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Biomedical subjects

M J Groot

Publications and source records attributed to M J Groot.

12 recordsLinked to original sources

Effects of phyto-oestrogens on veal calf prostate histology.

In veal calf production plant-based proteins are frequently included in milk replacer fed to the animals. Since soy products, which are mostly used, are known for their high levels of phyto-oestrogens, the effects of these feeds on the veal calf prostate were examined. Goal was to determine whether these compounds could interfere with histological screening for oestrogenic growth promoters. In a feeding experiment, four groups of veal calves fed plant-based protein-supplemented milk replacer (PBM), containing 5% soy concentrate, 5% soy isolate, 5% wheat gluten and 2% potato protein, for 4 weeks were compared to animals fed dairy-based control feed (DBM); animals treated with estradiol benzoate, diethylstilbestrol and ethinylestradiol served as positive controls. Daidzein and genistein levels measured in feed and urine showed high levels of genistein and daidzein in the soy isolate and soy concentrate supplemented feeds. Genistein and daidzein were also found in the urine of the animals that were fed these feeds. Haematoxylin-eosin-stained prostate sections of PBM-fed animals showed slight hyperplasia and some dilated tubules as compared to the DBM-fed group, but no metaplasia, which is used for screening for oestrogenic hormones. The positive controls showed extensive squamous metaplasia. Immunohistochemical staining for cytokeratin 5 (using RCK 103 monoclonal antibody) in basal cells showed a normal staining pattern of basal cells in the DBM-fed calves and extensive basal cell proliferation and squamous metaplasia in the oestrogen-treated positive control animals. PBM-fed calves showed no increase of basal cell staining but showed elongations of the basal cells in most animals, sometimes resulting in circular figures. It is concluded that the feeds examined in this study did not interfere with histological screening for oestrogens in male veal calves.

Animal Feed↗

Screening for estrogen residues in calf urine: comparison of a validated yeast estrogen bioassay and gas chromatography-tandem mass spectrometry.

Within the European Union, the control for residues of illegal hormones in food-producing animals is based on urine analysis for a few target analytes using gas chromatography/mass spectrometry and/or liquid chromatography-tandem mass spectrometry. Recently, we developed a robust yeast bioassay screening tool for estrogens, which was validated as a qualitative screening method in accordance with EC decision 2002/657/EC. In this study, we present long-term performance data and a comparison of urine data obtained with this bioassay, and data from an established gas chromatography-tandem mass spectrometry (GC/MS/MS) confirmatory analysis method. More than 120 calf urine samples from a controlled reference experiment were analysed using both protocols. According to the GC/MS/MS method, only the natural estrogens 17alpha-estradiol and estrone were present in the non-compliant samples. The bioassay was less sensitive than GC/MS/MS for the relatively weak estrogenic compound 17alpha-estradiol, in accordance with expectations. Assuming that application of the mass spectrometric method is considered beyond reasonable doubt, the bioassay performed very well: only 5.6% of the calf urine samples found compliant in GC/MS/MS were screened false suspect in the bioassay screening method. The bioassay results of non-compliant urine samples under routine conditions were as predicted, taking into account the relative estrogenicity of the natural estrogens 17alpha-estradiol and estrone vs. 17beta-estradiol. Only one sample was screened false negative for 17alpha-estradiol and estrone. Application of this fast and simple estrogen bioassay in routine surveillance and control can significantly reduce GC/MS/MS sample workload and allow higher percentages of animals to be screened for potential hormone abuse.

Animals↗

Histopathological effects of boldenone in cattle.

Histopathology of male cattle previously found positive for béta-boldenone in urine in the Netherlands and in Italy was studied. The animals were derived from practice and several weeks had passed after the finding of béta-boldenone before the animals were examined. The animals consisted of 34 male veal calves and one finishing bull. In the prostate gland hypersecretion, cyst formation (45%) and hyperplasia of the urethral epithelium was observed, in the bulbo-urethral gland similar alterations were present. The testis showed reduced development and degeneration of the germinal epithelium (70%), leading to debris and syncytial cell formation in the lumina. Stromal proliferation was evident. In some animals the liver was sampled and showed periportal fibrosis, bile duct proliferation and sometimes necrosis. The bull also showed degeneration of the germinal epithelium of the testis and absence of sperm production, the prostate gland showed some secretion and had an atrophic appearance. It is concluded that béta-boldenone may lead to degeneration of the germinal epithelium of the testis and hypersecretion and cyst formation in the prostate and bulbo-urethral gland, which alterations may heal in time.

Anabolic Agents↗

Hepatatis in growth promoter treated cows.

Adult female beef cattle found positive for stanozolol in the urine were investigated for liver pathology. In all the animals toxic hepatitis was found, including cholestasis, periportal fibrosis and inflammation, focal necrosis and blood filled lacunae. As no clinical data of the cows were available, apart from the history of illegal stanozolol abuse, it is not possible to attribute all changes to the illegal hormone treatment. Moreover, the cows have probably been treated with a cocktail, and apart from stanozolol more anabolic steroids may have been used. Management factors, viral and bacterial infections, former caesarean sections and especially feeding regime may also be responsible for the lesions described. Striking similarities with data from hepatotoxicity found in human body builders using similar agents, however, suggest a major role of stanozolol as causative agent.

Anabolic Agents↗

Pour on application of growth promoters in veal calves: analytical and histological results.

To investigate the possibilities for screening and confirmation methods when the 'pour on' method of application is used for administration of growth promoters, an animal experiment was performed using a cocktail of a combination of growth promoters derived from (illegal) practice. Two cocktails were used, cocktail A consisting of stanozolol and estradiol benzoate and cocktail B consisting of stanozolol, estradiol benzoate and beclomethasone dipropionate. The intended dose per animal was 110 mg stanozolol, 25 mg estradiol and 10 mg beclomethasone. The experiment was performed on 20 male veal calves, 16 treated and 4 vehicle treated controls and 3 female veal calves, 2 treated and 1 vehicle treated control. Half of the animals were shaven prior to the application of the drugs. The cocktails were administered using two types of vehicles: vehicle A; Miglyol 840 with butylated hydroxytoluene and vehicle B; di(ethyleneglycol) monobutylether. During a 28 day treatment period, one group of animals was treated once a week, another group of animals was treated once every two weeks and slaughtered. Preliminary results showed that pour on application of anabolic steroids markedly increased growth performance of veal calves, the animals treated with cocktail A performed better than the animals treated with cocktail B. Macroscopically, the thymus was reduced in weight and size in the B animals. The bulbo-urethral glands were enlarged in all treated animals. Histologically all treated animals showed squamous metaplasia in the prostate, bulbo-urethral gland and Bartholins glands. Moreover, a changed secretion pattern was observed in both the prostate and the bulbo-urethral gland. Severe cortical atrophy was observed in the thymus and to a lesser extent the adrenals of the beclomethasone treated animals. The recently discovered 16 beta-hydroxy-metabolite of stanozolol was detected in urine, in relatively high concentrations. This is the first report of the excretion of this metabolite in urine after pour on administration showing the prospect for detection of dermal treatment. Estradiol levels were remarkably elevated (up to 200 micrograms l-1) exceeding the endogenous levels (< 1 microgram l-1).

Administration, Topical↗

Combinations of growth promoters in veal calves: consequences for screening and confirmation methods.

This study investigates the influence of low dosages of beta-agonists combined with other growth promoters on screening and confirmation methods in male veal calves. Five groups of four calves were treated for 6 weeks with combinations of low dosages of beta-agonists (beta AG, clenbuterol, mabuterol and mapenterol, 0.4 microgram/kg each twice daily) in combination with 17 beta-estradiol (E2, 5 mg per animal per 14 days), methylthiouracil (MTU, 2.857 mg/kg bw twice daily) and dexamethasone (DEX, 4 mg per animal per 10 days during the last 20 days of treatment). Another group of four untreated animals served as controls. The weight and size of the thymus was reduced in the DEX group, the MTU group showed enlarged thyroids. Histologically the prostates showed vacuolar degeneration in the beta AG animals and metaplasia in the E2 group. Some vacuolization was also observed in the controls. The testis showed impaired development in all treatment groups, E2 leading to the most severe changes. DEX led to cortical atrophy of the thymus. MTU induced hyperplastic changes in the thyroid. These results indicate that comedication does not markedly affect the histological changes induced in the hormonal target tissues. For screening purposes histology of the prostate can be used as a marker for oestrogens, whereas the weight of the thymus and thyroid can be used as an indication for the use of corticosteroids and thyreostatics, respectively. Vacuolization in the prostate appeared no reliable indication for beta-agonists. Samples of urine, faeces, liver and eye (retina/choroid) were analysed for beta-agonists. E2 significantly increased the levels of all beta-agonists in the liver and faeces, whereas DEX significantly reduced these levels. These observations show that additional medication with different groups of growth promoters can markedly alter the excretion of beta-agonists and thus influence (regulatory) control.

2-Hydroxyphenethylamine↗

Cognition in test-anxious children: the role of on-task and coping cognition reconsidered.

This study investigated cognition in children with high, moderate, and low test anxiety under naturalistic test-taking conditions. Children with high test anxiety not only reported more negative self-evaluations and more off-task thoughts but also more on-task and coping thoughts. Consistent with previous research, no "task-facilitating" role for these on-task and coping cognitions was found. This finding lends support to the notion of "the power of non-negative thinking". The only facilitating role was for positive self-evaluations. It was hypothesized that positive cognition could be task-facilitating under certain conditions. Limited support was found for this idea.

Adaptation, Psychological↗

Yellow discoloration in veal calves: the role of hepatic copper.

Liver samples from four groups of calves were analysed chemically and histologically for copper and iron levels. Milk replacer-fed 'yellow' calves were compared with milk replacer-fed 'white' calves, concentrate and silage-fed 'pink' calves and concentrate and silage-fed young 'red' fattening bulls. In the milk replacer-fed calves high copper and low iron levels were measured in the liver, whereas in the concentrate and silage fed pink calves and fattening bulls lower copper and higher iron levels were found. The yellow calves appeared to be icteric and had chronic hepatitis. Their hepatic histopathology was characterised by fibrosis, cirrhosis, fatty change, increased amounts of stainable copper, necrobiosis and prominent cholestasis; some animals had intranuclear inclusion bodies in the hepatocytes. They had similar or lower hepatic copper levels than the white calves and varying iron levels, indicating that copper toxicity was not the primary cause of the hepatic damage.

Animal Feed↗

Combining two independent indirect methods as a new possibility for screening the illegal use of growth promotants.

In this study the possibilities of combined indirect screening methods for the growth promotants clenbuterol and estradiol-17 beta benzoate and the combination of both these agents were investigated. Three groups of 5 male goat kids (two months of age) were treated for 4 weeks; clenbuterol per os, 10 micrograms/day/kg body weight via milkreplacer twice a day; 2 mg estradiol-17 beta benzoate weekly per i.m. injection; combined clenbuterol per os+estradiol-17 beta benzoate per injection. Five animals were not treated and served as controls. Nuclear Magnetic Resonance (NMR) was used as an initial tool to monitor the urine composition. The analysis of NMR spectra showed, that most treated animals had increased L-lactate/creatine ratios (L/C-ratio) in their urine, compared to control animals. For screening, L-lactate/creatine ratios could be determined directly in urine--in a simple, quick and inexpensive manner--by using enzymatic methods and an autoanalyzer. Histologically clenbuterol induced degenerative changes in the urethral and glandular epithelium of the prostate. Changes consisted of vacuolization of the epithelium and necrosis with nuclear pyknosis and fragmentation. Estradiol-17 beta benzoate induced metaplastic changes in the glandular tissue of the goat prostate. In combination with estradiol-17 beta benzoate, characteristics of both agents could be observed. The histological methods and observed effects can be used directly for screening. The combination of independent histological and L/C-ratio methods looks very promising with respect to screening of growth promotants. Since in both cases effects of growth promotants are detected, it may be expected that this approach is not limited to goats or clenbuterol and estradiol alone.

Animals↗

Histological changes in the genital tract of female veal calves implanted with naturally occurring anabolic steroids.

The influence of implants composed of estradiol and testosterone on the genital tract of female veal calves was studied. Striking results were the metaplastic and hyperplastic lesions seen in Bartholin's glands (an estrogenic effect) and a preputial-like separation of the clitoral epithelium (an androgenic effect). Other changes were less pronounced or resembled those found in estrous animals. Similar though less distinct histological changes were observed in the genital tract of animals from a farm with a history of illegal administration of anabolic steroids suggesting illegal treatment. From these findings it is concluded that histological screening of Bartholin's glands for estrogenic influences and of the clitoris for androgenic activity can be useful in tracing illegal anabolic treatment of female veal calves with combination preparations.

Animals↗

[Histologic hormone studies, a practical evaluation].

Based on histological studies of the prostate or Bartholin's gland, chemical analysis of the urine, injection site or implant was conducted in 114 veal calves suspected of receiving hormonal treatment. In 65% of the samples the results of chemical analysis confirmed the histological findings.

Anabolic Agents↗

Influence of androgens on the genital tract of cyclic heifers.

Cyclic heifers were implanted with the synthetic androgen trenbolone acetate (TBA = 17 beta-hydroxy-19-norandrosta-4, 9, 11-trien-3-on-17 beta-acetate, C2OH2403) (for formulas see Fig. 1). The influence of this androgen on the macroscopic and microscopic morphology of the genital tract was studied. The most striking result was the induction of polycystic ovaries. The clitoris was markedly enlarged and the cervix showed an increased amount of mucus. Microscopically, extensive folding of the cervical epithelium, consisting of tall swollen columnar cells with basal nuclei was seen. The vagina showed an increase of PAS-positive granules in the superficial layer of the epithelium, while there was no increase in the height of the epithelium. These effects are due to androgenic influence on the female genital tract and are in no case specific to one particular product. They could be of help in the detection of the illegal use of agents with androgenic activity.

Androgens↗