scholarly journals Using a Commercially Available Enzyme Immunoassay to Quantify Testosterone in Avian Plasma

The Condor ◽  
2007 ◽  
Vol 109 (1) ◽  
pp. 181-186 ◽  
Author(s):  
Brian E. Washburn ◽  
Joshua J. Millspaugh ◽  
Dana L. Morris ◽  
John H. Schulz ◽  
John Faaborg

Abstract Abstract Using a commercially available testosterone enzyme immunoassay (EIA), we developed and validated an assay procedure for determining testosterone levels in small-volume (20 µL) avian plasma samples. We evaluated this EIA's utility by measuring plasma testosterone levels in Mourning Doves (Zenaida macroura), White-eyed Vireos (Vireo griseus), Red-eyed Vireos (Vireo olivaceus), and Indigo Buntings (Passerina cyanea). Standard biochemical validations (e.g., parallelism, recovery of exogenous testosterone) demonstrated that the assay accurately and precisely measured testosterone in avian plasma. We compared plasma testosterone levels in males and females of all four species and Indigo Buntings in various reproductive stages to physiologically validate the assay's ability to determine biologically important changes in testosterone levels. Plasma testosterone levels were higher in males compared to females in three of four species. Prebreeding and breeding male Indigo Buntings had higher circulating testosterone levels than postbreeding males. Testosterone levels in our study were similar to reported values for other passerine species using radioimmunoassay procedures. Our results suggest that this EIA procedure is very effective for determining testosterone levels in small-volume avian plasma samples and is sensitive enough to detect biologically important changes in the gonadal activity of birds. Thus, this assay has considerable utility for measuring testosterone in small birds (<15 g), from which only small volumes of plasma (20 µL) can be collected.

The Condor ◽  
2002 ◽  
Vol 104 (3) ◽  
pp. 558-563 ◽  
Author(s):  
Brian E. Washburn ◽  
Dana L. Morris ◽  
Joshua J. Millspaugh ◽  
John Faaborg ◽  
John H. Schulz

AbstractUsing a commercially available corticosterone I125 double-antibody radioimmunoassay, we developed and validated an assay procedure for determining corticosterone levels in small-volume (≤30 μL) avian plasma samples. We evaluated this procedure's utility by measuring plasma corticosterone levels in Indigo Buntings (Passerina cyanea), American Goldfinches (Carduelis tristis), Red-eyed Vireos (Vireo olivaceus), and Mourning Doves (Zenaida macroura). Standard biochemical validations (e.g., parallelism, recovery of exogenous corticosterone) demonstrated that the assay accurately and precisely measured corticosterone in avian plasma. We used a stress capture protocol to physiologically validate the assay's ability to determine biologically important changes in corticosterone levels. Males and females from four bird species exhibited a significant increase in plasma corticosterone in response to capture, handling, and restraint. Baseline and stress-induced corticosterone levels in our study were similar to reported values for other passerine species using other radioimmunoassay procedures. Our results suggest that this radioimmunoassay procedure is very effective for determining corticosterone levels in small-volume avian plasma samples and is sensitive enough to detect biologically important changes in the adrenocortical activity of birds. Thus, this assay has considerable utility for measuring stress levels and stress responses in small birds (<15 g), from which only small volumes of plasma (≤30 μL) can be collected.Utilización de un Radioinmunoensayo Disponible Comercialmente para la Cuantificación de Corticosterona en el Plasma de AvesResumen. Desarrollamos y validamos un proceso de ensayo para determinar los niveles de corticosterona en muestras de pequeño volúmen (≤30 μL) de plasma de aves utilizando un radioinmunoensayo para corticosterona I125 de doble anticuerpo disponible comercialmente. Evaluamos este procedimiento midiendo los niveles de corticosterona en Passerina cyanea, Carduelis tristis, Vireo olivaceus y Zenaida macroura. Validaciones bioquímicas estándares (e.g., paralelismo, recuperación de corticoesteroide exógeno) demostraron que el ensayo midió de modo exacto y preciso la corticosterona en el plasma de las aves. Utilizamos un protocolo de captura que producía estrés para validar fisiológicamente la habilidad del ensayo de detectar cambios biológicamente importantes en los niveles de corticosterona. Hembras y machos de las cuatro especies de aves mostraron un incremento significativo en los niveles de corticosterona en el plasma en respuesta a la captura, manipulación y retención. Los niveles basales e inducidos por el estrés de nuestro estudio fueron similares a valores reportados para otras especies paserinas que utilizaron otros procedimientos de inmunoensayo. Nuestros resultados sugieren que este procedimiento de radioinmunoensayo es muy efectivo para determinar los niveles de corticosterona en muestras de pequeño volúmen de plasma de aves y que es suficientemente sensible como para detectar cambios biológicamente importantes en la actividad adenocortical de las aves. De esta manera, este ensayo presenta considerable utilidad para medir los niveles y respuesta al estrés en aves pequeñas (<15 g) de las cuales sólo es posible colectar pequeños volúmenes de plasma (≤30 μL).


1978 ◽  
Vol 88 (4) ◽  
pp. 787-792 ◽  
Author(s):  
Anne Sundby ◽  
P. A. Torjesen

ABSTRACT Administration of 6000 IU HCG to 4 bulls was followed by an elevation of plasma testosterone lasting for 9–13 days. When HCG administration was repeated, the testosterone response was shortened to 4–6 days in 3 bulls due to the formation of antibodies against HCG. The appearance of HCG antibodies coincided with a sharp decrease in the plasma testosterone level, indicating that Leydig cells have to be under continuous HCG stimulation to maintain increased testosterone production. No antibody against bovine LH was detected in the plasma samples containing antibodies against HCG. In one bull the response following the second HCG injection was similar to the plasma testosterone pattern following the first. No antibodies against HCG were found in this bull. Five bulls received 750 IU HCG twice. Following the period with elevated plasma testosterone levels, subnormal levels were observed after both injections. One injection led to decreased levels without development of antibodies against HCG while the second HCG injection led to subnormal testosterone levels concomitant with measurable antibodies against HCG.


1971 ◽  
Vol 68 (3) ◽  
pp. 576-584 ◽  
Author(s):  
K. O. Nilsson ◽  
B. Hökfelt

ABSTRACT Metyrapone was administered either orally, 750 mg every four h, in a total of six doses, or intravenously 30 mg per kg body weight as a four h infusion. In three males with normal endocrine functions, metyrapone given orally or intravenously induced a fall in plasma testosterone and an elevation of androstenedione within 2–8 h. When metyrapone was administered to a patient given dexamethasone to suppress endogenous ACTH production, the androstenedione levels did not alter whereas the testosterone levels showed a slight, transient decrease. In two normal females metyrapone administration was followed by a marked increase in plasma androstenedione whereas testosterone showed only a minor, gradual increase. In one male patient with Addison's disease the basal plasma testosterone was normal whereas the level of androstenedione was low. Following metyrapone intravenously, there was a slight suppression of plasma testosterone but no change in the androstenedione concentration. In one patient with primary hypogonadism, two with secondary hypogonadism and two with Klinefelter's syndrome the plasma testosterone was low under basal conditions and did not change following metyrapone. Basal plasma androstenedione was within the range for normal males and increased markedly following metyrapone in all the cases.


1981 ◽  
Vol 96 (2) ◽  
pp. 273-280 ◽  
Author(s):  
Mridula Chowdhury ◽  
Robert Tcholakian ◽  
Emil Steinberger

Abstract. It has been suggested that treatment of intact male rats with oestradiol benzoate (OeB) causes an interference with testosterone (T) production by the testes by a direct inhibitory effect on steroidogenesis. To test this hypothesis, different doses (5, 10 or 25 IU) of hCG were administered concomitantly with 50 μg of OeB to adult intact or hypophysectomized male rats. The testicular and plasma testosterone, and serum hCG levels were determined. The sex accessory weights were recorded. In the intact OeB-treated group of animals, hCG stimulated both the secondary sex organs and plasma testosterone levels above the intact control group. However, in hypophysectomized animals, although plasma testosterone levels increased above that of intact controls, their secondary sex organ weights did not. Moreover, inspite of high circulating hCG levels, the testicular testosterone content and concentration remained suppressed in OeB-treated animals. The reason for such dichotomy of hCG action on OeB-treated animals is not clear at present.


2005 ◽  
Vol 12 (2) ◽  
pp. 135-141 ◽  
Author(s):  
Ilma Simoni Brum ◽  
Poli Mara Spritzer ◽  
Franyoise Paris ◽  
Maria Augusta Maturana ◽  
Franyoise Audran ◽  
...  

1991 ◽  
Vol 124 (4) ◽  
pp. 399-404 ◽  
Author(s):  
Wieland Kiess ◽  
Linda L. Liu ◽  
Nicholas R. Hall

Abstract. Sex-related differences in immune responsiveness are mediated at least in part by sex steroid hormones. Lymphocyte subset distribution in peripheral blood and natural killer cell function both have been reported to be under hormonal control. In order to gain more insight into sex steroid hormone action on the immune system, we have measured the lymphocyte subset distribution and natural killer cell activity in 18 men with idiopathic hypogonadotropic hypogonadism before treatment, and after hormonal treatment had normalized plasma testosterone levels. In untreated patients, the mean plasma testosterone concentrations were significantly lower than those in the treated men (3.0 ± 0.5 nmol/l vs 16 ± 1.7 nmol/l, p < 0.001). The percentage of peripheral CD3+ lymphocytes, CD8+ cells, the CD4+/CD8+ ratio, and the natural killer cell activity of peripheral mononuclear cells measured in a 51Cr release assay against target K 562 cells did not differ between patients with idiopathic hypogonadotropic hypogonadism and healthy adults, and most importantly, did not change during hormonal treatment which normalized plasma testosterone levels in the patients. In contrast, the percentage of peripheral CD4+ cells was significantly higher in untreated patients compared with normal adult subjects or patients with idiopathic hypogonadotropic hypogonadism after hormonal treatment that resulted in normal plasma testosterone levels (53 ± 2 vs 47 ± 2, p < 0.05). It should be noted that the percentage of peripheral CD 16+ cells was significantly lower in untreated men with low plasma testosterone levels than in normal controls. The percentage of CD16+ cells in peripheral venous blood rose significantly after hormonal treatment restored plasma testosterone levels to normal (6 ± 1 vs 11 ± 1, p < 0.001). In addition, the percentage of peripheral CD16+ cells correlated significantly with the plasma testosterone levels measured in men with idiopathic hypogonadotropic hypogonadism (r = 0.534, p < 0.001). In conclusion, both the percentage of peripheral CD4+ cells (T-helper lymphocytes) and peripheral CD16+ cells (non-T-non-B cells) are related to the plasma testosterone levels in men with idiopathic hypogonadotropic hypogonadism. These data suggest that in vivo human immune cells are under the regulatory influence of endogenous sex steroids.


1991 ◽  
Vol 6 (1-2) ◽  
pp. 185-191 ◽  
Author(s):  
P.S.P. Gupta ◽  
P.C. Sanwal ◽  
V.P. Varshney

1999 ◽  
Vol 202 (21) ◽  
pp. 3021-3028 ◽  
Author(s):  
T.C. Hoffman ◽  
G.E. Walsberg

We tested the hypothesis that birds can rapidly change the conductance of water vapor at the skin surface in response to a changing need for evaporative heat loss. Mourning doves (Zenaida macroura) were placed in a two-compartment chamber separating the head from the rest of the body. The rate of cutaneous evaporation was measured in response to dry ventilatory inflow at three ambient temperatures and in response to vapor-saturated ventilatory inflow at two ambient temperatures. At 35 degrees C, cutaneous evaporation increased by 72 % when evaporative water loss from the mouth was prevented, but no increase was observed at 45 degrees C. For both dry and vapor-saturated treatments, cutaneous evaporation increased significantly with increased ambient temperature. Changes in skin temperature made only a minor contribution to any observed increase in cutaneous evaporation. This indicates that Z. macroura can effect rapid adjustment of evaporative conductance at the skin in response to acute change in thermoregulatory demand.


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