Semiquantitative immunohistochemical detection of progesterone receptors in male accessory sex glands as a screening assay for anabolic steroid use in bulls

Abstract

We investigated the immunohistochemical expression of progesterone receptors (PRs) in the prostate and bulbourethral glands of thirty-two 10-14-mo-old Charolais bulls following treatment with a low dosage of estrogens. Animals were divided into 2 groups: 16 animals (group T) were treated for 71 d with a therapeutic dose of trenbolone acetate and estradiol by subcutaneous implant, 16 animals (group C) received no treatment. Urine samples were collected both at the beginning of the trial and 9 times during the study. A semiquantitative analysis of immunohistochemistry (IHC) was performed by counting the number of positive cells in 10 randomly selected high-power fields (hpf). Both groups showed no significant histologic lesions. IHC examination showed positive cells in the epithelium of both glands, with different patterns of distribution between groups. In group C, IHC-positive cells per hpf varied from 0 to 40 in the prostate and from 0 to 32 in the bulbourethral gland. In group T, positive cells varied from 0 to 85 per hpf in the prostate and from 0 to 75 in the bulbourethral gland. The treated group showed significantly higher median numbers of positively stained cells in both organs than the controls ( p < 0.001). Chemical analysis of the urine samples confirmed that the experimental treatment mimics continuous, low-dose administration of anabolic steroids. IHC quantification showed good sensitivity with a high predictive power to correctly classify treated animals and could be used as a preliminary screening test in bulls.

TMT‐Based Proteomics Profiling of Bovine Liver Underscores Protein Markers of Anabolic Treatments

“…Global untargeted approaches have been proposed to partially overcome such limitations and to extend the analytical spectrum. Despite strategies used by meat producers to reduce the amount of administered growth promoters to escape official controls, some overt effects on specific organs or tissues may occur that may be used to highlight illegal treatments . The analysis of proteomic profiles in tissues and body fluids has been introduced to provide comprehensive information about patho‐physiological processes, and to identify alterations among samples to be used as targets for medical treatments, or markers of the clinical status .…”

“…Despite strategies used by meat producers to reduce the amount of administered growth promoters to escape official controls, some overt effects on specific organs or tissues may occur that may be used to highlight illegal treatments. [12,13] The analysis of proteomic profiles in tissues and body fluids has been introduced to provide comprehensive information about pathophysiological processes, and to identify alterations among samples to be used as targets for medical treatments, or markers of the clinical status. [14,15] Strategies based on the measurement of proteomic changes that occur in response to anabolic drug administration have also been proposed as additional techniques to indirectly detect drug misuse in food-producing animals.…”