[Feed-related selenium poisoning in swine]. / Fütterungsbedingte Selen-Vergiftung beim Schwein.

The following case report describes a selenium toxicosis in a pig-fattening farm of two finisher groups. The diseased animals partly showed ataxia and paresis or intense lameness in connection with band-like ablation of the epidermis at the coronary band. Some of them suffered from alopecia. Foot-and-mouth disease and swine vesicular disease were excluded by serological tests. Dissection revealed a multifocal bilateral symmetric poliomyelomalacia. Histological changes in the claws ranged from severe cell-decay in the germinative layer to distinctive decay of the stratum corneum. Due to damage of the claw epidermis the corium was partly exposed. Feed analysis revealed 100-fold increased selenium content in the finishing premix from the feed mill and as a result 20- to 60-fold increased selenium values in feed samples from the farm-made finisher mixture. Selenium concentration in the blood of diseased animals was 4- to 10-fold higher than normal values, which confirmed the tentative diagnosis of a selenium toxicosis.

Serum testosterone and urinary excretion of steroid hormone metabolites after administration of a high-dose zinc supplement.

OBJECTIVES: To investigate whether the administration of the zinc-containing nutritional supplement ZMA causes an increase of serum testosterone levels, which is an often claimed effect in advertising for such products; to monitor the urinary excretion of testosterone and selected steroid hormone metabolites to detect potential changes in the excretion patterns of ZMA users. SUBJECTS: Fourteen healthy, regularly exercising men aged 22-33 years with a baseline zinc intake between 11.9 and 23.2 mg day(-1) prior to the study. RESULTS: Supplementation of ZMA significantly increased serum zinc (P=0.031) and urinary zinc excretion (P=0.035). Urinary pH (P=0.011) and urine flow (P=0.045) were also elevated in the subjects using ZMA. No significant changes in serum total and serum free testosterone were observed in response to ZMA use. Also, the urinary excretion pattern of testosterone metabolites was not significantly altered in ZMA users. CONCLUSIONS: The present data suggest that the use of ZMA has no significant effects regarding serum testosterone levels and the metabolism of testosterone in subjects who consume a zinc-sufficient diet.

[Effects of high selenium intake on selenium status, liver function and claw health of fattening bulls]. / Einfluss einer hohen Selenzufuhr auf den Selenstatus, die Leberfunktion und auf die Klauenqualität von Maststieren.

The effects of three dietary selenium (Se) levels (0.15, 0.35 and 0.5 mg/kg dry matter (dm) and of two Se-compounds (sodium selenite and Se-yeast) on the Se-status, liver function and claw health were studied using 36 fattening bulls in a two-factorial feeding trial that lasted 16 weeks. The claw health was assessed macroscopically and microscopically. Compared to the two control diets containing 0.15 mg Se/kg dm, the intake of the diets containing 0.35 and 0.50 mg Se/kg dm significantly (P < 0.05) increased the Se-concentration in serum, hair, liver and skeletal muscle. Compared to sodium selenite the intake of Se-yeast resulted in significantly (P < 0.05) higher Se-concentration in serum, liver and hair. Concerning the claw horn quality, there was no significant difference between the different groups; the animals receiving organic Se tended to have a better histological score (P = 0.06) at the coronary band than the groups fed with sodium selenite. The serum vitamin E level decreased significantly (P < 0.05) with increasing Se-intake, which had no influence (P > 0.1) on growth and liver function parameters. With the exception of the decrease of the serum vitamin E level indicating an oxidative stress caused by a high Se-intake, no negative effects of dietary selenium exceeding recommended levels for 4 months were observed.

Analytical strategies for the detection of non-labelled anabolic androgenic steroids in nutritional supplements.

Recent studies showed that non-hormonal supplements such as vitamins, minerals and amino acids can contain anabolic androgenic steroids not declared on the labels of the products. These undeclared substances (often prohormones of testosterone or 19-nortestosterone) can cause health risks to consumers and might lead to positive results in sports doping control, especially for the nandrolone metabolite norandrosterone. The analysis of nutritional supplements for anabolic steroids has proven to be rather difficult due to the different matrices in the various products. To conduct a broad-based analysis, a few robust methods capable of analysing various matrices are needed. To obtain a sensitive gas chromatography-mass spectrometry (GC-MS) analysis, a method including extraction and purification of the analytes followed by GC-MS analysis of the trimethylsilyl (TMS) derivatives of the steroids was developed. The limit of detection was improved by the addition of a mixture of 1-N,N-diisopropylamino-n-alkanes (DIPAs) to the final extract. In pure creatine monohydrate powder the limits of detection were demonstrated to be 0.1 ng microg (-1) for dehydroepiandrosterone (DHEA) and estr-4-ene-3beta,17beta-diol, 0.7 ng g(-1) for 5alpha-androstane-3beta,17beta-diol and androsta-1,4-diene-3,17-dione, 1 ng g(-1) for estr-5-ene-3beta,17beta-diol, estr-4-ene-3,17-dione, 19-nortestosterone, androst-4-ene-3, 17-dione and testosterone, and 2 ng g(-1) for androst-4-ene-3beta,17beta-diol and androst-5-ene-3beta,17beta-diol. The recovery (determined at 200 ng g(-1)) ranged from 32% for 19-nortestosterone to 92% for androst-5-ene-3beta,17beta-diol. During the investigation of different nutritional supplements, several analytical difficulties occurred. Aspects of homogenization, extraction, separation, derivatization and GC-MS measurement as well as strategies for the solution of problems arising were optimized. For quantitative measurements of the steroids in nutritional supplements, deuterated internal standards of the specific steroids or standard addition are necessary to compensate for matrix effects.

Analysis of non-hormonal nutritional supplements for anabolic-androgenic steroids - results of an international study.

Several recent studies have shown evidence of some nutritional supplements containing prohibited anabolic androgenic steroids, so-called prohormones, which were not declared on the label. Therefore, a broad-based investigation of the international nutritional supplement market was initiated to clarify the extent of this problem. From October 2000 until November 2001, 634 non-hormonal nutritional supplements were purchased in 13 countries from 215 different suppliers. Most supplements were bought in shops in the respective countries (578 samples = 91.2 %) and on the internet (52 samples = 8.2 %). 289 supplements were from prohormone-selling companies and 345 supplements came from companies which do not offer prohormones. After isolation from the supplement matrix 11 different anabolic androgenic steroids, mainly prohormones of testosterone and nandrolone, were analysed by gas-chromatography/mass spectrometry. Out of the 634 samples analysed 94 (14.8 %) contained anabolic androgenic steroids not declared on the label ("positive supplements"). We could not obtain reliable data for 66 samples (10.4 %) due to matrix effects. In relation to the total number of products purchased per country, most of the positive supplements were bought in the Netherlands (25.8 %), in Austria (22.7 %), in the UK (18.8 %) and the USA (18.8 %). According to the label, all positive supplements were from companies located in only five countries: the USA, the Netherlands, the UK, Italy and Germany. 21.1 % of the nutritional supplements from prohormone-selling companies contained anabolic androgenic steroids, whereas 9.6 % of the supplements from companies not selling prohormones were positive. The positive supplements showed anabolic androgenic steroid concentrations of 0.01 micro g/g up to 190 micro g/g. The administration of supplements containing nandrolone prohormones adding up to a total uptake of more than 1 micro g resulted in positive doping results for norandrosterone for several hours.

Regulatory science: a special update from the United States Food and Drug Administration: Preclinical issues and status of investigation of botanical drug products in the United States.

A recent survey was conducted across the therapeutic divisions within the CDER, U.S. FDA regarding the number of submissions related to botanical drug products over the past ten years. The overall number of botanical submissions as expressed in the parenthesis are as follows: 1990 (1), 1991 (4), 1992 (4), 1993 (5), 1994 (6), 1995 (5), 1996 (13), 1997 (16), 1998 (10). In the total of 64 counted, 50 of them are submitted in original IND and the rest (14) in pre-IND format. The therapeutic categories are focused on dermatological and topical (19), anti AIDS/antiviral (12), oncologic (13), neuropharmacologic (8), endocrine and metabolic (3), urologic (2), tobacco (2), and cardio-renal products (1). The regulatory actions taken on these submissions showed that 68% of them are evaluated as safe to proceed for the human trials, while the rest (32%) of submissions required agency's regulatory guidance. Among the submissions that required further guidance, 81% were deficient in preclinical pharmacology/toxicology information and the rest (19%) lacks information in other areas (chemistry, clinical protocols). Following agency's guidance, 93% of the submissions that were put on hold were allowed to proceed. In summary, a total of 94% of all the botanical INDs submitted to the agency were allowed to proceed without additional animal toxicity studies conducted. In conclusion, this survey indicates that the growing public interest in botanical supplements has prompted more formal evaluation of the efficacy/safety claims of these products.

Hoof horn abnormalities in Lipizzaner horses and the effect of dietary biotin on macroscopic aspects of hoof horn quality.

This study involved a macroscopic evaluation of hoof quality in 152 Lipizzaner horses (130 from Austria and 22 from other countries) and a controlled double blind trial of the effects of biotin on hoof horn growth and quality over 19 months in 42 stallions from the Spanish Riding School (SRS) in Vienna. Using a grading system that incorporated evaluation of horn wall, white line, sole and frog, the macroscopic study revealed the following: 90% of the Austrian Lipizzaners had soft white lines and crumbling, fissured horn at the bearing border of the walls; 39% of the stallions of the SRS, > 4-years-old, had medium to severe hoof horn changes. Daily administration of 20 mg biotin to a test group of horses (n = 26) and a placebo to a control group (n = 16) showed that after 9 months the test group had significantly improved compared to the beginning of the trial and the placebo group (P < 0.01). In the test group, further improvement was observed during the following 5 months and, subsequently, the same good level of hoof condition was maintained over 3 further years of observation. Growth rate of the horn wall was equal in the biotin and placebo group, being 7 mm/28 days, giving a wall renewal period of 11 months. Mean plasma biotin level of untreated horses was 350 ng/l; plasma levels of biotin supplemented horses were > 1000 ng/l. It was concluded that continuous dietary supplementation with biotin at a daily dose of 20 mg is indicated to improve and maintain hoof horn quality in horses with less than optimum quality hoof.

Histological and physical assessment of poor hoof horn quality in Lipizzaner horses and a therapeutic trial with biotin and a placebo.

This paper represents the second part of a study searching for factors which could be responsible for an inferior hoof horn quality of the Lipizzaner horses of the Viennese Spanish Riding School (SRS) noticed in the late 1980s. It includes an evaluation of a treatment with biotin and 42 Lipizzaner horses were tested in a double blind study. The following parameters were evaluated: 1) the histology of the hoof horn in samples from the bearing border 2) the tensile strength of the coronary horn in bearing border samples and 3) the influence of biotin upon the histological changes and the tensile strength after application of 20 mg/day of biotin during 38 months. The histological alterations of bearing border specimens were assessed by use of a grading system (Grade 0 = unchanged; Grade 1 = slight changes; Grade 2 = moderate changes; Grade 3 = severe changes). Initially more than two thirds of the horses showed moderate to severe changes: microcracks visible in the transition from the middle to the inner zone of the coronary horn; separation of the sole from the coronary horn in the region within the white zone. The tensile strength of the coronary horn, with a mean of 39 N/mm2, was 13 N/mm2 lower compared with the unchanged hooves of 10 Warmblood horses, which were investigated by Küng (1991). After 19 months of biotin treatment, the horn quality showed a small but significant improvement. This could be shown by comparing the histological tissue structure before and after treatment, as well as treated animals vs. a placebo group.(ABSTRACT TRUNCATED AT 250 WORDS)

Glycosylation of glycoprotein 55 encoded by the anaemia-inducing strain of Friend spleen focus-forming virus.

Normal rat kidney cells, non-productively infected with the anaemia-inducing variant of Friend spleen focus-forming virus (F-SFFVA), were metabolically labelled with [2-3H]mannose. The primary translation product of the viral envelope gene (env), representing a glycoprotein with an apparent molecular M(r) of 55,000 (gp55), was isolated from cell lysates by immunoaffinity chromatography and purified by preparative SDS/PAGE. Radiolabelled oligosaccharides, released from tryptic glycopeptides by treatment with endo-beta-N-acetylglucosaminidase H, were characterized chromatographically, by enzymic digestion and by acetolysis. The results revealed that F-SFFVA gp55 obtained from this source carried predominantly oligomannose type sugar chains with five to nine mannoses. As a characteristic feature, glycans with seven to nine mannoses contained, in part, an additional glucose residue. Although the amount of glucosylated species found was higher in F-SFFVA gp55 (about 25% of total endo-H-sensitive oligosaccharides) than in gp55 of the corresponding polycythaemia-inducing variant (F-SFFVP, 16.3%), the overall glycosylation pattern of the F-SFFVA env product closely resembled that of F-SFFVP gp55 [Strube et al. (1988) J Biol Chem 263:3762-71]. Hence, our results demonstrate that the different intracellular processing and transport of the primary F-SFFVA env product cannot be attributed to aberrant trimming of its oligomannose type glycans.

The long-term influence of biotin supplementation on hoof horn quality in horses.

The influence of dietary biotin in horses with brittle hoof horn and chipped hooves was investigated in a long-term study, which was performed over a period from one to six years. 97 horses received 5 mg of biotin per 100 to 150 kg of body weight, per os, daily; 11 horses were not supplemented with biotin and served as controls. The hooves of all horses were evaluated macroscopically every three to four months. Hoof horn specimens of the proximal wall were examined histologically and physically in 25 and 15 horses, respectively. The tensile strength of normal coronary horn was 60 N/mm2 or greater; it was reduced in areas of histological alterations, the lowest value being 20 N/mm2. The hoof horn condition of the biotin-supplemented horses improved after eight to 15 months of supplementation as determined by macroscopic and histologic examinations. The hoof horn condition of most control horses remained constant throughout the study. The growth rate of the coronary horn of horses supplemented with biotin and of control horses was the same. The hoof horn condition deteriorated in 7 of 10 horses after biotin supplementation was reduced or terminated. It was concluded that biotin should be continuously supplemented at the full dosage in horses with severe hoof horn alterations.