Fecal Steroid Research Articles

Parámetros sanguíneos y perfil de hormonas reproductivas de hembras de Choloepus hoffmanni en cautiverio

In wild animal species, the use of non-invasive and non-stressful procedures to determine hormone profiles, such as fecal steroid measurements, has considerably increased the comprehension of their reproductive physiology. Since there is limited information related to the reproductive hormone profiles of the two-toed sloth, Choloepus hoffmani, a study was conducted in captive specimens at the “Sloth Sanctuary” (Cahuita, Limón, Costa Rica), in order to determine: (i) the reliability of the fecal progesterone and estrogen extraction and its quantification with an AIA-360® analyzer, (ii) assess blood parameters in this species and (iii) evaluate if there is a correlation between fecal and plasmatic steroids. The study was performed over a three-month period, from November, 2013 to January, 2014, with a total amount of 208 fecal samples collected from five sexually mature females weighing 6.32 Kg in average. The average of the median concentrations of progesterone in feces of the five females was 124.21 ng/g, and 1 708.95 pg/g for estrogen. The average minimal and maximal values were 50.96 ng/g and 1 057.46 ng/g for progesterone and, 1 191.77 pg/g and 2 159.24 pg/g for estradiol. In plasma, progesterone median values were 1.26 ng/mL, showing a minimum of 0.32 ng/mL and 12.84 ng/mL as maximum values. The plasmatic estrogen levels were below the detection limit of the equipment (25 pg/mL). Although there was no strong statistical correlation between the fecal and plasmatic progesterone fluctuations, our data suggests that the plasmatic events are mostly reflected in feces two days afterwards. Also, the levels of progesterone were elevated during the first half of November and, subsequently, showed a successive and important reduction in all the females tested. Finally, our results demonstrated that fecal steroid extractions and their measurement in a AIA-360®, allowed the successful detection and represents an alternative non-invasive determination of hormone profiles in C. hoffmani.

EARLY ONSET OF THE REPRODUCTIVE SEASON IN CAPTIVE WHITE-TAILED DEER ( ODOCOILEUS VIRGINIANUS VERAECRUCIS) IN EASTERN MEXICO.

The white-tailed deer ( Odocoileus virginianus) inhabits a wide latitudinal range in the Americas. Deer species dwelling throughout wide latitudinal ranges have developed subspecies with variations in their reproductive seasonality. In northern subspecies of white-tailed deer, such as those from Canada and the United States, the breeding season occurs from October through December. Odocoileus virginianus veraecrucis is a subspecies that inhabits eastern Mexico, and because its reproductive season has not been studied, it is believed to be similar to that from northern subspecies. The objective of the study was to determine the onset of the breeding season and the profile of fecal steroid hormone metabolites throughout the year in captive white-tailed deer subspecies veraecrucis in Mexico. Two groups of adult deer were included: 1) six does and one buck at a Wildlife Conservation Unit, and 2) five does and one buck at a zoo. From each group of deer, representative fecal samples were collected on a weekly basis for 1 yr for fecal analysis of progesterone and estradiol in the does, and testosterone in the bucks. Data on antler casting, parturitions, and velvet shedding were recorded. Progesterone was high during pregnancy and low throughout the parturition period. Estradiol fluctuated throughout the year. Testosterone was high during the rut and low after antler casting. Antlers were cast in March and velvet was shed in August at both sites. Parturitions started in February at the zoo and in April at the Wildlife Unit. In captive white-tailed deer subspecies veraecrucis the breeding season started in July, and therefore earlier than what has been reported for subspecies from northern latitudes.

Rare sugars, d-allulose, d-tagatose and d-sorbose, differently modulate lipid metabolism in rats.

Rare sugars including d-allulose, d-tagatose, and d-sorbose are present in limited quantities in nature; some of these rare sugars are now commercially produced using microbial enzymes. Apart from the anti-obesity and anti-hyperglycaemic activities of d-allulose, effects of these sugars on lipid metabolism have not been investigated. Therefore, we aimed to determine if and how d-tagatose and d-sorbose modulate lipid metabolism in rats. After feeding these rare sugars to rats, parameters on lipid metabolism were determined. No diet-related effects were observed on body weight and food intake. Hepatic lipogenic enzyme activity was lowered by d-allulose and d-sorbose but increased by d-tagatose. Faecal fatty acid excretion was non-significantly decreased by d-allulose, but significantly increased by d-sorbose without affecting faecal steroid excretion. A trend toward reduced adipose tissue weight was observed in groups fed rare sugars. Serum adiponectin levels were decreased by d-sorbose relative to the control. Gene expression of cholesterol metabolism-related liver proteins tended to be down-regulated by d-allulose and d-sorbose but not by d-tagatose. In the small intestine, SR-B1 mRNA expression was suppressed by d-sorbose. Lipid metabolism in rats varies with rare sugars. Application of rare sugars to functional foods for healthy body weight maintenance requires further studies. © 2017 Society of Chemical Industry.

Effect of environment and castration on dominance rank and fecal steroids in male Japanese macaques (Macaca fuscata)

Effect of environment and castration on dominance rank and fecal steroids in male Japanese macaques (Macaca fuscata)

Validation of an Enzyme Immunoassay for Measuring Fecal Cortisol Metabolites of Bearded (Sapajus libidinosus) and Black (Sapajus nigritus) Capuchins

Fecal steroid analysis is a powerful noninvasive tool for behavioral endocrinology, but enzyme immunoassays (EIAs) require experimental validation before they are applied. We conducted a physiological validation of an in-house EIA measuring fecal cortisol metabolites (FCMs) by performing an adrenocorticotrophic hormone (ACTH) challenge, dexamethasone suppression, and control saline solution injection experiment with six male and five female bearded capuchins (Sapajus libidinosus). We also took advantage of a presumably stressful incident to perform a biological validation for females. In addition, we conducted high-performance liquid chromatography (HPLC) immunograms to characterize the FCMs measured in both sexes of bearded capuchin, and in a closely related species (S. nigritus, black capuchin monkeys). Male and female S. libidinosus showed FCM peaks after ACTH injections, and females also showed FCM peaks in the biological validation. Three of four individuals (two males and one female) had an FCM peak shortly after injection of dexamethasone and both sexes then showed prolonged low FCM levels after dexamethasone injection. We observed no effects of saline solution injections. The time to peak FCM excretion after ACTH injection or a stressful incident varied 1.5–8.5 h. HPLC results revealed no differences in FCM profiles between sexes or species and suggest that the EIA can also be used in male and female S. nigritus. Our results validate an in-house EIA for both sexes of S. libidinosus but show large individual variation in FCM excretion, which highlights the need for carefully planned feces collection in endocrinologial research.

Adrenal responses of large whales: Integrating fecal aldosterone as a complementary biomarker to glucocorticoids

Until now, physiological stress assessment of large whales has predominantly focused on adrenal glucocorticoid (GC) measures. Elevated GC concentrations in feces (fGC) are known to reflect stressful disturbances, such as fishing gear entanglement and human-generated underwater noise, in North Atlantic right whales (Eubalaena glacialis). However, there can be considerable variation in GC production as a function of sex and life history stage, which may confound the interpretation of fGC levels. Additionally, GC antibodies used in immunoassays can cross-react with other fecal metabolites (i.e., non-target steroids), potentially influencing fGC data. Here, aldosterone concentrations (fALD; aldosterone and related metabolites) were measured in fecal samples from right whales (total n=315 samples), including samples from identified individuals of known life history (n=82 individual whales), to evaluate its utility as a complementary biomarker to fGC for identifying adrenal activation. Concentrations of fALD were positively correlated with fGCs in right whales (r=0.59, P<0.001), suggesting concurrent secretion of these hormones by the adrenal gland. However, fALD levels were less influenced by concentrations of reproductive steroids in feces, minimizing the potential confounder of assay cross-reactivity in samples with highly skewed hormone ratios. Across different life history states for right whales, fALD concentrations showed similar patterns to those reported for fGC, with higher levels in pregnant females (35.9±7.6ng/g) followed by reproductively mature males (9.5±0.9ng/g) (P<0.05), providing further evidence of elevated adrenal activation in these groups of whales. The addition of fALD measurement as a biomarker of adrenal activation may help distinguish between intrinsic and external causes of stress hormone elevations in large whales, as well as other free-living wildlife species, providing a more comprehensive approach for associating adrenal activation with specific natural and anthropogenic stressors.

Wastewater contamination in Antarctic melt-water streams evidenced by virological and organic molecular markers

Human activities in the Antarctica including tourism and scientific research have been raised substantially in the last century with the concomitant impact on the Antarctic ecosystems through the release of wastewater mainly from different scientific stations activities. The aim of this study was to assess the wastewater contamination of surface waters and sediments of three melt-water streams (11 sites) by leaking septic tanks located in the vicinity of the Uruguayan Scientific Station in the Fildes Peninsula, King George Island, Antarctica, during summer 2015. For this purpose, we combined the analysis of fecal steroids in sediments by using gas chromatography and six enteric viruses in surface waters by quantitative and qualitative PCR. Coprostanol concentrations (from 0.03 to 3.31μgg−1) and fecal steroids diagnostic ratios indicated that stations C7 and C8 located in the kitchen stream presented sewage contamination. Rotavirus was the only enteric virus detected in five sites with concentration ranging from 1.2×105gcL−1 to 5.1×105gcL−1 being three of them located downstream from the leaking AINA and Kitchen septic tanks. This study shows for the first time the presence of both virological and molecular biomarkers of wastewater pollution in surface waters and sediments of three melt-water streams in the vicinity of a scientific station in the Antarctica. These results highlight the importance of the complementation of these biomarkers in two different matrices (surface waters and sediments) to assess wastewater pollution in an Antarctic environment related to anthropogenic activities in the area.

Evidence of massive river pollution in the tropical megacity Jakarta as indicated by faecal steroid occurrence and the seasonal flushing out into the coastal ecosystem

In the rapidly growing coastal megacities of developing and emerging countries, the implementation of new sewage treatment facilities is often not keeping pace with the enormous population growth, leading to a deterioration of the urban water resources. Only very few studies discuss faecal water contamination in these urban areas. Our study area Jakarta, the capital of Indonesia, is such a megacity. We investigated chemical markers of faecal contamination in water and sediments from the rivers and canals flowing through Jakarta. Moreover, also the spatial distribution of faecal markers in Jakarta Bay, the coastal ecosystem that receives all urban river discharges, was assessed. The concentrations of the faecal steroid coprostanol in river water ranged from 0.45 to 24.2 µg L−1, and in sediments from 0.3 to 400 µg g−1, reflecting the problem of inadequate sewage treatment capacities in the city. The steroid distribution in Jakarta Bay in May 2013 as compared to dry season data indicates a flushing out of particle-associated pollutants from the urban rivers far offshore during the precedent rainy season, where the city experienced a severe flood. This flushing out of particle-associated pollutants during times of heavy rainfall as observed in our study is a pollutant transport mechanism that is important for all tropical coastal systems. The associated impacts on sensitive coastal habitats might become more severe in the future, as a consequence of the expected climate change effects on monsoon variability.

Integrating trans-abdominal ultrasonography with fecal steroid metabolite monitoring to accurately diagnose pregnancy and predict the timing of parturition in the red panda (Ailurus fulgens styani).

Red pandas (Ailurus fulgens styani) exhibit a variable gestation length and may experience a pseudopregnancy indistinguishable from true pregnancy; therefore, it is not possible to deduce an individual's true pregnancy status and parturition date based on breeding dates or fecal progesterone excretion patterns alone. The goal of this study was to evaluate the use of transabdominal ultrasonography for pregnancy diagnosis in red pandas. Two to three females were monitored over 4 consecutive years, generating a total of seven profiles (four pregnancies, two pseudopregnancies, and one lost pregnancy). Fecal samples were collected and assayed for progesterone (P4) and estrogen conjugate (EC) to characterize patterns associated with breeding activity and parturition events. Animals were trained for voluntary transabdominal ultrasound and examinations were performed weekly. Breeding behaviors and fecal EC data suggest that the estrus cycle of this species is 11-12 days in length. Fecal steroid metabolite analyses also revealed that neither P4 nor EC concentrations were suitable indicators of pregnancy in this species; however, a secondary increase in P4 occurred 69-71 days prior to parturition in all pregnant females, presumably coinciding with embryo implantation. Using ultrasonography, embryos were detected as early as 62 days post-breeding/50 days pre-partum and serial measurements of uterine lumen diameter were documented throughout four pregnancies. Advances in reproductive diagnostics, such as the implementation of ultrasonography, may facilitate improved husbandry of pregnant females and allow for the accurate prediction of parturition.

Organic geochemical and palynological evidence for Holocene natural and anthropogenic environmental change at Lake Dojran (Macedonia/Greece)

In this study, we present lipid biomarker and palynological data for a sediment core from Lake Dojran (Macedonia/Greece), which covers the entire Holocene period. We analyzed vascular plant-derived n-alkanes, combustion-derived polycyclic aromatic hydrocarbons (PAHs), fecal steroids, and bacterial and archaeal glycerol dialkyl glycerol tetraether (GDGT) lipids in concert with microcharcoal and pollen assemblages to reconstruct climatic, environmental, and human impact in the Dojran catchment and the greater Dojran area. Overall, our results suggest a relationship between anthropogenic activity and environmental/climatic change since increased human impact corresponds to phases of higher humidity and high lake levels at Lake Dojran. During the early Holocene, the record reveals increasing temperatures and humidity and concurrent increasing vegetation cover and runoff/soil erosion, respectively. Following a thermal maximum during the middle early Holocene, temperatures decrease gradually until present. The middle-Holocene at Lake Dojran is characterized by relatively stable environmental conditions followed by greater climatic instability and strong anthropogenic overprint during the late-Holocene. The fecal stanol record reveals phases of increased human impact during the early Bronze Age, the late Bonze/early Iron Age, and the Middle Ages. A phase of low stanol and PAH concentrations from the late Iron Age until the early Middle Ages is either related to ecosystem changes and/or changes in settlement pattern since concurrent pollen data indicate intensified land use. Human impact re-intensified during the Middle Ages with some variability probably related to climatic variations of the ‘Medieval Warm Period’ and the ‘Little Ice Age’.

Steroid Biomarkers Revisited - Improved Source Identification of Faecal Remains in Archaeological Soil Material.

Steroids are used as faecal markers in environmental and in archaeological studies, because they provide insights into ancient agricultural practices and the former presence of animals. Up to now, steroid analyses could only identify and distinguish between herbivore, pig, and human faecal matter and their residues in soils and sediments. We hypothesized that a finer differentiation between faeces of different livestock animals could be achieved when the analyses of several steroids is combined (Δ5-sterols, 5α-stanols, 5β-stanols, epi-5β-stanols, stanones, and bile acids). We therefore reviewed the existing literature on various faecal steroids from livestock and humans and analysed faeces from old livestock breed (cattle, horse, donkey, sheep, goat, goose, and pig) and humans. Additionally, we performed steroid analyses on soil material of four different archaeological periods (sites located in the Lower Rhine Basin, Western Germany, dating to the Linearbandkeramik, Urnfield Period / Bronze Age, Iron Age, Roman Age) with known or supposed faecal inputs. By means of already established and newly applied steroid ratios of the analysed faeces together with results from the literature, all considered livestock faeces, except sheep and cattle, could be distinguished on the basis of their steroid signatures. Most remarkably was the identification of horse faeces (via the ratio: epi-5β-stigmastanol: 5β-stigmastanol + epicoprostanol: coprostanol; together with the presence of chenodeoxycholic acid) and a successful differentiation between goat (with chenodeoxycholic acid) and sheep/cattle faeces (without chenodeoxycholic acid). The steroid analysis of archaeological soil material confirmed the supposed faecal inputs, even if these inputs had occurred several thousand years ago.

The Changes of Fecal Steroid Hormones and Bacterial Composition in Different Gestation Stages of Meishan Sows

Currently, the importance of gut microbiota to the health of their host has been well discussed, yet the knowledge about the host hormonal effect on the gut microbiota is limited. In this study, a combination of the high-throughput 16S rRNA gene-based pyrosequencing and the Mass Spectrometry (MS)-based metabolomics techniques were used to investigate the stool microbial composition and microbial metabolites at different gestation day in Meishan sows. The results showed that, the stool steroid hormones including estradiol (E2), progesterone (P4) and cortisol (CORT) were increased at gestation day 90 (P<0.05), when compared to gestation day 30. In coincide with the changes of steroid hormones, the relative abundance of Burkholderiales and Selenomonadales of the stool microbiota at d90 of pregnancy was significantly increased than that of at d30 of pregnancy at the order level (P<0.05). However, no significant difference of the richness estimators (ACE and Chao), and the diversity indices (Shannon and Simpson) were found between the d30 and d90 of gestation. Further, the metabolomics profile revealed 13 metabolites changed greatly from the gestation day 30 to 90, including amino acid metabolic pathway, lipid and carbohydrate metabolic pathway (P<0.001). The results suggest that the interaction of the steroid hormones and the gut bacteria may account for the metabolic changes throughout the length of pregnancy.

Lactoferrin interacts with bile acids and increases fecal cholesterol excretion in rats.

Lactoferrin (LF) is a multifunctional cationic protein (pI 8.2-8.9) in mammalian milk. We previously reported that enteric-LF prevented hypercholesterolemia and atherosclerosis in a diet-induced atherosclerosis model using Microminipig, although the underlying mechanisms remain unclear. Because LF is assumed to electrostatically interact with bile acids to inhibit intestinal cholesterol absorption, LF could promote cholesterol excretion. In this study, we assessed the interaction between LF and taurocholate in vitro, and the effect of LF on cholesterol excretion in rats. The binding rate of taurocholate to LF was significantly higher than that to transferrin (pI 5.2-6.3). When rats were administered a high-cholesterol diet (HCD) containing 5% LF, LF was detected using ELISA in the upper small intestine from 7.5 to 60 min after the administration. Rats were fed one of the following diets: control, HCD, or HCD + 5% LF for 21 days. Fecal neutral steroids and hepatic cholesterol levels in the HCD group were significantly higher than those in the control group. The addition of LF to a HCD significantly increased fecal neutral steroids levels (22% increase, p < 0.05) and reduced hepatic cholesterol levels (17% decrease, p < 0.05). These parameters were inversely correlated (R = -0.63, p < 0.05). These results suggest that LF promotes cholesterol excretion via interactions with bile acids.

Excretion and measurement of corticosterone and testosterone metabolites in bank voles (Myodes glareolus)

The bank vole is a commonly used model species in behavioral and ecophysiological studies. Thus, presenting a validated method for noninvasive monitoring of corticosterone and testosterone secretion is of high relevance. Here, we evaluated the effect of time of day and an ACTH challenge test on measured fecal corticosterone (FCM) and testosterone (FTM) metabolites in both sexes. Furthermore, we performed radiometabolism experiments for both steroids and sexes to study metabolism and excretion of 3H-corticosterone and 3H-testosterone. FCM and FTM were analysed with a 5α-pregnane-3β,11β,21-triol-20-one enzyme immunoassay (EIA) and a testosterone (measuring 17β-hydroxyandrostanes) EIA, respectively. Males had significantly higher FCM levels than females and their main excretion route was via the feces (∼72%), whereas females excreted nearly equal portions in both feces and urine. For testosterone the main excretion route was via the feces in both sexes (∼80%). The time course of excretion was similar in both sexes, but for the first time a significant difference between injected steroids was found: Corticosterone was excreted faster than testosterone, both in urine (median of peak levels: 4h vs 6h) and feces (6h vs 8h). Several metabolites were present in the feces and the tested EIAs reacted with some of them. Time of day had a significant effect on measured fecal steroid metabolites. As expected, males had significantly higher FTM levels than females. ACTH administration significantly increased FCM values; peaks were observed 4–8h after injection. In conclusion, both tested EIAs proved suited for a noninvasive measurement of glucocorticoids and androgens in bank voles.

Validation of a field-friendly extraction and storage method to monitor fecal steroid metabolites in wild orangutans.

Measuring hormone metabolites from feces is the most often used method to assess hormonal status in wildlife. Although immediate freezing of fecal samples collected in the field is the best method to minimize the risk of degradation of hormones over time, this is often not possible in remote field sites. Therefore, alternative storage and preservation methods for fecal samples are required in these conditions. We conducted an experiment to investigate if fecal glucocorticoid (FGCM) and progesterone metabolite (pregnanediol-3-glucuronide; PdG) levels measured from samples that were extracted with a simple, field-friendly methodology correlate with those generated from frozen samples. We also evaluated whether storing fecal samples in alcohol is a suitable alternative to preserve FGCM and PdG concentrations long-term (i.e. over a 9-month period) at locations where fecal extraction is not feasible. Finally, we tested if the hormone concentrations in unpreserved fecal samples of orangutans change over 14h when stored at ambient conditions, representing the maximum duration between sample collection and return to the camp. FGCM and PdG levels measured from samples that were extracted with the field-friendly method showed strong correlations with those generated from frozen samples, and mean levels did not differ significantly between these methods. FGCM concentrations showed no significant change compared to control samples when fecal samples were stored for up to 6months in alcohol at ambient temperature and PdG concentrations even remained stable for up to 9months of storage. FGCM concentrations of fecal samples kept at ambient temperature for up to 14h post-defecation did not significantly differ compared to control samples frozen immediately after collection. These results provide the basis for the successful monitoring of the physiological status of orangutans living in remote natural settings, like those included in the Indonesian reintroduction programs.

Puberty, ovarian cycle, pregnancy, and postpartum ovulation in captive Sichuan golden monkeys (Rhinopithecus roxellana) based on changes in urinary and fecal gonadal steroid metabolites

The purpose of this study was to evaluate the reproductive status and clarify the reproductive physiology of captive Sichuan golden monkeys. The concentrations of urinary estradiol-3-glucuronide (E2G) and pregnanediol-glucuronide (PdG) or fecal estradiol-17β (E2) and PdG in two females, and fecal testosterone concentrations in a male, were measured continuously using enzyme immunoassays. On the basis of these hormone profiles, the follicular phase, luteal phase, and ovarian cycle were calculated to be 14.7 ± 4.8, 10.4 ± 2.8, and 25.1 ± 3.3 days, respectively. The first ovulation (puberty) in a female monkey was observed at 5.1 years old, and the first pregnancy was diagnosed at 6.4 years old. For the first 2 months of pregnancy (204 days), fecal E2 and PdG maintained constant high values and then increased until parturition. These profiles were similar to urinary E2G and PdG changes. During the last trimester of a twin pregnancy, fecal PdG was up to approximately three times higher compared with a single pregnancy. Therefore, fecal PdG levels in late pregnancy may be effective for the detection of a twin pregnancy. The first postpartum ovulation occurred 66 (fetal death and artificial rearing), 143 (fetal death), and 189 (natural suckling) days after parturition. The anovulation period of the natural suckling case was longer than the others. Conception and postpartum ovulation were detected between September and January. Fecal testosterone levels of the male were correlated with the fecal E2 level of the nonpregnancy period in exhibited together female. Our results reported that urinary (E2G and PdG) and fecal (E2 and PdG) hormone measurement is effective for monitoring the reproductive status, thereby expanding knowledge of the reproductive endocrinology of this endangered species.

Thiol oxidation of hemolymph proteins in oysters Crassostrea brasiliana as markers of oxidative damage induced by urban sewage exposure

Urban sewage is a concerning issue worldwide, threatening both wildlife and human health. The present study investigated protein oxidation in mangrove oysters (Crassostrea brasiliana) exposed to seawater from Balneário Camboriú, an important tourist destination in Brazil that is affected by urban sewage. Oysters were exposed for 24 h to seawater collected close to the Camboriú River (CAM1) or 1 km away (CAM2). Seawater from an aquaculture laboratory was used as a reference. Local sewage input was marked by higher levels of coliforms, nitrogen, and phosphorus in seawater, as well as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), linear alkylbenzenes (LABs), and fecal steroid in sediments at CAM1. Exposure of oysters to CAM1 caused marked bioaccumulation of LABs and decreased PAH and PCB concentrations after exposure to both CAM1 and CAM2. Protein thiol oxidation in gills, digestive gland, and hemolymph was evaluated. Lower levels of reduced protein thiols were detected in hemolymph from CAM1, and actin, segon, and dominin were identified as targets of protein thiol oxidation. Dominin susceptibility to oxidation was confirmed in vitro by exposure to peroxides and hypochlorous acid, and 2 cysteine residues were identified as potential sites of oxidation. Overall, these data indicate that urban sewage contamination in local waters has a toxic potential and that protein thiol oxidation in hemolymph could be a useful biomarker of oxidative stress in bivalves exposed to contaminants. Environ Toxicol Chem 2017;36:1833-1845. © 2016 SETAC.

Dix ans de collaboration entre la France et le Japon - Etudes sur la reproduction chez les macaques japonais

Our research interests are focused on the evolution of mating systems in primates and we aim at providing a better understanding of the adaptive signification of reproductive processes in female primates, using a comparative approach. Our Franco-Japanese collaboration (2006 - present) allowed us to study sociality and reproduction in a Catarrhine species, the Japanese macaque (Macaca fuscata), and on different but complementary topics (reproductive energetics, reproductive strategies, sexual signaling). We first investigated the links between energetics (i.e. nutritional status, body composition) and reproductive outcomes and our studies in captive Japanese macaques showed that, even in captivity with relaxed energetic constraints (e.g. constant food availability), seasonal breeding entails relatively high energy costs and females with higher energy status could invest more in reproductive activities and could afford to reproduce more rapidly. We also investigated the information that male Japanese macaques received from females that might enable them to discern periods with higher probabilities of conception and we developed a research program on the extent to which Japanese macaques, in common with people, have evolved a system of multimodal sexual signaling. We focused on the cues and signals available to males to make their mating decisions, and we provided information on the potential role of female behavior, female skin color, odours and vocalizations as sexual signals. We suggested, in particular, that females advertized their pregnancy through changes in behavioral, visual, and potentially auditory signals, and that males use these signals to adjust their mating behaviors and avoid wasting energy on non-reproductive copulations. This holistic study, combining fieldwork and laboratory experiments, and using a multidisciplinary approach (behavioral observations (sexual interactions), digital photography (visual scoring of female sexual skin color), hormonal (fecal ovarian steroids) and chemical data (components of female vaginal secretions)) allowed us to assess how different signals may combine to influence patterns of primate mate choices.

Creosote Bush (Larrea tridentata) Improves Insulin Sensitivity and Reduces Plasma and Hepatic Lipids in Hamsters Fed a High Fat and Cholesterol Diet.

Creosote bush, Larrea tridentata (Sesse y Moc. Ex DC, Zygophyllaceae) is a shrub found in the deserts of Northern Mexico and Southwestern United States. In traditional medicine, it is used to treat a variety of illnesses including type 2 diabetes. The present study aims to investigate the effects of creosote bush ethanolic extract on plasma and liver parameters associated with the metabolic syndrome in hamsters fed a high fat and cholesterol diet (HFD), comparing them with those induced by ezetimibe (EZ). Seven groups of six hamsters each were formed. Six groups were fed HFD for 2 weeks. The following 2 weeks, the HFD groups received: (1) only HFD, (2) HFD + 3 mg% EZ, (3) HFD + 0.2% creosote bush ethanolic extract, (4) only standard diet (Std Diet), (5) Std Diet + 3 mg% EZ, (6) Std Diet + 0.2% creosote bush ethanolic extract. The beneficial effects of creosote bush ethanolic extract in the HFD hamster model were a reduction of insulin resistance, associated with lower serum insulin and leptin, lower hepatic lipid peroxidation and higher liver antioxidant capacity. Plasma and liver lipids tended or were reduced to values closer to those of animals fed standard diet. A similar effect on lipids was induced by EZ, although with even lower hepatic cholesterol and total lipids concentrations. In general, the change from HFD to standard diet plus ethanolic extract induced the same but deeper changes, including a reduction in plasma glucose and an increase in the percentage of HDL cholesterol. Unlike creosote bush extract, EZ increased food consumption and neutral fecal steroids, with no significant effect on body weight, epididymal fat pads, liver peroxidation or antioxidant capacity. Also EZ did not modify serum insulin and leptin. However, insulin sensitivity improved to values similar to those induced by the extract. This suggests that the mechanism of action of creosote bush ethanolic extract is different to inhibition of cholesterol absorption or increase excretion. The ethanolic extract of L. tridentata could be useful in the treatment of the metabolic syndrome.

The occurrence of PAHs and faecal sterols in Dublin Bay and their influence on sedimentary microbial communities

The source, concentration, and potential impact of sewage discharge and incomplete organic matter (OM) combustion on sedimentary microbial populations were assessed in Dublin Bay, Ireland. Polycyclic aromatic hydrocarbons (PAHs) and faecal steroids were investigated in 30 surface sediment stations in the bay. Phospholipid fatty acid (PLFA) content at each station was used to identify and quantify the broad microbial groups present and the impact of particle size, total organic carbon (%TOC), total hydrogen (%H) and total nitrogen (%N) was also considered. Faecal sterols were found to be highest in areas with historical point sources of sewage discharge. PAH distribution was more strongly associated with areas of deposition containing high %silt and %clay content, suggesting that PAHs are from diffuse sources such as rainwater run-off and atmospheric deposition. The PAHs ranged from 12 to 3072ng/g, with 10 stations exceeding the suggested effect range low (ERL) for PAHs in marine sediments. PAH isomer pair ratios and sterol ratios were used to determine the source and extent of pollution. PLFAs were not impacted by sediment type or water depth but were strongly correlated to, and influenced by PAH and sewage levels. Certain biomarkers such as 10Me16:0, i17:0 and a17:0 were closely associated with PAH polluted sediments, while 16:1ω9, 16:1ω7c, Cy17:0, 18:1ω6, i16:0 and 15:0 all have strong positive correlations with faecal sterols. Overall, the results show that sedimentary microbial communities are impacted by anthropogenic pollution.