Plasma Sex Steroid Research Articles

Effects of the sea louse Lepeophtheirus salmonis on temporal changes in cortisol, sex steroids, growth and reproductive investment in Arctic charr Salvelinus alpinus

Groups of mature (5+ year old) Arctic charr Salvelinus alpinus held in sea water were exposed for 34 days to either a high (mean +/-s.e. 0.15 +/- 0.01 sea lice Lepeophtheirus salmonis g(-1) fish mass) (HI), medium (0.07 +/- 0.00 sea lice g(-1) fish mass) (MI) or no [control (C)] sea-lice infection during early stages of gonad development (June to July). Infection with sea lice resulted in increased plasma cortisol concentrations and this was related to intensity of infection; females tended to have higher cortisol concentrations than males at high infection intensities (HI group: female c. 130 ng ml(-1); male c. 80 ng ml(-1)). Plasma osmolality (C c. 330, MI c. 350 and HI c. 415 mOsm) and chloride concentrations (C c. 135, MI c. 155 and HI c. 190 mM) increased significantly with infection intensity, indicating osmoregulatory problems in infected fish. A strong positive relationship between plasma osmolality and cortisol concentration was recorded. Plasma sex-steroid concentrations were influenced negatively by sea-lice infection, particularly in the HI group, and were inversely related to plasma cortisol concentrations. The most heavily infected fish postponed the initiation of reproductive development until exposed to fresh water and timing of ovulation tended to be delayed in these fish. Growth rate and condition were negatively influenced by sea-lice infection and growth rate was inversely related to plasma cortisol concentrations. Sea-lice infection resulted in mortality among females in the HI group, and the proportion of maturing females was lower in the MI group (46%) than in the controls (85%). Egg production in the MI and HI groups was c. 50 and 30% of the C group. Egg size, embryonic survival and fry mass did not differ across groups. Sea lice influence reproductive development and egg production in S. alpinus, and consequently these parasites may influence populations via sublethal effects on broodfish, affecting growth and condition, and their reproductive output.

Influence of the lunar cycle on plasma melatonin, vitellogenin and sex steroids rhythms in Senegal sole, Solea senegalensis

The effect of the moon light cycle on plasma melatonin rhythms was examined in Senegal sole ( Solea senegalensis) exposed to natural outdoor or artificial indoor lighting conditions. Furthermore, in a second experiment, the effect of the lunar cycle on vitellogenin and sex steroids (Testosterone, T; Estradiol, E2; 11-ketotestosterone, 11kt) was studied using mature individuals during reproductive season. In the first experiment, during full moon, plasma melatonin peaked at night in covered tanks (deprived of night illumination) from both outdoor (133.2 ± 12.8 pg ml − 1 ) and indoor (190.6 ± 41.5 pg ml − 1 ) groups. However, for fish in the open tanks, exposed to approximately 0.3 lx of illumination, nocturnal plasma melatonin was significantly reduced ( p < 0.05), approaching to mid-light (ML) values, (79.6 ± 7.1 and 81.8 ± 14.0 pg ml − 1 , for outdoor and indoor groups, respectively). During new moon a similar pattern was observed in outdoor group: fish in the covered tank showed higher melatonin values than those in the open tank, which were exposed to the near undetectable night illumination. In the second experiment, plasma sex steroid concentrations were significantly higher during the full moon compared to the new moon. In the case of females, E2 concentration reduced from 2.4 ± 0.6 to 0.4 ± 0.1 ng ml − 1 between full and new moon samplings, while T decreased from 0.3 ± 0.0 to 0.2 ± 0.0 ng ml − 1 . Vitellogenin, however, did not show such differences between moon phases. In males, 11kt exhibited a plasma concentration of 14.3 ± 2.1 ng ml − 1 during full moon and 4.7 ± 0.7 ng ml − 1 during new moon while T values were 2.6 ± 0.4 and 1.0 ± 0.1 ng ml − 1 for full and new moon, respectively. In conclusion, these findings pointed out the high sensitivity to moon light of the Senegal sole, which could be using the melatonin signalling to synchronize their reproduction rhythms to the lunar cycle.

Relationship of plasma sex steroids to the mating season of copperheads at the north‐eastern extreme of their range

A multi-year radio-telemetric study of the copperhead Agkistrodon contortrix (Serpentes: Viperidae) was conducted at the north-eastern extreme of its range to determine the relationship of plasma sex steroids of males to the mating season. Blood samples were collected in situ approximately every 2 weeks (repeat-test group) from radio-telemetered males during the 7-month active season (April–October) from 2001 to 2003 and assayed for concentrations of testosterone (T) and progesterone (P4). Blood samples were also obtained from a large number of incidental males (single-test group) for the analysis of seasonal levels of T and P4. The profiles of T and P4 showed a peak in August–September that corresponded to the single mating season (late July to late September). Both T and P4 had similar seasonal profiles, but absolute levels of these steroids were significantly different, with concentrations of T four- to fivefold greater. The mating season of the population we investigated differs from other (e.g. southern) populations, which show two mating seasons (late summer/early fall and spring) before the period of ovulation in mid- to late spring. When a mating season is absent in spring, inseminated females are obligated to store sperm over winter until ovulation in the spring. In studies of A. contortrix that document two mating seasons, peak levels of T in males are coincident with both of these periods. In contrast, we found that peak levels of T and P4 in males coincided with the occurrence of the single mating season, and levels were basal in spring. These results further support the idea that plasma sex steroids influence the expression of sexual behaviour in A. contortrix and other snakes. Nonetheless, confirmation of this view awaits experimental studies.

Distinguishing Ovarian Maturity of Farmed White Sturgeon (Acipenser transmontanus) by Fourier Transform Infrared Spectroscopy: A Potential Tool for Caviar Production Management

Fourier transform infrared spectroscopy (FT-IR, 4000-400 cm(-1)) was applied to blood plasma of farmed white sturgeon (N = 40) to differentiate and predict the stages of ovarian maturity. Spectral features of sex steroids (approximately 3000 cm(-1)) and vitellogenin (approximately 1080 cm(-1)) were identified. Clear segregation of maturity stages (previtellogenesis, vitellogenesis, postvitellogenesis, and follicular atresia) was achieved using principal component analysis (PCA). Progression of oocyte development in the late phase of vitellogenesis was also monitored using PCA based on changes in plasma concentrations of sex steroid and lipid content. The observed oocyte polarization index (PI, a measure of nuclear migration) was correlated with changes in plasma sex steroid levels revealed by FT-IR PCA results. A partial least squares (PLS) model predicted PI values within the range 0.12-0.40 (R = 0.95, SEP = 2.18%) from differences in spectral features. These results suggest that FT-IR may be a good tool for assessing ovarian maturity in farmed sturgeon and will reduce the need for the invasive ovarian biopsy required for PI determination.

Effects of Shortened Photoperiod on Gonadotropin-Releasing Hormone, Gonadotropin, and Vitellogenin Gene Expression Associated with Ovarian Maturation in Rainbow Trout

Reproductive activities of salmonids are synchronized by changes in photoperiod, which control the endocrine system via the brain-pituitary-gonadal axis. Gonadotropin-releasing hormone (GnRH) in the brain regulates synthesis and release of the pituitary gonadotropins (GTHs; FSH and LH). FSH and LH in turn stimulate the production of sex steroids for oocyte growth and maturation-Inducing steroid hormones for oocyte maturation and ovulation, respectively, in female salmonids. To clarify effects of long-term photoperiod manipulations on the reproductive activity of salmonids from early recrudescence to ovulation, we Investigated the gene expression profiles of GnRH, GTHs, and vitellogenin (VTG), and plasma sex steroids in female rainbow trout (Oncorhynchus mykiss). In addition, the percentages of eyed embryos and hatched alevins were examined together with the number of ovulated eggs to evaluate the effects of photoperiod regimes on egg quality. During late summer, the mRNA levels of GnRHs, GTHalpha, and LHbeta, and the plasma level of a maturational steroid (17alpha,20beta-dihydroxy-4-pregnen-3-one; 17,20beta-P) were significantly elevated by a gradually shortened photoperiod under constant temperature, in accordance with accelerated sexual maturation. The percentages of eyed embryos and hatched alevins from fish ovulated in August were comparable to those of control fish observed in December. These results clearly indicate that syntheses of GnRHs, LH, VTG, and 17,20beta-P are effectively accelerated by a programmed long-short photoperiod regime in early recrudescent female rainbow trout, without a marked deterioration in egg quality.

Endocrine effects of the tapewormLigula intestinalisin its teleost host, the roach (Rutilus rutilus)

The effects of parasite infection by the cestode Ligula intestinalis on the reproductive function and endocrine system of wild roach Rutilus rutilus were evaluated. Gonad maturation, plasma vitellogenin, plasma steroid concentrations (i.e. progesterone, 11-keto-testosterone and 17-beta-estradiol) and brain aromatase activity were investigated in relation with parasitization. A low prevalence (8%) of ligulosed roach and a moderate impact of parasitization (mean parasitization index of 8.8%) were found in the studied population. Inhibition of gonad maturation generally resulted from infestation but 5% of the ligulosed roach nevertheless reached maturity. Main sex steroid plasma content was depleted in both genders. Male 11-keto-testosterone, female 17-beta-estradiol and progesterone plasma concentrations of both genders were, respectively, 27, 5 and 3 times lower in ligulosed fish when compared to their non-infected counterparts. Progesterone levels were negatively correlated with the parasitization index in females. Brain aromatase activity of infected roach was reduced to 50% of that of the non-infected fish. These results demonstrate significant negative effects on the reproductive function of wild roach infected by the tapeworm L. intestinalis collected from a site with low contamination.

Older triploid fish retain impaired reproductive endocrinology in the European sea bass Dicentrarchus labrax

This paper reports on an evaluation of growth, gonadal development and reproductive endocrinology of older triploid (3n) European sea bass Dicentrarchus labrax in comparison with their diploid (2n) counterparts throughout their fifth and seventh annual cycle of life. While older triploids retained impaired reproductive endocrinology, a sexually related dimorphic growth was observed with 3n females attaining the largest sizes. Comparisons of some body indexes showed that 3n females had a significantly lower hepato-somatic index (I(H)) than 2n females but a significantly higher viscero-somatic index (I(F)). In contrast, both male and female triploids showed significantly lower gonado-somatic index (I(G)) than diploids. Accordingly, diploids produced mature gametes but triploids did not, demonstrating that despite the longer time given to triploids for gonadal development, they could not reproduce. Furthermore, older triploids had lower levels of plasma sex steroids (testosterone, T; 11-ketotestosterone, 11-KT and oestradiol-17beta, E(2)) and luteinizing hormone (LH) than their 2n counterparts with 3n females showing drastic effects of triploidization on their reproductive endocrinology. Vitellogenin (VTG) was undetectable in 3n females. Gonadal content of steroid hormones and Sparus aurata-type gonadotropin-releasing hormone (sbGnRH) in the brain and pituitary were also lower in triploids compared with diploids. These results suggest that older 3n D. labrax retain functional sterility in both sexes, and 3n females might reach larger sizes than 3n males and their 2n counterparts in this species.

Regulation of Steroidogenic Acute Regulatory Protein Transcription in Largemouth Bass by Orphan Nuclear Receptor Signaling Pathways

The steroidogenic acute regulatory (StAR) protein mediates the rate-limiting step of mitochondrial transport of cholesterol for steroid biosynthesis. To investigate the regulation of this protein in lower vertebrates, we cloned the StAR coding region from large-mouth bass for analysis. Induction of the mRNA corresponded with increasing levels of plasma sex steroids in vivo. Cultures of largemouth bass ovarian follicles were exposed to dibutyryl cAMP (dbcAMP), a potent signaling molecule for steroidogenesis. StAR mRNA expression was significantly up-regulated by dbcAMP signaling, suggesting that the 5' regulatory region of the gene is functionally conserved. To further analyze its transcriptional regulation, a 2.9-kb portion of the promoter was cloned and transfected into Y-1 cells, a steroidogenic mouse adrenocortical cell line. The promoter activity was induced in a dose-responsive manner upon stimulation with dbcAMP; however, deletion of 1 kb from the 5' end of the promoter segment significantly diminished the transcriptional activation. A putative retinoic acid-related receptor-alpha/rev-erb alpha element was identified between the -1.86- and -2.9-kb region and mutated to assess its potential role in dbcAMP regulation of the promoter. Mutation of the rev-erb alpha element significantly impeded dbcAMP-induced activation. Chromatin immunoprecipitation and EMSA results revealed rev-erb alpha and retinoic acid-related receptor-alpha enrichment at the site under basal and dbcAMP-induced conditions, respectively. These results implicate important roles for these proteins previously uncharacterized for the StAR promoter. Altogether these data suggest novel regulatory mechanisms for dbcAMP up-regulation of StAR transcription in the distal part of the largemouth bass promoter.

Acute experimental autoimmune encephalomyelitis induces sex dimorphic changes in neuroactive steroid levels

Incidence, progression and severity of the multiple sclerosis, an inflammatory, demyelinating disease of the central nervous system (CNS) are affected in a sex-depending way. Physiological situations characterized by changes in sex steroid plasma levels, such as menstrual cycle, menopause or pregnancy, affect the disease course, suggesting that these molecules might exert a role in this disease. In order to understand better this possible relationship, we have here assessed the levels of neuroactive steroids present in different CNS regions of male and female rats affected by acute experimental autoimmune encephalomyelitis (EAE). In addition, we compared these levels with those present in plasma. Data obtained by liquid chromatography–tandem mass spectrometry indicate that the levels of neuroactive steroids show sex and regional differences in control and EAE nervous system and that a clear difference is also observed between CNS and plasma levels. In particular, among neuroactive steroids here considered, the levels of progesterone metabolites (i.e., dihydroprogesterone, tetrahydroprogesterone and isopregnanolone) and testosterone metabolites (i.e., dihydrotestosterone and 5α-androstane-3α17β-diol), show sex dimorphic and region-specific changes in the CNS. Moreover, some changes observed in the CNS were not detected in plasma. These findings might represent an interesting background to design therapies and possibly sex-specific therapies for multiple sclerosis based on neuroactive steroids or synthetic ligands able to interact with classical and non-classical steroid receptors.

Reproductive biology of middle Mississippi River shovelnose sturgeon: insights from seasonal and age variation in plasma sex steroid and calcium concentrations

Shovelnose sturgeon Scaphirhynchus platorynchus are endemic to the Mississippi River drainage and are commercially harvested for roe in several states. The status of shovelnose sturgeon populations throughout much of its range is unknown or declining; Mississippi River stocks are experiencing recruitment overfishing and may be at risk of collapse. Restoration of shovelnose sturgeon populations will require additional information on their reproductive biology, including age at maturity and reproductive status and cycles. The objectives of this study were to determine the effects of reproductive stage, fish age, and season on plasma 17-β estradiol (E2) and calcium (Ca2+) concentrations in female shovelnose sturgeon and plasma testosterone (T) concentrations in male shovelnose sturgeon from the middle Mississippi River (MMR). The relationship was also assessed between plasma vitellogenin (VTG) and Ca2+concentrations for female shovelnose sturgeon. Plasma E2 and Ca2+concentrations in females and T concentrations in males differed among reproductive stages, consistent with results of prior research on shovelnose sturgeon and other sturgeon species. VTG and Ca2+concentrations were strongly correlated in female shovelnose sturgeon, indicating that Ca2+can be used as a surrogate for VTG assays for identification of vitellogenic females, as has been shown for other sturgeon species. Age at maturity was estimated at 10 years for males and 9 years for females based on T and E2 profiles, consistent with other recent age at maturity estimates in the MMR and Wabash River determined by gonadal examination. Peaks in plasma sex steroid and Ca2+concentrations during April and October possibly reflected the spring spawning season and provide additional evidence in support of recent documentation of autumn spawning by shovelnose sturgeon in the MMR. Additional research is needed to elucidate autumn spawning by shovelnose sturgeon in the MMR.

Naturally-induced endocrine disruption by the parasite Ligula intestinalis (Cestoda) in roach ( Rutilus rutilus)

Fish represent the most frequently used vertebrate class for the investigation of endocrine disruption (ED) in wildlife. However, field studies are complicated by exposure scenarios involving a variety of anthropogenic and natural influences interfering with the endocrine system. One natural aspect rarely considered in ecotoxicological studies is how parasites modulate host physiology. Therefore, investigations were carried out to characterise the impacts of the parasitic tapeworm Ligula intestinalis on plasma sex steroid levels and expression of key genes associated with the reproduction in roach ( Rutilus rutilus), a sentinel species for wildlife ED research. Parasitisation by L. intestinalis suppressed gonadal development in both genders of roach and analysis of plasma sex steroids revealed substantially lower levels of 17β-oestradiol (E2) and 11-ketotestosterone (11-KT) in infected females as well as E2, 11-KT, and testosterone in infected males. Consistently, in both, infected females and males, expression of the oestrogen dependent genes such as vitellogenin and brain-type aromatase in liver and brain was reduced. Furthermore, parasitisation differentially modulated mRNA expression of the oestrogen and androgen receptors in brain and liver. Most prominently, liver expression of oestrogen receptor 1 was reduced in infected females but not in males, whereas expression of oestrogen receptor 2a was up-regulated in both genders. Further, insulin-like growth factor 1 mRNA in the liver was increased in infected females but not in males. Despite severe impacts on plasma sex steroids and pituitary gonadotropin expression, brain mRNA levels of gonadotropin-releasing hormone (GnRH) precursors encoding GnRH2 and GnRH3 were not affected by L. intestinalis-infection. In summary, the present results provide basic knowledge of the endocrine system in L. intestinalis-infected roach and clearly demonstrate that parasites can cause ED in fish.

Plasma sex steroids and SHBG in patients with breast cancer and their relation to tumor oestrogen-dependency.

In order to evaluate the possible role of sex steroids in breast cancer and their relation to the presence of oestradiol receptors (ER), ER were quantified in 112 breast adenocarcinomas and 14 benign tumours. The sex hormones of the patients were determined in plasma: oestradiol (E2), progesterone, testosterone, androstenedione, as well as SHBG and gonadotrophins. The ER were evaluated in cytosolic and nuclear fractions. Steroids and gonadotrophins were determined by RIA. SHBG was determined by following a procedure of saturation with tritiated dihydrostestosterone. The mean levels of E2 in postmenopausal women with breast cancer was found to be significantly higher than of the control group. However, no significant variations for the rest of the steroids were seen among the patients with breast cancer and the control group or benign tumour. Nor was any relation found between the presence of ER and the hormonal steroids studied or SHBG. It is suggested that the possible influence of the sex steroids in the etiopathogenesis of breast cancer might be due to the greater production accompanied by a parallel metabolic clearance that would maintain their plasma levels.

Delayed sexual maturation through gonadotropin receptor vaccination in the rainbow trout Oncorhynchus mykiss

In fish, gonadotropin hormones FSH-GTH1 and LH-GTH2 are less specific for their cognate receptors than in mammals. The respective reproductive functions of fish LH and FSH are thus difficult to establish. We aimed to study the effect of specific antagonists of the two gonadotropin receptors on trout sexual maturation in both sexes by targeting specific regions of LH and FSH receptors, Lhr and Fshr. Filamentous phages displaying Lhr specific or Fshr specific decapeptides from the extracellular hormone binding domain were engineered. Recombinant phages were used as receptor-specific antagonistic vaccines. Male and female trouts were immunized with anti-LHR, anti-FSHR, anti-FSHR + LHR or adjuvant alone, through multiple injections over 8–24 weeks, starting at different stages of sexual maturation. The consequences of immunization on gonadal development were evaluated by determining gonad growth, by histological analysis of testis and ovaries at the end of the vaccination period and by measuring blood plasma sex steroids using radioimmunoassay. We show for the first time in fish that the anti-receptor vaccinations could have specific antagonistic effects on the development of the reproductive functions; while the anti-FSHR affected the sexual maturation of prepubertal males and delayed sperm production, the anti-LHR blocked vitellogenesis in females. In maturing males, the combined anti-FSHR + LHR vaccine inhibited spermatogenesis and affected steroidogenesis. In that case, the effects of the vaccine on spermatogenesis were transient and reversible when immunization was stopped. Such an immunological strategy to specifically and transiently inhibit a receptor provides a promising approach for discovering their specific functions; it could also lead to a new technology for controlling the onset of puberty in aquaculture species.

Behavioural and endocrine effects of chronic cola intake

Consumption of cola beverages is very high worldwide. The health effects of cola intake are not clear, although epidemiological studies point toward associations with obesity, kidney diseases and osteoporosis. Experimental studies are surprisingly rare. In this study, we substituted drinking water of adult male Wistar rats with three different cola beverages for 3 months. Behavioural phenotyping, measurement of sex steroids in plasma and oxidative stress in testes were performed at the end of the study. A light-dark box showed increased locomotor activity and anxiety in all groups with cola intake. A subtle anti-depressive effect was seen in the forced swim test. Chronic cola intake increased both oestradiol and testosterone levels suggesting an additional mechanism of action beyond the known effects of caffeine on adenosine receptors.

Activity of the pituitary–gonadal axis is increased prior to the onset of spawning migration of chum salmon

The activity of the pituitary-gonadal axis (PG axis) in pre-migratory and homing chum salmon was examined because endocrine mechanisms underlying the onset of spawning migration remain unknown. Pre-migratory fish were caught in the central Bering Sea in June, July and September 2001, 2002 and 2003, and in the Gulf of Alaska in February 2006. They were classified into immature and maturing adults on the basis of gonadal development. The maturing adults commenced spawning migration to coastal areas by the end of summer, because almost all fish in the Bering Sea were immature in September. In the pituitaries of maturing adults, the copy numbers of FSHbeta mRNA and the FSH content were 2.5- to 100-fold those of the immature fish. Similarly, the amounts of LHbeta mRNA and LH content in the maturing adults were 100- to 1000-fold those of immature fish. The plasma levels of testosterone, 11-ketotestosterone and estradiol were higher than 10 nmol l(-1) in maturing adults, but lower than 1.0 nmol l(-1) in immature fish. The increase in the activity of the PG-axis components had already initiated in the maturing adults while they were still in the Gulf of Alaska in winter. In the homing adults, the pituitary contents and the plasma levels of gonadotropins and plasma sex steroid hormones peaked during upstream migration from the coast to the natal hatchery. The present results thus indicate that the seasonal increase in the activity of the PG axis is an important endocrine event that is inseparable from initiation of spawning migration of chum salmon.

Spermatogenesis, epididymis morphology and plasma sex steroid secretion in the male lizard Podarcis sicula exposed to diuron

The present study investigates the effects of diuron, a substituted urea-based herbicide, in the male lizard Podarcis sicula utilizing quantitative and qualitative morphological features of the reproductive system and endocrinological analysis. Besides the control group, lizards were divided into three groups ([a–c]) ( n=6/group) and placed for 3 weeks in terraria on polluted soil substrate sprayed with 3.75 L/ha of herbicide Toterbane 50F (50% diuron). Each terrarium was supplemented either with drinking water contaminated by herbicide (i.e. 1.08 μg/mL of diuron; group [a]), or with food contaminated by herbicide (i.e. 5.4 mg of diuron; group [b]), or with drinking water and food contaminated as described above (group [c]). None of the animals exposed to the contaminant showed any signs of general toxicity or death during the course of the experiments. Severe testicular effects are evidenced in all herbicide-treated groups, although, such effects are of a greater magnitude in lizards exposed to contaminated water (groups [a] and [c]). The main degenerative changes observed include: (1) a significant decrease in the mean gonadosomatic index of 55% in group [a] ( P<0.001), 21% in group [b] ( P<0.01) and 34% in group [c] ( P<0.001) compared with control group; (2) a significant shrinking ( P<0.001) of seminiferous tubule diameter (more than 60% of the control) in groups [a] and [c], and about 18% in group [b] ( P<0.01); (3) a significant decrease in the crude numbers of spermatogonia of 92% in group [a] ( P<0.001), 27% in group [b] ( P<0.01) and 62% in group [c] ( P<0.001) compared with control group. A complete loss of meiotic and mature germ cells in groups [a] and [c], and a reduction of primary spermatocytes, secondary spermatocytes and spermatids (more than 27% of the control) and a decrease of spermatozoa (more than 90% of the control) in group [b]; and (4) an hypertrophy of interstitial connective tissue which contains numerous lymphocytes, neutrophils and monocytes. The decrease and/or loss of germ cells seems to be related to an induction of inflammation (necrosis) rather than to apoptotic processes. Indeed, this hypothesis is supported by a TUNEL-assay, which failed to reveal any apoptotic cells either in the seminiferous epithelium or in the interstitial space in the testis of all exposed groups. Also the epididymis appears affected by diuron exposure. In particular, in experimental groups [a] and [c] it is regressed with abundant connective tissue and low epithelial cells without secretory granules, whereas in group [b] it appears partially regressed, with some secretory granules still present. At the same time, an impairment of the plasma sex-hormone levels is observed in treated lizards, as evidenced by RIA analysis. Testosterone values significantly decreased by 43% in group [a] ( P<0.001), 34% in group [b] ( P<0.01) and 52% in group [c] compared with control group. Instead, 17 β-estradiol plasma content is undetectable in all diuron-exposed lizards. Taken together, the results presented here indicate that diuron exposure resulted in direct male reproductive toxicity and reveal that this lizard is suitable as a laboratory reptile species for toxicological investigations.

Brain and gonadal aromatase activity and steroid hormone levels in female and polymorphic males of the peacock blenny Salaria pavo

In the peacock blenny Salaria pavo large males with well-developed secondary sexual characters establish nests and attract females while small “sneaker” males mimic female sexual displays in order to approach the nests of larger males and parasitically fertilize eggs. These alternative reproductive tactics are sequential, as sneakers irreversibly switch into nesting males. This transition involves major morphologic and behavioral changes and is likely to be mediated by hormones. This study focuses on the role of aromatase, an enzyme that catalyses the conversion of androgens into estrogens, in the regulation of male sexual polymorphism in S. pavo. For this, sex steroid plasma levels and aromatase activity (AA) in gonads, whole brain and brain macroareas were determined in sneakers, transitional males (i.e. sneakers undergoing the transition into nesting males), nesting males and females collected in the field. AA was much higher in ovarian tissue than in testicular tissue and accordingly circulating estradiol levels were highest in females. This supports the view that elevated AA and estradiol levels are associated with the development of a functional ovary. Transitional males are in a non-reproductive phase and had underdeveloped testes when compared with sneakers and nesting males. Testicular AA was approximately 10 times higher in transitional males when compared with sneakers and nesting males, suggesting high AA has a suppressive effect on testicular development. Nesting males had significantly higher plasma levels of both testosterone (T) and 11-ketotestosterone when compared with the other male morphs and previous studies demonstrated that these androgens suppress female-like displays in sneakers. In the brain, AA was highest in macroareas presumably containing hypothalamic nuclei traditionally associated with the regulation of reproductive behaviors. Overall, females presented the highest levels of brain AA. In male morphs AA increased from sneakers, to transitional males, to nesting males in all brain macroareas. These results suggest that the transition into the nesting male tactic is accompanied both by an increase in testicular androgen production and by a higher conversion of androgens into estrogens in the brain. The increase in androgen production is likely to mediate the development of male secondary sexual characters while the increase in brain AA may be related to the behavioral changes associated with tactic transition.

Effects of cadmium chloride on the development of rainbow trout Oncorhynchus mykiss early life stages

The sub‐chronic (28–56 days) effects of exposure to low concentrations of cadmium (Cd; 0·05, 0·25, 0·50 and 2·50 μg l−1) shortly following fertilization on embryos, larvae and juvenile rainbow trout Oncorhynchus mykiss were examined. Premature hatching occurred at lower concentrations (0·05 and 0·25 μg l−1 Cd), however, delayed hatching was seen in the 2·50 μg l−1 Cd group, with &gt;90% of hatching occurring on the last day of the hatching period. Larval growth was negatively affected by Cd exposure in a concentration‐dependent manner. Larvae exposed to 2·50 μg l−1 Cd were 13·9 ± 0·8% shorter in total length (LT) and weighed 22·4 ± 3·5% (mean ±s.e.) less than controls at the end of the exposure period. Plasma sex steroid concentrations (oestradiol in juvenile females and 11‐ketotestosterone in juvenile males) were elevated (four‐ to 10‐fold over controls) in exposed fish in both males and females, following 28 days of exposure to 0·05, 0·25 and 0·50 μg l−1 Cd, respectively. These results suggest that environmentally realistic concentrations (in the μg l−1 range) of Cd can affect the development of O. mykiss impacting embryos, larvae and juvenile fish.

Age-related changes in ovarian characteristics, plasma sex steroids and fertility during pubertal development in captive female Murray cod Maccullochella peelii peelii

Age-related changes in ovarian development characteristics and plasma sex steroids in female Murray cod were examined throughout their second, third and fourth years of life to better understand the physiological and endocrine processes associated with puberty in this species in captivity. Spawning performance of 2+ and 3+ year old females was also assessed to identify ontogenetic differences in egg fertility. Puberty was acquired in 38% of 1+ year old females and 100% of age 2+ females. By age 3+, all females had developed full (adult) reproductive function. Ovarian development in pubertal fish was characterised by a rapid transition between cortical alveoli and lipid droplet oogenic phases, coinciding with significantly lower plasma 17β-oestradiol in age 2+ females ( p < 0.05). Mean mature oocyte diameter (2.44 mm), post-fertilisation viability (30.80%) and hatchability (0.99%) of eggs from age 2+ females were significantly reduced relative to age 3+ adults (2.81 mm, 84.89% and 23.58%, respectively). Ovaries of pubertal Murray cod exhibited both vitellogenic and ovulatory capacities, yet functional abnormalities during secondary oocyte growth are likely to have contributed to poor egg fertility and consequently, evaluations of age-at-first maturity based on the presence of advanced ovarian stages may overestimate the reproductive potential of younger broodstock populations.

Using Blood Plasma for Monitoring Organochlorine Contaminants in Juvenile White Sturgeon, Acipenser transmontanus, from the Lower Columbia River

Organochlorine (OC) pesticide concentrations in blood plasma samples from 88 juvenile white sturgeon collected from the lower Columbia River were measured and compared to plasma sex steroid and OC tissue levels previously measured in corresponding fish. Significant squared correlation coefficients between summation operator DDT concentrations in sturgeon plasma and gonads and livers were 0.37 and 0.32, respectively. Significant negative correlations between plasma testosterone concentration and plasma Sigma DDT concentration in male fish (r(2)=0.26), plasma 17beta estradiol concentration and plasma Sigma DDT concentration in female fish (r(2)=0.38) and condition factor and plasma Sigma DDT concentration in all fish were found (r(2)=0.17). These results suggest that blood plasma may be a suitable nondestructive method for monitoring adult sturgeon population for persistent OC contaminants.