunica albuginea and homogenising the left testis in phosphate-buffered saline containing 10% dimethyl sulfoxide. Homogenisation-resistant spermatid nuclei were counted with a hemacytometer.
Litter observations:
not specified
Postmortem examinations (parental animals):
GROSS PATHOLOGY: Yes
- Necropsy was performed at study end on all animals.
- Organs weighed were the heart, right kidney, liver, lung, right testis, and thymus.

HISTOPATHOLOGY: Yes
- Complete histopathology was performed on control, and 16 mg/m3 mice. In addition to gross lesions and tissue masses, the following tissues were examined: adrenal gland, bone with marrow, brain, clitoral gland, esophagus, gallbladder (mice only), heart and aorta, large intestine (cecum, colon, and rectum), small intestine (duodenum, jejunum, and ileum), kidney, larynx, liver, lung and mainstem bronchi, lymph nodes (mandibular, mediastinal, mesenteric, and bronchial), mammary gland, nose, ovary, pancreas, parathyroid gland, pituitary gland, preputial gland, prostate gland, salivary gland, skin, spleen, stomach (forestomach and glandular), testis (with epididymis and seminal vesicle), thymus, thyroid gland, trachea, urinary bladder, and uterus.
- The lungs of mice in all remaining exposure groups and the thymus in 8 mg/m3 mice were also examined.
Postmortem examinations (offspring):
not specified
Statistics:
Analysis of Continuous Variables:
Two approaches were employed to assess the significance of pairwise comparisons between exposed and control groups in the analysis of continuous variables. Organ and body weight data, which historically have approximately normal distributions, were analyzed with the parametric multiple comparison procedures of Dunnett (1955) and Williams (1971, 1972). Hematology, clinical chemistry, urinalysis, urine concentrating ability, cardiopulmonary, immunotoxicologic, cell proliferation, tissue concentrations, spermatid, and epididymal spermatozoal data, which have typically skewed distributions, were analyzed using the nonparametric multiple comparison methods of Shirley (1977) and Dunn (1964). Jonckheere’s test (Jonckheere, 1954) was used to assess the significance of the dose-related trends and to determine whether a trend-sensitive test (Williams’ or Shirley’s test) was more appropriate for pairwise comparisons than a test that does not assume a monotonic dose-related trend (Dunnett’s or Dunn’s test). Average severity values were analyzed for significance with the Mann-Whitney U test (Hollander and Wolfe, 1973). Treatment effects were investigated by applying a multivariate analysis of variance (Morrison, 1976) to the transformed data to test for simultaneous equality of measurements across exposure concentrations. (for more information see publication)
Reproductive indices:
not specified
Offspring viability indices:
not specified
Clinical signs:
no effects observed
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, non-treatment-related
Description (incidence):
- One male exposed to 16 mg/m3 died before the end of the study.

Please also refer to the field "Attached background material" below.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- final mean body weights and body weight gains of 8 and 16 mg/m3 males and 4 mg/m3 or greater females were significantly less than those of the chamber controls.
- in male mice, necropsy body weights were statistically significantly lower at 16 mg/m3.
- in female mice, necropsy body weight was statistically significantly lower in all treatment groups.

Please also refer to the field "Attached background material" below (file: "Reproductive funtion sperm measures and oestrus cycle.pdf")
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
The incidences of inflammation of the lung were increased in mice exposed to 2 mg/m3 or greater. Mice exposed to 2 mg/m3 or greater had epithelial hyperplasia of the lung. The severities of these lesions generally increased with increasing exposure concentration. Inflammation was characterized by multiple foci of a mixed cellular infiltrate oriented around blood vessels and bronchioles. The infiltrate was composed primarily of macrophages with abundant cytoplasm and fewer lymphocytes and neutrophils. The infiltrate extended into the surrounding perivascular interstitium and often filled adjacent alveoli. Hyperplasia involved alveolar and, to a lesser extent, bronchiolar epithelium. This change involved the distal airways and associated alveolar ducts and alveoli. Normally flattened epithelium was replaced with larger cuboidal cells.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
effects observed, treatment-related
Description (incidence and severity):
The epididymal spermatozoal motility of males exposed to 8 or 16 mg/m3 was statistically significant decreased. However, the effect on sperm motility was not very pronounced without a clear dose-response relationship. The lowest value was observed in the 8 mg/m3 group (mid dose) and was less pronounced in the high dose group.

Please also refer to the field "Attached background material" below.
Reproductive performance:
not specified
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
- The mouse that died early appeared thin.
- There were no other clinical findings related to vanadium pentoxide exposure.

ORGAN WEIGHTS (PARENTAL ANIMALS)
- Other organ weight changes were considered related to body weight decreases.

HISTOPATHOLOGY - NON-NEOPLASTIC
Lymphoid depletion of the thymus was observed in some males and females in the 16 mg/m3 groups (males: chamber control, 0/9; 8 mg/m3, 0/8; 16 mg/m3, 2/7; females: 0/9, 0/9, 1/10). This lesion may have been associated with the body weight decreases and debilitation
of the mice.

REPRODUCTIVE FUNCTION: OESTROUS CYCLE (PARENTAL ANIMALS)
No significant differences were noted in oestrous cycle parameters between exposed and chamber control females.


Atrophy of the secondary reproductive organs observed at 16 mg/m3 is also described (no supporting data provided). These findings are most probably secondary to the observed general toxicological and body weights effect in the high dose group and can thus not be attributed to a direct vanadium induced toxicological finding.

Please also refer to the field "Attached background material" below.

Dose descriptor:
NOAEC
Remarks:
reproductive toxicity
Effect level:
> 16 mg/m³ air
Based on:
test mat.
Sex:
female
Basis for effect level:
other: no effects on estrus cycle parameters determined
Dose descriptor:
NOAEC
Remarks:
reproductive toxicity
Effect level:
4 mg/m³ air
Based on:
test mat.
Sex:
male
Basis for effect level:
other: decreased epididymal spermatozoal motility at an above 8 mg/m3
Critical effects observed:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Anogenital distance (AGD):
not specified
Nipple retention in male pups:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
not specified
Remarks on result:
not measured/tested
Critical effects observed:
not specified
Critical effects observed:
not specified
Reproductive effects observed:
not specified
Conclusions:
Groups of 10 male and 10 female B6C3F1 mice were exposed to V2O5 aerosols (whole body exposure) at concentrations of 0, 1, 2, 4, 8 or 16 mg/m3 (equivalent to 0, 0.56, 1.12, 2.25, 4.50, and 8.99 mg V/m³) by inhalation, 6 hours/d, 5 d/wk, for 3 months (91-92 days). Effects on fertility and sexual function of male and female mice were evaluated after inhalation exposure.
The results showed that epididymal spermatozoal motility was significantly decreased in males exposed to 8 and 16 mg/m3. Female oestrus cycle parameters were not affected.
Endpoint:
fertility, other
Remarks:
90- day toxicity study with evaluation of reproductive organs
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-09-18/19 (first exposure) to 1990-12-18/19 (necropsy date)
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Well reported study. Test procedure in accordance with generally accepted scientific standards and described in sufficient detail.
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
no guideline followed
Principles of method if other than guideline:
Groups of 10 male and 10 female rats were exposed to particulate aerosols of vanadium pentoxide at concentrations of 0, 1, 2, 4, 8, or 16 mg/m3 (equivalent to 0, 0.56, 1.12, 2.25, 4.50, and 8.99 mg V/m³) by inhalation, 6 hours per day, 5 days per week for 3 months. Effects on fertility and sexual function of male and female rats were evaluated after 90 days of inhalation exposure.
GLP compliance:
yes
Limit test:
no
Justification for study design:
not applicable
Specific details on test material used for the study:
not specified
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Taconic Farms, Inc. (Germantown, NY)
- Age at study initiation: average age: 6 or 7 weeks old on the first day of the study (4 weeks old on receipt)
- Weight at study initiation: range of mean body weights in the exposure groups: 133-137 g (males) and 108-111 g (females)
- Housing: housed individually; stainless steel wire mesh (Lab Products, Inc., Maywood, NJ),), changed weekly
- Diet: ad libitum, except during exposure periods; NIH-07 open formula pelleted diet (Zeigler Brothers, Inc., Gardners, PA); feed was also
withheld during urine collection
- Water: ad libitum; tap water (City of Chicago municipal supply) via automatic watering system; water was withheld for the second urine collection
- Acclimation period: quarantined for 12 or 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 23.9± ca.2°C (75° ± 3° F)
- Humidity (%): 55% ± 15%
- Air changes (per hr): 15/hour
- Photoperiod (hrs dark / hrs light): 12 hours/day
Route of administration:
inhalation: aerosol
Type of inhalation exposure (if applicable):
whole body
Mass median aerodynamic diameter (MMAD):
>= 1 - <= 1.3 µm
Remarks on MMAD:
GSD: 2.1 - 3.0
Vehicle:
air
Details on exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- For the 3-month studies, vanadium pentoxide aerosol generation was based on the principle of pneumatic dispersion and consisted of two major components: a screw feeder (Model 310, Accurate, White Water, WI) that metered vanadium pentoxide powder at a constant rate and a Jet-O-Mizer jetmill (Fluid Energy Corp., Harfield, PA) that used compressed air to disperse the metered powder and form the aerosol.
- Aerosol leaving the jetmill passed through a one-stage impactor and a vertical elutriator to eliminate or deagglomerate the large particles before entering a plenum and manifold distribution system. The aerosol delivery system consisted of three holding chambers that diluted the aerosol in three stages. A metered amount of diluted aerosol was removed and mixed with conditioned air at the inlet to each exposure chamber to achieve the appropriate exposure concentration. The electrical charge buildup on the aerosol particles was neutralized by mixing the aerosol with high concentrations of bipolar ions, which were generated using a Pulse Gun (Static Control Services, Palm Springs, CA) air nozzle. For the 3-month study, a transvector air pump was installed at the aerosol inlet to each exposure chamber to provide additional control of the aerosol flow rate and improve stability of the chamber concentration.
- The stainless-steel inhalation exposure chambers (Lab Products, Inc., Maywood NJ), were designed so that uniform aerosol concentrations could be maintained throughout the chambers when catch pans were in place. The total active mixing volume of each chamber was 1.7 m³.

CHAMBER ATMOSPHERE CHARACTERIZATION
- The particle size distribution in each chamber was determined prior to the start, during the first week of the 3-month study, and monthly during the 3-month study.
- For the 3-month study, a 10-stage Quartz Crystal Microbalance-based cascade impactor was used to separate the aerosol particles into sequential size ranges; the mass median aerodynamic diameter was calculated from the corresponding mass fraction of particles at each stage.

OTHER
- The uniformity of aerosol concentration in the inhalation exposure chambers without animals was evaluated before each of the studies began; concentration uniformity with animals present in the chambers was also measured. During the 3-month study, minor excursions in chamber uniformity values were observed in one or more exposure chambers, but these excursions had no impact on the studies.
- The stability of vanadium pentoxide in the exposure system was tested with XRD analysis. XRD analyses indicated no detectable build-up of degradation products at a detection limit of approximately 1%.
Details on mating procedure:
not specified
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
- Chamber aerosol concentrations were monitored with real-time aerosol monitors (RAMs) that used a pulsed-light-emitting diode in combination with a silicon detector to sense light scattered over a forward angular range of 45° to 95° by particles traversing the sensing volume. The instruments respond to particles 0.1 to 20 μm in diameter.
- During the 3-month study, an individual monitor was used for each exposure chamber. The voltage output of the online monitors was read and recorded, and the calibration curve was applied to the voltages measured by the RAM to convert the measured voltages to exposure chamber concentrations.
- Each RAM was calibrated daily during the 3-month study by correlating the measured voltage with vanadium pentoxide concentrations determined by gravimetric analysis of glass fiber filters.
Duration of treatment / exposure:
3 months (91-92 days)
Frequency of treatment:
6 hours per day, 5 days per week
Details on study schedule:
not specified
Dose / conc.:
1 mg/m³ air (analytical)
Remarks:
SD. 0.0; target concentration: 1 mg/m³; equivalent to 0.56 mg V/m³
Dose / conc.:
2 mg/m³ air (analytical)
Remarks:
SD. 0.0; target concentration: 2 mg/m³; equivalent to 1.12 mg V/m³
Dose / conc.:
4 mg/m³ air (analytical)
Remarks:
SD. 0.1; target concentration: 4 mg/m³; equivalent to 2.25 mg V/m³
Dose / conc.:
7.9 mg/m³ air (analytical)
Remarks:
SD. 0.2; target concentration: 8 mg/m³; equivalent to 4.50 mg V/m³
Dose / conc.:
15.9 mg/m³ air (analytical)
Remarks:
SD. 0.4; target concentration: 16 mg/m³; equivalent to 8.99 mg V/m³
No. of animals per sex per dose:
10 male and 10 female rats
Control animals:
yes
Details on study design:
- Dose selection rationale: based on decreased survival in the 32 mg/m3 males and body weight decreases in 32 mg/m3 males and females in a 16-day study, an exposure concentration of 32 mg/m3 was considered too high for use in a 3-month study. Therefore, the exposure concentrations selected for the 3-month inhalation study in rats were 0, 1, 2, 4, 8, and 16 mg/m3.
- Rationale for animal assignment (if not random): randomly into groups of approximately equal initial mean body weights.
Positive control:
not specified
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
Clinical findings were recorded weekly.

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: initially, weekly, and at the end of the study

FOOD CONSUMPTION AND COMPOUND INTAKE: No data
WATER CONSUMPTION AND COMPOUND INTAKE: No data
OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of the study
- Anaesthetic used for blood collection: rats were anesthetized with a 70% CO2:30% air mixture
- Animals fasted: no data
- How many animals: mainly 8-10 per exposure group (except week 13, highest dose group: N=3 for males and 6 for females)
- Parameters checked: automated hematocrit; manual hematocrit; hemoglobin concentration; erythrocyte, reticulocyte, nucleated erythrocyte, and platelet counts; erythrocyte morphology; mean cell volume; mean cell hemoglobin; mean cell hemoglobin concentration; leukocyte count and differentials

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the study
- Animals fasted: no data
- How many animals: mainly 8-10 per exposure group (except week 13, highest dose group: N=3 for males and 6 for females)
- Parameters checked: urea nitrogen, creatinine, total protein, albumin, alanine aminotransferase, alkaline phosphatase, creatine kinase, sorbitol dehydrogenase, bile acids

URINALYSIS: Yes
- Time schedule for collection of urine: during week 12 (females) or 13 (males)
- Metabolism cages used for collection of urine: for 16 hours immediately following exposure
- Animals fasted: yes
- Parameters checked: urinalysis: volume, specific gravity; urine concentrating ability: volume, specific gravity

IMMUNOLOGY: No data

NEUROBEHAVIOURAL EXAMINATION: No data
Oestrous cyclicity (parental animals):
- At the end of the 3-month studies, samples were collected for vaginal cytology evaluations from rats exposed to 4, 8, or 16 mg/m3.
- For 12 consecutive days prior to scheduled terminal sacrifice, the vaginal vaults of the females were moistened with saline, if necessary, and samples of vaginal fluid and cells were stained. Relative numbers of leukocytes, nucleated epithelial cells, and large squamous epithelial cells were determined and used to ascertain estrous cycle stage (i.e., diestrus, proestrus, estrus, and metestrus).
Sperm parameters (parental animals):
- At the end of the 3-month studies, samples were collected for sperm motility evaluations from core study rats exposed to 0, 2, 4 and 8 mg/m3.
- Male animals were evaluated for sperm count and motility. The left testis and left epididymis were isolated and weighed. The tail of the epididymis (cauda epididymis) was then removed from the epididymal body (corpus epididymis) and weighed. Test yolk was applied to slides and a small incision was made at the distal border of the cauda epididymis. The sperm effluxing from the incision were dispersed in the buffer on the slides, and the numbers of motile and nonmotile spermatozoa were counted for five fields per slide by two observers.
- Following completion of sperm motility estimates, each left cauda epididymis was placed in buffered saline solution. Caudae were finely minced, and the tissue was incubated in the saline solution and then heat fixed at 65° C. Sperm density was then determined microscopically with the aid of a hemacytometer.
- To quantify spermatogenesis, the testicular spermatid head count was determined by removing the tunica albuginea and homogenising the left testis in phosphate-buffered saline containing 10% dimethyl sulfoxide. Homogenisation-resistant spermatid nuclei were counted with a hemacytometer.
Litter observations:
not specified
Postmortem examinations (parental animals):
GROSS NECROPSY/ ORGAN WEIGHTS
- Necropsies were performed on all animals.
- Organs weighed were the heart, right kidney, liver, lung, right testis, and thymus..

HISTOPATHOLOGY: Yes.
- Complete histopathology was performed on control, 8 and 16 mg/m3 rats. In addition to gross lesions and tissue masses, the following tissues were examined: adrenal gland, bone with marrow, brain, clitoral gland, esophagus, gallbladder (mice only), heart and aorta, large intestine (cecum, colon, and rectum), small intestine (duodenum, jejunum, and ileum), kidney, larynx, liver, lung and mainstem bronchi, lymph nodes (mandibular, mediastinal, mesenteric, and bronchial), mammary gland, nose, ovary, pancreas, parathyroid gland, pituitary gland, preputial gland, prostate gland, salivary gland, skin, spleen, stomach (forestomach and glandular), testis (with epididymis and seminal vesicle), thymus, thyroid gland, trachea, urinary bladder and uterus.
- The lung and nose of rats in all remaining exposure groups were also examined.
Postmortem examinations (offspring):
not specified
Statistics:
Analysis of Continuous Variables:
Two approaches were employed to assess the significance of pairwise comparisons between exposed and control groups in the analysis of continuous variables. Organ and body weight data, which historically have approximately normal distributions, were analyzed with the parametric multiple comparison procedures of Dunnett (1955) and Williams (1971, 1972). Hematology, clinical chemistry, urinalysis, urine concentrating ability, cardiopulmonary, immunotoxicologic, cell proliferation, tissue concentrations, spermatid, and epididymal spermatozoal data, which have typically skewed distributions, were analyzed using the nonparametric multiple comparison methods of Shirley (1977) and Dunn (1964). Jonckheere’s test (Jonckheere, 1954) was used to assess the significance of the dose-related trends and to determine whether a trend-sensitive test (Williams’ or Shirley’s test) was more appropriate for pairwise comparisons than a test that does not assume a monotonic dose-related trend (Dunnett’s or Dunn’s test). Average severity values were analyzed for significance with the Mann-Whitney U test (Hollander and Wolfe, 1973). Treatment effects were investigated by applying a multivariate analysis of variance (Morrison, 1976) to the transformed data to test for simultaneous equality of measurements across exposure concentrations. (for more information see publication)
Reproductive indices:
not specified
Offspring viability indices:
not specified
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
- Abnormal breathing, thinness, lethargy, abnormal posture, and ruffled fur were observed in rats exposed to 16 mg/m3. Abnormal breathing, marked by shallow, rapid respiration, was first observed during and immediately following exposure periods; this was observed in all 16 mg/m3 rats by week 2 and in all 8 mg/m3 rats by week 4. By week 9, abnormal breathing was also observed in 16 mg/m3 rats during non exposure periods. Some rats in the 16 mg/m3 groups had diarrhea and nasal/eye discharge. Abnormal posture was observed in two males exposed to 8 mg/m3, and one of these rats was thin with ruffled fur and nasal/eye discharge.
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
- 7 males and 3 females exposed to 16 mg/m3 died during the study

Please also refer to the field "Attached background material" below.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- Final mean body weights and body weight gains of males exposed to 4 mg/m3 or greater and of females exposed to 16 mg/m3 were significantly less than those of the chamber controls
- in males exposed to vanadium pentoxide, there was a statistically significant effect on necropsy body weight at 8 mg/m³
- in females exposed to vanadium pentoxide, necropsy body weight was statistically significantly lower at 16 mg/m3

Please also refer to the field "Attached background material" below
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
- Microcytic erytrocytosis was observed in males and females
- Exposure of rats to vanadium pentoxide affected the circulating red cell mass
- In general, the erythrocyte counts reflected similar and proportional, but not significant, decreases.
- There were exposure concentration-related decreases in the mean cell volumes and mean cell hemoglobin values on day 23, which are consistent with a developing erythrocyte microcytosis.
- The mean cell hemoglobin concentration was minimally decreased in 16 mg/m3 females
- Microscopic evaluation of the red blood cell morphology detected increased polychromasia, anisochromia, and hypochromia in the 16 mg/m3 groups
- At week 13, there was evidence of an exposure-related leukopenia, demonstrated by decreases in leukocyte counts in 16 mg/m3 males and females; leukopenia appeared to be related to decreased lymphocyte counts. This alteration in lymphocyte counts suggests a physiological response and would be consistent with a stress-related corticosteroid- induced lymphopenia. Rats are considered a steroid-sensitive species and corticosteroid-induced lymphopenia may be related to lympholysis in blood and altered distribution (Jain, 1986).
- The 16 mg/m3 males and females demonstrated microscopic evidence of lymphoid depletion in numerous lymphoid organs. Since there was an exceptional effect on body weights of 16 mg/m3 rats, it is possible that these animals were under significant stress that then lead to increased endogenous corticosteroid concentrations and subsequent decreases in tissue and blood lymphocytes.
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Lungs:
- Incidence if several nonneoplastic lesions of the lung and nose were increased in animals exposed to 2 mg/m3 or greater
- There were significant increases in the incidences of epithelial hyperplasia of the lung in males and females exposed to 2 mg/m3 or greater.The epithelial hyperplasia occurred in the distal airways and associated alveolar ducts and alveoli. In the distal airways, the epithelium was composed of ciliated cuboidal to columnar cells crowded together in multiple layers, sometimes forming folds and small papillary projections. More distal and including the alveolar ducts and alveoli, the epithelium tended to be a single layer. The epithelial cells were larger and rounded and appeared morphologically similar to bronchioles. Goblet cells sometimes occurred within the epithelium of some of the smaller airways. Alveoli often contained one or two cells that were very large and occasionally binucleate.
- The incidences of inflammation or fibrosis were significantly increased in males exposed to 2 mg/m3 or greater and females exposed to 4 mg/m3 or greater.
- In one female exposed to 16 mg/m3, a single focus of squamous metaplasia was observed within an area of hyperplasia.
- Exudate of the bronchiole was diagnosed in many males and females exposed to 8 or 16 mg/m3 and was characterized by a mucoid material with admixed inflammatory cells within the lumens of some of the hyperplastic airways. The inflammation was characterized by accumulations of alveolar macrophages with abundant foamy cytoplasm in the alveoli adjacent to airways lined by hyperplastic epithelium. Mononuclear cells were also observed within the interstitium.
- Foci of fibrosis were found in the same areas and sometimes extended into surrounding alveoli. These foci were irregularly shaped and consisted of loosely arranged, delicate collagen fibers, fibroblasts, and mononuclear cells.

Nose:
- The incidences of hyperplasia and metaplasia of the nasal respiratory epithelium were significantly increased in males exposed to 8 or 16 mg/m3 and in females exposed to 4 mg/m3 or greater. There were significantly increased incidences of inflammation of the nose in males and females exposed to 16 mg/m3. The hyperplasia and metaplasia involved the respiratory epithelium covering the ventral portion of the nasal septum, the vomeronasal organ, and, to a lesser extent, the ventral lateral walls of the anterior portion of the nasal cavity. There were cellular crowding and epithelium thickening (hyperplasia) in some areas with loss of goblet cells and one or more layers of flattened nonkeratinizing squamous epithelium (metaplasia) in others.
- Inflammation was characterized by an infiltrate primarily of lymphocytes and lesser numbers of neutrophils within the affected mucosa and subjacent submucosa. Occasionally, neutrophils predominated, and an exudate was present.

Other:
- Depletion of lymphocytes in the spleen, thymus, and lymph nodes, atrophy of metaphyseal bone of the femur, and atrophy of the secondary reproductive organs were observed in 16 mg/m3 males and females and hypospermia of the testis and atypical cells of the epididymis were observed in 16 mg/m3 males. These lesions may have been associated with the marked body weight loss and general debilitation of these rats.
Histopathological findings: neoplastic:
not specified
Other effects:
effects observed, treatment-related
Description (incidence and severity):
- Significant exposure-related changes in pulmonary function were observed in male and female rats exposed to 4, 8, or 16 mg/m3; these results indicate that more restrictive lesions were present in groups exposed to 4 mg/m3 or greater, evidenced by reduced lung compliance, changes in breathing measurements, impaired capacity to diffuse carbon monoxide, reduced static and dynamic lung volumes, and exaggerated flows.
- Pulmonary function changes indicate an obstructive disease in the 16 mg/m3 groups, evidenced by changes in breathing mechanics, static lung volumes, and forced expiratory maneuvers. Expiratory resistance, an indicator of bronchoconstriction, and end expiratory and residual volume were increased, while dynamic lung volume was decreased.
- Together, the pulmonary function changes indicate that a restrictive disease was present in male and female rats exposed to 4 mg/m3 or greater, while an obstructive lung disease may have been present only in the 16 mg/m3 groups.
- The pulmonary lavage data indicate an inflammatory response in the lungs of exposed rats. Exposure concentration-related increases were observed in the total numbers of cells, lymphocytes, neutrophils, and protein recovered in pulmonary lavage fluid from rats exposed to vanadium pentoxide at concentrations up to 8 mg/m3.
Reproductive function: oestrous cycle:
effects observed, treatment-related
Description (incidence and severity):
- there was an increases in oestrus cycle length at 8 (statistically significant) and 16 mg/m3 (not statistically significant). However, this effect was highest in the mid dose group (8 mg/m3), thus showing no clear relation to dose and treatment.
- the number of cycling females in the 16 mg/m3 group was reduced.

Please also refer to the field "Attached background material" below.
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
not specified
CLINICAL CHEMISTRY
- At all time points, evidence of a hepatocellular effect was demonstrated by increases in serum alanine aminotransferase activities in 8 and 16 mg/m3 males and females. However, sorbitol dehydrogenase activities, another marker of hepatocellular injury or leakage, were not affected, and there were no histopathologic liver changes consistent with hepatocellular injury. This suggests that the increased alanine aminotransferase activities were not related to hepatocellular injury. Studies have shown that corticosteroids can induce increases in liver alanine animotransferase activities (Rosen, 1959; Rosen et al., 1959). Thus, stress-related corticosteroid-induced increases in liver alanine aminotransferase activities may, in part, help explain increased serum alanine aminotransferase activities.
- At all time points, there were generally exposure concentration-related decreases in serum alkaline phosphatase activities and total protein and albumin concentrations in males and females. At week 13, urea nitrogen concentrations were increased in 16 mg/m3 males and females, suggesting a possible effect on renal clearance. However, creatinine concentrations, another marker of renal clearance, were minimally decreased on day 23 in 8 mg/m3 males and at week 13 in 16 mg/m3 males and females; these decreases would be consistent with the decreased body weights observed in these groups. Since alkaline phosphatase activity and total protein, albumin, and urea nitrogen concentrations can be affected by altered nutritional status, the changes in these variables may have been related secondarily to body weight decreases and altered food intake.
- Significant increases and decreases in bile acid concentrations and creatine kinase activities in various exposure groups at various time points were not considered to be toxicologically relevant.

URINALYSIS
- After 12 (females) or 13 (males) weeks of exposure, the baseline (water-replete) overnight urine collection demonstrated decreased urine volumes and increased urine specific gravities in the 8 mg/m3 male and 16 mg/m3 female groups, suggesting these animals were able to concentrate their urine; the 16 mg/m3 male group was not tested. These findings also suggest that these exposure groups may have been in a partially dehydrated state prior to the water deprivation studies.
- The 4-hour urine collection following a 16-hour water deprivation period demonstrated no differences in urine volume or specific gravity. This suggests that animals in all exposed groups maintained their ability to concentrate urine.
- For baseline and water deprivation test samples, microscopic evaluation of the urine demonstrated slight increases in formed elements in 8 mg/m3 males (baseline: casts, epithelial cells, erythrocytes, and leukocytes; water-deprived: casts) and 16 mg/m3 females (baseline and water deprived: leukocytes) (data not presented). While changes in formed urine elements can be indicative of various renal effects, the alterations in this study were not excessive and possibly reflected the hydration status of the animals; they were not considered to be toxicologically relevant.

ORGAN WEIGHTS
- Other organ weight differences were considered to be related to body weight decreases

GROSS PATHOLOGY
- The carcasses of males and females exposed to 16 mg/m3 were very thin, and the spleens and thymuses appeared disproportionately small
- Lungs of 4 mg/m3 or greater males and 8 and 16 mg/m3 females varied from red to pale or mottled.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
- in males exposed to vanadium pentoxide there was a statistically significant effect on necropsy body weight at 8 mg/m³ and no effect on male reporductive organ weights. Spermatid measurements (heads and counts) were not statistically significant effected as well as epididymal sperm motility and concentration.
- atrophy of the secondary reproductive organs in male rats at 16 mg/m3 is described in the study.
- a statistically significant reduction of absolute testes weight, which was not reflected in the relative testis weights, is given for the 16 mg/m3 group, and may thus be related most probably to the lower necropsy body weight. At 8 mg/m3, no such effects were observed on absolute and relative testes weights which were in the same order of magnitude as in control animals and the other treatment groups.
- a hypospermia of the testis and atypical cells of the epididymis at 16 mg/m3 is described in the study (data for 16 mg/m3 not shown) where only a very mild effects without statistical significance is evident in the 8 mg/m3 group, and is most probably an unspecific effect related to general toxicity at this dose level.

Please also refer to the field "Attached background material" below.

Dose descriptor:
NOAEC
Remarks:
reproductive toxicity
Effect level:
4 mg/m³ air
Based on:
test mat.
Sex:
female
Basis for effect level:
other: increased oestus cycle at and above 8 mg/m3 and decreased no. of cycling females at 16 mg/m3
Dose descriptor:
NOAEC
Remarks:
reproductive toxicity
Effect level:
> 16 mg/m³ air
Based on:
test mat.
Sex:
male
Basis for effect level:
other: male fertility parameters were not affected at the highest dose level
Critical effects observed:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Anogenital distance (AGD):
not specified
Nipple retention in male pups:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
not specified
Remarks on result:
not measured/tested
Critical effects observed:
not specified
Reproductive effects observed:
not specified
Conclusions:
Groups of 10 male and 10 female F344/N rats were exposed to V2O5 aerosols (whole body exposure) at concentrations of 0, 1, 2, 4, 8 or 16 mg/m3 (equivalent to 0, 0.56, 1.12, 2.25, 4.50, and 8.99 mg V/m³) by inhalation, 6 hours/d, 5 d/wk, for 3 months (91-92 days). Effects on fertility and sexual function of male and female rats were evaluated after inhalation exposure.
The results did not indicate effects on male sexual functions. In female rats exposed to 8 mg/m3, oestrous cycles were significantly longer than in animals of the chamber control group, and the number of cycling females in the 16 mg/m3 group was reduced.

Effect on fertility: via oral route
Quality of whole database:
Several studies with oral exposure are available, but the reliability is low and the results are not considered relevant for human exposure.
Effect on fertility: via inhalation route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEC
4 mg/m³
Study duration:
subchronic
Species:
rat
Quality of whole database:
One reliable study (RL1) with inhalation exposure of rat and mice with V2O5, and several studies with RL3 and apprpriate routes of exposure are available. Additional studies with exposure by a non-relevant route (ip) were disregarded.
4 mg/m3 V2O5 corresponds to 2.25 mg V/m3
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

An extensive literature/data search and evaluation programme on animal and human data relating to possible adverse effects of vanadium substances on reproductive toxicity has recently been conducted. As an outcome, some data are available for the soluble vanadium substances via the oral and inhalation route. Two NTP studies with inhalation exposure of V2O5 in mice and rats were conducted under GLP and although not specific reproduction toxicity study they are regarded as key studies, because effects on male and female reproductive organs were evaluated. The other studies can be regarded as studies of low reliability. The studies show supportive evidence of a potential effect of vanadium substances on male and female fertility.

In the two 3-months studies (NTP) with inhalation exposure of V2O5 in male and female rats and mice, animals (10/sex/group) were exposed to 0, 1, 2, 4, 8, or 16 mg V2O5/m3 (0, 0.56, 1.12, 2.25, 4.50, 8.99 mg V/m3) by whole-body inhalation for 6h/d, 5d/week for 3 months. In the highest dose (16 mg/m3), 3/10 females died during the study and necropsy body weight was statistically significantly lower. There was a statistically significantly increases in estrus cycle length at 8 and 16 mg/m3. However, this effect was highest in the lower dose group of 8 mg/m3, thus showing no clear relation to dose and treatment. In contrast, no deaths occurred in female mice. Final body weights started to be reduced in the exposure group of 4 mg/m³, but there was no statistically significant effect on estrus cycle length.

In male rats, severe toxicity was present in the high dose group with 7/10 males found dead during the study, and final body weights from animals in dose groups 4, 8 and 16 mg V2O5/m3 were significantly reduced. In male rats there was a statistically significant effect on necropsy body weight at 8 mg/m3, and no effects on male reproductive organ weights. Spermatid measurements (heads and counts) were not statistically significantly affected as well as epididymal sperm motility and concentration. Atrophy of the secondary reproductive organs in male rats at 16 mg/m3 is described, but at this dose level clear signs of general toxicity were shown. In addition, hypospermia of the testis and atypical cells of the epididymis at 16 mg/m3 are described, while only a very mild effects without statistical significance is evident in the 8 mg/m3 group and is most probably an unspecific effect related to general toxicity at this dose level. Thus, the findings in male rats can be considered secondary to the observed general toxicological and body weights effect in the high dose group and can thus not be attributed to a direct vanadium induced toxicological finding. In male mice, 1/10 males died during the study in the highest dose group. Necropsy body weights were statistically significantly lower at 16 mg/m3, and there was a statistically significant effect on sperm motility at 8 and 16 mg/m3. However, the effect on sperm motility was not very pronounced without a clear dose-response relationship. The lowest value was observed in the 8 mg/m3 group (mid dose) and was less pronounced in the high dose group.

In conclusion, the effects in female animals were observed only in a single species (rat) and not in the other species (mice). The effects in male animals were also observed only in a single species, but in mice, and not in the other species (rat). For both effects, there was no clear relation to dose and treatment and thus the findings must be regarded of low evidence for a vanadium induced effect on male and female fertility. Taking the results together, a dose level of 4 mg V2O5/m3 may be regarded as a NOEC for male and female fertility based on these studies.

 

In addition, there is one other study (Morgan A.M. and El-Tawil O. 2003) with ammonium metavanadate treatment via drinking water in rat available in which male and female animals were paired and effects on fertility evaluated. This study is only of RL3, because of limitations with respect to cohabitation, dose level (only one) and reporting. Sprague-Dawley rats were treated with ammonium metavanadate (200 ppm in drinking water). Male rats (n=10) were treated with 10 mg NH4VO3/kg bw/d corresponding to 4.35 mg V/kg bw/d (calculated with default factors according to ECHA (2012)) for 70 days, and 20 females with 11.43 mg NH4VO3/kg bw/d, (4.97 mg V/kg bw/d) for 61 days (14 days premating, during mating, till weaning of pups (21 days of age).

The results of this mechanistic study conducted only with one dose level show that mating and fertility indices were reduced in treated males mated with untreated females (group 1) and treated females mated with untreated males (group 2) (mating index: control: 100%, treated males 1: 65%; treated females: 70%; fertility index: 95%, 46.15%, 71.43%, respectively). In treated males, reduced weight of testes, epididymis, prostate gland, seminal vesicles, (p<0.05), with no reduction in body weight between control and treated males were observed. In females, the estrus cycle was disturbed, total number corpora lutea was reduced (control: 220, group 1: 54; group 2: 94), and signs of dystocia (no. of dams: 0, 1, 4) and delayed birth date (no. of dams: 0, 3, 5) were observed. No information on general toxicity is presented.

 

There is another rat study (Domingo et al. 1986) conducted with an appropriate route of exposure (oral) and different dose levels sodium vanadate compared to a control group (0, 5, 10 or 20 mg/kg bw/d sodium vanadate per os equivalent to 2.1, 4.2 and 8.4 mg vanadium/kg/day, respectively) available, but only of RL3. Females were treated for 14 days and males for 60 days before mating. Females continued to receive treatment with vanadium until study termination. Parameters evaluated on day 14 sacrifice were the nos. of corpora lutea, total implantations, living and dead fetuses and resorptions.

Fertility was not affected by treatment of male and female rats with sodium vanadate and there were no signs of maternal toxicity (no data presented). At interim sacrifice of females on day 14 of gestation, no significant adverse effects were observed on nos. of corpora lutea, total implantations, living and dead fetuses and resorptions. Thus, at 20 mg/kg bw/dsodium vanadate per os no effects could be determined on fertility of rats.

 

The reproductive toxicity of sodium metavanadate (Llobet 1993) was also studied in male Swiss mice exposed doses of 0, 20, 40, 60 and 80 mg/kg bw per day (equivalent to 8.4, 16.8, 25.2 and 33.6 mg vanadium/kg bw/day) given in the drinking water for 64 days. To evaluate the fertility of the vanadium-treated animals, males were mated with untreated females. A significant decrease in the pregnancy rate was observed at 60 and 80 mg/kg per day. However, the test substance did not reduce fertility in male mice at 20 and 40 mg/kg per day. Decreased body and epididymis weights were only observed in the 80 mg/kg per day group, while testicular weights were not altered by the treatment. Sperm count was significantly decreased at 60 and 80 mg/kg per day, but the sperm motility was unaffected. Histopathological examination revealed that testes were normal and that epididymis of treated male mice contained normal appearing sperm. In this reproduction toxicity study in mice, at 40 mg/kg bw/dsodium metavanadate per os (drinking water) no effects could be determined on fertility of rats.

 

Four additional references (Fortoul T.I. et al. 2007, Mussali-Galante et al. 2005, Bizarro-Nevares et al. 2016, Rodriguez-Lara V et al. 2016) are available with studies conducted with one dose of V2O5 by inhalation (1.4 mg V2O5/m3 equivalent to 0.79 mg V/m³) with a reliability of 3 conducted by the same group of researchers. The study design described in the publications is similar, but different mechanistic endpoints are addressed in the papers. The results of these publications support the evidence of a vanadium related effects on male reproductive organs at a certain exposure level. Supportive evidence might be taken also from several studies with ip injection of divanadium pentaoxide or sodium metavanadate to mice, rats or Guinea-pigs. The studies support the mechanistic evidence of a vanadium induced effect of male reproductive organs but were not taken into consideration.

Developmental Neurotoxicity

Several studies are available with exposure of suckling pups via the milk of their mothers. All studies show deficiencies with respect to methods (e.g. only one dose level, purity of the applied compound not given, no documentation of general unspecific toxicity etc.) and reporting of results and are therefore only of reliability score 3 and should thus be considered only as supportive. In these mechanistic studies, exposure of lactating mothers was conducted by IP injections in order to reach high amounts of secretion into the milk. IP injection is in general a non-relevant route of exposure, but in this case exposure of suckling pups by the milk of their mothers is the relevant route of exposure for this end point. Because of the IP administration to dams, exposure of sucklings by the milk of their mothers might be higher than under realistic exposure considerations and thus represents a worst case scenario.

 

These non-regulatory mechanistic studies were conducted with only one effective dose level of 3 mg NaVO3/kg body weight/day (1.25 mg V/kg bw/day) in comparison to a control group. In 6 of 7 studies conducted either in suckling pups, directly exposed pups or adult rats, effects were observed to a different degree on behaviour and motor coordination in open field tests. The effect on negative geotaxis investigated in 3 studies was not consistent and rotarod performance was affected in pups (exposed via milk or directly), but not in adult rats. In addition, histochemistry and immunohistochemistry investigations were conducted in these studies. There were occasions of a negative effect of vanadium treatment on myelin fibre density or demyelination in different brain areas in suckling pubs as well as in adult rats. Astrogliosis was observed in 4 studies in pubs with varying degree, and oligodendrocytes were shown to be sensitive against vanadium treatment with oligodendrocyte progenitor cells (in vivo and in vitro) being more sensitive to vanadium exposure than astrocytes or mature oligodendrocytes. Lipid peroxidation was shown in adult rats, but could not be detected in suckling pubs. In addition, a few other single endpoints were evaluated.

In conclusion, there is limited evidence that exposure of suckling pubs to vanadium via the milk of their mothers may result in neurotoxic effects evidenced by behavioural changes, myelin damage and astrogliosis under the conditions of these studies. However, based on the available information, no conclusion can be drawn with respect to effect or no effect levels and to realistic exposure scenarios of breastfed children.

The NTP testing program conducted sub-chronic drinking water studies on VOSO4 and NaVO3 as follows:

- Genetic toxicology studies, i.e. the Salmonella gene mutation assays, with NaVO3 and VOSO4 - negative

- 14 days with Harlan Sprague-Dawley rats and B6C3F1/N mice (Dose: R&M: 0, 125, 250, 500, 1000, 2000 mg/L)

- 90-d oral toxicity studies (dosed feed: NaVO3; dosed water: VOSO4) with Harlan Sprague-Dawley rats and B6C3F1/N mice (dose: rats and mice: 0, 31.3, 62.5, 125, 250, or 500 ppm) - ongoing

- Organ systems toxicity, i.e. 28-d immunotoxicity study of NaVO3 (dosed-water) with female B6C3F1/N mice (dose: 0, 31.3, 62.5, 125, 250, or 500 ppm) - ongoing

- post-natal developmental toxicity study: gestation day 6 (GD 6) until postnatal day 119 (PND 119) with Harlan Sprague-Dawley rats (dose: 0, 31.3, 62.5, 125, 250, or 500 ppm) - ongoing

The results of the 14-day dose range finding experiments have already been published (Roberts et al. 2015) and also the first results of the 90-day repeated dose toxicity study and the post-natal developmental toxicity study have been shown as poster on a toxicological conference (Roberts et al. 2019). The publication of the fully study report is expected for end 2020 (personal communication with NTP staff).

These studies will address issues for which to date equivocal or no data at all exist. All hereto available information has been included in the dossier as robust study summaries. Only upon availability of the results from these studies, it will be possible to render a more meaningful decision on whether or not testing for reproductive toxicity is required. Therefore, for the time being this data requirement should be waived in consideration of animal welfare.

Effects on developmental toxicity

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
no data available
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
only summary results but no details or individual data presented
Qualifier:
no guideline followed
Principles of method if other than guideline:
Evaluation of the effects of vanadate (V5+) when administered once daily by gavage as sodium orthovanadate to mice throughout organogenesis.
GLP compliance:
no
Limit test:
no
Species:
mouse
Strain:
Swiss
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Animals were obtained from Letica (Barcelona, Spain).
- Weight at study initiation: 26-29g
- Housing: mice were housed in solid-bottom plastic cages with stainless steel wire lids.
- Diet: ad libitum, standard laboratory chow
- Water: ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 45 +/- 5
- Photoperiod: 12 hours dark/light cycle
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
- The test substance was dissolved in deionised water.

Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No details are given.
Details on mating procedure:
After acclimatisation, females were mated overnight with adult males of the same strain. The morning on which a copulation plug was detected was considered as day 0 of gestation. At this time, animals were randomly assigned to either the control or the vanadate-treated group.
Duration of treatment / exposure:
10 days
Frequency of treatment:
once daily on days 6-15 of pregnancy
Duration of test:
till day 18 of pregnancy
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
7.5 mg/kg bw/day (nominal)
Dose / conc.:
15 mg/kg bw/day (nominal)
Dose / conc.:
30 mg/kg bw/day (nominal)
Dose / conc.:
60 mg/kg bw/day (nominal)
No. of animals per sex per dose:
14-20 dams per dose group
Control animals:
yes
Details on study design:
- Dose selection rationale: The choice of the dosage levels was based on data from a previous study on the developmental toxicity of vanadyl sulfate in mice (Paternein, J.L.; et al. 1990).
Maternal examinations:
CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: general appearance of pregnant mice were monitored daily.

BODY WEIGHT: Yes
- Time schedule for examinations: body weight of pregnant mice were monitored daily.

FOOD CONSUMPTION: Yes
- Time schedule for examinations: Food consumption of pregnant mice were monitored daily.

WATER CONSUMPTION: No data

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 18 with an overdose of ether.
- All dams were evaluated for body weight, liver and kidney weights.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of resorptions: Yes
- Number of dead and live fetuses: Yes
Fetal examinations:
- External examinations: all live fetuses were dissected from the uterus and evaluated for body weight, sex and external abnormalities.
- Soft tissue examinations: 1/3 of the fetuses from each litter were placed in Bouin's fluid to be examined for soft tissue abnormalities.
- Skeletal examinations: 2/3 of the fetuses from each litter were cleared and stained with alizarin red S before examination for skeletal malformations and variations.
- Head examinations: No data
Statistics:
Homogeneity of variance was analysed by Barlett's test. If variances were homogenous, a one-way analysis of variance (ANOVA) was used to test all dose groups simultaneously. The Kruskal-Wallis test was used when variances were not homogenous. Differences between control and orthovanadate-treated groups were analysed by Student's t-test. Incidence data were analysed using the chi-square test. The level of significance for all analyses was p<0.05.
Indices:
no details given
Historical control data:
no data
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
- In the high dose group, 17 dams (from 19 pregnant females dosed) died during the treatment period. Consequently, the two remaining dams were not included in the teratological evaluation of orthovanadate.
- At 30 mg/kg/day, 4 dams (from 18 pregnant females does) were found dead during the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- Maternal weight gain was significantly reduced below control values in the 30 mg/kg/day dose group on gestational days 6-15.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
- Food consumption exhibited a significant decrease in the 15 and 30 mg/kg/day dose groups on gestational days 0-18.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
- There was an increase in relative kidney weight at 30 mg/kg/day, which were statistically significant versus controls.
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Maternal toxic effects:yes

Details on maternal toxic effects:
Doses of 30 and 60 mg/kg/day of sodium orthovanadate resulted in maternal toxicity.
- Exposure to 7.5 or 15 mg/kg/day of the test substance did not result in any maternal death.
- At scheduled termination on gestation day 18, there were no significant decreases in body weight, gravid uterine weight, corrected body weight and change in corrected body weight.
- There was a decrease in absolute and relative liver weight in the 15 mg/kg/days dose group and an increase in relative kidney weight at 30 mg/kg/day, which were statistically significant versus controls. However, the decreases in liver weight were not dose-related and therefore they were not attributed to treatment.
Number of abortions:
not specified
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
no effects observed
Details on maternal toxic effects:
- Evaluation of gestational parameters for the mice indicated no treatment-related effects on number of total implantations per litter, number of live and dead/resorbed fetuses per litter, sex ratio, fetal body weights and the number of stunted fetuses.
Dose descriptor:
NOAEL
Effect level:
7.5 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake
mortality
Abnormalities:
not specified
Fetal body weight changes:
not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
- Treatment-related changes were found during the examination of the incidence and type of skeletal anormalies: significant decreases in the number of ossified sacrococcygeal vertrebrae, as well as the number of ossified forelimb and hindlimb proximal phalanges were seen at 30 mg/kg bw/d.
Visceral malformations:
no effects observed
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes. Remark: skeletal anormalities

Details on embryotoxic / teratogenic effects:
- Sodium orthovanadate did not induce significant incidence of gross and visceral malformations or variations in mouse fetuses.
- Treatment-related changes were found during the examination of the incidence and type of skeletal anormalies at 30 mg/kg bw/d.
- No significant increases in the number of fetuses with reduced ossification of occipital and parietal bones or sternebrae were observed.
Dose descriptor:
NOAEL
Effect level:
15 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
skeletal malformations
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
At 30 and 60 mg sodium orthovandate/kg body weight, deaths occurred among the dams (4/18 and 17/19). Body weight gain was significantly reduced (approximately 20%) at 15 mg/kg b.w. No differences were reported in final body weight, gravid uterine weight, or corrected body weight.
NOAEL for maternal toxicity: 7.5. mg/kg. b.w./day
NOAEL for foetotoxicity 15 mg/kg b.w./day
Executive summary:

Sodium orthovanadate in deionised water was administered once daily by gavage on gestational days 6 -15 to mice at doses of 0, 7.5, 15, 30 and 60 mg/kg body weight/day. Dams were killed on day 18 of pregnancy, and fetuses were examined for external, visceral and skeletal defects. Maternal toxicity was observed at the highest doses levels, as evidenced by a significant number of deaths (60 and 30 mg/kg body weight/day) and reduced weight gain and food consumption (30 and 15 mg/kg body weight/day). Embryolethality and teratogenicity were not observed at maternally toxic doses and below, but fetal toxicity was evidenced by a significant delay in the ossification process of some skeletal districts at 30 mg/kg body weight/day. The NOAEL for maternal toxicity was 7.5 mg/kg body weight/day and 15 mg/kg body weight/day represented a NOAEL for developmental toxicity in mice under the conditions of this study.

Although this publication shows some deficiencies with respect to reporting of methods and results it was conducted in a suficient number of female mice and can be regarded as relevant and approriate.

Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
no data available
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well reported study. The relevance of the study is limited since all developmental effects were associated by pronounced maternal toxicity. Purity of the test substance not stated.
Guideline:
other: no information available if a guideline was followed
GLP compliance:
no
Limit test:
no
Species:
mouse
Strain:
Swiss
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: adult mice were obtained from Panlab (Barcelone, Spain).
- Weight at study initiation: 25-30 g
- Diet: ad libitum, commercial chow
- Water: ad libitum
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 45 +/- 5
- Photoperiod: 12 hours dark/light cycle
NO further details are given.
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: no data


Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No details are reported.
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/2; for mating, one male and two females were placed into a breeding cage.
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
No further details are given.
Duration of treatment / exposure:
on gestational days 6-15
Frequency of treatment:
once daily
Duration of test:
till day 18 of gestation
Remarks:
Doses / Concentrations:
0 mg/kg body weight/day
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
37.5 mg/kg body weight/day
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
75 mg/kg body weight/day
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
150 mg/kg body weight/day
Basis:
nominal conc.
No. of animals per sex per dose:
Three groups of 22 sperm-positive females were randomly assigned to each group.
Control animals:
yes
Details on study design:
- Dose selection rationale: the doses are approximately 1/12, 1/6 and 1/3 of the acute oral LD50 of VOSO4 * 5H2O for adult mice (LLobet, J.M.; Domingo, J.L., 1984).
Maternal examinations:
CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: mice were observed daily for mortality and morbidity throughout the study.

BODY WEIGHT: Yes
- Time schedule for examinations: body weights were computed for the pre-treatment, treatment and post-treatment periods from daily records of these parameters.

FOOD CONSUMPTION: Yes
- Time schedule for examinations: food consumption was computed for the pre-treatment, treatment and post-treatment periods from daily records of these parameters.

WATER CONSUMPTION: No data

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 18 by over-exposure to ether.
- Organs examined: the uteri were excised and weighed.
- Dams were examined for visceral gross pathology.

OTHER: Yes
- Maternal liver, kidney, spleen and placenta were analysed for the concentration of vanadium. Vanadium concentrations were determined by atomic absorption spectrophotometry.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of resorptions: Yes
- Other: post-implantation losses were calculated for each litter as follows: (No. of implants - No. of live fetuses)/No. of implants x 100. The number of stunted fetuses (under 2/3 average body weight) was also recorded.
Fetal examinations:
The fetuses were taken by caesarean section.
- External examinations: fetuses were removed from their placentas and examined externally. Sex, weight and length were recorded.
- Soft tissue examinations: after the external observation was completed, approximately 1/3 of the remaining fetuses from each group were fixed in Bouin's solution and examined for soft tissue abnormalities.
- Skeletal examinations: remaining fetuses were fixed with Alizarin red solution and examined for skeletal abnormalities.
- Head examinations: No / No data
- Other: 3 fetuses from each dam were used for whole body analyses of vanadium.
Statistics:
Test groups were compared to the control group at a level of significance of P<0.05. Continuous data (e.g. maternal body weight, body weight gain, food consumption etc.) were analysed using one-way analysis of variance with significant F values analysed further using Student's t-test or the Mann-Whitney U test. Statistical comparison of the vanadium analyses were made by Student's t-test (one tailed), with a Bonferroni correction for multiple comparisons between treatment groups. A probability of P<0.05 was used as the criterion of statistical significance. Incidence data were compared using chi-square contingency tables (2x4). Each test group was compared to the control group when chi-square was significant.
Indices:
no details given
Historical control data:
no data
Details on maternal toxic effects:
Maternal toxic effects:yes. Remark: body weight gain, organ weights, food consumption

Details on maternal toxic effects:
- No deaths or treatment-related clinical signs of toxicity were observed in the study. 2 animals died in the low-dose group, but these deaths appeared to be related of dosing accidents.
- There were no treatment-related gross necropsy findings.
- Mean body weight gain during the pre-treatment interval, days 0-6 of gestation, was comparable between treated and control groups. However, there were significant decreases in body weight gain during the exposure period in treated animals. The reduction in maternal weight gain during this period was dose-related and was observed at all dose levels. The body weight gain during the post-exposure interval indicated that the treated animals gained less weight than the controls, but these differences were not statistically significant.
- No significant differences occurred between vanadium-treated groups and the control group in food consumption during pregnancy. Although an apparent decrease in the mean food consumption was observed for the 75 and 150 mg/kg body weight/day groups from day 0 to day 18 of gestation, this decrease was not statistically significant.
- Maternal organ weights at termination, including absolute and corrected body weight, absolute liver weight, absolute kidney weight and placenta weight, were affected by treatment. Absolute maternal liver weight (g), but not relative liver weight (% body weight), was significantly decreased at 75 and 150 mg/kg body weight/day, whereas absolute maternal kidney weight was significantly decreased at 150 mg/kg/day.
- Gravid uterine weights were significantly lower at all dose levels relative to controls.
- There was no effect of vanadium on the number of total implantations, live and dead fetuses, late resorptions per litter, or on the percentage post-implantation losses or the sex ratio. The number of early resorptions per litter exhibited a dose-related increase for all vanadium-treated groups.
- Vanadium could not be detected in control mice. In contrast, it was detected in both maternal and fetal tissues of the treated animals. All these values were significant. However, a significant dose-related increase of vanadium concentrations was observed only in the dam kidney of treated animals.
Dose descriptor:
LOAEL
Remarks:
(lowest dose level)
Effect level:
37.5 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes. Remark: external defects, skeletal variations

Details on embryotoxic / teratogenic effects:
- Mean fetal body weights and mean fetal body lengths were significantly lower in the treated groups when compared to the control group.
- There was a significant incidence of stunted fetuses at the 75 mg/kg/day group (P<0.05) and at the 150 mg/kg/day group (P<0.001) relative to controls.
- Cleft palate and micrognathia were the most significant external malformations observed in the 150 mg/kg/day group.
- The total number of external defects was statistically significant for all vanadium-treated groups relative to controls.
- Hydrocephaly in fetuses from dams given 75 or 150 mg/kg/day of test substance was the only incidental abnormality observed after visceral examination.
- Increased incidence of skeletal variations was seen in all vanadium-treated groups. Decreased ossification of the supraoccipital bone and decreased ossification of the carpus and tarsus were the most common anormalies found. The increase in the total number of skeletal defects was significantly different from the controls in all the treated groups. In addition, this increase was dose-dependent.
Dose descriptor:
LOAEL
Remarks:
(lowest dose level)
Effect level:
37.5 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: embryotoxicity
Dose descriptor:
LOAEL
Remarks:
(lowest dose level)
Effect level:
37.5 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: fetotoxicity
Dose descriptor:
LOAEL
Remarks:
(lowest dose level)
Effect level:
37.5 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: teratogenicity
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
The present study involved oral administration of vanadyl sulphate pentahydrate to mice during organogenesis. Maternal toxicity in terms of decreased weight gain and reduction in body weight appeared to be produced in pregnant mice treated with vanadyl sulphate pentahydrate at doses of 37.5, 75 or 150 mg/kg/day. Embryo-/fetotoxicity was incidenced by an increased number of early resorptions and by lower fetal weights and lengths, as well as by increased incidence of poorly ossified skeletal elements at all dose levels. A significantly increased number of major malformations and minor anormalies indicated teratogenicity.
The NOEL for maternal toxicity, embryotoxicity, fetotoxicity and teratogenicity was <37.5 mg/kg/day of vanadyl sulphate pentahydrate, as signs of maternal and developmental toxicity were observed at this dose.
Executive summary:

Vanadium, as vanadyl sulphate pentahydrate, was evaluated for its embryotoxic, fetotoxic and teratogenic potential in Swiss mice. The compound was administered by gavage to pregnant mice at doses of 0, 37.5, 75 or 150 mg/kg body weight/day on days 6 -15 of pregnancy. On gestation day 18, all live fetuses were examined for external, visceral and skeletal malformations and variations.

Maternal toxicity was observed in all vanadium-treated animals, evidend by reduced weight gain, reduced body weight on gestation day 18 (corrected for gravid uterine weight) and decreased absolute liver and kidney weights at 75 and 150 mg/kg body weight/day. The number of total implants, live and dead fetuses, late resorptions, sex ratio and post-implantation losses were not significantly different between the vanadium-treated mice and the controls. However, there was a significant increase in the number of early resorptions per litter at all dose levels. Fetotoxicity was evidend by lower fetal weights and lengths and the presence of developmental variations. Malformation incidence also increased by administration of vanadium. Thus, the NOEL for maternal toxicity, embryo-/fetotoxicity and teratogenicity for vanadyl sulphate pentahydrate under these test conditions can be expected below 37.5 mg/kg body weight/day in Swiss mice.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
no data available
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: No data regarding maternal toxicity was reported
Remarks:
Material and methods described only briefely and only summary results but no details or individual data presented
Qualifier:
no guideline followed
Principles of method if other than guideline:
In order to obtain an overall understanding of vanadium toxicity the present study was undertaken to further examine the adverse effects of sodium metavanadate in rats when given throughout organogenesis.
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Rats were obtained from Interfauna (Barcelona, Spain).
- Age at study initiation: adult rats
- Weight at study initiation: not less than 250g
- Housing: The animals were kept individually in Makrolon cages.
- Diet: ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23
- Humidity (%): 40-60 :
- Photoperiod: 12 hours dark/light cycle

No further details are given.
Route of administration:
other: intragastrically
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The sodium metavanadate solutions were prepared to give any dose in a volume of 1 mL/250 g body weight.

DIET PREPARATION
not applicable
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
no data
Details on mating procedure:
The females were caged with males overnight and examined the following morning for the copulating plug or for spermatozoa by vaginal lavage. The day of vaginal plugs or spermatozoa detection was defined as day 1 of pregnancy.
Duration of treatment / exposure:
from gestation day 6 to 14
Frequency of treatment:
daily
Duration of test:
8 days
Remarks:
Doses / Concentrations:
5.0 mg test item/kg
Basis:
nominal in water
Remarks:
Doses / Concentrations:
10 mg tets item/kg
Basis:
nominal in water
Remarks:
Doses / Concentrations:
20 mg test item/kg
Basis:
nominal in water
No. of animals per sex per dose:
Groups of 20 pregnant rats.
Control animals:
other: yes; distilled water
Details on study design:
No further details on study design.
Maternal examinations:
na data
Ovaries and uterine content:
Cesarean sections were performed on the 20th day of gestation and the following examination made and compared with the control values:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of resorptions: Yes
- Number of live and dead fetuses: Yes
- Other: placental weights
Fetal examinations:
- External examinations: Yes: average fetus body weight, fetal body length and fetal tail length. All fetuses were examined for abnormalities and sexed.
- Soft tissue examinations: Yes: One half of the fetuses from each litter were fixed in Bouin's solution and subsequently examined for visceral abnormalities by razor-blade sectioning.
- Skeletal examinations: Yes: The remaining fetuses from each litter were stained with alizarin red S and examined for skeletal anormalies.
- Head examinations: No data
Statistics:
The magnitude of the differences between the groups was calculated either by Student's t-test or Mann-Whitney U test. The litter was the treatment unit on which statistical analyses were based.
Indices:
The incidence of abnormalities in fetuses from treated dams compared to controls was determined.
Historical control data:
no data
Details on maternal toxic effects:
Maternal toxic effects:no data

Details on maternal toxic effects:
The number of apparently non-pregnant animals increased in the group receiving the highest dose of NaVO3. The number of litters decreased when 20 mg/kg/day NaVO3 were administered. The administration of vanadium had no significant adverse effects on the number of corpora lutea or the number of implantations.
The doses of 10 and 20 mg/kg/day NaVO3 caused an increase of the number of resorptions and number of dead fetuses, although no significant effect on the resorption rate could be demonstrated.
The average weights of placentas were similar.
Dose descriptor:
NOAEL
Effect level:
5 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
The incidence of abnormalities in fetuses from treated dams was remarably higher than the incidence in the control group, especially in the group given 20 mg/kg/day NaVO3. The differences between the number of male and female fetuses were not significant.
Slight differences in body weight, body length and tail length were observed between fetuses from treated dams and controls.
Visceral and skeletal examinations of fetuses did not reveal significant abnormalities in any group. However, the incidence of haemorrhage in facial area (18.4%) in fetuses from dams given 20 mg/kg/day NaVO3 was significantly higher than the incidence in the controls (0.0%). Moreover, in some fetuses of the same group, additional abnormalities were observed (hydrocephaly, 1.0%). In the 5 and 10 mg/kg/day groups, a greater number of abnormal fetuses than in the control group were also observed.
Remarks on result:
not determinable because of methodological limitations
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
In this non-guideline study on developmental toxicity, no significant maternal toxicity was determined. The number of apparently non-pregnant females increased in the high dose group with the number of litters beeing 14, 14, 12, 8 in the control and treated groups, respectively. There were no effects on nos. of corpora lutea and implantations. An increased number of abnormal foetuses was observed from 5 mg/kg bw/d onwards without dose-response relation. At 10 and 20 mg/kg/d, an increase of the number of resorptions (not dose-related) and number of dead foetuses was observed although no significant effect on the resorption rate could be demonstrated. No skeletal abnormalities were determined, and the incidence of visceral abnormalities (hydrocephaly, haemorrhages) in foetuses from treated dams at 20 mg/kg/d was remarkably higher compared to the control group. With respect to haemrorhages there were also higher incidences in the low dose group than in the control group.

Overall, there is no clear evidence of direct developmental toxicity following exposure to sodium metavanadate under the condition of this study.
Possibly no observed effect level for embryonic developent: 10 mg NaVO /kg bw.

In conclusion, the study gives no clear evidence of direct developmental toxicity (WHO 2001)
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
not specified
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
poster only, no full description of methods and results
Qualifier:
no guideline followed
Principles of method if other than guideline:
In this study, time-mated F0 Hsd:Sprague-Dawley rats were exposed to sodium metavanadate at concentrations of 0, 31, 63, 125, 250, and 500 mg/L via drinking water beginning on gestation day 6. Groups of male and female F1 animals were exposed during gestation, lactation and 13-weeks post-weaning (5 animals for biological sampling). The following parameters were investigated: mortality, body weight, water consumption, compund intake, and analysis of vanadium in plasma and urine.
GLP compliance:
not specified
Remarks:
not specified in the publication
Limit test:
no
Specific details on test material used for the study:
not specified
Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
not specified
Route of administration:
oral: drinking water
Vehicle:
unchanged (no vehicle)
Details on exposure:
not specified
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Time-mated animals
Duration of treatment / exposure:
F0 group: gestation day 6 upto weaning of offspirng
F1 group: gestation day 6 upto weaning and 13- weeks post-weaning
Frequency of treatment:
daily
Dose / conc.:
31 mg/L drinking water
Dose / conc.:
63 mg/L drinking water
Dose / conc.:
125 mg/L drinking water
Dose / conc.:
250 mg/L drinking water
Dose / conc.:
500 mg/L drinking water
No. of animals per sex per dose:
F0 group: 16 females
F1 group: 15 males / 15 females
Control animals:
yes
Details on study design:
- Dose selection rationale:
drinking water concentrations were selected on 14-day data.

Time-mated F0 rats were exposed to the test item via drinking water beginning on gestation day 6.
Groups of male and female F1 animals were exposed during gestation, lactation and 13-weeks post-weaning (5 animals for biological sampling).
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Cage side observations checked: surivival

DETAILED CLINICAL OBSERVATIONS: Not specified

BODY WEIGHT: Yes
- Time schedule for examinations: gestational, lactational F0 body weights

FOOD CONSUMPTION AND COMPOUND INTAKE: Not specified

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Not specified

WATER CONSUMPTION AND COMPOUND INTAKE: Yes

OPHTHALMOSCOPIC EXAMINATION: Not specified

CLINICAL PATHOLOGY: Yes (still under evaluation)
NEUROBEHAVIOURAL EXAMINATION: Not specified
IMMUNOLOGY: Not specified

ORGAN WEIGHTS: Yes (still under evaluation)
HISTOPATHOLOGY: Yes (still under evaluation)
Ovaries and uterine content:
not specified
Fetal examinations:
F1 body weights and pup surivival

Measurement of vanadium concentration in plasma and urine
- at the end of the 13 week post weaning exposure period, 5 rats per sex per group, were placed in metabolism cages (24 hours) for collection of urine.
- after 24 hours, rats were euthanized and blood was collected.
- speciation of vanadium in biological fluids was not possible, thus total vanadium was measured.
- samples were thawed, processed by acid digestion with heat, spiked with internal standard (praseodymium) and diluted with DI H2O for analysis.
- samples were analysed using inductively-coupled plasma-mass spectrometry on a Thermo X-Series, ThermoFisher Scientific (Waltham, MA).
Statistics:
mean and standard deviation
Indices:
Fertility and fecunditiy
Historical control data:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
- moribundity resulted in removal of F0 animals during parturition and throughout lactation in the 250 and 500 mg/L groups.

Please also refer to the field "Attached background material" below (Table 4 in the attached document).
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
- there were lower body weights in dams during gestation and lactation that was proportional to sodium metavanadate exposure.

Please also refer to the field "Attached background material" below (Figure 4 in the attached document).
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
- there was lower water consumption in the 250 and 500 mg/L groups during gestation and lactation.
- due to lower water consumption, chemical consumption in the higher groups was slightly less than dose-proportional.

Please also refer to the field "Attached background material" below (Table 5 in the attached document).
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
MORTALITY:
- there was no impact on surivival during gestation.
Please also refer to the field "Attached background material" below (Table 4 in the attached document).

Number of abortions:
not specified
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
not specified
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
not specified
Remarks on result:
not determinable
Abnormalities:
not specified
Fetal body weight changes:
not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
not specified
Changes in litter size and weights:
effects observed, treatment-related
Description (incidence and severity):
- body weight effects were only seen in pups at 500 mg/L.
- at the end of the study for F1 pups (13-weeks post weaning), lower body weights were seen in males at ≥ 125 mg/L (-10 to -20%) and in females at 500 mg/L (-12 %).

Please also refer to the field "Attached background material" below (Figure 4 in the attached document).
Changes in postnatal survival:
effects observed, treatment-related
Description (incidence and severity):
- lower pup surivival was observed in 500 mg/L pups from postnatal day 1 - 10; surivival after postnatal day 10 was similar between exposed groups and controls.

Please also refer to the field "Attached background material" below (Table 4 in the attached document).
External malformations:
not specified
Skeletal malformations:
not specified
Visceral malformations:
not specified
Other effects:
effects observed, treatment-related
Description (incidence and severity):
- there was lower water consumption in the 250 and 500 mg/L groups during post-weaning for male and female F1 offspring.
- due to lower water consumption, chemical consumption in the higher groups was slightly less than dose-proportional.

Please also refer to the field "Attached background material" below (Table 5 in the attached document).
Details on embryotoxic / teratogenic effects:
ANALYSIS FOR TOTAL VANADIUM IN PLASMA AND URINE:
Please also refer to the field "Attached background material" below (table 7 as well as figure 5 in the attached document).
Remarks on result:
not determinable
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
In this study, time-mated F0 Hsd:Sprague-Dawley rats were exposed to sodium metavanadate at concentrations of 0, 31, 63, 125, 250, and 500 mg/L via drinking water beginning on gestation day 6. Groups of male and female F1 animals were exposed during gestation, lactation and 13-weeks post-weaning (5 animals for biological sampling). Dams/pups exposed to the substance at 250 and 500 mg/L in drinking water exhibited moribundity at birth, failure to thrive, and higher moribundity during lactation. Lower body weights were observed in dams during gestation and lactation, and in pups continuing until study termination. 13 weeks post-weaning.
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
not specified
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
poster only, no full description of methods and results
Qualifier:
no guideline followed
Principles of method if other than guideline:
In this study, time-mated F0 Hsd:Sprague-Dawley rats were exposed to vanadyl sulfate at concentrations of 0, 21, 42, 84, 168, and 335 mg/L via drinking water beginning on gestation day 6. Groups of male and female F1 animals were exposed during gestation, lactation and 13-weeks post-weaning (5 animals for biological sampling). The following parameters were investigated: mortality, body weight, water consumption, compund intake, and analysis of vanadium in plasma and urine.
GLP compliance:
not specified
Remarks:
not specified in the publication
Limit test:
no
Specific details on test material used for the study:
not specified
Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
not specified
Route of administration:
oral: drinking water
Vehicle:
unchanged (no vehicle)
Details on exposure:
not specified
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Time-mated animals
Duration of treatment / exposure:
F0 group: gestation day 6 up to weaning of offspirng
F1 group: gestation day 6 up to weaning and 13- weeks post-weaning
Frequency of treatment:
daily
Dose / conc.:
21 mg/L drinking water
Dose / conc.:
42 mg/L drinking water
Dose / conc.:
84 mg/L drinking water
Dose / conc.:
168 mg/L drinking water
Dose / conc.:
335 mg/L drinking water
No. of animals per sex per dose:
F0 group: 16 females
F1 group: 15 males / 15 females
Control animals:
yes
Details on study design:
- Dose selection rationale: drinking water concentrations were selected on 14-day data.

Time-mated F0 rats were exposed to the test item via drinking water beginning on gestation day 6.
Groups of male and female F1 animals were exposed during gestation, lactation and 13-weeks post-weaning (5 animals for biological sampling).
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Cage side observations checked: surivival

DETAILED CLINICAL OBSERVATIONS: Not specified

BODY WEIGHT: Yes
- Time schedule for examinations: gestational and lactational F0 body weights

FOOD CONSUMPTION AND COMPOUND INTAKE: Not specified

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Not specified

WATER CONSUMPTION AND COMPOUND INTAKE: Yes

OPHTHALMOSCOPIC EXAMINATION: Not specified

CLINICAL PATHOLOGY: Yes (still under evaluation)
NEUROBEHAVIOURAL EXAMINATION: Not specified
IMMUNOLOGY: Not specified

ORGAN WEIGHTS: Yes (still under evaluation)
HISTOPATHOLOGY: Yes (still under evaluation)
Ovaries and uterine content:
not specified
Fetal examinations:
F1 body weights and pup surivival

Measurement of vanadium concentration in plasma and urine
- at the end of the 13 week post weaning exposure period, 5 rats per sex per group, were placed in metabolism cages (24 hours) for collection of urine.
- after 24 hours, rats were euthanized and blood was collected.
- speciation of vanadium in biological fluids was not possible, thus total vanadium was measured.
- samples were thawed, processed by acid digestion with heat, spiked with internal standard (praseodymium) and diluted with DI H2O for analysis.
- samples were analysed using inductively-coupled plasma-mass spectrometry on a Thermo X-Series, ThermoFisher Scientific (Waltham, MA).
Statistics:
mean and standard deviation
Indices:
Fertility and fecunditiy
Historical control data:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
- there was lower water consumption in the 168 and 335 mg/L groups during gestation and lactation (335 mg/L only); all others were within 10 % of controls.
- due to lower water consumption, chemical consumption in the higher groups was slightly less than dose-proportional.
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
MORTALITY:
- there was no impact on F0 dam survival during gestation or lactation.

BODY WEIGHT AND WEIGHT CHANGES:
- there were no impacts on body weight during gestation or lactation for F0 females.

Please also refer to the field "Attached background material" below (Tables 2 and 3 in the attached document).

ANALYSIS FOR TOTAL VANADIUM IN PLSMA AND URINE:
Please also refer to the field "Attached background material" below (table 6 as well as figure 5 in the attached document).
Number of abortions:
not specified
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
not specified
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
not specified
Details on maternal toxic effects:
MORTALITY:
- there was no impact on F0 dam survival during gestation or lactation.

BODY WEIGHT AND WEIGHT CHANGES:
- there were no impacts on body weight during gestation or lactation for F0 females.

Please also refer to the field "Attached background material" below (Tables 2 and 3 in the attached document).
Remarks on result:
not determinable
Abnormalities:
not specified
Fetal body weight changes:
not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
not specified
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
External malformations:
not specified
Skeletal malformations:
not specified
Visceral malformations:
not specified
Other effects:
effects observed, treatment-related
Description (incidence and severity):
- post-weaning, there was lower water consumption for F1 males ≥ 83.8 mg/L and females in the 335 mg/L group.
- due to lower water consumption, chemical consumption in the higher groups was slightly less than dose-proportional.
Details on embryotoxic / teratogenic effects:
CHANGES IN POSTNATAL SURVIVAL
- there was no impact on F1 survival during lactation or post-weaning.

CHANGES IN LITTER SIZE AND WEIGHTS
- no impact on F1 body weights during lactation or post-weaning.
- terminal F1 body weights were within 5 % of controls.

Please also refer to the field "Attached background material" below (Tables 2 and 3 in the attached document).

ANALYSIS FOR TOTAL VANADIUM IN PLSMA AND URINE:
Please also refer to the field "Attached background material" below (table 6 as well as figure 5 in the attached document).
Remarks on result:
not determinable
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
In this study, time-mated F0 Hsd:Sprague-Dawley rats were exposed to vanadyl sulfate at concentrations of 0, 21, 42, 84, 168, and 335 mg/L via drinking water beginning on gestation day 6. Groups of male and female F1 animals were exposed during gestation, lactation and 13-weeks post-weaning (5 animals for biological sampling). Vanadyl sulfate, up to 335 mg/L in drinking water, was well tolerated in time-mated rats during gestation and lactation, and their pups during lactation and up to 13-weeks post weaning.
Effect on developmental toxicity: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
15 mg/kg bw/day
Species:
mouse
Quality of whole database:
Data of the developmental toxicity are available for very soluble tetra- and pentavalent substances via the oral route.
15 mg/kg bw/d sodium orthovanadate corresponds to 4.2 mg V/kg bw/d.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Developmental toxicity – animal data

An extensive literature/data search and evaluation programme on animal and human data relating to possible adverse effects of vanadium substances on developmental toxicity has recently been conducted. As an outcome, some data are available for very soluble tetra- and pentavalent substances (VOSO4 and Na3VO4) via the oral route.

One study (Sanchez et al. 1991) with administration of different dose levels of sodium orthovanadate of 0, 7.5, 15, 30 and 60 mg/kg body weight/day per gavage to pregnant mice on gestation days 6-15 was identified. In this study, maternal toxicity was observed at the highest doses levels, as evidenced by a significant number of deaths (60 and 30 mg/kg body weight/day) and reduced weight gain and food consumption a 30 and 15 mg/kg body weight/day. Embryo-lethality and teratogenicity were not observed at maternally toxic doses and below, but foetal toxicity was evidenced by a significant delay in the ossification process of some skeletal districts at 30 mg/kg body weight/day. The NOAEL for maternal toxicity was 7.5 mg/kg body weight/day and 15 mg/kg body weight/day represented a NOAEL for developmental toxicity in mice under the conditions of this study. However, it needs to be considered that foetal toxicity was observed at maternal toxic dose levels and can thus be regarded as secondary. Thus, the dose level of 30 mg/kg bw/d represents the LOEL for developmental effects and 15 mg/kg bw/d the NOEL for developmental effects at maternally toxic dose levels (reduced body weight gain and food consumption). These values correspond to 8.3 mg V/kg bw/d (LOEL) and 4.2 mg V/kg bw/d (NOEL).

One supportive developmental toxicity study (Paternain et al. 1990) with administration of different dose levels of vanadyl sulphate to Swiss mice was identified. Vanadyl sulphate was evaluated for its embryotoxic, fetotoxic and teratogenic potential in Swiss mice. The compound was administered by gavage to pregnant mice at doses of 0, 37.5, 75 or 150 mg/kg body weight/day on days 6 -15 of pregnancy. Maternal toxicity was observed in all vanadium-treated animals, evident by reduced weight gain, reduced body weight on gestation day 18 (corrected for gravid uterine weight) and decreased absolute liver and kidney weights at 75 and 150 mg/kg body weight/day. The number of total implants, live and dead foetuses, late resorptions, the sex ratio and the post-implantation losses were not significantly different between the vanadium-treated mice and the control. However, there was a significant increase in the number of early resorptions per litter at all dose levels. Fetotoxicity was evident by lower fetal weights and lengths and the presence of developmental variations. Malformation incidence also increased by administration of vanadium. Thus, the NOEL for maternal toxicity, embryo-/fetotoxicity and teratogenicity for vanadyl sulphate pentahydrate under these test conditions can be expected below 37.5 mg/kg body weight/day (11.72 mg V/kg bw/d) in Swiss mice.

In a study with intragastric administration of sodium metavanadate (Paternain et al. 1987), an oral NOAEL of 20 mg/kg bw/d was determined at the high dose level corresponding to 8.4 mg V/kg bw/d. There is no clear evidence of direct developmental effects in foetuses of dams exposed during gestational day 6-14 to different dose levels. However, no information on maternal toxicity was reported in the study. Additional supportive studies with administration of V2O5 by non-relevant routes of exposure (ip or iv) (Altamirano-Lozano, M. et al. 1993, Altamirano et al. 1991, Wide, 1984), or other vanadium compounds, e.g. ammonium metavandate (Morgan and El-Tawil, 2003, Carlton et al 1982) or sodium metavandate (Domingo et al. 1986, Gomez et al., 1992) could be found, but were not further considered.

From the study results it can be concluded that fetotoxic effects occurred in absence of unspecific parental toxicity, and therefore there is evidence that vanadium compounds induce developmental toxicity.

 

 

The NTP testing program conducted sub-chronic drinking water studies on VOSO4 and NaVO3 as follows:

- Genetic toxicology studies, i.e. the Salmonella gene mutation assays, with NaVO3 and VOSO4 - negative

- 14 days with Harlan Sprague-Dawley rats and B6C3F1/N mice (Dose: R&M: 0, 125, 250, 500, 1000, 2000 mg/L)

- 90-d oral toxicity studies (dosed feed: NaVO3; dosed water: VOSO4) with Harlan Sprague-Dawley rats and B6C3F1/N mice (dose: rats and mice: 0, 31.3, 62.5, 125, 250, or 500 ppm) - ongoing

- Organ systems toxicity, i.e. 28-d immunotoxicity study of NaVO3 (dosed-water) with female B6C3F1/N mice (dose: 0, 31.3, 62.5, 125, 250, or 500 ppm) - ongoing

- post-natal developmental toxicity study: gestation day 6 (GD 6) until postnatal day 119 (PND 119) with Harlan Sprague-Dawley rats (dose: 0, 31.3, 62.5, 125, 250, or 500 ppm) - ongoing

The results of the 14-day dose range finding experiments have already been published (Roberts et al. 2015) and also the first results of the 90-day repeated dose toxicity study and the post-natal developmental toxicity study have been shown as poster on a toxicological conference (Roberts et al. 2019). The publication of the fully study report is expected for end 2020 (personal communication with NTP staff).

These studies will address issues for which to date equivocal or no data at all exist. All hereto available information has been included in the dossier as robust study summaries. Only upon availability of the results from these studies, it will be possible to render a more meaningful decision on whether or not testing for developmental toxicity in rodents is still required. Therefore for the time being this data requirement should be waived in consideration of animal welfare.

Nevertheless, a testing proposal is added for a pre-natal developmental toxicity study in rabbits with sodium meta vanadate, being the source substances for the soluble vanadium substances read-across group.

Justification for classification or non-classification

An extensive literature/data search and evaluation programme on animal and human data relating to possible adverse effects of relevant vanadium substances on fertility and developmental toxicity has been conducted. There is only equivocal evidence for effects on fertility for the soluble vanadium substances read-across group and no evidence for the poorly-soluble vanadium substances read-across group. Due to ongoing (NTP study expected for 2021) and planned testing (see testing proposal), a definitive conclusion for classification and labelling cannot be made at this point in time. The registrant will update the dossier without undue delay, as soon as the above mentioned information will become available.

Additional information