Effects of dietary phytoestrogens on plasma testosterone and triiodothyronine (T3) levels in male goat kids

<p>Abstract</p> <p>Background</p> <p>Exposure to xenoestrogens in humans and animals has gained increasing attention due to the effects of these compounds on reproduction. The present study was undertaken to investigate the influence of low-dose dietary phytoestrogen exposure, i.e. a mixture of genistein, daidzein, biochanin A and formononetin, on the establishment of testosterone production during puberty in male goat kids.</p> <p>Methods</p> <p>Goat kids at the age of 3 months received either a standard diet or a diet supplemented with phytoestrogens (3 - 4 mg/kg/day) for ~3 months. Plasma testosterone and total and free triiodothyronine (T₃) concentrations were determined weekly. Testicular levels of testosterone and cAMP were measured at the end of the experiment. Repeated measurement analysis of variance using the MIXED procedure on the generated averages, according to the Statistical Analysis System program package (Release 6.12, 1996, SAS Institute Inc., Cary, NC, USA) was carried out.</p> <p>Results</p> <p>No significant difference in plasma testosterone concentration between the groups was detected during the first 7 weeks. However, at the age of 5 months (i.e. October 1, week 8) phytoestrogen-treated animals showed significantly higher testosterone concentrations than control animals (37.5 nmol/l vs 19.1 nmol/l). This elevation was preceded by a rise in plasma total T₃that occurred on September 17 (week 6). A slightly higher concentration of free T₃was detected in the phytoestrogen group at the same time point, but it was not until October 8 and 15 (week 9 and 10) that a significant difference was found between the groups. At the termination of the experiment, testicular cAMP levels were significantly lower in goats fed a phytoestrogen-supplemented diet. Phytoestrogen-fed animals also had lower plasma and testicular testosterone concentrations, but these differences were not statistically significant.</p> <p>Conclusion</p> <p>Our findings suggest that phytoestrogens can stimulate testosterone synthesis during puberty in male goats by increasing the secretion of T₃; a hormone known to stimulate Leydig cell steroidogenesis. It is possible that feedback signalling underlies the tendency towards decreased steroid production at the end of the experiment.</p

Exogenous estradiol improves shell strength in laying hens at the end of the laying period

Background: Cracked shells, due to age related reduction of shell quality, are a costly problem for the industry. Parallel to reduced shell quality the skeleton becomes brittle resulting in bone fractures. Calcium, a main prerequisite for both eggshell and bone, is regulated by estrogen in a complex manner. The effects of estrogen, given in a low continuous dose, were studied regarding factors involved in age related changes in shell quality and bone strength of laying hens. A pellet containing 0.385 mg estradiol 3-benzoate (21-day-release) or placebo was inserted subcutaneously in 20 birds each of Lohmann Selected Leghorn (LSL) and Lohmann Brown (LB) at 70 weeks of age. Eggs were collected before and during the experiment for shell quality measurements. Blood samples for analysis of total calcium were taken three days after the insertion and at sacrifice (72 weeks). Right femur was used for bone strength measurements and tissue samples from duodenum and shell gland were processed for morphology, immunohistochemical localization of estrogen receptors (ER alpha, ER beta), plasma membrane calcium ATPase (PMCA) and histochemical localization of carbonic anhydrase (CA). Results: Estrogen treatment increased shell thickness of both hybrids. In addition, shell weight and shell deformation improved in eggs from the brown hybrids. The more pronounced effect on eggs from the brown hybrid may be due to a change in sensitivity to estrogen, especially in surface epithelial cells of the shell gland, shown as an altered ratio between ER alpha and ER beta. A regulatory effect of estrogen on CA activity, but not PMCA, was seen in both duodenum and shell gland, and a possible connection to shell quality is discussed. Bone strength was unaffected by treatment, but femur was stronger in LSL birds suggesting that the hybrids differ in calcium allocation between shell and bone at the end of the laying period. Plasma calcium concentrations and egg production were unaffected. Conclusions: A low continuous dose of estrogen improves shell strength but not bone strength in laying hens at the end of the laying period

Age-related changes in the shell gland and duodenum in relation to shell quality and bone strength in commercial laying hen hybrids

Abstract Background During the production period of laying hens, the number of cracked eggshells increases and the skeleton becomes brittle. Both these problems are related to ageing of the hen and cause economic problems for egg producers and impaired animal welfare. This study investigated key factors in the shell gland and duodenum related to eggshell quality and bone strength in laying hens during the production period. Five Lohmann Selected Leghorn (LSL) and five Lohmann Brown (LB), common hybrids in commercial egg production, were euthanized at 21, 29, 49 and 70 weeks (wk) of age. Blood samples for analysis of total calcium were taken at euthanization. Right femur and humerus were used for bone strength measurements and tissue samples from shell gland and duodenum were processed for morphology, immunohistochemical localisation of oestrogen receptors (ERα, ERβ), plasma membrane calcium ATPase (PMCA) and histochemical localisation of carbonic anhydrases (CA). Eggs were collected for shell quality measurements. Results At age 49 week, shell and bone strength had both deteriorated, but the hens were then able to maintain the level until 70 week of age and femur bone strength even improved. The main physiological findings associated with the effects seen at 49 week were reduced gland density and a shift in balance between ERα and ERβ in the shell gland, which coincided with a reduction in CA activity in the duodenum. Somewhat surprisingly, capillary density and capillaries with CA activity both increased in the shell gland over time, the latter possibly mediated via ERβ. These findings were independent of hybrid. PMCA was found in both shell gland and duodenum, but appeared unrelated to the age-related changes in shell and bone quality. Conclusions In hens around half-way through the production period, both shell quality and bone strength had deteriorated. Decreased gland density and a shift in the balance between ERα and ERβ in the shell gland, co-occurring with a dramatic drop in duodenal CA activity, are suggested as possible factors involved in age-related changes in shell and bone quality

CA in spider silk glands.

<p>CA activity and Azure blue staining of histological sections from (A) the sac of a <i>Tegenaria sp</i>. major ampullate gland, (B) <i>E. australis</i> minor ampullate gland, (C) <i>A. diadematus</i> aggregate gland duct, (D) <i>Tegenaria sp</i>. tubuliform gland, and (E) the third limb of the duct of an <i>A. diadematus</i> major ampullate gland. Black precipitates represent CA activity (arrow heads). In (A) the glandular lumen is labeled and the dotted arrow points towards the duct. Nuclei are indicated by (N) and the lumen by (Lu). Scale bar, (A) 50 µm and (B–E) 20 µm.</p

pH, bicarbonate, and carbon dioxide in major ampullate glands.

<p>Photograph of a major ampullate gland in which measured pH and HCO₃⁻ values and calculated pCO2 values at different locations are indicated. See Table 1 for details of ISM measurements. Scale bar, 1 mm.</p

CT forms amyloid-like fibrils at low pH.

<p>(A) ThT fluorescence for CT at pH 6.0, 5.7, 5.5, 5.3, and 5.0, and NT at pH 5.0, in sodium acetate buffer. (B) Transmission electron micrograph showing fibrils formed from CT incubated at pH 5.5. Scale bar, 200 nm. (C) Congo red stained fibrils formed by CT at pH 5.5, viewed under crossed polarizers. Green birefringence is visible. Scale bar, 50 µm.</p

CA in spider silk glands.

<p>CA activity and Azure blue staining of histological sections from (A) the sac of a <i>Tegenaria sp</i>. major ampullate gland, (B) <i>E. australis</i> minor ampullate gland, (C) <i>A. diadematus</i> aggregate gland duct, (D) <i>Tegenaria sp</i>. tubuliform gland, and (E) the third limb of the duct of an <i>A. diadematus</i> major ampullate gland. Black precipitates represent CA activity (arrow heads). In (A) the glandular lumen is labeled and the dotted arrow points towards the duct. Nuclei are indicated by (N) and the lumen by (Lu). Scale bar, (A) 50 µm and (B–E) 20 µm.</p