Progestagen Concentrations Research Articles

Lymphoplasmacytic endometritis and ovarian adenoma in a pinscher bitch associated with hydrometra

Hydrometra is the accumulation of sterile fluid in the uterine lumen, asymptomatic and abnormally observed in bitches, often identified accidentally during ovariosalpingohysterectomy (OSH). The aim of this study is to report a case of hydrometra in a seven-year-old nulliparous female Pinscher dog, which was brought to the veterinarian due to a gradual increase in abdominal volume. Prior to the consultation, the dog had received a dose of medroxyprogesterone acetate (50 mg). Abdominal ultrasound revealed normal configurations of abdominal organs, except for the uterus, which displayed a lumen filled with anechoic, non-cellular content. During OSH, a considerable enlargement of uterine horns was observed, along with circulatory changes in the organ, such as congestion. The uterine horns were incised longitudinally, revealing clear, translucent, and non-viscous intrauterine fluid. Histopathological examination confirmed mild lymphoplasmacytic endometritis in the uterus and the presence of ovarian adenoma. Therefore, it is believed that ovarian adenoma and hydrometra may result from elevated circulating exogenous progestagen concentrations. Thus, the importance of veterinary medical monitoring is emphasized to prevent and identify possible alterations in the early stages of hydrometra.

Vaginal temperature and collar activity variations in relation to fecal progestagens of captive moose

BackgroundTiming of reproductive events can be crucial for a species’ population growth and stability. Accurate detection of reproductive phenology presents a challenge to scientists studying wild species, including moose (Alces alces). Currently, there are several established methods for monitoring reproductive activity and events in domestic ruminants, including the use of biologging devices. The main objective of this study was to determine whether female moose display a distinct thermal and activity pattern associated with luteal activity during the estrous cycle, which could be used to determine the onset of their breeding season. We deployed biologging devices and collected fecal samples from 12 captive female moose on the Kenai Peninsula, Alaska, USA to explore variation in vaginal temperature and collar activity and the relationship between these variables and fecal progestagen concentrations. Fecal samples were collected from mid-August to mid-October and analyzed using radioimmunoassay to determine the concentration of fecal progestagens to classify luteal activity.ResultsCaptive female moose displayed an identifiable thermal pattern during the onset of luteal activity from mid-September to mid-October, associated with the initial estrous cycle of their breeding season. In contrast, we did not observe a distinct pattern in activity during this period. Recurring patterns in both vaginal temperature and activity were identified between mid-October and mid-November, however, which were likely associated with subsequent estrous cycles but not included in our fecal sampling period.ConclusionsThis study supports that female moose display an identifiable pattern in vaginal temperature which is associated with luteal activity of the initial estrous cycle of the breeding season. An identifiable pattern was observed for both vaginal temperature and activity registrations at the presumed timing of subsequent estrous cycle of the breeding season.

Noninvasive monitoring of steroid hormone production and activity of zoo-housed banteng (Bos javanicus)

This study describes patterns of steroid hormone production and activity for banteng (Bos javanicus), an endangered member of the Bovidae family. Using validated assays, concentrations of fecal progestagens, androgens, and glucocorticoids were quantified for four females and one male at the Saint Louis Zoo. A commercial activity monitor was also validated for assessing movement. The devices were then used to characterize activity in relation to season, reproductive status, and fecal steroid concentrations. General linear mixed models assessed differences in activity and steroid concentrations among individuals, in regards to reproductive status and season. Ovulatory cycle patterns, changes in activity around estrus and parturition, and events correlated with increased glucocorticoid production were also documented. Cycle lengths were 24.7 ± 0.4 days, and cycle lengths varied among individuals. Females cycled year-round, but luteal progestagen concentrations, along with glucocorticoids and male androgens, increased during the summer. Activity also increased in the summer. Progestagen concentrations were greater in pregnant females, and the gestation length of one pregnancy was 254 days. Pregnant females were less active overall, but activity increased the day before parturition. Activity was also greater preceding the onset of the luteal phase. The majority of glucocorticoid concentrations were in the range of baseline concentrations. However, a small number of elevated concentrations were correlated with husbandry and veterinary events. This study is the first to validate non-invasive methods for monitoring reproduction, welfare, and activity of banteng. Our results may contribute to the improved management of captive populations.

Predictors of helminth parasite infection in female chacma baboons (Papio ursinus).

Helminth parasite infection can impose major consequences on host fitness. Several factors, including individual characteristics of hosts, environmental conditions, and patterns of coinfection, are thought to drive variation in parasite risk. Here, we report on four key drivers of parasite infection—phase of reproduction, steroid hormone profiles, rainfall, and patterns of coinfection—in a population of wild female chacma baboons (Papio ursinus) in South Africa. We collected data on reproductive state and hormone profiles over a 3-year span, and quantified helminth parasite burdens in 2955 fecal samples from 24 female baboons. On a host level, we found that baboons are sensitive to parasite infection during the costliest phases of the reproductive cycle: pregnant females harbored higher intensities of Protospirura eggs than cycling and lactating females; lactating and cycling females had a higher probability of Oesophagostomum infection than pregnant females; and cycling females exhibited lower Trichuris egg counts than pregnant and lactating females. Steroid hormones were associated with both immunoenhancing and immunosuppressive properties: females with high glucocorticoid concentrations exhibited high intensities of Trichuris eggs but were at low risk of Oesophagostomum infection; females with high estrogen and progestagen concentrations exhibited high helminth parasite richness; and females with high progestagen concentrations were at high risk of Oesophagostomum infection but exhibited low Protospirura egg counts. We observed an interaction between host reproductive state and progestagen concentrations in infection intensity of Protospirura: pregnant females exhibited higher intensities and non-pregnant females exhibited lower intensities of Protospirura eggs with increasing progestagen concentrations. At a population level, rainfall patterns were dominant drivers of parasite risk. Lastly, helminth parasites exhibited positive covariance, suggesting that infection probability increases if a host already harbors one or more parasite taxa. Together, our results provide a holistic perspective of factors that shape variation in parasite risk in a wild population of animals.

Comparisons of Serum Progesterone and Progestagen Concentrations in Normal and Abnormal Bottlenose Dolphin (Tursiops truncatus) Pregnancies

Although previous studies have described progesterone profiles during pregnancy in the bottlenose dolphin (Tursiops truncatus), most of these focused on normal pregnancy (NORM) or compared NORM to only one or two abnormal pregnancy types, such as abortion (AB) or perinatal loss (PNL). Hormonal pregnancy biomarker analysis from reproductive events with different outcomes may reveal differences in concentrations so that we are able to identify high risk pregnancies. The aim of this study was to describe longitudinal profiles of circulating progesterone and progestagens during reproductive events in the female bottlenose dolphin, including NORM, failure to thrive, PNL, AB, early loss (EL), and false pregnancy (FP). Progesterone differed from NORM during EL at EARLY (month post conception [MPC] 1–4), AB at MID (MPC 5–8), and FP at LATE (MPC 9–12) stages. Progestagens differed from NORM during AB and FP at MID and LATE stages and during MPC 12 in PNL and MPC 4 in EL. Progestagens may be better at predicting poor reproductive outcome in the bottlenose dolphin and a suite of hormone tests, including progesterone and progestagens, should be incorporated into existing clinical diagnostic and management practices in this species. Furthermore, analysis of multiple hormonal pregnancy biomarkers from a single sample may enable pregnancy diagnosis for wild animals.

Post fetal death development of endometrial cups in a Jenny donkey (Equus asinus)

SummarySerial ultrasound examinations of the gravid uterus of a Jenny donkey (Equus asinus) carrying a transferred extraspecies horse (Equus caballus) conceptus between 11 and 33 days of gestation demonstrated that the embryo died before Day 33 even though the conceptus persisted in the uterus and the horse chorionic girdle invaded the maternal endometrium to form a circle of typically large horse endometrial cups in the right uterine horn. The high concentrations of horse CG secreted by these cups with its higher than usual ratio of FSH‐like to LH‐like biological activities of 1.4 compared to the 0.1 ratio of normal donkey CG hyper‐stimulated the donkey’s ovaries to develop multiple large accessory follicles and resulting secondary corpora lutea which secreted high concentrations of progestagens that persisted in her bloodstream for the next 90 days. This case illustrated that, in all likelihood, girdle invasion can happen after the death of the embryo indicating autonomy of the chorionic girdle.

Placentation and hormonal maintenance of pregnancy in the impala (Aepyceros melampus)

IntroductionThe impala is a widely distributed African ungulate. Detailed studies of the placenta and ovaries in impala undertaken in the 1970s did not address the endocrine functions of the placenta. MethodsThe uteri of 25 pregnant impala estimated to be between 49 and 113 days of the 190 day gestation were examined grossly, histologically and immunohistochemically. ResultsA single corpus luteum was present in either maternal ovary but the conceptus was always situated in the right uterine horn. The fetal membranes extended to the tips of both uterine horns. The amnion was in intimate contact with, but not fused to, the allantochorion. Placentation was typically ruminant with fetal macrocotyledons attached to the rows of maternal caruncles. The fetal villi were highly branched, especially in the centre of each placentome where the attenuated maternal epithelium lining the placental crypts was absent in some places. Both the corpus luteum and the uninucleate trophoblast cells of the interplacentomal allantochorion stained strongly for 3-β hydroxysteroid dehydrogenase, and progestagen concentrations in allantoic and amniotic fluids increased significantly as gestation progressed, with a tendency to do likewise in maternal serum. Binucleate trophoblast cells stained positively for bovine placental lactogen, but neither the placenta nor the maternal corpus luteum showed evidence of oestrogen synthesis. DiscussionDespite exhibiting the same basic type of placentation, both the gross and histological structure of the impala placenta, along with its immunohistochemical properties, demonstrates that great variation exists across ruminant placentas.

UNDERSTANDING PROLACTIN REGULATION AND DETERMINING THE EFFICACY OF CABERGOLINE AND DOMPERIDONE TO MITIGATE PROLACTIN-ASSOCIATED OVARIAN CYCLE PROBLEMS IN ZOO AFRICAN ELEPHANTS (LOXODONTA AFRICANA ).

Perturbations in serum prolactin secretion, both over- and underproduction, are observed in zoo African elephants (Loxodonta africana) that exhibit abnormal ovarian cycles. Similar prolactin problems are associated with infertility in other species. Pituitary prolactin is held under constant inhibition by a hypothalamic-derived neurotransmitter, dopamine; thus, regulation by exogenous treatment with agonists or antagonists may be capable of reinitiating normal ovarian cycles. This study tested the efficacy of oral administration of cabergoline (agonist) and domperidone (antagonist) as possible treatments for hyperprolactinemia or chronic low prolactin, respectively. Hyperprolactinemic (overall mean prolactin, >30 ng/ml), acyclic elephants were administered oral cabergoline (2 mg, n = 4) or placebo (dextrose capsule, n = 4) twice weekly. Overall mean prolactin concentration decreased in treated females compared with controls (32.22 ± 14.75 vs 77.53 ± 0.96 ng/ml; P = 0.01). Interestingly, overall mean progestagen concentrations also increased slightly (P < 0.05) in treated females (0.15 ± 0.01 ng/ml) compared with controls (0.07 ± 0.01 ng/ml), but no reinitation of normal cyclic patterns was observed. Chronic low prolactin (overall mean prolactin, <10 ng/ml), acyclic females were orally administered domperidone (2 g/day, n = 4) or placebo (dextrose capsule, n = 4) for 4 wk, followed by 8 wk of no treatment (four cycles) to simulate the prolactin pattern observed in normal cycling elephants. Overall mean prolactin concentrations increased (P = 0.005) during domperidone treatment (21.77 ± 3.69 ng/ml) compared with controls (5.77 ± 0.46 ng/ml), but progestagen concentrations were unaltered. Prolactin regulation by dopamine was confirmed by expected responses to dopamine agonist and antagonist treatment. Although prolactin concentrations were successfully reduced by cabergoline, and domperidone initiated the expected cyclic prolactin pattern, neither treatment induced normal ovarian activity.

A preliminary study of the heterogeneity in endometrial morphology and glycosylation in the uterine horns of the non-pregnant impala (Aepyceros melampus)

Placentation commences only in the right uterine horn in impala (Aepyceros melampus). To investigate possible differences in morphology or glycosylation between the two horns, right and left uterine horns from six non-pregnant, wild impala were examined morphometrically and histochemically using a panel of 23 lectins and an avidin-biotin revealing system. The presence of ovarian 3ß hydroxysteroid dehydrogenase (3ßHSD) and aromatase was also investigated using immunocytochemistry. There were few detectable differences in morphology and glycosylation between right and left uterine horns in five of the specimens, but the sixth had deep clefts and plentiful exocrine secretions in the right horn, and not the left. Heavily glycosylated clusters of supranuclear granules were present in the epithelial cells, which had many classes of O-linked glycans. The serum progestagen was not markedly different, however, from that of the other specimens. In five of the six specimens, the height of luminal epithelium was greater on the right than that on the left, and the height of the gland epithelium was also greater on the right side in four of these. The 3ßHSD and aromatase activities were present in the ovaries and were similar in impala with or without progestagen concentrations >1 ng/ml in peripheral blood. No corpus haemorrhagicum or corpus luteum could be discerned. These findings indicate there are morphological and biochemical differences between right and left uterine horns in the impala and further studies are needed on both impala and other species in which placentation commences only in one uterine horn, to establish the cyclical hormone changes which induce them.

Occurrence and removal of progestagens in municipal wastewater treatment plants from different regions in China

Progestagens discharged from municipal wastewater treatment plants (WWTPs) have increasingly gained attention due to their potential risks to the aquatic organisms. However, limited information is available on the occurrence and removal of various progestagens in WWTPs in different cities of China. This work investigated the occurrence and removal of 11 progestagens in 21 WWTPs from 19 Chinese cities. Results showed that progestagens are widely distributed in the investigated WWTPs, with higher influent concentrations of total progestagens in northern WWTPs. The concentration of progestagens in WWTP influent were closely correlated with influent quality, service population and daily service volume of the WWTPs. Additionally, progesterone (PGT) and dydrogesterone (DDT) were two predominant progestagens in influent, effluent and excess sludge. Up to 5 of 11 progestagens showed high aqueous removal efficiencies (median removal efficiency >90%), whereas megestrol acetate (MTA), chlormadinone acetate (CMA), drospirenone (DSP) and levonorgestrel (LNG) had a removal efficiency of below 50%. Specially, the behaviors of progestagens along the anaerobic-anoxic-oxic of a WWTP were further explored and the aerobic tank is the main contributor to the removal of progestagens. Finally, in the effluent of these 21 WWTPs, daily mass loadings of the total progestagens ranged from 0.51 to 10.4 g d−1. Notably, LNG exhibited high potential risk to the fish base on risk quotient.

Effect of time and temperature on stability of progestagens, testosterone and cortisol in Asian elephant blood stored with and without anticoagulant.

The value of biological samples collected in the field is compromised if storage conditions result in analyte degradation, especially in warmer climates like Thailand. We evaluated the effects of time and temperature on immunoactive steroid hormone stability in Asian elephant (Elephas maximus) blood stored with and without an anti-coagulant before centrifugation. For each elephant (5 male, 5 female), whole blood was aliquoted (n = 2ml each) into 13 red top (without anticoagulant) or purple top (with anticoagulant) tubes. One tube from each treatment was centrifuged immediately and the serum or plasma frozen at -20°C (Time 0, T0). The remaining 12 aliquots were divided into stored temperature groups: 4°C, room temperature (RT, ~22°C), and 37°C, and centrifuged after 6, 24, 48 and 62h of storage. Serum and plasma concentrations of progestagens in females, testosterone in males and cortisol in both sexes were quantified by validated enzyme immunoassays. Steroid concentration differences from T0 were determined by a randomized complete block ANOVA and Dunnett's tests. The only evidence of hormone degradation was cortisol and testosterone concentrations in serum stored at 37°C. Testosterone concentrations declined by 34% at 48h and 52% at 62h, cortisol was decreased by 19% after 48h and 27% after 62h at 37°C, respectively. None of the other aliquots displayed significant changes over time at any temperature. In conclusion, steroids appear to be stable in blood for nearly 3days at room or refrigeration temperatures before centrifugation; steroids in samples with ethylenediaminetetraacetic acid were particularly stable. However, warmer temperatures may negatively affect steroids stored without anti-coagulant, perhaps due to red blood cell metabolism. Thus, under field conditions with no access to cold or freezer temperatures, collection of plasma is a better choice for elephants up to at least 62h before centrifugation.

Endocrine-disrupting compounds in the Xiangjiang River of China: Spatio-temporal distribution, source apportionment, and risk assessment

Endocrine-disrupting compounds (EDCs) were seasonally investigated in the surface water of the Xiangjiang River (south China) in order to understand their spatio-temporal distribution, source apportionment, and ecological risks. The occurrence of 21 EDCs were determined with liquid chromatography-tandem mass spectrometry in the water samples collected along the river over four seasons, and the results were statistically analyzed. The concentrations of progestagens, androgens, estrogens ranged from not detected (ND) to 98.3 ng L−1; while the concentrations of alkylphenols ranged from 0.8 to 3.1 × 103 ng L−1; and that of caffeine ranged from 0.1 to 49.8 ng L−1. The detection frequencies of bisphenol A, 4-tert-octylphenol, 4-n-nonylphenol, estrone, and 17β-estradiol were 95–100% during the four sampling campaigns. The seasonal and spatial variation trend of EDCs in the Xiangjiang River was noticeable. The concentration of EDCs in Yueyang section (downstream) was the highest in winter, while the concentration in Yongzhou (upstream) section was the lowest in spring. The concentration of EDCs in the Xiangjiang River was significantly correlated with the levels of the total organic carbon, water temperature, and dissolved oxygen. Source analysis indicated that untreated sewage was the major source of EDCs. Furthermore, the potential risks of EDCs in the surface water to aquatic organisms were assessed with the risk quotient method (European Commission, 2003), and the results indicated the highest ecological risk of 17β-estradiol in the Xiangjiang River.

Prolonged luteal lifespan and pseudopregnancy in Asian elephants (Elephas maximus)

Pseudopregnancy is a physiological occurrence in mammals which have copulation induced ovulation, but is rarely described in spontaneous ovulating species. In this study, three cases of prolonged luteal lifespan are reported in non-pregnant Asian elephants (Elephas maximus). Case 1 was a 25-year-old female that had produced three calves previously; Case 2 was a nulliparous and 32-year-old at the start of the pseudopregnancy episode; and Case 3 occurred in a 49-year-old nulliparous elephant. Serum progesterone metabolite concentrations remained elevated for 10 months in Case 1. Urinary progestagens were high for >16 months in Case 2 and for five months in Case 3. In Case 1, multiple persistent corpora lutea were visualized monthly by ultrasonography. In all three cases, uterine leiomyoma were present and progestagen concentrations decreased spontaneously. In Case 1, the elephant became pregnant 3 years later, whilst with Case 2, the female resumed estrous cycling normally, and for the Case 3 female, there was continuation with another prolonged luteal phase before ovarian function was purposely suppressed. These examples indicate that persistently elevated progestagen concentrations may not always be indicative of pregnancy in elephants. The reasons for prolonged luteal lifespan are not understood, although serum prolactin concentrations quantified in the Case 1 female were elevated compared to values from previous reports and two other herd mates. Furthermore, all three elephants had varying degrees of uterine pathologies. It is believed that the resulting damage to the endometrium may have led to a reaction similar to implantation, which includes prolactin secretion. Prolactin may exert luteotropic properties and is thought to initiate luteal rescue during pregnancy in elephants.

Longitudinal profiles of relaxin and progestagens during pregnancy, pregnancy loss and false pregnancy in the killer whale (Orcinus orca)

The circulating pattern of immunoreactive relaxin and progestagens based on monthly and gestational stage (early, mid, late) profiles were determined during pregnancies that resulted in live calves (LIVE, n = 30), stillbirths (STILLB, n = 3), abortions (ABORT, n = 5) and presumptive false pregnancies (FALSE, n = 8), and during the follicular (n = 34) and luteal phase (n = 58). Monthly LIVE relaxin concentrations steadily increased during gestation, but values did not significantly exceed those of the luteal phase until 9 months prior to parturition, peaking during the final month at 2356 ng/ml. Relaxin surged (P < 0.05) during the final week of gestation (36,397 ng/ml), undergoing a 3 and 9-fold increase compared with concentrations in the preceding two weeks, respectively. Monthly relaxin production did not differ among each reproductive state with the exception of months-13–16 where concentrations were higher (P < 0.001) for STILLB than LIVE. Relaxin concentration was reduced (P < 0.0001) by 849% in placental versus maternal serum collected within 1 day of labor. Mid- and late-pregnancy progestagen concentrations were lower for FALSE (P < 0.001) compared with STILLB and LIVE. Late pregnancy progestagen concentrations were reduced for FALSE (P < 0.05) and ABORT (P < 0.02) compared with LIVE and STILLB. Monthly progestagen production in ABORT tended to be lower than LIVE across a range of gestational months (Months 2, 7, 8, 11) but this difference only became significant during months 14 and 15. Results indicate that relaxin is primarily produced by the CL during pregnancy, and that concentrations could not be used to differentiate from non-pregnant females until the final 6 months of gestation. In addition, as would be expected from a primarily CL product, relaxin cannot be used to detect abnormal pregnancies. Conversely, progestagens, which are produced by both the placenta and CL can be used to differentiate FALSE from normal pregnancy and may be useful indicators of fetal health in the killer whale.

Urofecal steroid profiles of captive Blue-fronted parrots (Amazona aestiva) with different reproductive outcomes

Despite Psitaciformes (parrots) being the third largest nonpasserine order (398 species), it currently ranks second in number of threatened species (28%) according to the Internatinal Union for Conservation of Nature (IUCN) criteria. Since most of the literature concerning reproductive endocrinology in avian species derives from domestic and song birds, it is puzzling that advances in reproductive science for the Psitaciformes order lags far behind, in spite of the growing threats against them. In order to expand our knowledge of Neotropical parrots (Psittacidae), we examined annual changes in urofecal sex steroid metabolites of Blue-fronted amazon pairs (Amazona aestiva) exhibiting successful (nestlings) and unsuccessful breeding (infertile or no eggs). Urofecal samples were collected over a year from eight breeding pairs housed under the same environmental and management conditions. Fecal androgen and progestagen concentrations were determined in males and females, respectively, by enzyme immunoassays previously validated for this species. All eggs were registered between late winter and mid-spring, and egg-laying intervals varied between females (range: 1–8 days; average 3.60 ± 0.51 days). Similar profiles of urofecal progestagens were observed in reproductively successful females and females producing infertile eggs, with progestagen peaks preceding egg laying events (1.77 ± 0.50 days). In contrast, non-laying females had no rises in progestagens during the year. Successful and unsuccessful males did not displayed distinct annual patterns of androgen production, and apart from the peaks during the breeding season, more than half of the individuals intriguingly presented significant increases from late summer to early autumn, a period without reproductive activity. Finally, we noticed that samples with progestagen levels exceeding 40 ng/g had very high probability (>97.5%) to be from females in pre-laying or laying phases, suggesting a feasible application of this characteristic to noninvasively discriminate the reproductive status in amazon females with an accuracy and sensitivity of 94.55% and 58.13%, respectively. Our findings confirmed that urofecal progestagens and androgens are good indicators of the gonadal condition in Blue-fronted amazons, but there is still much to be done for their extensive use in artificial insemination or selection of the most suitable breeding birds for the season.

Relationships between POPs, biometrics and circulating steroids in male polar bears (Ursus maritimus) from Svalbard

The aim of this study was to determine the effects of persistent organic pollutants (POPs) and biometric variables on circulating levels of steroid hormones (androgens, estrogens and progestagens) in male polar bears (Ursus maritimus) from Svalbard, Norway (n = 23). Levels of pregnenolone (PRE), progesterone (PRO), androstenedione (AN), dehydroepiandrosterone (DHEA), testosterone (TS), dihydrotestosterone (DHT), estrone (E1), 17α-estradiol (αE2) and 17β-estradiol (βE2) were quantified in polar bear serum by gas chromatography tandem mass spectrometry (GC-MS/MS), while POPs were measured in plasma. Subsequently, associations between hormone concentrations (9 steroids), POPs (21 polychlorinated biphenyls (PCBs), 8 OH-PCBs, 8 organochlorine pesticides (OCPs) and OCP metabolites, and 2 polybrominated diphenyl ethers (PBDEs)) and biological variables (age, head length, body mass, girth, body condition index), capture date, location (latitude and longitude), lipid content and cholesterol levels were examined using principal component analysis (PCA) and orthogonal projections to latent structures (OPLS) modelling. Average concentrations of androgens, estrogens and progestagens were in the range of 0.57–83.7 (0.57–12.4 for subadults, 1.02–83.7 for adults), 0.09–2.69 and 0.57–2.44 nmol/L, respectively. The steroid profiles suggest that sex steroids were mainly synthesized through the Δ-4 pathway in male polar bears. The ratio between androgens and estrogens significantly depended on sexual maturity with androgen/estrogen ratios being approximately 60 times higher in adult males than in subadult males. PCA plots and OPLS models indicated that TS was positively related to biometrics, such as body condition index in male polar bears. A negative relationship was also observed between POPs and DHT. Consequently, POPs and body condition may potentially affect the endocrinological function of steroids, including development of reproductive tissues and sex organs and the general condition of male polar bears.

Non-invasive endocrine monitoring indicates seasonal variations in gonadal hormone metabolites in dholes (Cuon alpinus).

To date, there is no information on reproductive endocrinology of dholes (Cuon alpinus). The objectives of the present study were as follows: (i) to characterize longitudinal profiles of gonadal steroids; and (ii) to examine the relationship between gonadal hormones and sexual behaviours in dholes. Three breeding pairs and two bachelor males were included in the study. Among these, four animals (2 males and 2 females; 4 years old) were imported from The Netherlands to Thailand 3 months before the study onset; the remaining individuals (3 males and 1 female; 5-7 years old) were native born. Faecal samples were collected 3-7 days/week for 12 months, extracted and assessed for gonadal hormone metabolites using a validated enzyme immunoassay. Observations of behaviour were conducted in 30min sessions, 3-5 days/week. For the three breeding males, testosterone was elevated (P<0.05) from October to January in the two imported males, whereas the concentration of steroid metabolites was high from April to June and from September to November in the native male. However, there was no clear seasonal pattern of reproductive hormone in the bachelor group. Oestrogen metabolite level of imported females was elevated for 9-12 days in January, followed by a rise in progestagen concentration. For native females, oestrogen metabolites were above the basal values in April and September, each of which was followed by a rise in progestagen concentration that remained elevated for 77 and 112 days, respectively. Sexual behaviours, including solicitation, mounting and copulations, were observed during the oestrogen peak in all females. Our findings indicate that reproductive seasonality of dholes may depend on the animals' origin and social group.

Altered adrenal and gonadal steroids biosynthesis in patients with burn injury

IntroductionBurn injury inevitably leads to changes in the endogenous production of cytokines, as well as adrenal and gonadal steroids. Previous studies have reported gender-related differences in outcome following burn injury, which suggests that gonadal steroids may play a role. The aim of this study was to assess alterations in concentration of endogenous steroids in patients with burn injury. MethodsFor this single-center, prospective descriptive study, high-sensitivity liquid chromatography tandem mass spectrometry (LC-MS/MS)-based steroid quantification was used to determine longitudinal profiles of the concentrations of endogenous steroids in plasma from sixteen adult male patients with burn injury (14.5–72% of total body surface area). Steroids were extracted from plasma samples and analyzed using multiple reaction monitoring acquisition, with electrospray ionization on a triple quadruple mass spectrometer. Total protein concentration was measured in the samples using spectrophotometry. ResultsSteroid and total protein concentration distributions were compared to reference intervals characteristic of healthy adult men. Concentrations of the following steroids in plasma of burn injured patients were found to correlate positively to the area of the burn injury: cortisol (r=0.84), corticosterone (r=0.73), 11-deoxycortisol (r=0.72), androstenedione (r=0.72), 17OH-progesterone (r=0.68), 17OH-pregnenolone (r=0.64) and pregnenolone (r=0.77). Concentrations of testosterone decreased during the acute phase and were up to ten-times lower than reference values for healthy adult men, while concentrations of estrone were elevated. By day 21 after injury, testosterone concentrations were increased in younger, but not older, patients. The highest concentrations of estrone were observed on day 3 after the injury and then declined by day 21 to concentrations comparable to those observed on the day of the injury. ConclusionBurn injury alters endogenous steroid biosynthesis, with decreased testosterone concentrations and elevated estrone concentrations, during the first 21days after the injury. Concentrations of glucocorticoids, progestagens and androgen precursors correlated positively with the area of burn injury. The finding of increased estrone following burn injury needs to be confirmed in a larger hypothesis-driven study.

Characterization and longitudinal monitoring of serum progestagens and estrogens during normal pregnancy in the killer whale (Orcinus orca)

The secretory patterns of progestagens and estrogens were characterized throughout 28 normal pregnancies until two month post-partum in eleven killer whales. HPLC analysis of serum from different reproductive stages (luteal phase, EARLY, MID, and LATE pregnancy) identified three major immunoreactive progestagen peaks; progesterone (P4), 5α-pregnane-3,20-dione (5α-DHP) and pregnanediol, with 5α-DHP approximately half of that for P4 in the luteal phase, and EARLY, but approximately 2/3 of P4 during MID and LATE pregnancy. At birth, 5α-DHP was the only significant (>10% immunoreactivity) immunoreactive progestagen detected in placental (umbilical cord) serum. Maternal recognition of pregnancy appears to occur between day 21 and 28 post-ovulation when a significant deviation in progestagen concentrations between conceptive and non-conceptive cycles was detected. Progestagen concentrations during pregnancy displayed a bimodal pattern with significant peaks (P<0.05) in EARLY (indexed month post-conception [IMPC] 2, 3, 4) and MID (IMPC 9, 10) before decreasing (P<0.05) over an 11day interval to luteal phase concentrations on the day of parturition. Among estrogens, estriol was secreted in the highest concentrations but only estrone (free and conjugated) and estradiol increased (P<0.001) during pregnancy, with peaks observed during the final month of gestation, and an influence (P<0.05) of fetal sex on estradiol production was detected. Collective findings indicate that P4 derived from the corpus luteum is the major biologically active progestagen during the luteal phase and pregnancy, and that 5α-DHP production, possibly from both luteal and placental sources, increases during the second half of pregnancy.

Occurrence and removal of progestagens in two representative swine farms: Effectiveness of lagoon and digester treatment

A total of 21 progestagens were screened in animal wastes and environmental samples from two representative swine farms and surrounding environments of South China using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) to assess the effectiveness of simple lagoon (and digester) treatment. The results showed that 11, 8 and 8 of 21 target progestagens were detected with the minimum concentration of 2.31 ng/L and maximum of 6150 ng/L in the water samples, with the minimum of 1.36 ng/L and maximum of 98.3 ng/L in the suspended particles, and with the minimum of 1.57 ng/g dry weight (dw) and maximum of 3310 ng/g dw in the solid samples, respectively. Trace levels (a few ng/L or ng/g levels) of dydrogesterone, 5α-dihydroprogesterone, norgestrel and progesterone were found in samples from nearby surface waters and vegetable fields impacted by animal wastes. The residual progestagens at the reported levels may still pose potential risks to aquatic organisms such as fish in the receiving aquatic environments. This finding suggests that swine wastewater and feces could lead to contamination of some detectable progestagens in the surrounding environments. Significant reduction in total progestagen concentrations were observed from the fresh swine wastewaters to the fish ponds, indicating effective removal of these compounds by the lagoon (and digester) treatment. In addition, the biogas digesters provided high removal of the progestagens in the waste streams. This low-cost and eco-friendly treatment system should be promoted in developing countries with concentrated animal operations.