Plasma Sex Steroid Research Articles

Ex Vivo 13C-Metabolic Flux Analysis of Porcine Circulating Immune Cells Reveals Cell Type-Specific Metabolic Patterns and Sex Differences in the Pentose Phosphate Pathway.

In general, females present with stronger immune responses than males, but scarce data are available on sex-specific differences in immunometabolism. In this study, we characterized porcine peripheral blood mononuclear cell (PBMC) and granulocyte energy metabolism using a Bayesian 13C-metabolic flux analysis, which allowed precise determination of the glycolytic, pentose phosphate pathway (PPP), and tricarboxylic acid cycle (TCA) fluxes, together with an assessment of the superoxide anion radical (O2•-) production and mitochondrial O2 consumption. A principal component analysis allowed for identifying the cell type-specific patterns of metabolic plasticity. PBMCs displayed higher TCA cycle activity, especially glutamine-derived aspartate biosynthesis, which was directly related to mitochondrial respiratory activity and inversely related to O2•- production. In contrast, the granulocytes mainly utilized glucose via glycolysis, which was coupled to oxidative PPP utilization and O2•- production rates. The granulocytes of the males had higher oxidative PPP fluxes compared to the females, while the PBMCs of the females displayed higher non-oxidative PPP fluxes compared to the males associated with the T helper cell (CD3+CD4+) subpopulation of PBMCs. The observed sex-specific differences were not directly attributable to sex steroid plasma levels, but we detected an inverse correlation between testosterone and aldosterone plasma levels and showed that aldosterone levels were related with non-oxidative PPP fluxes of both cell types.

Effects of different hormonal treatments on spermatogenesis advancement in hatchery-produced greater amberjack Seriola dumerili (Risso 1810)

In earlier studies, wild-caught greater amberjack Seriola dumerili (Risso, 1810) males reared in sea cages showed gametogenesis impairment and low sperm production and quality. Here, we (a) examined if F1 hatchery-produced males reared in sea cages also exhibit reproductive dysfunctions and (b) evaluated the effects of gonadotropin releasing hormone agonist (GnRHa) administration through injections (GnRHainj) or sustained-release implants (GnRHaimpl), and human chorionic gonadotropin (hGC) injections on spermatogenesis/spermiation enhancement. Fish were given a hormone treatment just prior to the spawning season, and were transferred to land-based tanks, according to an established spawning induction protocol. Blood samples (n = 6) were obtained on Days 0, 7 and 13 after treatment. Testis samples were obtained on Days 0 (n = 4) and 13 (n = 2 per treatment). The fish prior to their transfer from the sea cages to the land-based tanks, exhibited a low gonadosomatic index, altered sex steroid hormone profile and high density of testicular apoptotic cells. After transfer to tanks, there was a general depression of sex steroid plasma levels parallel to an increase in cortisol concentrations. Despite the negative effect on steroidogenesis by the transfer from the sea, the hormonal treatments increased the number of fish from where sperm could be obtained, as well as testis growth, and reduced testicular apoptosis. Treatment with hCG resulted in the most significant changes in spermatogenesis, while GnRHaimpl appeared to induce less intense, but likely longer-lasting effects. The study indicated that F1 hatchery-produced males also exhibited reproductive dysfunctions as wild-caught captive-reared greater amberjack, and that the observed positive effects of the hormone treatments on spermiation/spermatogenesis were likely mediated by factors other than sex steroid hormones.

Associations of urinary non-persistent endocrine disrupting chemical biomarkers with early-to-mid pregnancy plasma sex-steroid and thyroid hormones

Background/ObjectivesPregnant women are exposed to numerous endocrine disrupting chemicals (EDCs) that can affect hormonal pathways regulating pregnancy outcomes and fetal development. Thus, we evaluated overall and fetal sex-specific associations of phthalate/replacement, paraben, and phenol biomarkers with sex-steroid and thyroid hormones. MethodsIllinois women (n = 302) provided plasma for progesterone, estradiol, testosterone, free T4 (FT4), total T4 (TT4), and thyroid stimulating hormone (TSH) at median 17 weeks gestation. Women also provided up-to-five first-morning urine samples monthly across pregnancy (8–40 weeks), which we pooled to measure 19 phthalate/replacement metabolites (reflecting ten parent compounds), three parabens, and six phenols. We used linear regression to evaluate overall and fetal sex-specific associations of biomarkers with hormones, as well as weighted quantile sum and Bayesian kernel machine regression (BKMR) to assess cumulative associations, non-linearities, and chemical interactions. ResultsIn women of relatively high socioeconomic status, several EDC biomarkers were associated with select hormones, without cumulative or non-linear associations with progesterone, FT4, or TT4. The biomarker mixture was negatively associated with estradiol (only at higher biomarker concentrations using BKMR), testosterone, and TSH, where each 10% mixture increase was associated with −5.65% (95% CI: −9.79, −1.28) lower testosterone and −0.09 μIU/mL (95% CI: −0.20, 0.00) lower TSH. Associations with progesterone, testosterone, and FT4 did not differ by fetal sex. However, in women carrying females, we identified an inverted u-shaped relationship of the mixture with estradiol. Additionally, in women carrying females, each 10% increase in the mixture was associated with 1.50% (95% CI: −0.15, 3.18) higher TT4, whereas in women carrying males, the mixture was associated with −1.77% (95% CI: −4.08, 0.58) lower TT4 and −0.18 μIU/mL (95% CI: −0.33, −0.03) lower TSH. We also identified select chemical interactions. ConclusionSome biomarkers were associated with early-to-mid pregnancy hormones. There were some sex-specific and non-linear associations. Future studies could consider how these findings relate to pregnancy/birth outcomes.

Effect of prolonged feeding of broodstock diet with increased inclusion of essential n-3 fatty acids on maturing and spawning performance in 3-year-old Atlantic salmon (Salmo salar)

Atlantic salmon (Salmo salar) broodstock recruits are normally fed a specialized diet with a higher content of essential nutrients for a limited time period prior to fasting and transfer to freshwater. Typically, this period lasts for about six months, but may vary among producers. Reduced use of marine ingredients in commercial salmon diets during the last decades has affected the content of essential nutrients, such as n-3 long chained polyunsaturated fatty acids (LC-PUFA), minerals and vitamins. Furthermore, to minimize the risk of losses and implement new breeding achievements faster, breeding companies have shortened the production cycle of broodstock from 4 to 3 years, which may affect the number of fish that are large enough to mature. In the present study, we have extended the broodstock feeding period from 6 to 15 months prior to the freshwater transfer giving a higher content of n-3 LC-PUFA (higher inclusion of marine oils) from February to December (Phase 1), and thereafter a diet with a higher energy content to ensure growth towards the spring and maturation (Phase 2). Four sea cages with approximately 80.000 salmon postsmolt, two sea cages with males and two with females, were given a control diet and an experimental diet. Samples were taken in Phase 1 at start (1.7 kg), mid (3.4 kg) and end Phase 1/start of Phase 2 (8.3 kg), and end of Phase 2 (13.4 kg). The fish were thereafter fasted, and selected fish transferred to landbased freshwater tanks where light and temperature were used to manipulate the spawning time of the fish in two groups (early or late). Due to disease in the facility, measures of egg quality and hatching were only obtained from the early group. During the trial and spawning period, biometrical measurements were recorded, and samples of liver, gonad, fillet and red blood cells (RBC) were collected for fatty acid composition and blood plasma for analysis of lipid and health-related parameters. Samples were also collected for gonadal transcriptomic analysis by microarray and qPCR (end Phase 2) and plasma steroids (end Phase 2, mid maturation and spawning). Males fed the test diet had a larger body size compared to the control group at the end of Phase 2, while no differences were observed between dietary groups for the females. Total mortality in the trial was lower in the test group compared to the control, losses were caused mainly by sea lice treatments, loser fish or cardiomyopathy syndrome (CMS). The dietary LC-PUFA levels in the test diet were reflected in the tissues particularly during Phase 1, but only different in the fillet samples and eggs at the end of Phase 2 and at spawning. Plasma sex steroids content increased at mid maturation and showed lower levels of androgens and estrogens in females fed the test diet compared to the control. At the end of Phase 2, transcriptional analysis showed upregulation of steroidogenic enzymes, although not reflected in changes in plasma steroids in Phase 2, indicating changes to come during maturation. The differences in LC-PUFA content in tissues and plasma steroids did not appear to affect fecundity, sperm quality, egg survival or hatching rate, but the test group had larger eggs compared to the control in the early spawner-group. Prolonged feeding of n-3 LC-PUFA to pre-puberty Atlantic salmon broodstock appears to be important for higher survival in challenging sea cage environments and has an effect on sex steroid production that, together with high energy diet during early maturation, cause the test group to produce larger eggs.

Reproductive cycles of alligator snapping turtles (Macrochelys temminckii)

The alligator snapping turtle (Macrochelys temminckii) is a species for which captive propagation and reintroduction programs are well established; however, little is known about its reproductive behavior and physiology. In this study, we measured monthly plasma sex steroid hormone concentrations of androgen (T + DHT) estradiol-17B (E2), and progesterone (P4), and used ultrasonography to monitor annual reproductive cycles of a captive population of alligator snapping turtles that is maintained under semi-natural conditions in southeastern Oklahoma. Concurrently, we used automated radio telemetry to measure the relative activity levels of male and female alligator snapping turtles and examine these activity patterns in the context of their reproductive cycles. We also measured monthly concentrations of the glucocorticoid (GC) corticosterone (CORT). Seasonal variation was only detected for T in males, but was observed for T, E2, and P4 in females. Vitellogenesis began in August and ended in April and coincided with elevated E2. Ovulation took place 10–29 April and the nesting period lasted from 11 May – 3 June. Males exhibited greater relative activity levels than females in the fall, winter, and early spring, which coincided with the period when mature sperm would be available for mating. Females were more active than males during the peri-nesting period in the spring. Seasonal changes in CORT were detected and did not differ between males and females. CORT concentrations were elevated in the late spring and summer, coincident with the foraging season, and depressed in the fall, and winter, and at their nadir in the early spring.

Reproductive Maturation of Meagre Argyrosomus regius (Asso, 1801) Reared in Floating Cages.

The meagre Argyrosomus regius (Asso, 1801) is a promising aquaculture species that shows reproductive dysfunctions when reared in tanks. The aim of this study was to assess the capacity of meagre, reared in cages under routine farming conditions, to mature gonads and reproduce spontaneously. Meagre adults, reared in a fish farm located in the Gulf of Taranto (Italy), were sampled from March to July 2021. The gonadosomatic index and sex steroid plasma concentrations increased from March-April to June, and then decreased in July. In March-April, most of the females showed perinucleolar or cortical alveoli oocytes as the most advanced stages in the ovaries, and most of the males had testes at early spermatogenesis stage. In June, most of the sampled females had oocytes at late vitellogenesis or early post-vitellogenesis stages, and males had seminiferous tubules filled with spermatozoa. In July, most of the females had signs of previous spawning, and males showed scarce amounts of luminal spermatozoa. The present study demonstrated the capacity of meagre, reared in sea cages under commercial conditions, to carry out gametogenesis and spontaneously release gametes. Meagre reproduction, under routine farming conditions, may represent an opportunity for the expansion of meagre aquaculture production.

Nonlethal Tools to Identify Mass Ovarian Follicular Atresia in Burbot

AbstractSkipped spawning, or variation in spawning periodicity, occurs in many annual spawning fish species and is an important consideration for population management. We assessed plasma sex steroid concentrations and measured gonad size and ovarian follicle diameter as metrics to nonlethally identify mass ovarian follicular atresia, which may contribute to skipped spawning in Burbot Lota lota. We maintained wild fish in captivity and exposed them to increasing water temperatures during a 3-wk period before the spawning season to induce mass ovarian follicular atresia. We collected ovarian follicles, blood plasma, and gonadal sonograms from fish weekly between January 28, 2018, and March 25, 2018. We histologically analyzed ovarian follicles to confirm stage of maturity. We measured concentrations of plasma sex steroids testosterone (T) and estradiol-17β (E2) by radioimmunoassay. We measured gonad diameter and circumference by ultrasonography and ovarian follicle diameter by image analysis. Mean plasma T concentration decreased from 8.94 ng/mL during late vitellogenesis to 1.83 ng/mL during atresia, suggesting that plasma T concentrations may be used to identify mass ovarian follicular atresia. We do not recommend using plasma E2 concentrations to identify mass ovarian follicular atresia because E2 concentrations rapidly decreased during the completion of vitellogenesis and the initiation of atresia in Burbot; therefore, plasma E2 may not accurately identify mass ovarian follicular atresia. Mean gonad diameter measured by ultrasonography decreased from 4.05 cm during late vitellogenesis to 3.65 cm during atresia. Mean diameter of ovarian follicles decreased during the final week of the study, suggesting that ovarian follicle diameter may be used to identify advanced mass ovarian follicular atresia. The nonlethal tools assessed—plasma sex steroid concentrations, ultrasonography, and ovarian follicle diameter—enable fisheries biologists to determine the occurrence and frequency of mass ovarian follicular atresia among Burbot in Lake Roosevelt and may be applied to other Burbot populations.

Variation in immuno-reproductive milieu of testis in Clarias magur from pre-spawning to spawning phase: An indication towards non-canonical role of immune elements in testes

Immune mechanisms are major players in ensuring the normal functioning of testicular functions. However, apart from their role in active defence against pathogens, prior studies have also suggested a possibility for reproduction-related (non-immune) functions of certain immune elements. This study employs a comparative transcriptomics approach followed by network analysis for tracking the variation in the immuno-reproductive milieu of Clarias magur testis in spawning versus pre-spawning phase. The results show a significant modulation of both reproduction and immune-relevant genes in spawning versus pre-spawning phase. The functional enrichment of the upregulated reproduction-relevant gene network also shows immune-related biological processes which indicates a probability of involvement of these candidates in spermatogenesis-related events for switching from pre-spawning to spawning phase. The upregulated immune network is highly dense with 40 hubs, 10 cluster sub-networks and 142 functionally enriched pathways in comparison to its downregulated counterpart with only 5 hubs, 1 cluster and 1 enriched pathway. These findings indicate that the synchronisation in modulation of both reproductive and immune-related factors is critical for progression of testicular events guiding the switch from pre-spawning to spawning phase. The reproductive phase-dependent variation in plasma sex steroid levels and the selected genes for quantitative PCR also corroborated this hypothesis. The study also serves as a preliminary screening step for probable immune candidates that may be involved in reproductive functions of testis in addition to defence.

Growth, sexual transition, and maturation of blacktip grouper Epinephelus fasciatus under long-term artificial rearing: Puberty and its associated physiological and endocrine changes

The blacktip grouper Epinephelus fasciatus is considered a valuable marine fish for aquaculture. This study was conducted to obtain information on its age of puberty, growth performance, gonadal development, sexual transition pattern, and plasma sex steroid levels from juvenile stage to maturity in specimens reared in captivity. The stages of gonadal development were identified by histological analysis, and plasma sex steroid levels were measured using an enzyme-linked immunosorbent assay. Our results show that maturation and spawning in blacktip groupers occurred at almost 3 years of age; although some females matured at 2 years. Plasma 17β-estradiol (E2) levels increased rapidly to high levels during puberty at 3 years of age, especially during the late stage of vitellogenesis at the tertiary yolk stage, and reduced at the ripe and ovulated stages prior to spawning. The gonadal somatic index also showed an increase consistent with the rise in plasma E2 levels. Plasma 11-ketotestosterone (11-KT) levels increased dramatically during sex change and in males. The significant decrease in plasma E2 levels and remarkable increase in plasma 11-KT were the key factors for the progress of sex change in these fish. Interestingly, there were the concomitant appearances of spermatocytes in ovaries of immature females at 1 year 3 months of age. The earliest appearance of sex-transitioning and male fish in blackfish groupers was observed at 2 years, although no mature females were observed below this age. This indicated that the sex change and male transformation in these fish took place without them undergoing the mature female stage. In conclusion, this species had the ability to reach puberty at 2 years of age, but most individuals matured and were functional at 3 years old. Puberty might be affected by age rather than size. In addition, although this species had a protogynous hermaphrodite pattern, when individuals born in the same year were reared together, some individuals were capable of directly becoming males without experiencing a mature female stage. This was due to the lack of socialization in captivity wherein individuals of various ages do not coexist, unlike that in the wild.

Long-term monitoring of egg-laying cycle using ultrasonography reveals the reproductive dynamics of circulating sex steroids in an oviparous catshark, Scyliorhinus torazame

The many diverse reproductive strategies of elasmobranchs (sharks, skates and rays) from lecithotrophic oviparity to matrotrophic viviparity have attracted significant research attention. However, the endocrine control of elasmobranch reproduction is less well-documented largely due to their reproductive characteristics, such as a long reproductive cycle, and/or repeated internal fertilization using stored sperm in oviparous species. In the present study, for the first time, we succeeded in non-invasive monitoring of the continuing egg-laying cycle of the cloudy catshark Scyliorhinus torazame using portable ultrasound devices. Furthermore, long-term simultaneous monitoring of the egg-laying cycle and measurement of plasma sex steroids revealed cycling patterns of estradiol-17β (E2), testosterone (T) and progesterone (P4). In particular, a decline in T followed by a reciprocal surge in plasma P4 were consistently observed prior to the appearance of the capsulated eggs, implying that P4 is likely associated with the ovulation and/or egg-case formation. While the cycling pattern of E2 was not as apparent as those of T and P4, threshold levels of E2 (>5 ng/mL) and T (>1 ng/mL) appeared to be crucial in the continuation of egg-laying cycle. The possibility to trace the dynamics of plasma sex steroids in a single individual throughout the reproductive cycles makes the catshark a useful model for regulatory and mechanistic studies of elasmobranch reproduction.

Estropause, Sex Hormones and Metal Homeostasis in the Mouse Brain.

Alterations in brain metal ion homeostasis have been reported with aging and are implicated in the pathogenesis of neurodegenerative diseases. To assess whether age-related changes in hypothalamic-pituitary-gonadal (HPG) hormones might be involved in modulating brain metal ion homeostasis, we treated 7.5-month intact, sham-ovariecomized and ovariectomized C57B6SJL mice with vehicle or leuprolide acetate (for 9-months) to differentiate between whether sex steroids or gonadotropins might modulate brain metal ion concentrations. Unlike other aging mammals, there was no increase in plasma luteinizing hormone (LH) and follicle-stimulating hormone (FSH) concentrations following estropause in mice, suggesting there was sufficient residual production by the follicle depleted ovary, of sex steroids like estrogens and protein hormones like the inhibins, in order to suppress pituitary LH/FSH production. Castration on the other hand induced significant increases in circulating LH and FSH. Modulation of plasma sex steroid and gonadotropin levels did not significantly alter the concentrations of brain metals tested (Fe, Zn, Cu, Mn, Co, Ni, Al, Li), although there was a tendency for a decrease in all brain metals following ovariectomy (low estrogens and progesterone, high gonadotropins), a response that was reversed with leuprolide acetate treatment (low sex steroids, low gonadotropins). Brain Cu concentration was the only metal correlated with plasma LH (−0.37, n = 30, p < 0.05) and FSH (−0.42, n = 29, p < 0.01). This study demonstrates that sex hormones do not markedly alter brain metal ion homeostasis, unlike previously reported studies of circulating metal ion homeostasis. The role of gonadotropins in regulating metal ion homeostasis does however warrant further study.

Preterm infant circulating sex steroid levels are not altered by transfusion with adult male plasma: a retrospective multicentre cohort study

ObjectiveTo determine if plasma transfusions with male donor plasma to very preterm infants affect circulatory levels of sex steroids.Design and patientsRetrospective multicentre cohort study in 19 infants born at gestational...

Characterization of ovarian culture in vitro and sex steroids in vivo by recombinant eel gonadotropin treatments in the eel Anguilla japonica

In the present study, we investigated the effects of recombinant eel gonadotropins (rec-GTHs) on maturation induction in immature ovarian culture in vitro and sex steroid hormones in vivo in the Japanese eel Anguilla japonica. To study the in vitro effects of rec-GTHs on estradiol-17β (E2) production in immature ovarian tissues, ovarian tissues were incubated with different doses of rec-follicle-stimulating hormone (rec-FSH) or rec-luteinizing hormone (rec-LH). The results revealed that the E2 levels in the rec-FSH (0.1, 0.5, or 1 μg/mL)- and rec-LH (0.1 or 0.5 μg/mL)-treated groups were significantly higher than those in the female eels from the control group. Furthermore, to investigate the in vivo effects of rec-GTHs on the gonadosomatic index (GSI) and plasma sex steroid hormone levels, the eels were injected intraperitoneally with eel’s ringer (control), salmon pituitary extract (SPE; for female eels), human chorionic gonadotropin (hCG; for male eels), rec-FSH, rec-LH, and rec-FSH + rec-LH once a week. The results revealed that except for the SPE and the hCG groups, none of the groups exhibited a significant difference in GSI values. However, in vivo plasma E2 levels increased at the end of 4 weeks after rec-FSH treatment in female eels. Based on these results, it is suggested that rec-GTHs may have a positive effect on sexual maturation in female eels; however, further studies on complementary rec-protein production systems and additional glycosylation of rec-hormones are needed to elucidate hormone bioactivity in vivo and in vitro.

Effects of gonadotropin-releasing hormone agonist administered in microparticles on sperm quality and quantity, and plasma sex steroid levels in northern pike

Artificial reproduction of northern pike Esox lucius is impeded by the likelihood of obtaining only a small volume of sperm of inconsistent quality. A controlled-release hormone delivery system has the potential to enhance sperm production while avoiding multiple injections The objective of this study was to investigate the effects of mammalian gonadotropin-releasing hormone agonist (mGnRHa) incorporated into poly(lactic-co-glycolic acid) (PLGA) microparticles on milt production, spermatozoon characteristics, and secretion of 17β-estradiol (E2), 11-keto testosterone (11-KT), and testosterone in northern pike. Fish were divided into four groups and injected with 2 mg/kg BW carp pituitary extract (CPE), 20 µg/kg BW mGnRHa in PLGA microparticles, or 20 µg/kg BW mGnRHa plus 20 mg/kg BW metoclopramide (MET) in PLGA microparticles (PLGA + MET), along with a control group injected with 1 ml/kg 0.9% NaCl. At 48 h postinjection, the volume of milt produced was significantly greater in groups treated with CPE and PLGA + MET than in other groups. At 96 h postinjection, all hormone-treated groups exhibited significantly higher spermatozoon average velocity than recorded in the control group. Spermatozoon motility was significantly increased (P < 0.05) in the CPE and PLGA groups compared to baseline values. All treated groups showed significantly lower levels of 11-KT after the hormone injection compared to baseline values and to controls. Plasma testosterone levels increased in all hormone-treated groups. The use of PLGA microparticles, with or without metoclopramide, is suitable for use as a carrier of hormone treatments to regulate spermiation in mature northern pike.

Duration of chilling phase, but not thermal condition, influence the gonad maturation of male and female domesticated pikeperch (Sander lucioperca)

AbstractThis study was designed to determine the influence of the thermal level and duration of chilling period on the initiation and progress of gonadogenesis and gametogenesis under controlled conditions in pikeperch (Sander lucioperca). A set of 120 fish was distributed into eight tanks (15 fish per tank). The effects of two temperatures (6 or 12°C) in combination with two durations of chilling period, either short (S; 75 days at 6°C and 125 days at 12°C) or long (L; 135 days at 6°C and 185 days at 12°C), were investigated for the effect on growth, gonadosomatic index (GSI), hepatosomatic index (HSI), gamete developmental stage and sex‐steroid levels (17 beta‐estradiol‐E2 and testosterone‐T). These zootechnical and physiological parameters were selected as they reflect the progressive advancement of gametogenesis in percids including pikeperch. The results show that the temperature did not affect the reproductive parameters but down‐regulated the HSI and the condition factor in females. In addition, the exposure to a longer chilling period, whatever the temperature, allowed for (1) further advancement of gonadogenesis in females (GSI = 6.5% vs. 11%), (2) further advancement of gametogenesis (0% vs. 60% of oocytes at late vitellogenesis, 15% vs. 80% of spermatozoa in males) and (3) the maintenance or increase of plasma sex steroids in both sexes (e.g. 10 ng/mL vs. 27 ng/mL of T in females). Together with already published data, these results suggest that, depending on the features of fish populations and photothermal programmes, the maintenance of temperature at either 6 or 12°C does not necessarily impact pikeperch gonad maturation. However, the duration of the chilling period is an important factor for the optimization of gonadogenesis and gametogenesis achievement before reproduction.

Seasonal changes of plasma sex steroids in captive longtooth grouper Epinephelus bruneus (Bloch) in subtropical regions

Longtooth grouper (LG), Epinephelus bruneus, which is a protogynous hermaphrodite fish, is a highly valued grouper in Japan. Recently, LG farming and larviculture have started in subtropical waters but the reproductive information is scarce. We conducted two experiments to estimate the breeding season for LG held in a sea cage at Amami Island, Japan. Seasonal changes of plasma sex steroid (E2 and 11-KT) levels were measured in Experiment 1. Blood samples were collected from April to July and September 2018 and February 2019. E2 levels of the females increased from February (0.43 ± 0.10 ng/ml) and were significantly higher between April (0.97 ± 0.14 ng/ml) and May (0.66 ± 0.07 ng/ml) compared with those of June, July, and September. Eleven-KT levels of males were also higher between February and June. Spermiation responses indicating maturity of testis were examined throughout the year in Experiment 2. Spermiation was also observed between February and June. These results suggested that the breeding season of LG in subtropical regions was between April and May, with a residual period in June, which was earlier than those of temperate sites of Mainland Japan.

Accuracy of histology, endoscopy, ultrasonography, and plasma sex steroids in describing the population reproductive structure of hatchery‐origin and wild white sturgeon

Hatchery-origin white sturgeon Acipenser transmontanus in the lower Columbia River, Canada are approaching puberty, and describing the reproductive structure of the population is critical to determine if they are capable of contributing to spawning events in the wild, a key management uncertainty. Few studies have compared the accuracy of available tools (histology, ultrasound, endoscopy, and plasma sex steroids) used to assign sex and stage of maturity within the same population of prepubertal and post-pubertal sturgeon. Population reproductive structure was described using these tools in 332 hatchery-origin and 75 wild individuals over 2 years (2017 and 2018). True sex was determined using histological analysis of gonadal tissue, which is 100% accurate at assigning sex and stage of maturity in fish when germ cells are present in the biopsy. All hatchery-origin fish assessed had not reached puberty and were pre-meiotic males (n = 158) or pre-vitellogenic females (n = 174). Assignment of true sex using histology was 97% in hatchery-origin and 94% in wild fish as several biopsies did not contain germ cells. Fish with gonadal biopsies that did not contain germ cells and intersex fish (n = 3) were not included in further analyses of other tools. Accuracy in assigning sex to both the hatchery-origin (98%) and wild (100%) fish was highest using endoscopy (an otoscope). The other tools evaluated were less accurate, with 69% accuracy in hatchery-origin and 74% accuracy in wild fish for plasma sex steroids and 57% accuracy in hatchery-origin and 70% accuracy in wild fish for ultrasonography. Based on these results, endoscopy was the most reliable tool for assigning sex in both prepubertal and post-pubertal fish and can be easily complimented with histology when determining stage of maturity or describing population reproductive structure.

Sex steroids are correlated with environmental factors and body condition during the reproductive cycle in females of the lizard Sceloporus torquatus

Reproduction is regulated by multiple factors that influence physiology and behavior to ensure the continuity of species. However, more work is needed to examine the complex relationships between environmental factors and endocrine transducers that modulate reproductive cycles, particularly in lizards. Here, we aimed to characterize the variation in plasma sex steroid levels in different stages of the reproductive cycle in the lizard Sceloporus torquatus and assess whether sex steroid levels were related to environmental factors (temperature, photoperiod, precipitation, and relative humidity) and body condition. Plasma concentrations of estradiol (E2) and progesterone (P4) from blood samples were quantified by enzyme-linked immunosorbent assay (ELISA) and radioimmunoanalysis (RIA), respectively. Our results indicate that sex steroid concentrations were positively related to follicular development but negatively related to temperature and precipitation. E2 increased as the follicles grew, and its concentrations were highest in the preovulatory phase. P4 showed a similar pattern and persisted during pregnancy. Changes in body condition were non-significant and mainly unrelated to the reproductive stage and plasma sex steroids. Our findings indicate that sex steroids change depending on the season and reproductive stage. We observed high concentrations of E2 and P4 in the late vitellogenic and preovulatory stages, probably because of their role in promoting vitellogenesis and ovulation. Additionally, we observed that follicular development is correlated with temperature and photoperiod. To better understand the mechanisms underlying reproduction, future studies of captive populations where environmental factors can be manipulated are needed.

Health status of fathead minnow (Pimephales promelas) populations in a municipal wastewater effluent-dominated stream in the Canadian prairies, Wascana Creek, Saskatchewan.

Their unique hydrological and climatic conditions render surface water systems in the southern Canadian Prairies at an elevated risk from exposure to contaminants released from municipal wastewater effluents (MWWEs). The aim of this study was to characterize the potential health effects and their underlying molecular mechanisms in populations of fathead minnow (Pimephales promelas; FHM) in Wascana Creek, an effluent dominated stream in Southern Saskatchewan, Canada. Studies were conducted during the spawning season in 2014 and 2015 to assess responses in terms of overall health, reproductive functions, plasma sex steroid hormone levels, and expression of selected genes along the hypothalamus-pituitary-gonadal axis. FHM downstream of the effluent fallout had lower gonadosomatic indices and significantly greater hepatosomatic indices compared to upstream populations. In both male and female FHMs, significantly greater occurrence and severity of gonadal degradation and delayed maturation were observed in downstream fish compared to upstream fish. Downstream males also displayed lower scores of secondary sexual characteristics and a decreasing trend in plasma 11-ketotestosterone levels. Interestingly, no indications of exposure to estrogenic compounds, such as occurrence of testicular oocytes were observed, which was in accordance with the lack of presence of key biomarkers of estrogenic exposure, such as induction of vitellogenin. In general, expression of the majority of transcripts measured in FHMs downstream of the effluent fallout was significantly downregulated, which supports observations of the general deterioration of the health and reproductive status of these fish. Chemical analysis indicated that 10 pharmaceuticals and personal care products (PPCPs) were present at the downstream site, some at sufficiently great concentrations that may present a risk to aquatic organisms. With continuous exposure to a diverse number of stressors including high nutrient and ammonia levels, the presence of a variety of PPCPs and other contaminants, Wascana Creek should be considered as an ecosystem at risk.

수컷 자바리 Epinephelus bruneus에 있어서 hCG 투여에 따른 혈중 성호르몬 및 정자 활성도 변화

This study compared the milt volume, spermatocrit, and sex steroid hormone levels in the blood, and sperm motility and velocity in male longtooth grouper (Epinephelus bruneus), with the application of human chorionic gonadotropin (hCG). The experimental group was injected with 500 IU hCG/kg body weight, whereas the control group was injected with saline solution (0.9% NaCl). The results indicated the former showed significantly higher levels of milt volume and spermatocrit than the latter, whereas sperm motility and velocity showed no significant differences between the two groups.<BR> Regarding the analysis of sex steroid hormone levels in the blood, the plasma testosterone (T) and 11-ketotestosteorne (11-KT) levels of the group with hCG was significantly higher than those of the control two and seven days after the injection respectively. Therefore, hCG administration increased the plasma sex steroid hormone levels (T and 11-KT) and sperm volume of male longtooth grouper, whereas it had no effect on the sperm quality (i.e., motility and velocity). In follow-up research, the fertilization and hatching rates based on the sperm analysis will be compared and investigated to provide basic data for breeding in this species.