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We determined changes in plasma hormone concentrations in gilts after treatment with a progesterone agonist, Altrenogest (AT), and determined the effect of exogenous gonadotropins on ovulation and plasma hormone concentrations during AT treatment. Twenty-nine cyclic gilts were fed 20 mg of AT/(day X gilt) once daily for 15 days starting on Days 10 to 14 of their estrous cycle. The 16th day after starting AT was designated Day 1. In Experiment 1, the preovulatory luteinizing hormone (LH) surge occurred 5.6 days after cessation of AT feeding. Plasma follicle-stimulating hormone (FSH) increased simultaneously with the LH surge and then increased further to a maximum 2 to 3 days later. In Experiment 2, each of 23 gilts was assigned to one of the following treatment groups: 1) no additional AT or injections, n = 4; 2) no additional AT, 1200 IU of pregnant mare's serum gonadotropin (PMSG) on Day 1, n = 4); 3) AT continued through Day 10 and PMSG on Day 1, n = 5, 4) AT continued through Day 10, PMSG on Day 1, and 500 IU of human chorionic gonadotropin (hCG) on Day 5, n = 5; or 5) AT continued through Day 10 and no injections, n = 5. Gilts were bled once daily on Days 1-3 and 9-11, bled twice daily on Days 4-8, and killed on Day 11 to recover ovaries. Termination of AT feeding or injection of PMSG increased plasma estrogen and decreased plasma FSH between Day 1 and Day 4; plasma estrogen profiles did not differ significantly among groups after injection of PMSG (Groups 2-4). Feeding AT blocked estrus, the LH surge, and ovulation after injection of PMSG (Group 3); hCG on Day 5 following PMSG on Day 1 caused ovulation (Group 4). Although AT did not block the action of PMSG and hCG at the ovary, AT did block the mechanisms by which estrogen triggers the preovulatory LH surge and estrus.

The ovarian cycles of 48 culled dairy cows were monitored by assaying plasma progesterone concentrations. Twenty-four cows received a subcutaneous implant of 300 mg trenbolone acetate (Finaplix; Hoechst) at the beginning of the study. Of the implanted cows, two were pregnant, six continued to cycle although their peak progesterone concentrations were significantly lower than in control cows (5.65 +/- 0.71 compared with 8.19 +/- 0.47 ng/ml; P less than 0.01). Prolonged periods (13 to 92 days) of low progesterone concentrations (less than 1 ng/ml) occurred in 12 of the implanted cows. In six of seven cows in which normal cycles had not resumed by the time of slaughter two to five ovarian follicles of diameter greater than or equal to 10 mm were found post mortem. Persistent luteal function (greater than 35 days) occurred in the absence of pregnancy or gross uterine pathology in five of the implanted cows, two of which had recovered spontaneously by the time they were slaughtered. There was no difference in plasma luteinising hormone concentrations between the implanted and control cows. It is concluded that trenbolone acetate affected the ovarian cycle of the cows in several ways through changes other than the modification of tonic luteinising hormone secretion.

The effects of the anabolic steroid trenbolone acetate (TBA) on adrenal function and hepatic enzyme activity have been studied in growing female rats. Treatment with TBA resulted in a decrease in the peak of plasma corticosterone concentration which occurred during the afternoon. The enzymes tyrosine aminotransferase (TAT) and phosphoenol-pyruvate carboxykinase were measured in the livers of treated and control rats. Activities of both enzymes were maximal at 22.00 h, but that of TAT was reduced in TBA-treated rats. After injection of ACTH, TBA-treated rats showed a smaller increase in plasma corticosterone than did controls. Treatment with TBA did not affect the induction of TAT activity after corticosterone treatment. All TBA-treated rats grew significantly faster than controls and the possible relevance of this reduced adrenal function to the increased growth rate is discussed.

Two trials were conducted to evaluate the effectiveness of a synthetic progestogen, allyl trenbolone, for control of estrus and ovulation in gilts. Estrus was effectively synchronized with 33 of 38 gilts exhibiting estrus 2 to 7 days after last progestogen feeding. There was no effect on fertilization rate in gilts free of ovarian abnormalities. Ovulation rate was higher in gilts fed the progestogen in both Trial 1 (P less than .01) and Trial 2 (P less than .10). Progestogen treatment caused an increase (P less than .01) in the incidence of cystic follicles which resulted in a decrease (P less than .01) in conception rate in Trial 1.

A synthetic progestogen (altrenogest) was fed to 24 Yorkshire X Duroc boars to determine effects on body growth, serum testosterone, and testicular characteristics. Boars from six litters (blocks) were allotted randomly to one of four treatment groups at 12 wk of age. Treatment groups were: controls, altrenogest fed for 3 wk, altrenogest fed for 6 wk and altrenogest fed for 9 wk. Treatment began at 15 wk of age at a daily dose of 20 mg X boar-1. Although there were no differences among treatment groups for gain and feed intake, boars fed altrenogest for 6 and 9 wk tended to consume more feed and were less (P less than .05) efficient than boars fed altrenogest for 3 wk or controls. Boars fed altrenogest for 3, 6 and 9 wk maintained lower (P less than .05) peripheral serum testosterone concentrations than controls from 15 to 24 wk of age. However, serum testosterone increased after altrenogest withdrawal in the 3- and 6-wk treatment groups but did not reach control concentrations by 24 wk. Boars fed altrenogest for 9 wk maintained serum testosterone below 1 ng/ml during the treatment period. Despite lower concentrations of serum testosterone in altrenogest-treated boars, backfat thickness was similar to controls, perhaps suggesting a slight anabolic effect of altrenogest on nutrient partitioning. Testicular weights and volumes at 24 wk decreased (P less than .001) linearly with increased duration of altrenogest feeding. Serum testosterone and estradiol in testicular venous effluent at castration were lower (P less than .01) in altrenogest-treated boars than in controls. These data demonstrate that feeding altrenogest inhibits both testicular growth and steroidogenesis of boars without altering body growth or backfat thickness.

Crossbred (Chester White X Yorkshire X Duroc) boars were used to evaluate the effects of feeding a progestogen (altrenogest) on body growth, endocrine function (determined during feeding and after withdrawal of altrenogest), carcass composition, boar odor and testicular function (determined after a 30-d withdrawal from altrenogest). Boars from 18 litters were assigned at 12 wk of age to three treatments: 1) 18 control boars; 2) 18 boars fed altrenogest (20 mg/day) for 6 wk from 15 to 21 wk of age, followed by 30 d with no treatment; and 3) 18 boars castrated at 2 wk of age (barrows). Daily gains were greater (P less than .05) in boars fed altrenogest than in barrows through 21 wk of age but were lower (P less than .05) than those of control boars and barrows during the 30-d withdrawal period. Boars fed altrenogest weighed less (P less than .05) than control boars and barrows at 25 wk of age (at slaughter). Both groups of boars were similar in percentage of muscle and had less (P less than .05) backfat than barrows, whereas control boars had the largest (P less than .05) loineye areas. Based on evaluations by a trained sensory panel, intensity of boar odor in fat samples was similar for both groups of boars and was greater (P less than .05) than that for barrows. Weights of accessory reproductive glands and weight and sperm content of testes and epididymides were reduced (P less than .05) in boars fed altrenogest.(ABSTRACT TRUNCATED AT 250 WORDS)

The ability of zeranol and trenbolone acetate (trenbolone) to alter testis function, weight gain and carcass traits of young bulls was studied. In Exp. 1, the effects of age at initial zeranol implantation was determined. After a 235-d experimental period, sequential implantation (56-d intervals) beginning at 100 or 150 d of age had reduced testis growth (P less than .01), sperm production (P less than .01) and serum testosterone concentration in response to gonadotropin releasing hormone (GnRH; P less than .01). The 200-d age group was partially suppressed, while the 250-d age group was not affected. Body weights were similar to controls in all groups. In Exp. 2, bulls previously implanted with zeranol at 175 and 231 d of age received single implants of zeranol, trenbolone or trenbolone plus zeranol at approximately 300 d of age. At slaughter (135 d later), body weight and carcass characteristics in all treatments were similar to controls. However, trenbolone reduced sperm production (P less than .05), zeranol reduced sperm production and testes weight (P less than .05), but trenbolone plus zeranol was similar to controls. Mean testosterone response to GnRH was suppressed in all implant groups on d 65 (P less than .01), but only in trenbolone or trenbolone plus zeranol groups on d 112 (P less than .05). Results indicate that zeranol suppresses spermatogenesis and testosterone production if implanted before approximately 200 d of age. Reduction of endogenous testosterone without alteration of weight gain or carcass characteristics may be of benefit if behavioral or masculinity traits of bulls are altered. Also, it appears that no benefit is derived from implanting bulls with both trenbolone and zeranol.

In vivo covalent binding to rat liver DNA was assayed with 2 major metabolites of trenbolone, alpha-OH-trenbolone (alpha-TB) and beta-OH-trenbolone (beta-TB), in male Sprague-Dawley rats. DNA purified by ultracentrifugation in cesium trifluoroacetate showed protein contamination less than 0.4% of DNA by weight. The mean results obtained for the covalent binding indexes were 2.4 +/- 0.3 for alpha-TB and 5.3 +/- 2 for beta-TB. The covalent binding of alpha-TB was linear as a function of dose, which was not the case for beta-TB. The higher affinity of beta-trenbolone for DNA contaminating proteins was not sufficient to explain the difference. When these results are compared with the genotoxicity tests already performed with trenbolone by several authors (almost all completely negative), it may be concluded that the very low levels of binding could not be regarded as responsible for a direct genotoxic action of trenbolone.

The current methods for the control of the illegal use of anabolics in cattle breeding are far from satisfactory in terms of specificity and sensitivity. Here we describe a method for the simultaneous detection of diethylstilbestrol, zeranol and trenbolone in plasma and edible cattle tissues. The method is rapid (total analysis time 30 min), specific and sensitive (ppt level). The technique employed is a tandem mass spectrometric method: collisionally activated decomposition mass-analysed ion kinetic energy (CAD MIKE) spectrometry. Detection involves the injection of the crude ethanol extracts of plasma and tissues and selection/collision of molecular ions of each targeted compound. Confirmation of the presence of particular compounds is made by matching the complete CAD spectra so obtained with those of the pure compounds.

The influence of varying doses of human chorionic gonadotropin (hCG) on the preovulatory luteinizing hormone (LH) surge, estradiol-17 beta (E2) and progesterone (P4) was studied in synchronized gilts. Altrenogest (AT) was fed (15 mg X head-1 X d-1) to 24 cyclic gilts for 14 d. Pregnant mares serum gonadotropin (PMSG; 750 IU) was given im on the last day of AT feeding. The gilts were then assigned to one of four groups (n = 6): saline (I), 500 IU hCG (II), 1,000 IU hCG (III) and 1,500 IU hCG (IV). Human chorionic gonadotropin or saline was injected im 72 h after PMSG. No differences in ovulation rate or time from last feeding of AT to occurrence of estrus were observed. All gilts in Groups I and II expressed a preovulatory LH surge compared with only four of six and three of six in Groups III and IV, respectively. All groups treated with hCG showed a rapid drop (P less than .01) in plasma levels of E2 11, 17, 23 h after hCG injection when compared with the control group (35 h). The hCG-treated gilts exhibited elevated P4 concentrations 12 h earlier than the control group (3.1 +/- .5, 3.4 +/- .72, 3.1 +/- .10 ng/ml in groups II, III and IV at 60 h post-hCG vs .9 +/- .08 ng/ml in group I; P less than .05). These studies demonstrate that injections of ovulatory doses of hCG (500 to 1,500 IU) had three distinct effects on events concomitant with occurrence of estrus in gilts: decreased secretion of E2 immediately after hCG administration, failure to observe a preovulatory LH surge in some treated animals and earlier production of P4 by newly developed corpora lutea.

The ultrastructural changes in hepatocytes of steers were studied in animals implanted with 140 mg trenbolone acetate, alone or in combination with 20 mg oestradiol. The mean volume of the nucleus in the hepatocytes of the animals implanted with trenbolone acetate alone was significantly greater than that in control animals (P less than 0.001) and in animals implanted with the combined preparation (P less than 0.05). It is suggested that the increase indicates an androgen-mediated response in the nuclei which is not seen when oestradiol is included in the implant. Similarly, the peroxisome volume density was elevated significantly above that in the controls by trenbolone acetate (P less than 0.001); although the inclusion of oestradiol in the implant significantly reduced this level (P less than 0.001) it was still greater than the control values (P less than 0.05). No mitochondrial abnormalities were observed.

A technique of high-performance thin-layer chromatography (HPTLC plates) by which to identify the anabolic substance of trenbolone acetate (TBA) and its metabolite trenbolone is described in this paper. Plasma, bile, urine, faeces, liver, and meat can be used for testing. The sensitivity of the method is 1 ng. Additional reliability criteria are enumerated. Semiquantitative TBA determination has proved to be suitable in random sampling for residual analysis.

This experiment was conducted to compare the negative effects of charcoal-extracted porcine follicular fluid (pFF) and the positive effects of purified porcine follicle-stimulating hormone (pFSH) on growth of follicles and on plasma hormone concentrations. Twenty gilts were fed altrenogest for 18 days (20 mg.day-1.gilt-1) to suppress spontaneous growth of large follicles (greater than 6 mm in diameter). Gilts, assigned at random to receive pFF and pFSH administered in a 2 x 2 factorial arrangement, were injected 9 times at 8-h intervals starting 48 h before the last feeding of altrenogest and ending 8 h before slaughter (24 h after the last feeding of altrenogest). Blood was collected periodically through vena cava catheters. Treatment groups and mean number of medium follicles (3 to 6 mm in diameter)/gilt at necropsy were 1) 20 ml of charcoal-extracted porcine serum i.v. + 4 ml saline i.m., 30.8; 2) 20 ml of pFF i.v. + saline i.m., 0.2; 3) serum i.v. + 8 micrograms of pFSH (USDA-pFSH-B1)/kg BW in saline i.m., 59.0; and 4) pFF i.v. + pFSH in saline i.m., 36.2. Injections of pFF decreased (p less than 0.01) and injections of pFSH increased the number of medium follicles, and the interaction of pFF and pFSH was not significant. Plasma FSH decreased (p less than 0.01) during pFF treatment of saline-injected gilts at a rate of 0.29 ng.ml-1.h-1. During pFSH treatment, plasma FSH increased (p less than 0.05) at statistically identical rates of 0.33 and 0.32 ng.ml-1.h-1 in serum- and pFF-injected gilts.(ABSTRACT TRUNCATED AT 250 WORDS)

Two heifers were implanted with 300 mg of the radiolabeled anabolic steroid, trenbolone acetate (TBA). After a 60 day slaughter and a 60 day removal followed by 76 day slaughter, total 3H-content in various tissues was 0.5--25 ng/g equivalents of TBA. Radioimmunoassay of the tissues showed that only 1--5% of the total residue present was TBA, its main metabolite trenbolone (TBOH), and TBOH glucuronide, plus up to 5% of other organic-soluble material. Of the radioactivity remaining about half was directly water-soluble, and the insoluble residue could be made water-soluble by treatment with the proteolytic enzymes pepsin and trypsin. These 2 portions were purified with Sephadex G-25 to give a low and high molecular weight fraction. Raney nickel reduction of sulfur bonds in either fraction released up to 50% of radioactivity into the organic phase. TLC showed that the latter contained 2 components which had characteristics similar to TBOH and its metabolites, and thus were at least partly drug-related metabolites. In vitro experiments with bovine liver also showed a small but definite protein binding. It is proposed that in dealing with these covalently bound residues, priority be given to the reactive drug intermediate and the type of binding to macromolecules rather than to the presence of the bound residue itself.

The effects of the nature of energy intake, of the slaughter weight and of the use of hormones on the quality of veal meat were studied. Concerning the colour of meat, the use of hormones has no effect either on the pigment content or on the ultimate pH, but the nature of energy intake and the slaughter weight can affect these two parameters. With the increase of the slaughter weight and the substitution of lipids by starch in the diet, the probability to get a redder meat increases. For a given carcass weight, the use of hormones goes with a significant decrease of the tenderness of meat.

Fertility of 104 gilts artificially inseminated (AI) at a predetermined time (scheduled AI) after estrous synchronization with altrenogest (15 mg X gilt-1 X d-1 for 18 d) was compared with that of 103 gilts checked for estrus (estrus checked) and inseminated after altrenogest. Scheduled-AI gilts were inseminated once on d 5, 6 and 7 after the last altrenogest feeding (d 0). Estrus-checked gilts were exposed to a boar twice daily at 0830 and 1630 h and inseminated after the second and third estrous detection period following first detected estrus. Percentage of gilts assigned to treatment that farrowed (72.8 vs 67.3%), total pigs farrowed (11 +/- .4 vs 11.3 +/- .4) and pigs born alive (10.1 +/- .4 vs 10.5 +/- .4) were similar for estrus-checked and scheduled-AI gilts, respectively. We conclude that scheduled AI can be used with estrous synchronization for gilts and may have advantages in breeding herd management and the use of AI in swine.

Altrenogest (ALT), a synthetic progestogen, and exogenous gonadotropins were administered to the Swine Leukocyte Antigen (SLA) inbred miniature pig that typically has a reduced ovulation rate and small litter size. In Study 1, 15 adult female pigs were either controls (Group I, n = 5), fed 15 mg ALT/day for 14 days (Group II, n = 5), or fed ALT and treated with pregnant mare's serum gonadotropin (1200 IU, i.m.) and human chorionic gonadotropin (500 IU, i.m.) 24 h and 104 h, respectively, after ALT withdrawal (Group III, n = 5). In Study 2, three groups of pigs (I, n = 24; II, n = 11; III, n = 11) were treated similarly to those in Study 1 except that pigs in Study 2 (Groups II and III) were mated. Ovaries of all animals in both studies were examined laparoscopically or by laparotomy 5-8 days after estrus to assess ovarian activity. The uteri of Group II and III animals in Study 2 were flushed with tissue culture medium, which was evaluated for the presence and quality of embryos. No pig demonstrated estrous behavior during ALT administration. Not all animals demonstrated typical estrous behavior after ALT withdrawal. When the data from both studies were combined, 2 of 16 (12.5%) of group II and 6 of 16 (37.5%) of Group III pigs failed to demonstrate standing estrus. Following ALT withdrawal, females in Groups II and III returned to estrus in 5.6 +/- 0.3 and 5.8 +/- 0.3 days, respectively (p greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Lactating mares were assigned as controls or fed altrenogest (.044 mg.kg body wt-1.d-1) for 15 d after foaling. Mares (n = 6) fed altrenogest were inseminated during the first estrus after treatment and mares (n = 6) in the control group were inseminated during the second postpartum estrus. Ovulation during the estrus in which mares were inseminated occurred 26 +/- 1 d postpartum for treated mares and 36 +/- 1 d postpartum for control mares. The percentage of mares conceiving was not different for control (67%) and alternogest-treated (100%) mares. No differences were observed in tone and size of the uterus or size of the ovulatory follicle between treated and control groups. Uterine cultures and biopsies collected on d 7 and 15 postpartum were similar between treatment and control groups in bacterial populations or endometrial epithelial cell height. Blood was collected on d 7, 11, 15, 19 and 23 postpartum, and concentrations of estradiol-17 beta in serum were determined by radioimmunoassay. Mean concentrations of estradiol-17 beta across days were 10 +/- .8 and 12 +/- .6 pg/ml for control and treated mares, respectively. Concentrations of serum estradiol-17 beta were higher (P less than .05) in treated mares on d 23 postpartum. Daily milk yields, determined by the weigh-suckle-weigh method, and milk composition were similar between treatment groups on each collection day. Altrenogest can be used to predictably delay estrus in the postpartum mare without altering fertility, yield and composition of milk, or foal growth.

Twenty wether sheep were allocated to seven groups and received implants near the base of one ear with pellets containing: for group 1, (OE) 20 mg oestradiol-17 beta alone; for group 2, (TBA/OE) 20 mg oestradiol-17 beta intimately mixed with 140 mg trenbolone acetate; for group 3, (T/OE) 20 mg oestradiol-17 beta mixed with 200 mg testosterone; for group 4, (P/OE) 20 mg oestradiol-17 beta mixed with 200 mg progesterone; for group 5, (TBA/OE2) 20 mg oestradiol-17 beta in one ear and 140 mg trenbolone in the other ear; for group 6, (TBA/OE1) 20 mg oestradiol-17 beta and 140 mg trenbolone as separate pellets in one ear; group 7 sheep received implants of carrier material and served as controls. The concentrations of steroids were measured in plasma samples collected from both jugular veins during the 16-week period after implantation. The absorption of oestradiol-17 beta was slower and more sustained from the pellets in which it was mixed with other steroids (groups 2, 3 and 4) than from the pellets containing oestradiol-17 beta alone (groups 1, 5 and 6). The concentration of each steroid in plasma was higher in the jugular vein ipsilateral to the implant than in the vein on the opposite side. The difference between the concentrations in the two veins was used to calculate the biological half-lives of the steroids; for oestradiol-17 beta and trenbolone the mean values ranged from 1.8 to 6.8 min and from 3 to 4 min, respectively, and for testosterone and progesterone the mean values were 4.7 and 3.5 min, respectively.

1. The metabolism of trenbolone acetate, 17 beta-acetoxyestra-4,9,11-trien-3-one (TBA), an anabolic compound used as a growth promoter, was compared in rat and cow. 2. [6,7-3H] TBA was injected i.v. into rats and a heifer, and bile was collected for 24 h. In both species, the bile was the major route of excretion. TBA undergoes an extensive hydrolysis to 17 beta-hydroxyestra-4,9,11-trien-3-one and the unchanged compound was not detected, but subsequent major metabolic pathways are different in the two species. 3. In the rat, oxidation of the 17 beta-hydroxyl to the 17-oxo group and hydroxylation in the 16 alpha-position are the major routes. The three major metabolites are 17 beta-hydroxyestra-4,9,11-trien-3-one, 16 alpha, 17 beta-dihydroxyestra-4,9,11-trien-3-one and 16 alpha-hydroxyestra-4,9,11-trien-3, 17-dione. 4. In the heifer, 17 alpha-epimerization is the major pathway and the main metabolite is the 17 alpha-hydroxyestra-4,9,11-trien-3-one. 5. In both species, estra-4,9,11-trien-3,17-dione and the other metabolites, resulting either from hydroxylation in 1, 2, 6 beta, 16 alpha or 16 beta positions, or from aromatization of the A ring, were minor products. 6. Overall, 60% of the 3-oxotriene structures identified in the rat bile were 17 beta-hydroxylated and the remainder were 17-keto metabolites, whereas in the heifer bile 90% were 17 alpha-hydroxylated compounds. 7. Thus, in bovine species, the major pathway is similar to those of testosterone or 17 beta-estradiol which are mainly excreted as their 17 alpha-epimers. The epimerization strongly decreases the biological potency, as with the natural 17 beta-hormones, and leads to detoxication of tissue residues.

1. Tissue composition and skeletal muscle cathepsin D (EC3.4.23.5) activity weremeasured in wether lambs treated with trenbolone acetate (TBA) and oestradiol-17β (Oe) in combination and female lambs treated with TBA or zeranol. Muscle and liver protein fractional synthesis rates and plasma leucine flux were measured in the female lambs.2. Male castrate lambs treated with TBA plus Oe showed increased growth rate, improvedfood conversion efficiency, decreased muscle RNA concentration and decreased total cathepsin D activity in muscle.3. Female lambs treated with TBA or zeranol showed increased weight gain, improved food conversion efficiency, decreased muscle RNA and DNA concentrations and decreased free cathepsin D activity in muscle. Mixed muscle protein fractional synthesis rate was decreasedafter TBA treatment. Plasma leucine flux, not corrected for oxidation or food intake, wasnot increased by TBA or zeranol treatment.4. Treatment of female lambs with TBA or zeranol caused increased growth rate. This increased growth rate is probably due in part to decreased muscle protein degradation, since evidence was obtained that muscle protein synthesis is decreased by TBA and zeranol treatment.

Plasma disposition and urinary recovery of sulfamethazine (SMZ), its N4-acetylated metabolite (N4AcSMZ), and two of its hydroxylated metabolites [5-hydroxysulfamethazine (5OHSMZ) and 6-hydroxymethylsulfamethazine (6CH2OHSMZ)] were determined in male and female rats, in castrated males, and in rats pretreated with various steroid hormones. Male rats had a 2-fold higher SMZ plasma clearance than females, castrates, and males treated with flutamide (a testosterone antagonist). When castrated male rats were treated with testosterone or trenbolone, SMZ plasma clearance returned to normal values. Higher SMZ plasma clearance rates in the presence of androgens went together with higher urinary recoveries of the 6CH2OHSMZ metabolite. In hepatic microsomes of male rats lower apparent KM values, and higher Vmax values for 6CH2OHSMZ and 5OHSMZ formation were found than in microsomes of female rats. Castration or treatment of male rats with flutamide markedly reduced microsomal SMZ hydroxylation rates. Pretreatment of male or female rats with phenobarbital or triacetyl-oleandomycin had no effect on microsomal SMZ hydroxylation, whereas a continuous infusion with bovine somatotropin in male rats caused a marked decrease in SMZ hydroxylation rate. Finally, SMZ hydroxylation to 6CH2OHSMZ and 5OHSMZ in hepatic microsomes from male rats was strongly inhibited by monoclonal antibodies against P-4502C11. These results suggest that the male-specific P-4502C11 enzyme plays an important role in the hydroxylation of SMZ to 6CH2OHSMZ and 5OHSMZ in rats. SMZ seems a useful model compound to assess hormone effects on oxidative biotransformation (in vivo and in vitro) in rats.

Most studies related to research on steroids in main edible tissues (muscle, liver or kidney) have focused on measurement of parent or major metabolite residues. In order to evaluate the estradiol content in bovine edible tissues, a multi-step extraction procedure was developed in conjunction with parallel metabolism studies of [14C]-17beta-estradiol in cattle (1-2). Various classes of free estradiol and conjugates were separated: estradiol -17beta and -17alpha, estradiol-17-fatty acid esters, estradiol 17-glycoside, estradiol 3-glucuronide, estradiol-17-glycoside and 3- glucuronide (diconjugates) were separated. No sulphates conjugated forms have been found at the detection level of the method. The quantification was realized by calibration with deuterated 17beta -estradiol -d3 standard and was validated at the ng x kg(-1) (ppt) level. Muscle, liver, kidney and fat samples from control or Revalor S single (licensed implantation) or multi-implanted steers have been assayed. The results show a wide variation between animals, but both the highest value and the mean of total estradiol content in each group proportionally increase from untreated to multi-implanted animals. In accordance with international rules, a calculation of the daily food supply of estradiol by such edible tissues in comparison with the acceptable daily intake was performed.

Twenty-four gilts and 24 barrows were divided into two groups of 12 of each gender. One group was implanted with 300 mg of trenbolone acetate, and the other one was used as a control. Treatment had no effect on fat thickness in gilts but significantly reduced fatness in barrows. The fatty acid composition of the backfat showed a tendency toward greater unsaturation in treated barrows. Key words: Pig, trenbolone acetate, anabolic

The relative effectiveness of implanting 300 mg trenbolone acetate alone or in combination with either 15, 30 or 45 mg hexoestrol was studied in three 90-day experiments using 64 Friesian steers. In experiment 1 hexoestrol was shown to improve live-weight gain and efficiency of feed conversion in steers implanted with trenbolone acetate. In experiments 2 and 3 trenbolone acetate in combination with 30 mg hexoestrol gave a better growth response than when combined with either 15 or 45 mg. However, in experiment 3 trenbolone acetate plus 15 mg hexoestrol was shown to improve live-weight gain by about 36 per cent compared with untreated controls. In experiment 2 small differences between treated groups in mean values for plasma urea, serum albumin, plasma glucose and free fatty acids were recorded.

The development of a radioimmunoassay method for the detection of trienbolone is described. It is dependent on antibody raised against a bovine serum albumin conjugate of trienbolone 17-hemisuccinate in sheep.

In order to study the fate and residues of trenbolone acetate in edible tissues, two groups of six animals from two ruminant species (ewes and calves) were implanted with [3H]trenbolone acetate. The distribution of extractable radioactive residues was measured in liver, kidney and muscle. We found that the largest proportion of residues was not extractable and thus was considered as covalently bound residues. The proportion of the main extractable metabolites (17 alpha-trenbolone, trendione, 17 beta-trenbolone) was measured. The evaluation of the distribution of trenbolone acetate metabolites directly soluble in water showed that unknown metabolite(s) were predominant. The covalent binding to nucleic acids was measured. It was so low that it was not detectable. The results are discussed in light of the data presented in the scientific report on anabolic agents in animal production from the European scientific working group.