ssniff Complete feeds with low phytoestrogen content for rats and mice / nude rats and nude mice*

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1 ssniff Complete feeds with low phytoestrogen content for rats and mice / nude rats and nude mice* Complete diets for all development and life stages has producing various low-phytoestrogen diets (free from soya or/and alfalfa for rats and mice as standard feeds for years. Nevertheless, these special diets with considerably reduced phytoestrogen content should be reserved for certain scientific questions dealing with this subject directly or indirectly (i.e. endocrinological studies on the effect of phytoestrogens, estrogens, bone metabolism etc.. However, it can be not recommended using these low-phytoestrogen diets as standard feedingstuffs for breeding and maintaining rats and mice and for most conventional research projects for the following reasons: Since the introduction of laboratory animal diets, phytoestrogens have been contained in these feed mixtures and also occur in the natural feed of rats and mice. Consequently if there were to be a change from conventional laboratory animal feed to low-phytoestrogen diets, the reproducibility of experimental data would have to be queried (see historical data/toxicology. Phytoestrogens evidently develop a large number of positive effects, at least in humans, (see inter alia the review paper by Stetchell and Cassidy That is why soya products are increasingly used in human food. Although some of the effects observed ex vivo and also in vivo require verification by systematic, broad-based examinations, it can still be derived despite this that phytoestrogens might also be able to contribute to reducing chronically progressive diseases in laboratory animals too. In an extensive study various laboratory animal diets were examined for their influence on the sexual development of rats (Odum et al. 01. According to this study similar effects were ascertained for instance on the uterus with AIN76A, that contains no phytoestrogens, and Purina 5001, with a very high phytoestrogen concentration (about 10 µg genistein and 150 µg daidzein/g feed. The authors come to the conclusion that other (unknown substances in the diets may be responsible for the findings surveyed. This study shows that the endocrinological processes and reactions in the organism are very complex and partly not predictable. Moreover, in a recently published study with rats it could be demonstrated, that diets free from phytoestrogens evidently induced a dramatic impairment of the breeding performance and affected the health status of the offspring (Souzeau et al. 05. ssniff complete feeds low in phytoestrogens are designed for rats/mice maintenance, mice breeding, rat breeding, as well as for nude mice and rats (breeding/maintenance. They are generally intended for the ad libitum intake, however, from case to case it may be necessary to restrict the daily feed consumption, e.g. under certain study conditions or in animals susceptible to get obese. Moreover, further diets with reduced phytoestrogen content - also with increased energy density - are available for the breeding of rats and mice.* Phytoestrogen reduced diets for breeding and maintenance of rats/mice and nude rats/nude mice * Article V155 V135 V115 V5 Product R/M-H R-Z M-Z NM/NR Crude protein, CP [%] Crude fat, CL [%] Cude fibre, CF [%] [g/mj] * Further special diets on request

2 ssniff R/M-H Low-Phytoestrogen Complete feed for rats and mice - maintenance, soy-free This diet was developed for the maintenance of rats and mice. The feed contains not any soybean and alfalfa product and is therefore characterized by very low concentrations of the phytoestrogens genistein, daidzein and coumestrol. Accordingly, that type of diet is particularly suitable for studies, in which the effects of estrogens and similar substances or other sex hormones should be investigated. Dry matter.0 Crude protein (N x Crude fat 3.3 Crude fibre 4.4 Crude ash 6.0 N free extracts 55.1 Starch 36.1 Sugar 3.1 Gross (GE.5 Metabolizable ( *. 57 % from 9 % 34 % from Phosphorus 0.70 Sodium 0. Magnesium 0.21 Potassium 0.6 C :0 C : C : C 1: C 1: C 1: C 1: C : descending order of feedingstuffs (FMV minerals, vegetable oils, vitamins, trace elements. * calculated according to the pig formula, V Meal, single ground V mm pellets V mm pellets, autoclavable (vitamin fortified; not shown Lysine 1.01 Methionine 0.3 Met+Cys 0.75 Threonine 0.2 Tryptophan 0.25 Arginine 1.01 Histidine 0.46 Valine 1.0 Isoleucine 0. Leucine 1.71 Phenylalanine 1.05 Phe+Tyr 1. Glycine 0.93 Glutamic acid 3.77 Aspartic acid 1.1 Proline 1.43 Alanine 1.09 Serine 1.01 Phone:+49-( Fax: +49-( mg Vitamin K (as menadione 5 mg 22 mg 19 mg 100 µg Nicotinic acid 0 mg Pantothenic acid 39 mg 6 mg Biotin 460 µg Choline-Chloride 2,530 mg density [MJ /kg] and protein/energy ratio [g CP/MJ ] mg 0 mg 101 mg 14 mg

3 ssniff R-Z Low-Phytoestrogen Complete feed for rats - breeding, soy-free This complete diet meets ideally the increased energy and protein requirements for reproduction and growth of high performance rats. The feed contains not any soybean and alfalfa product and is therefore characterized by very low concentrations of the phytoestrogens genistein, daidzein and coumestrol. Accordingly, that type of diet is particularly suitable for studies, in which the effects of estrogens and similar substances or other sex hormones should be investigated. Dry matter.2 Crude protein (N x Crude fat 4.3 Crude fibre 4.2 Crude ash 6.1 N free extracts 52.2 Starch 34.0 Sugar 3.0 Gross (GE. Metabolizable ( * % from 11 % 37 % from Phosphorus 0.70 Sodium 0.19 Magnesium 0. Potassium 0.67 C :0 C : C : C 1:0 0.0 C 1:1 0.2 C 1: C 1: C : descending order of feedingstuffs (FMV vegetable oils, minerals, vitamins, trace elements. * calculated according to the pig formula, V Meal, single ground V mm pellets V mm pellets, autoclavable (vitamin fortified; not shown Lysine 1.22 Methionine 0.44 Met+Cys 0.3 Threonine 0.97 Tryptophan 0.2 Arginine 1.13 Histidine 0.51 Valine 1.25 Isoleucine 1.03 Leucine 1.95 Phenylalanine 1.21 Phe+Tyr 2. Glycine 1.05 Glutamic acid 3.96 Aspartic acid 2.1 Proline 1.53 Alanine 1.26 Serine mg Vitamin K (as menadione 5 mg 22 mg mg 100 µg Nicotinic acid 0 mg Pantothenic acid 40 mg 7 mg Biotin 460 µg Choline-Chloride 2,30 mg density [MJ /kg] and protein/energy ratio [g CP/MJ ] 9 mg 7 mg 101 mg 14 mg Phone:+49-( Fax: +49-(

4 ssniff M-Z Low-Phytoestrogen Complete feed for mice - breeding, soy-free This diet was designed for breeding mice (reproduction, growth. Accordingly, the protein content (amino acid concentration and energy density (fat addition are markedly increased. In that feed neither soybean nor alfalfa products are used, resulting very low concentrations of the phytoestrogens genistein, daidzein and coumestrol. Accordingly, that type of diet is particularly suitable for studies, in which the effects of estrogens and similar substances or other sex hormones should be investigated. Dry matter.4 Crude protein (N x Crude fat 5.4 Crude fibre 4.0 Crude ash 6.1 N free extracts 50.2 Starch 32.5 Sugar 2.9 Gross (GE 17.2 Metabolizable ( * % from 14 % 37 % from Phosphorus 0.72 Sodium 0.1 Magnesium 0. Potassium 0.67 C :0 C :0 0.6 C : C 1: C 1: C 1: C 1: C : descending order of feedingstuffs (FMV minerals, vegetable oils, vitamins, trace elements. * calculated according to the pig formula, V Meal, single ground V mm pellets V mm pellets, autoclavable (vitamin fortified; not shown Lysine 1.31 Methionine 0.46 Met+Cys 0.7 Threonine 1.03 Tryptophan 0.29 Arginine 1.1 Histidine 0.53 Valine 1.31 Isoleucine 1.09 Leucine 2.05 Phenylalanine 1.27 Phe+Tyr 2.33 Glycine 1.09 Glutamic acid 4.01 Aspartic acid 2.33 Proline 1.57 Alanine 1.32 Serine mg Vitamin K (as menadione 5 mg 22 mg 19 mg 100 µg Nicotinic acid 117 mg Pantothenic acid 3 mg 6 mg Biotin 450 µg Choline-Chloride 2,290 mg density [MJ /kg] and protein/energy ratio [g CP/MJ ] 5 mg 75 mg 9 mg 14 mg Phone:+49-( Fax: +49-(

5 ssniff M-Z Ereich, low phytoestrogen content Complete feed for mice - breeding, high energy, extruded (autoclavable This feed was designed for the breeding of mice. It is therefore characterized by a high protein content with excellent protein quality (amino acid composition and by a markedly increased energy density. The diet is ideal compsed for animals loosing weight during breeding and some transgenic mouse lines. Moreover, the feed can be used for endocrinological studies or for (tox studies dealing with endocrine disrupters, because it is free from soybean and alfalfa products resulting in very low concentrations of the phytoestrogens genistein, daidzein and coumestrol. Dry matter 9.0 Crude protein (N x Crude fat 6.0 Crude fibre 3.3 Crude ash 6.3 N free extracts 49.6 Starch 33.4 Sugar 2. Gross (GE 17.3 Metabolizable ( * % from 15 % 37 % from Phosphorus 0.71 Sodium 0.21 Magnesium 0.21 Potassium 0.4 C : C : C 1:0 0. C 1: C 1: C 1: C : descending order of feedingstuffs (FMV minerals, vegetable oils, amino acids, vitamins, trace elements. * calculated according to the pig formula, Lysine 1.55 Methionine 0.56 Met+Cys 0.97 Threonine 1.02 Tryptophan 0.29 Arginine 1.1 Histidine 0.54 Valine 1.30 Isoleucine 1.0 Leucine 2.10 Phenylalanine 1.29 Phe+Tyr 2.36 Glycine 1.09 Glutamic acid 4.04 Aspartic acid 2.31 Proline 1.61 Alanine 1.33 Serine 1.23 Vitamin A 30,000 IU 1,0 IU 0 mg Vitamin K (as menadione 25 mg 101 mg 3 mg 34 mg 10 µg Nicotinic acid 2 mg Pantothenic acid 65 mg 11 mg Biotin 730 µg Choline-Chloride 2,330 mg 0 mg density [MJ /kg] and protein/energy ratio [g CP/MJ ] 15 mg 79 mg 105 mg mg 2.4 mg 2.4 mg NM M-Z Ereich M-Z Extr. M-Z M-Z auto V16-3 Extrudate (autoclavable 1 Phone:+49-( Fax: +49-(

6 ssniff NM/NR Low-Phytoestrogen Complete feed for nude rats and mice - maintenance / breeding, soy-free This diet was designed for the breeding and maintenance of nude mice and rats. The protein content (amino acid concentration and energy density (fat addition are adjusted ideally to the higher requirements of the mice/rat lines. In that feed neither soybean nor alfalfa products are used, resulting very low concentrations of the phytoestrogens genistein, daidzein and coumestrol. Accordingly, that type of diet is particularly suitable for studies, in which the effects of estrogens and similar substances or other sex hormones should be investigated. Dry matter.5 Crude protein (N x Crude fat 5.7 Crude fibre 3. Crude ash 5.9 N free extracts 4.1 Starch 31.0 Sugar 2. Gross (GE 17.5 Metabolizable ( * % from 14 % 40 % from Phosphorus 0.70 Sodium 0.1 Magnesium 0.19 Potassium 0.66 C :0 C : C : C 1: C 1: C 1: C 1: C : descending order of feedingstuffs (FMV minerals, vegetable oils, vitamins, trace elements. * calculated according to the pig formula, V50-0 Meal, single ground V mm pellets V mm pellets, autoclavable (vitamin fortified; not shown Lysine 1.53 Methionine 0.52 Met+Cys 0.96 Threonine 1.1 Tryptophan 0.33 Arginine 1.32 Histidine 0.59 Valine 1.49 Isoleucine 1.25 Leucine 2.30 Phenylalanine 1.45 Phe+Tyr 2.66 Glycine 1.22 Glutamic acid 4.23 Aspartic acid 2.73 Proline 1.6 Alanine 1.50 Serine mg Vitamin K (as menadione 5 mg 23 mg mg 105 µg Nicotinic acid 1 mg Pantothenic acid 39 mg 7 mg Biotin 450 µg Choline-Chloride 2,170 mg density [MJ /kg] and protein/energy ratio [g CP/MJ ] mg 75 mg 99 mg 15 mg Phone:+49-( Fax: +49-(