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Description of key information

Oral:
NOAEL (subchronic, rat) = 3000 mg/kg bw/day (males)
NOAEL (subchronic, rat) = 3300 mg/kg bw/day (females)
NOAEL (subacute, rat) = 5110 mg/kg bw/day (males)
NOAEL (subacute, rat) = 5221 mg/kg bw/day (females)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion
Dose descriptor:
NOAEL
3 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat

Additional information

Within the Vinasses category, data on repeated dose toxicity are only available for one subgroup of Vinasses, e. g. Vinasses, residue of fermentation. However, all Vinasses subgroups share a common origin and are therefore constituted of the same components determining their toxicological properties. Thus, read-across is performed based on a category approach (A detailed justification is attached in IUCLID section 13).

 

Oral

Vinasses, residue of fermentation were tested for subchronic oral toxicity in Wistar rats following OECD guideline 408 and in compliance with GLP (Appel, 2003). The test material was fed ad libitum to groups of 5 male and female rats at 2, 6 and 15% in the diet for 13 weeks. The corresponding mean dose levels over the study period were 1000, 3000 and 8000 mg/kg bw/day in males and 1200, 3300 and 9600 mg/kg bw/day in females. A negative control group fed a plain diet, and a positive control group receiving additional ammonium (derived from (NH4)2S04); approx. 0.6% in diet, corresponding to 320 and 380 mg/kg bw/day in males and females, respectively) were included.

There were no unscheduled deaths during the test. No clinical signs of toxicity were observed. A slight reduction in growth and slightly reduced food consumption (during the first week of the study) were observed in the high-dose group, which were probably related to the high level of ammonium nitrogen in the test substance. Water consumption was generally increased in a dose-related manner in treated animals.

No effects were observed at neurobehavioural and ophthalmoscopic examinations as well as in haematological values. Clinical chemistry parameters were generally similar among the groups. However, albumin was decreased in females of the ammonium-control group and of the high-dose group and the albumin/globulin ratio was decreased in males of the ammonium-control group and in females of the high-dose group. These changes were probably related to the high level of ammonium nitrogen in the test substance. The decrease in chloride in females of the high-dose group was considered treatment-related.

Urinary volume and density were similar among the treated groups. Urinary pH was lower and urinary crystals were increased in males of the ammonium-control group and of the high dose group. These changes were considered related to the high level of ammonium nitrogen in the test substance.

Some changes were found in the absolute and/or relative weight of kidneys, brain, liver, testes and uterus. All changes were considered either chance findings or related to the ammonium- level in the test substance, except for the change in relative liver weights, which was considered treatment-related.

Macroscopic and microscopic examination at autopsy did not reveal any treatment-related changes. Measurement of fertility parameters did not show any treatment-related changes.

Based on the overall results of the study, the NOAEL is considered to be 3000 mg/kg bw/day in male and 3300 mg/kg bw/day in female rats.

In an earlier subchronic study, rats (10 per sex) were dosed orally by gavage for 94 days with two types of Vinasses, residue of fermentation at 625, 1250 and 2500 mg/kg bw and 2500 mg/kg bw, respectively (van Eeken, 1985). A control group (20 animals per sex) was treated with vehicle (physiological saline). Observations and examinations included mortality, clinical signs, body weight, clinical pathology, organ weights and histopathology.

No clinical signs or mortality occurred in relation to the test substance. Two animals died due to a technical error during dosing. No effects were observed in body weight (gain), haematological and clinical chemistry parameters as well as at necropsy and histopathological examinations. Absolute and relative kidney weights were slightly increased (only statistically significant in the relative kidney weight). Since no morphological changes were noted at microscopic examination and no changes in clinical haematology was noted, this change of weight was considered not toxicologically significant.

Based on the study results, the NOAEL was considered to be 2500 mg/kg bw/day for both male and female rats which corresponds to the highest dose tested.

In addition, reliable subacute oral toxicity studies are available.

In a GLP-study study following the US FDA Toxicological Principles for the Safety Assessment of Direct Food Additives and Color Additives Used in Food (1982), groups of Sprague-Dawley rats (10 per sex per dose) were fed diets containing Vinasses, residue of fermentation at 500, 5000 and 50000 ppm ad libitum for 4 weeks (Richard, 1993). The corresponding mean dose levels over the study period were 50, 498 and 5110 mg/kg bw/day in males and 48, 486 and 5221 mg/kg bw/day in females. A negative control group fed a plain diet as well as additional satellite groups for immunological examinations consisting of 8 males per dose were included.

No mortalities occurred and no treatment-related clinical signs were noted in any group. The food consumption, body weight gain and food efficiency were similar in control and treated groups. The water consumption was higher in the 50000 ppm group when compared to the control group. This could be attributed to the exogenous supply of sodium in the 50 000 ppm preparations: i.e. a supply of 6.86 g of sodium per kg of mixture.

No abnormalities were noted at ophthalmological examination of the animals of the 50000 ppm group.

No treatment-related haematological findings were noted. Regarding blood chemistry, the only relevant finding was a higher level of aspartic acid, hydroxyproline, glutamic acid, alanine and methionine in the males of the 50000 ppm group compared to the controls; this was considered to be related to the composition of the test material.

No effects in urinalysis parameters were observed.

Changes in organ weights included: a slight increase in relative kidney weight in males given 50000 ppm; a slight decrease in absolute and relative adrenal glands weight in males given 5000 ppm; a slight decrease in absolute adrenal glands weight in males given 50000 ppm; a slight decrease in absolute and relative thymus weight in females given 5000 ppm. Changes were considered to be of no toxicological importance.

Macroscopic and microscopic examinations were considered as being of no toxicological importance. Changes observed were reported to be those which are commonly recorded as spontaneous findings in the laboratory rat.

In the satellite groups, no treatment-related changes were noted in the immune function tests, except for a lower response to a T-dependent antigen and a slight decrease in B-lymphocyte proliferation in response to lipopolysaccharide at the 5000 ppm concentration. The relationship of these findings to the treatment was considered to be very doubtful.

Based on the study results, the NOAEL is considered to be 50000 ppm corresponding to 5110 and 5221 mg/kg bw/day in male and female rats, respectively, which corresponds to the highest dose tested.

In another subacute GLP-study, following OECD guideline 407, groups of Wistar rats (5 per sex per dose) were daily given Vinasses, residue of fermentation at 0.75, 3 and 10 mL/kg bw/day by gavage for 28 days (Til, 1992).

Two female rats of the high-dose group died in the course of the study. These deaths are probably due to faulty dosing accompanied by regurgitation and aspiration of the test substance, rather than to toxicity. Diarrhoea occurred in all rats at the end of the 4-week study. Slightly decreased body weights were observed in males receiving 10 mL/kg bw/day, accompanied by decreased food intake and food efficiency. Water intake was increased in this group in both sexes.

Haematology showed increased haemoglobin concentration, packed cell volume and red blood cell count in males and females of the high-dose group. No further noticeable changes were observed. Fasting blood glucose levels were increased in the high-dose group in both sexes, while plasma blood sodium concentration was decreased in females of this group. No further toxicologically relevant changes were noted.

Urinary volume showed dose-related increases in the mid- and high-dose groups in both sexes. Urinary density was slightly decreased in the high-dose group, and increased in the low-dose group in both sexes.

In the high-dose group, the relative kidney weight was increased in both sexes and that of adrenals in males.

At autopsy, no treatment-related gross alterations were observed. Microscopic examination of the adrenals revealed hypertrophy of the zona glumerosa in males of the high-dose group. Histopathological changes referable to treatment were not seen in any other organs examined.

No obvious signs of toxicity were noted in the mid-dose animals. Therefore the, NOAEL was considered to be 3 mL/kg bw/day in both male and female rats.

In an earlier study, three groups of five male and five female Wistar rats were given one of three concentrates of Vinasses, residue of fermentation at 5000 mg/kg bw by gavage for 13 days (van Eeken, 1974). A control group was treated with the vehicle (water) only. No mortalities occurred and no effects were observed on clinical signs, body weight (gain), food consumption, organ weights and at macroscopical examination.

Furthermore, various subgroups of Vinasses, namely (sugar) beet and (sugar) cane Vinasses, are listed in the Catalogue of feed materials (Annex to Commission Regulation (EU) No 242/2010). Several feeding studies in different species have investigated the use of these Vinasses as protein source likely to be used in replacement of soybean meal in livestock feed mixtures.

Overall, no effects on growth and slaughter performance and no indications for toxicologically relevant changes were reported after feeding Vinasses, residue of fermentation containing biomass of bakers yeast (Saccharomyces cerevisiae) to young female goats at 2526 and 4995 mg/kg bw/day for 6 months (Ringdorfer, 2009); to dairy cows at 640 and 1296 mg/kg bw/day for 12 weeks (Urdl and Schauer, 2009); to bulls at 2135 mg/kg bw/day for 356 days (Leitgeb, 2010); to pigs at 10428 mg/kg bw/day for 55 days and at 5245 mg/kg bw/day for further 45 days (Windisch and Schedle, 2010).

In chicks fed Vinasses, residue of fermentation containing biomass of bakers yeast (Saccharomyces cerevisiae) at 8, 16 and 24 % in diet for 36 days, a significant decrease in growth and slaughter performance was observed at the highest concentration with slight evidence of dose-dependency (Windisch and Leitgeb, 2009). The corresponding Vinasses mean dose levels over the study period were 6438, 13197 and 20780 mg/kg bw/day. The decrease in daily body weight gain was correlated with a decrease in daily feed intake. The absolute weight of abdominal fat, heart, liver stomach and individual body parts determined at the end of the study was also significantly decreased at the highest dose level. However, except for breast, the corresponding relative weights were not reduced and even a slight dose-dependent increase was observed.

No effects were reported for bulls fed Vinasses at 14% in the diet for 10 days (Stemme et al., 2005). In the same study, pigs fed Vinasses at 16 and 43% in the diet for 36 days showed no effects, except for osmotically driven diarrhoea in animals from the 43% group, related to an unexpected high sulfate content in the Vinasses used.

 

Dermal

This information is not available.

 

Inhalation

This information is not available.

Justification for classification or non-classification

The available data on the repeated dose toxicity of the substance via the oral route are conclusive but not sufficient for classification according to the DSD (67/548/EEC) and CLP (1272/2008/EC) criteria. A specific target organ toxicity (STOT) cannot be defined.