Circulating oestrogen concentrations during pregnancy in the African elephant (Loxodonta africana)

Oestrone, oestradiol-17 beta and oestriol were measured in plasma samples from non-pregnant and pregnant African elephants shot in the wild. Enzymic hydrolysis of plasma showed that approximately 90 and 96% of the total (i.e. conjugated plus unconjugated) concentrations of oestrone and oestradiol-17 beta, respectively were represented by conjugated hormones. Unconjugated oestrogens remained low (less than 50 pg ml) in all samples, with no distinction between non-pregnant and pregnant animals. Levels of total oestrone during pregnancy varied between 160 and 594 pg/ml but were not significantly different from non-pregnant values. Total oestradiol-17 beta concentrations were significantly elevated during pregnancy (P less than 0 X 01) and, despite considerable individual variation (193-1428 pg/ml), were consistently higher than non-pregnant values after 6 months of gestation. The elevated levels of oestradiol-17 beta resulted in a reversal of the total oestradiol-17 beta: oestrone concentration ratio at about 6 months of pregnancy. Concentrations of total oestriol did not exceed 103 pg/ml. An indirect method of measurement indicated that oestradiol-17 beta sulphate was probably the most abundant circulating oestrogen during pregnancy in the African elephant.     

Androgens and FSH affect androgen receptor and aromatase distribution in the porcine ovary

In the present study the authors investigated whether androgens could interact with FSH to induce aromatase and androgen receptor expression in porcine granulosa cells. Dissected whole porcine follicles (small, medium, and large) were incubated for 8 hours in M199 medium supplemented with testosterone (10(-7) M), FSH (100 ng/ml) or both those hormones. After incubation, the follicles were fixed and immunostained to visualise androgen receptor and aromatase. In cultures of granulosa cells isolated from small and large follicles, oestrogen secretion was measured by appropriate RIA. Incubation of follicles with testosterone and FSH increased aromatase immunoreactivity in preantral and early antral (i.e. small) follicles. The immunostaining for androgen receptor was slightly higher in medium follicles, while such hormonal stimulation had no effect on small and large follicles. Moreover, granulosa cells isolated from small follicles cultured with both testosterone and FSH produced more estradiol than control cultures (40 pg vs. 100 pg/10(5) cells). The level was relatively close to that obtained in the culture of control granulosa cells isolated from large preovulatory follicles (105 pg/10(5) cells). These results indicate that testosterone acts synergistically with FSH to increase aromatase expression in the small porcine follicles.     

Total unconjugated oestrogen and progesterone concentrations in peripheral blood during the oestrous cycle of the dog.

From the beginning of pro-oestrus to the end of metoestrus, daily peripheral blood samples were withdrawn from six bitches. Further samples were obtained during anoestrus. Oestrogen values rose from the onset of pro-oestrus to attain a peak value of 25-3 +/- 4-8 pg/ml on Day 1 of oestrus. Progesterone concentrations began to rise 2 days before the oestrogen peak, and reached their highest levels of 18-9 +/- 1-0 ng/ml 16 days after the end of oestrus. After oestrus, an oestrogen rise was detected which reached a peak at Day 18 of metoestrus. The duration of oestrus was unrelated to the length of time that oestrogen levels were raised, or the maximum values were attained. No significant oestrogen levels were detected during anoestrus.     

Reduction in serum IL-6 after vacination of breast cancer patients with tumour-associated antigens is related to estrogen receptor status

Elevated serum IL-6 concentrations have been associated with poor prognosis in a variety of cancers, and decreases in serum IL-6 concentrations have been reported after chemotherapy. We have demonstrated that serum IL-6 concentrations are elevated in breast cancer patients [normal women 0.7 +/- 2.5 pg/ml (n=36), breast cancer patients 38.3 +/- 138.7 pg/ml (n = 111)]. After vaccination of breast cancer patients with a combination of tumour-associated antigens and biological adjuvants (IL-2 and GM-CSF), the concentration of IL-6 decreased significantly (P<0.05) to 8.1 +/- 14.6 pg/ml (n=85). Other studies have shown that oestrogen suppresses IL-6 production in oestrogen receptor positive breast cancer cells. We have demonstrated that the decrease in IL-6 associated with vaccination is related to the oestrogen receptor status of the tumours from breast cancer patients, as a decrease in IL-6 from 124.0 +/- 267.5 pg/ml (n=26) to 6.2 +/- 11.0 pg/ml (n=34) only occurs in patients with oestrogen receptor negative tumours. The IL-6 concentration in breast cancer patients with oestrogen receptor positive tumours remained unchanged (9.5 pg/ml before vaccination, and 9.3 pg/ml after vaccination). These results suggest that postmenopausal women with oestrogen receptor negative breast cancers, who do not respond well to either hormonal therapy with tamoxifen or adjuvant chemotherapy, may have a significant response to vaccination with autologous tumour-associated antigens.     

Simultaneous determination of urinary androgen glucuronides by high temperature gas chromatography-mass spectrometry with selected ion monitoring

An efficient procedure is described for the simultaneous determination of 9 androgen glucuronides including androsterone, etiocholanolone, 11-ketoandrosterone, 11-ketoetiocholanolone, 11beta-hydroxyandrosterone, 11beta-hydroxyetiocholanolone, and dehydroepiandrosterone (DHEA) in 3-glucuronide form and dihydrotestosterone (DHT) and testosterone in 17-glucuronide form from urine specimens. The method involves solid-phase extraction of the urinary steroids using Serdolit PAD-1 resin, with subsequent conversion to methyl ester-trimethylsilyl (Me-TMS) ether derivatives for the direct analysis by gas chromatography-mass spectrometry (GC-MS) using high temperature MXT-1 (Silcosteel-treated stainless steel) capillary column. Upon split injection of Me-TMS steroids at 330 degrees C into the MXT-1 capillary column initially maintained at 300 degrees C then programmed to 322 degrees C at 2 degrees C/min, each androgen glucuronide was well separated in excellent peak shape. The characteristic ions at m/z 217 constituting the base peaks in the electron-impact (20 eV) mass spectra for most steroids permitted their sensitive detection by GC-MS with selected-ion monitoring (SIM), whereas base peak ion at m/z 271 was used for the SIM of dehydroepiandrosterone-3-glucuronide. The detection limits for SIM of most of the steroids were 15 pg except for the 3-glucuronides of 11-ketoandrosterone and 11-ketoetiocholanolone, which could be detected down to 20 pg. The SIM responses were linear with correlation coefficients varying from 0.981 to 0.993 in the concentration range of 20 to 3000 ng/ml for the androgens studied. When applied to urine samples, the present method allowed rapid screening for the 7 androgens in their glucuro-conjugated forms simultaneously with good overall precision and accuracy within the normal concentration ranges of 15.1 to 3124.6 ng/ml.     

Plasma oestrogen levels in rainbow trout Salmo gairdneri Richardson

Levels of oestrogens in plasma of mature fall-spawning rainbow trout Salmo gairdneri were determined using a radio-immunoassay. No significant difference was found in oestrogen concentration between the sexes (male =2.5 ng/ml; female =4.4 ng/ml); between individual variability was great. Four blood samplings over a 24-h period via cardiac puncture of males revealed no diel variation or change in estrogen levels due to the stress of bleedings. Although no gonado-somatic index-estrogen relationship could be demonstrated for either sex, there was correlation between oestrogen and androgen levels in the female.     

Testicular endocrine function, seasonality and semen quality of the stallion

To gain further information on gonadal function of the stallion, concentrations of testicular steroids in blood plasma (bpl) and seminal plasma (spl) and their distribution in the ejaculate were determined. Blood and semen samples from a total of 11 stallions were collected from November to July. Estrone (E1), estrone sulfate (E1S), estradiol-17beta (E2beta) and testosterone (T) were determined in bpl and spl, and in addition androstenedione (A), dehydroepiandrosterone (DHEA) and 5alpha-dihydrotestosterone (5alpha-DHT) were measured in spl. At certain points of time, aliquots of an ejaculate were centrifuged, washed and the distribution of E1, E1S, E2beta and T into seminal plasma and the sperm fraction was assessed. Hormone assay was by RIA, partly after prior separation by HPLC. Mean concentrations (X(g) x DF) were as follows: E2beta (bpl) 31.1 (1.16), (spl) 24.2 (1.42) pg ml(-1); E1 (bpl) 143.3 (1.21), (spl) 117.7 (1.53) pg ml(-1); E1S (bpl) 157.3 (1.44), (spl) 2.92 (1.42) ng ml(-1); T (bpl) 570.6 (1.43), (spl) 23.1 (1.68) pg ml(-1); A (spl) 17.9 (1.39) pg ml(-1); DHEH (spl) 12.4 (1.51) pg ml(-1); 5alpha-DHT (spl) 9.7 (1.29) pg ml(-1). Except for E2beta and A in seminal plasma, a seasonal pattern was established for all other steroids with lowest mean values occurring from November to April. From the semen parameters determined, only motility was correlated to season. There was a higher correlation among oestrogen in blp than in spl and the only correlation identified between oestrogenic and androgenic steroids was between T and E2beta in blp. In spl, T was correlated with A and 5alpha-DHT. T was the dominant free steroid in bpl while it was E1 in spl; T and E1S concentrations were about 23- and 54-fold lower in spl compared to bpl with E1S, however, showing the highest absolute values in both fluids. In the fractionated ejaculate an association of free oestrogens, particularly E2beta, with spermatozoa was observed.     

Effect of oestradiol — 17β and oxytocin treatment on prostaglandin F alpha release in the anoestrous ewe

The concentration of prostaglandin Fα in the vena caval blood at a point anterior to the entry of uterine venous blood was determined by radioimmunoassay in nine anoestrous ewes. At all times the concentrations were much greater than those in jugular blood. The ewes were given oestradiol-17β or oxytocin or oestradiol followed 24 hours later by oxytocin.Oxytocin alone had negligible effect on prostaglandin release. Oestradiol alone increased the concentration of prostaglandin in the posterior vena cava about threefold to 160 pg/ml. When oxytocin was given to oestrogen primed ewes the concentration in the posterior vena cava rose dramatically to 1000 pg/ml within 5 minutes after which it declined to pre-treatment values: repetition of oxytocin treatment in one hour was much less effective.     

The effects of stress and of ACTH administration in hormone profiles, oestrus and ovulation in pigs

Hormone concentrations and oestrous cycle patterns were studied in five chronically cannulated gilts. During oestrous cycles that were unaffected by stress, plasma oestrogen concentrations remained at basal luteal phase levels (10 to 30 pg/ml) until plasma progesterone had decreased to less than 2 ng/ml. The pre-oestrus surge of oestrogen ranged from 40 to 80 pg/ml. Plasma corticoid concentrations varied randomly and were not related to oestrogen, progesterone concentrations, or the stage of the oestrous cycle. There was, however, evidence of a positive relationship between elevated corticoid levels and observed stressful events. The stress of surgery or illness acting during the follicular phase of the oestrous cycle delayed the onset of oestrus, and corticoid levels were frequently elevated on these occasions. Elevated plasma corticoid concentrations in response to ACTH treatment were associated with either a change in the timing of or a suppression of the pre-oestrus LH peak. Altering the timing of the LH peak resulted in the formation of large partially luteinized ovarian cysts, while suppressing LH interfered with follicular development and led to small ovarian cysts. These experiments suggest that stress acting via the adrenal gland may play a role in the aetiology of infertility in sows.     

Further evidence for the usefulness of the salivary testosterone radioimmunoassay in the assessment of androgenicity in man in basal and stimulated conditions

Since correct assessment of testicular function and androgenic status in humans requires multiple sampling, a sensitive and accurate radioimmunoassay (RIA) of testosterone (T) was established for male and female saliva samples. This easily collected biological fluid, which contains nonprotein-bound T, may represent an attractive alternative or a complement to total plasma T assays. In saliva samples from 5 normal males, a clear circadian rhythm was observed, and morning concentrations (135 +/- 31 pg/ml) were significantly higher (p less than 0.02) than evening samples (85 +/- 23 pg/ml). In 11 normal females, morning saliva levels were 12.8 +/- 1.8 pg/ml. The levels of T in male saliva, in response to both exogenous T administration (100 mg i.m.) and HCG stimulation (2 X 2,000 IU i.m.), accurately reflected the changes observed in plasma T, and the magnitude of increase in T levels was clearly greater in saliva than in plasma samples during the intramuscular administration of the long-acting T preparation. In males, significant correlations were observed between salivary and plasma T concentrations in morning samples (r = 0.61, p less than 0.01), following HCG stimulation (r = 0.89, p less than 0.05) and during T administration (r = 0.87, p less than 0.05). In women, the correlation at 8 a.m. was also significant (r = 0.82, p less than 0.05).     

Concentrations of oestradiol-17 beta in plasma and corpora lutea throughout pregnancy in the tammar, Macropus eugenii

Oestradiol-17 beta concentrations were measured by radioimmunoassay in peripheral blood samples from 10 tammar wallabies after their pouch young were removed to terminate embryonic diapause. Oestradiol concentrations rose from 8.3 +/- 1.2 pg/ml on Days 3 and 4 to peak of 15.8 +/- 2.9 pg/ml on Day 5, coincident with an increase in 'progesterone' concentrations, and then fell to 10.5 +/- 2.7 pg/ml on Day 7. No changes in oestradiol concentrations were associated with parturition. Five females came into oestrus and mated 9.8 +/- 6.1 h post partum; peak concentrations of plasma oestradiol (20.9 +/- 2.1 pg/ml) occurred around the time of mating. None of the females that did not mate up to the end of the experiment at Day 30 had a rise in plasma oestradiol concentrations. Corpora lutea contained 20-100 pg oestradiol during pregnancy. The highest ovarian oestradiol content (greater than 1200 pg) was measured in whole ovaries containing Graafian follicles from full-term pregnant females. The rise in oestradiol concentrations at Day 5 may be important in the termination of diapause. The post-partum increase in plasma oestradiol concentrations coincides with oestrus. The source of this oestrogen appears to be the preovulatory follicle.     

Steroid metabolism by endometrial and conceptus tissues during early pregnancy and pseudopregnancy in gilts

Endometrial and conceptus tissues were obtained on Days 10.5, 11, 12, 16 and 25 of pregnancy and Day 25 of pseudopregnancy of gilts and incubated for 6 h in Minimal Essential Medium (5 ml) containing 35 ng [3H]progesterone. Metabolism of [3H]progesterone to oestrone, oestradiol and oestriol was determined by gas and high-pressure liquid chromatography and successive recrystallizations with unlabelled standards. Conceptuses collected between Days 10.5 and 12 were spherical, tubular or filamentous and incubated with 500 mg endometrium and [3H]progesterone. Production of oestrone by spherical conceptuses was not detected, but was 44-47 pg/tubular conceptus and 21 pg/filamentous conceptus. A similar trend was observed for oestradiol. Conceptus tissues from Days 16 and 25 (chorion) were most active in producing oestrone (123 and 520 pg/mg tissue, respectively) and oestradiol (277 and 876 pg/mg tissue, respectively). Endometrial oestrogen production was less than that for conceptus tissue for oestrone and oestradiol on Days 16 and 25 of gestation. Coincubations of endometrium and conceptus tissues had lower oestrogen production than conceptus alone. Endometrium from Day 25 of pseudopregnancy metabolized [3H]progesterone to several non-polar metabolites, but no oestrogens were detected. An unidentified phenolic metabolite of [3H]progesterone was detected in higher quantities than either oestrone or oestradiol; 445 to 461 pg/conceptus at the tubular stage. These results indicate temporal changes in the conversion of [3H]progesterone to oestrogens by conceptus and endometrial tissue from pregnant gilts, but not endometrium from pseudopregnant gilts.     

Determination of free and total (free plus protein-bound) melatonin in plasma and cerebrospinal fluid by high-performance liquid chromatography with fluorescence detection

A simple, sensitive and accurate method for the estimation of free and total (free plus protein-bound) melatonin (MLT) in human plasma and cerebrospinal fluid (CSF) is described. Via Chem-Elut cartridges, free and total MLT (the latter obtained after a deproteinization step) were quantified in dichloromethane-extracted samples and analyzed in one chromatographic run by high-performance liquid chromatography (HPLC) with fluorimetric detection. The column used was an Extrasil ODS-2 (3 microm, 150 x 4.6 mm I.D.), while the mobile phase consisted of 75 mM sodium acetate-acetonitrile (72:28, v/v) (pH 5.0). Repeatability and reproducibility of the method were 3.24 and 9.4%, respectively. The recovery of melatonin from plasma and CSF was 99.9+/-4.0% for non-deproteinized samples and 93.2+/-4.8% for deproteinized samples. The detection limit of the assay was 0.5 pg/ml. In human plasma, the mean+/-SD concentrations in the darkness period were 23.18+/-7.44 pg/ml for free melatonin and 82.5+/-36.48 pg/ml for total melatonin, while the lowest concentrations detected during daytime were 2.23+/-2.22 and 7.40+/-5.68 pg/ml, respectively. Detection of MLT in CSF was 5.01+/-2.31 and 28.55+/-6.95 pg/ml for the free and total fraction, respectively.     

Does prolactin affect steroid secretion by isolated rainbow trout ovarian cells?

The in vitro secretion of progesterone (P(4)), androgen (A) and estradiol (E(2)) by follicular cells, isolated monthly from the rainbow trout ovaries during the whole annual cycle, was studied. Cells were cultured as monolayers in control and prolactin (PRL) supplemented media. E(2) secretion showed two distinct maxima in September and January: 4959+/-220 pg/ml and 3166+/-121 pg/ml, respectively, i.e. during vitellogenesis and before the spawning time. PRL had a significant (16%) suppressive effect on E(2) secretion when the level of secreted steroid was at its highest (4167+/-193 pg/ml) at the end of vitellogenesis and by 32% (2157+/-124 pg/ml), before ovulation. Increased P(4) levels observed in February (988+/-69 pg/ml) and March (2008+/-74 pg/ml) may be connected with the need for a substrate for the synthesis of 17alpha20betaOH-P (MIS). At this time, the secretion of P(4) was also suppressed by PRL and was reduced to 1395+/-78 pg/ml. Our results indicate that PRL may play a role in fish reproduction.     

In vitro effects of glycosylated insulin and glucagon in perfused liver of the rat.

Using perfused liver of the rat, the hepatic uptake of glycosylated insulin (GI) and glucagon (GG) and its effects on hepatic glucose output were investigated. Insulin and glucagon were glycosylated in ambient high glucose concentration, and GI80 or GG80 (insulin or glucagon incubated with 0.08% glucose), GI350 or GG350 (incubated with 0.35% glucose), and GI1000 or GG1000 (incubated with 1% glucose) were prepared. The liver was perfused with the medium containing 1000 microU/ml insulin and 200 pg/ml glucagon or 200 microU/ml insulin and 1000 pg/ml glucagon. The fractional uptake of insulin or glucagon by perfused liver was not significantly altered by the glycosylation. In the liver perfused with 1000 microU/ml insulin and 200 pg/ml glucagon, glucose output was not changed by the glycosylation of the hormones, while in the liver perfused with 200 microU/ml insulin and 1000 pg/ml glucagon, GI1000 reduced its biological activity, as reflected by insulin-mediated decrease in glucose output. These results suggest that in the liver insulin incubated with markedly high concentration of glucose reduces its biological activity at a physiological concentration in the presence of high concentration of glucagon.     

A precursor role for DHA in a feto-placental unit for oestrogen formation in the mare.

Plasma levels of total oestrogens and dehydroepiandrosterone (DHA) were measured by radioimmunossay in samples taken from various blood vessels in both maternal and fetal compartments in 11 Pony mates. High concentrations of oestrogens (greater than 100 ng/ml of plasma), expressed as oestrone equivalents, were found in the fetal circulation. On both the fetal and maternal sides, oestrogen concentrations were lower in blood going to than from the placenta. DHA concentrations, on the other hand, were higher in blood flowing to the placenta from the fetus. The fetal gonads were seen as the source of DHA, which was present in remarkably high concentrations (greater than 800 ng/ml of plasma) in venous samples from fetal ovaries and fetal testes. A precursor role in placental oestrogen formation is suggested for DHA secretion by the fetal gonads.     

Spontaneous uterine activity and plasma progesterone and oestrogens in the ewe with live and stillborn lambs

Peripheral plasma progesterone and oestrogen concentrations were measured during late pregnancy and the parturient period in 12 ewes producing live lambs and three others producing stillborn lambs. Progesterone values started declining by 10 days before lambing but at minus 24 hours were still 6.1 +/- 3 ng/ml in the ewes bearing live lambs; during the last 24 hours progesterone was significantly lower in the ewes producing stillborn lambs. Oestrogens reached a maximum level of 550 +/- 280 pg/ml at the time of delivery and declined rapidly to basal values shortly after lambing. Oestrogens did not rise at lambing in the ewes producing stillborn lambs. In seven of the 12 ewes bearing live lambs, uterine activity, as determined by intrauterine pressure changes, was recorded throughout parturition and compared with the plasma values for progesterone and oestrogens. It was found that there was a highly significant positive correlation between uterine activity and plasma oestrogen concentrations, and a highly significant negative correlation between uterine activty and both plasma progesterone concentrations and the progesterone: oestrogen ratio.     

Determination of plasma melatonin levels by enzyme-linked immunosorbent assay (EIA) in turbot (Scophtalmus maximus L.) and tench (Tinca tinca L.)

An enzymoimmunoassay (EIA) kit for plasma melatonin (MLT) measurements was employed in tench (Tinca tinca) and in turbot (Scophtalmus maximus). Tench and turbot plasma samples were purified with a C18 reversed phase extraction columns because this kit is designed for human serum measurements. The lowest detection limit of the technique was 11.48 pg/well with a sensitivity at 50% binding of 100 pg/well. Intra-assay and inter-assay CV (%) were always less than 5% (n=8), and 9% (n=6) in tench plasma samples, and less than 5% (n=8) and 13% (n=5) in turbot plasma samples, respectively. Correlation coefficients between EIA and RIA measurements in tench and turbot plasma samples were 0.93 and 0.89 (p<0.001) respectively. Diurnal and nocturnal plasma melatonin mean levels were 14.7+/-2.1 pg/ml and 87.4+/-11 pg/ml in tench (n=15), and 3.5+/-0.4 pg/ml and 28.1+/-2.1 pg/ml in turbot (n=15). These species showed a melatonin circadian rhythm as in other animals studied. The results suggest that the commercial kit used in this experiment could be a suitable and alternative method to RIA for plasma MLT determinations in tench and turbot although it is necessary to increase volumes (1ml) and concentrate daytime samples.     

Plasma steroid hormone profiles and reproductive biology of the epaulette shark, Hemiscyllium ocellatum.

Examination of the reproductive biology of the oviparous epaulette shark, Hemiscyllium ocellatum, was conducted on a wild population. Male sharks were found to reach maturity at between 55-60 cm total length (TL) and female sharks mature around 55 cm TL. Blood samples collected from mature male and female sharks were analyzed for sex steroid hormones to examine seasonal hormone patterns. Plasma samples were analyzed via radioimmunoassay techniques with female samples measured for estradiol, progesterone, and androgen concentrations, and male samples measured for androgen concentrations. Male androgen concentrations showed a single broad peak from July to October with maximum hormone concentrations (60 ng/ml) occurring in August. Male androgen concentrations were lowest in December-February (<20 ng/ml), and appeared to correlate with reproductive activity and water temperature. Female androgen concentrations were an order of magnitude lower than those for males and showed peaks in June (6 ng/ml) and December (8 ng/ml). Estradiol concentrations in females peaked during the months of September-November (0.5 ng/ml) coinciding with the egg laying period. Progesterone concentrations ranged up to 0.5 ng/ml prior to the mating season. Observations of ova size and egg production showed eggs develop in pairs and ova are ovulated at a size of 25-27 mm. Females lay eggs from August to January. Males were observed with swollen claspers from July through December, with the highest amount of sperm storage in the epididymis occurring between August through November. Our observations indicate that epaulette sharks in the waters near Heron Island mate from July through December. J. Exp. Zool. 284:586-594, 1999.     

Concurrent levels of 11-ketotestosterone in fish surface mucus, muscle tissue and blood

A rapid, reproducible method is described for extracting and comparing levels of the ether‐soluble fish androgen 11‐ketotestosterone (11‐KT) in blood serum, muscle tissue and surface mucus. Because widely different volumes of extract were recovered after centrifugation from the three sources, it was important to express androgen levels as pg 11‐KT/mg of total soluble protein (TSP). For six male and four female sexually‐staged freshwater Koi (Cyprinus carpio), the method yielded similar pg 11‐KT/mg TSP ratios in blood serum and extracts of muscle tissue and surface mucus, with the strongest correlation between blood serum and surface mucus. While male Koi were distinguishable from females based on the magnitude of 11‐KT levels, reproductive stage and gonadosomatic index levels were not correlated with the 11‐KT levels of either sex. Similar pg 11‐KT/mg TSP ratios were also found for autologous muscle tissue and surface mucus extracts of 37 captured and sexed wild marine fishes representing seven genera. However, high 11‐KT levels were not restricted to mature males. Collectively, results suggest that surface mucus collection (followed by 11‐KT assay) is a useful alternative to more invasive methods of determining systemic hormone levels in fish. Without knowledge of seasonal variation in levels of this and other sex hormones, however, reliance on 11‐KT levels alone may lead to spurious identification of gender, let alone reproductive stage.