Fate and residues of trenbolone acetate in edible tissues from sheep amd calves implanted with tritium-labeled trenbolone acetate

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.     

Body weight and tissue gain in lambs fed an all-concentrate diet and implanted with trenbolone acetate or grazed on alfalfa

Targhee x Hampshire lambs (average BW 23 +/- 1 kg) were used in two experiments to determine the effects of finishing on concentrate with an anabolic implant or forage grazing after concentrate feeding on growth, organ and viscera weights, and carcass tissue accretion. In Exp. 1 and 2 lambs were penned by sex and assigned for slaughter at initial (23 kg), intermediate (37 kg), or end BW (ewes, 47.7; wethers 50.4 kg). From 23 to 37 kg BW, lambs were fed all-concentrate diets in drylot (DL) or grazed on alfalfa (ALF). Experiment 1 was a 2 x 2 factorial with 28 lambs; factors were wether vs ewe lambs and unimplanted vs DL implanted with trenbolone acetate-estradiol benzoate. There were no differences in organ and viscera weights due to implant status. However, ADG (P < .03) and lean gain (P < .02) were greater for implanted than for unimplanted wethers (507 vs 357 g and 1,314 vs 656 g, respectively). Ewes did not respond to the implant. Fat accretion was not affected by implantation. Experiment 2 was a 2 x 3 factorial with 42 lambs; factors were wether vs ewe lambs and drylot during growing and finishing phases (DL-DL) vs drylot during growing and alfalfa grazing during finishing (DL-ALF) vs alfalfa grazing during growing and finishing phases (ALF-ALF). In Exp. 2, ADG of DL-DL lambs was greater (P < .01) than ADG of DL-ALF or ALF-ALF lambs. Lambs on ALF-ALF had smaller (P < .05) livers and rumen/reticulum weights but heavier (P < .04) kidney, omasum, small and large intestine, and cecum weights than those on DL. In Exp. 2, DL-ALF and ALF-ALF lambs had overall hindsaddle lean gain equal to those on DL-DL with less mesenteric fat and 100 g less separable fat. Finishing lambs on alfalfa reduced fat accretion without decreasing lean accretion, whereas trenbolone acetate implants for lambs fed concentrate increased BW gain and lean accretion without affecting fat accretion.     

Growth and hormonal response of intact and castrate male cattle to trenbolone acetate and estradiol

Effects of castration and anabolic implants on weight gain, rib soft tissue composition and serum hormones were studied in cattle using a completely random design with a 2 x 2 factorial arrangement. Half of 16 bulls and 16 steers (Angus or Angus x Brahman) aged 9 mo and weighing 290 kg were treated with an implant (200 mg trenbolone acetate and 24 mg estradiol). Half of each group were not treated with an implant. A growing diet was fed for 95 d and half the animals in each group were slaughtered. Animals in the treated groups were reimplanted with trenbolone acetate and fed a finishing diet for 84 d and slaughtered. Percentage dry matter, fat and protein were determined on soft tissue from the 9-10-11th rib. Two blood samples were collected from each animal every 2 wk. Serum was assayed for five hormones. During the growing phase, untreated and treated bulls and treated steers gained more weight and had leaner rib sections that untreated steers (P less than .05); after the finishing phase, there were no differences among groups. Untreated steers had lower insulin-like growth factor (IGF-I) and higher cortisol concentrations during both phases of growth than untreated bulls did (P less than .05). Treatment with implants increased IGF-I concentrations in steers during both phases and reduced cortisol during the finishing phase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Administration of estradiol, trenbolone acetate, and trenbolone acetate/estradiol implants alters adipogenic and myogenic gene expression in bovine skeletal muscle

Twenty crossbred yearling steers (421 kg) were used to evaluate the effects of implanting with trenbolone acetate (TBA; 120 mg), estradiol-17β (E(2); 25.7 mg), and a combination (120 mg of TBA and 24 mg of E(2)) on adipogenic and myogenic mRNA concentrations. Animals were blocked by BW and within each block were assigned to 1 of 4 treatments. Animals were housed and fed in individual pens with 5 animals per treatment. All animals were weighed weekly, and muscle biopsy samples were taken from the LM of each steer on d 0 (before implantation), 7, 14, and 28. Total RNA was isolated from each sample and real-time quantitative PCR was used to measure the quantity of C/EBPβ, PPARγ, stearoyl CoA desaturase (SCD), myogenin, and 3 isoforms of bovine myosin heavy chain (MHC) mRNA. Total BW gain from the 28-d period was adjusted to d 0 by use of covariant analysis, and steers in the implant groups tended (P = 0.09) to have increased BW gain compared with nonimplanted control steers. Analysis of the gene expression of MHC showed that neither implant nor day (P > 0.20) had a significant effect on the expression of type I or IIX MHC mRNA There was also no treatment effect (P > 0.20) on MHC-IIA and myogenin, but increasing days on feed increased (P = 0.05) the expression of MHC-IIA mRNA. Relative mRNA abundance of C/EBPβ, PPARγ, and SCD increased (P < 0.05) during days of feed but PPARγ decreased (P < 0.05) with the treatment of combined TBA/E(2) implant. Results of this study indicate that implanting with TBA, E(2), or both increased BW gain and decreased adipogenic gene expression of finishing steers without significantly affecting the concentration of type I, IIA, or IIX MHC mRNA. Increasing days on feed increased both MHC-IIA and adipogenic gene expression in bovine skeletal muscle biopsy samples. We conclude that administration of steroidal implants had no effect on the proportion of the 3 MHC mRNA isoforms but decreased C/EBPβ, PPARγ, and SCD mRNA in bovine skeletal muscle.     

The effect of estradiol, trenbolone acetate, or zeranol on growth rate, mammary development, carcass traits, and plasma estradiol concentrations of beef heifers.

The effect of a single implantation (on d 1) with one or two long-acting, biodegradable estradiol implants (1E or 2E) on plasma estradiol concentrations in beef heifers was determined. The growth rates of these (2E) heifers, and of heifers repeatedly implanted with trenbolone acetate (TBA) or zeranol (Z) on d 1, 84, 168, and 252 of the trial, were compared to growth rates of controls. Trenbolone acetate alone was compared to TBA + 2E, and 2E was compared to 1E. At a mean age of 84 d (d 1 of experiment), 81 Hereford x Friesian heifers were allocated at random to the following treatments: Control (n = 15); TBA (n = 15); 1E (n = 12); 2E (n = 15); Z (n = 13); or TBA + 2E (n = 11). Mean live weight (kg) prior to slaughter on d 368 and hot carcass weight (kg) for heifers assigned to treatment Groups 1 to 6, respectively, were 366 and 200, 391 and 212, 374 and 201, 386 and 207, 387 and 210, and 391 and 208 (residual SD = 30.3 and 20.2). Heifers assigned to both the 2E and Z treatments were heavier on d 368 (P less than .05) and had longer teats on d 279 (P less than .05), less pelvic fat (P less than .05), and heavier kidneys (P less than .005) than control heifers. Heifers assigned to the TBA treatment had shorter teats on d 279 (P less than .001) but greater final live weight (P less than .05) and carcass weight than control heifers. Heifers given TBA alone had more pelvic fat (P less than .05) and lighter kidneys (P less than .05) than those given TBA + 2E. Mean estradiol concentrations in both the ipsilateral and contralateral jugular veins of heifers assigned to the 2E and TBA + 2E treatments, and in the ipsilateral jugular veins of heifers given 1E, were greater (P less than .05) than those in control heifers; concentrations did not decline during the experiment.     

Feedlot performance of beef heifers implanted with Synovex-H: effect of melengestrol acetate, ovariectomy or active immunization against GnRH

The effects on anabolic steroid implantation on feedlot performance and carcass composition and quality were examined in control (CNTRL), ovariectomized (OVX) or melengestrol acetate-fed (MGA) beef heifers or heifers actively immunized against GnRH (Anti-GnRH). Heifers (n = 112) were assigned randomly to a 2 x 4 factorial experiment. The two classes were made up of heifers not implanted and those implanted with Synovex-H. The four treatments were 1) CNTRL, 2) MGA, 3) OVX and 4) Anti-GnRH. Heifers were housed in individual pens and fed a high-energy diet for the 4-mo study. Synovex-H increased final live weight (P less than .005), carcass weight (P less than .005), ADG (P less than .0001) and feed efficiency (P less than .005) but did not alter carcass quality and yield grade (P greater than .05). Synovex-H increased deposition of protein (P less than .0001) and reduced deposition of fat (P less than .0001). Oral administration of MGA had no significant effect on feedlot performance or carcass quality. For heifers not implanted, active immunization against GnRH, but not ovariectomy, depressed ADG (P less than .05) and increased fat deposition (P less than .05) while reducing protein deposition (P less than .05). These effects of active immunization were reversed by concurrent administration of Synovex-H. Feedlot performance and carcass composition of heifers were improved by administration of anabolic steroids. When heifers were housed singly, neither ovariectomy, active immunization against GnRH nor oral administration of MGA improved feedlot performance of heifers implanted with Synovex-H.     

Skeletal muscle protein metabolism and serum growth hormone, insulin, and cortisol concentrations in growing steers implanted with estradiol-17 beta, trenbolone acetate, or estradiol-17 beta plus trenbolone acetate.

Skeletal muscle protein degradation, measured by urinary N tau-methylhistidine excretion, and circulating concentrations of growth hormone (GH), insulin (INS), and cortisol (CT) were monitored in steers before and after implantation with estradiol-17 beta (E2; 24 mg) and trenbolone acetate (TBA; 300 mg). Yearling crossbred steers (n = 43) were randomly assigned to four treatment groups in a 2 x 2 factorial arrangement: nonimplanted controls (C); TBA; E2; and TBA plus E2 (TBA+E2). A subgroup (Block 1) of 16 steers was bled on d -12, 31, and 72 after implanting. Deposition of skeletal muscle protein was markedly increased (P less than .001) by E2 and TBA+E2 treatment. This response occurred mainly within the first 40 d after implantation and declined (P less than .001) in concert with decreasing (P less than .01) concentration of serum E2. Anabolic steroid treatment did not affect the rate of skeletal muscle protein breakdown. There was no apparent relationship between reduced serum CT concentration (linear effect; P less than .01) in TBA-treated steers and skeletal muscle protein degradation rate. Blood concentration and pulse activity of INS were not affected by anabolic steroid administration. Both TBA- and TBA+E2-implanted steers displayed a linear decrease (P less than .05) in serum GH concentration over time, which was similar to C. Lowered mean GH concentration resulted from a reduction (TBA main effect; P less than .05) in pulse amplitude of GH. Unlike TBA, TBA+E2, and C, only E2 maintained serum GH concentrations over time. Although increased muscle protein deposition was evident in TBA+E2-treated steers, an obvious causal relationship between this response and circulating GH, INS, and CT was not revealed. These results do not support the concept that combined androgenic agent and estrogen administration effectively reduce bovine muscle protein degradation by static modulation of circulating endogenous anabolic and antianabolic hormones.     

The use of a progesterone-releasing device (CIDR-B) or melengestrol acetate with GnRH,LH, or estradiol benzoate for fixed-time AI in beef heifers

The objective of this experiment was to compare two progestins and three treatments for synchronizing follicular wave emergence and ovulation in protocols for fixed-time AI in beef heifers. On d 0 (beginning of the experiment), Angus and Angus-Simmental cross beef heifers at random stages of the estrous cycle either received a CIDR-B device (n = 257) or were started on 0.5 mg x anima(-1) x d(-1) melengestrol acetate (MGA; n = 246) and were randomly assigned to receive i.m. injections of 100 microg GnRH, 12.5 mg porcine LH (pLH), or 2 mg estradiol benzoate (EB) and 50 mg progesterone (P4). The last feeding of MGA was given on d 6 and on d 7, CIDR-B devices were removed and all heifers received 500 microg cloprostenol (PG). Consistent with their treatment groups on d 0, heifers were given either 100 microg GnRH or 12.5 mg pLH 48 h after PG (and were concurrently inseminated) or 1 mg EB 24 h after PG and were inseminated 28 h later (52 h after PGF). Estrus rate (combined for both progestins) in heifers receiving EB (92.0%) was greater (P < 0.05) than that in heifers receiving GnRH and pLH (combined) and a CIDR-B device (62.9%) or MGA (34.3%). Although the mean interval from PG treatment to estrus did not differ among groups (overall, 47.8 h; P = 0.85), it was less variable (P < 0.01) in MGA-fed heifers (SD = 2.5 h) than in CIDR-B-treated heifers (SD = 8.1 h). Pregnancy rates (determined by ultrasonography approximately 30 d after AI) did not differ (P = 0.30) among the six treatment groups (average, 58.0%; range, 52.5 to 65.0%). Although fixed-time AI was done, pregnancy rates were greater in heifers detected in estrus than in those not detected in estrus (62.6 vs 51.9%; P < 0.05). In conclusion, GnRH, pLH, or EB treatment in combination with a CIDR-B device or MGA effectively synchronized ovulation-for fixed-time AI, resulting in acceptable pregnancy rates in beef heifers.     

Use of trenbolone acetate and estradiol in intact and castrate male cattle: effects on growth, serum hormones, and carcass characteristics

The effects of anabolic implant on growth, carcass characteristics, and serum hormones were examined in 30 young bulls and steers fed a growing diet then a finishing diet. In a 2 X 3 factorial arrangement, steers and bulls received an implant of trenbolone acetate (TBA), TBA and estradiol-17 beta (E2), or no implant. Blood samples were taken serially (every 20 min for 6 h) at intervals during the growing and finishing phases. Percentage of DM, fat, protein, and ash and Warner-Bratzler shear test were measured and taste panel evaluations of the 9-10-11 rib section were obtained. Treatment with TBA and E2 increased weight gain in steers but not in bulls. There were no differences in feed efficiency, serum growth hormone (GH), and cortisol concentrations between bulls and steers or between treated groups and controls in bulls or steers, although during the finishing phase mean GH concentrations in treated steers were twofold higher than in controls and were similar to those in the bull groups. Serum insulin-like growth factor-I (IGF-I) increased twofold during the growing phase, then remained at that level. Steers implanted with TBA and E2, which had the highest gains among the steer groups, had the highest serum GH and IGF-I. Longissimus steaks from bulls treated with TBA alone or TBA and E2 were comparable to steaks from steers in the shear test. Taste panelists found steaks from TBA- and E2-treated bulls to be similar in tenderness and connective tissue to steaks from steers.(ABSTRACT TRUNCATED AT 250 WORDS)     

A simple semiquantitative determination of trenbolone acetate and trenbolone in biological material from farm animals

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.     

Lipogenesis and adipose tissue cellularity in steers switched from alfalfa hay to high concentrate diets

An experiment was conducted to 1) evaluate the effects of diet (alfalfa hay vs high concentrate) on adipose tissue cellularity and rates of in vitro lipogenesis and 2) determine if there was a relationship between in vitro lipogenic rates from acetate and lactate and rates of L- or D-lactate disappearance from plasma. Number of adipose cells/g of tissue decreased with time on experiment; however, hay-fed steers had fewer, but larger cells/g of subcutaneous adipose tissue compared with concentrate-fed steers (.78 +/- .04 vs 1.20 +/- .13 X 10(-6)/g, respectively). These results, however, are likely due to a higher (approximately 25%) intake of dry matter and metabolizable energy by the hay-fed steers. Carcass data obtained at slaughter (460 kg) indicated that the concentrate-fed steers had as much or more adipose tissue compared with the hay-fed steers. Characteristics describing D- or L-lactate disappearance from plasma were not highly correlated with lactate utilization for fatty acid synthesis. Utilization of acetate as a substrate for fatty acid synthesis in vitro was correlated (r = .64) with the rate of lactate utilization for fatty acid synthesis.     

Growth, carcass and palatability traits of intact males and steers implanted with zeranol or estradiol early and throughout life

This study was conducted to assess the impact of implanting intact beef males with protein anabolic agents at varying intervals throughout life. Ninety-six intact males were assigned to three implant treatments: 1) not implanted, 2) implanted at 9 wk of age, weaning and at 56-d intervals thereafter with a 36-mg zeranol implant or 3) estradiol implant at 9 wk of age and 68 d post-weaning. During the 118-d, post-weaning growing period, eight animals per treatment (one replication) were castrated. After a 114-d finishing period, cattle were slaughtered (average age of 13 to 14 mo). Feedlot performance, carcass and palatability data were obtained. Average daily gains and feed efficiency did not differ (P greater than .05) between zeranol and estradiol-implanted intact males. Regardless of implant treatment, steers had lighter carcass weights (P less than .05) and higher (P less than .01) quality grades than intact males. Implanting either intact males or steers with zeranol or estradiol resulted in higher (P less than .05) numerical yield grades. Quality grades were higher in zeranol-implanted cattle than the non-implanted or estradiol-implanted cattle. Intact males implanted with zeranol were similar in carcass fatness to zeranol-implanted steers. No differences (P greater than .05) in tenderness or connective tissue were detected. Implanting intact males early and throughout life with zeranol made them similar to steers in fatness, while estradiol implantation had few effects on carcass and palatability traits of intact males or steers.     

Induction of estrus in ovariectomized cows and heifers: effects of estradiol benzoate and gonadotropin releasing hormone

Three experiments were conducted with ovariectomized (OVX) cows and heifers to investigate potential neuroendocrine mechanisms controlling estrous behavior. In Exp. 1, 10 OVX cows were treated with either 125 micrograms estradiol benzoate and 10 cc saline (125 micrograms EB + SAL), 125 micrograms EB and 500 micrograms gonadotropin releasing hormone (125 micrograms EB + GnRH), 250 micrograms EB and 10 cc SAL (250 micrograms EB + SAL), 250 micrograms EB and 500 micrograms GnRH (250 micrograms EB + GnRH) or 500 micrograms EB and 10 cc SAL (500 micrograms EB + SAL) in a replicated 5 X 5 Latin-square design. During the 48 h following EB injection, 2-h observation blocks were alternated with 2-h non-observation blocks. During each 2-h observation block, 14 behavioral interactions were monitored. The percentage of cows in estrus was lower for cows receiving 125 micrograms EB as compared with those given the higher doses. However, the cows receiving 125 micrograms EB + SAL did not differ in their estrous response from those receiving 125 micrograms EB + GnRH. The interval from injection to the onset of estrus and the duration of estrus were similar for all treatments. In Exp. 2, 10 OVX heifers were subjected to the same treatments and observation procedures utilized in Exp. 1. The results of Exp. 2 were similar to those of Exp. 1. In Exp. 3, 10 OVX cows were treated with either 300, 600, 1,200, 2,400 or 4,800 micrograms EB in a replicated 5 X 5 Latin-square design.(ABSTRACT TRUNCATED AT 250 WORDS)     

Lipogenesis in acute and 48-hour cultures of bovine intramuscular and subcutaneous adipose tissue explants

In this study, the interactions among breed of cattle, adipose tissue site and specific incubation conditions were investigated. Subcutaneous and i.m. adipose tissues were obtained from 10 Angus and 9 Santa Gertrudis steers immediately postmortem. Adipose tissue explants were incubated acutely for 2 h immediately at slaughter or after being cultured 48 h with or without 1 mU/ml insulin and 30 mg/ml bovine serum albumin; the incorporation of 14C-labeled acetate and glucose (5 mM, plus 5 mM unlabeled lactate) into lipid fractions was measured. AT the same chronological age, Angus steers had a more youthful lean maturity score, higher USDA marbling score and higher USDA quality grade (P less than .05) than did carcasses from Santa Gertrudis steers. The lower marbling score of the Santa Gertrudis steers was paralleled by smaller i.m. adipocytes (P less than .05) relative to Angus steers. Pentose cycle reductase and NADP-malate dehydrogenase activities were greater in Angus i.m. adipose tissue than in Santa Gertrudis i.m. adipose tissue, which would provide more reducing equivalents (NADPH) and glycerol for fatty acid biosynthesis and triacylglycerol esterification. Correspondingly, Angus i.m. adipose tissue exhibited a greater rate of lipogenesis from acetate and glucose (P less than .05) than did Santa Gertrudis i.m. adipose tissue in acute incubations. The presence of insulin resulted in higher rates of lipogenesis from acetate in Angus s.c. adipose tissue than in Santa Gertrudis s.c. adipose tissue after 48 h of explant culture. These data indicate that i.m. and s.c. adipose tissues exhibit aspects of lipid metabolism unique to each tissue and suggest that breed-related differences in adipose tissues may explain the divergent responses to insulin observed in different laboratories.     

Increased luteinizing hormone secretion and ovarian function in heifers actively immunized against estrogen and progesterone

This study was designed to test the effects of active immunization against estrogen and progesterone on patterns of luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion, ovarian characteristics and growth rate of heifers. Heifers were randomly assigned to four treatments: 1) control injection (n = 10); 2) ovariectomy (n = 9); 3) immunization against estrogen (anti-E, n = 10); and 4) immunization against estrogen and progesterone (anti-E+P4, n = 10). Three booster immunizations were administered at 1, 1.5 and 6 mo after primary immunization. Progesterone antibody binding was 40% (34 fmol at 1:600 final dilution) in the anti-E+P4 heifers, and estradiol-17 beta binding was 35% (30 fmol) and 60% (52 fmol at 1:100 final dilution) in the anti-E+P4 and anti-E heifers, respectively, after the final immunization. Anti-E+P4 heifers had more pulses of LH and higher basal concentrations of LH than anti-E or control heifers (P less than .05). Concentrations of LH in anti-E+P4 heifers did not increase to concentrations found in ovariectomized heifers (P less than .05). Immunization against steroids did not alter the secretion of FSH. The number of large follicles (greater than 15 mm diameter) in anti-E+P4 and anti-E heifers was greater than in control heifers (P less than .05). Ovarian weight was increased in anti-E+P4 heifers (P less than .05). Average daily gain was not different among groups (P greater than .05). It was concluded that active immunization against estrogen and progesterone in heifers increased LH secretion and stimulated ovarian function.