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The present work investigated the effects of two different natural mixtures on aryl hydrocarbon receptor (AhR) and oestrogen receptor (ER)beta protein levels, as well as on the activity of cytochrome P450 (CYP) 1A1 and CYP2B. Consequently, the authors observed the effects of these mixtures on gonadotropine-stimulated steroid secretion by ovarian follicles. The natural mixtures that were studied were 'Mjosa' extracted from burbot liver, which contains a high level of PBDEs, and 'Marine mix', extracted from Atlantic cod liver, which contains a high level of polychlorinated biphenyls (PCBs). Follicular cells were exposed in vitro to 'Marine mix' and 'Mjosa mix' at doses of 3.6 and 1.4 microg ml(-1), respectively. Media were collected and used for steroid analysis and cell viability assays. Cells were used to estimate aromatase activity (CYP19), AhR and ER protein levels, and CYP1A1 and CYP2B1 activity. Western blot analysis indicated down-regulation of AhR by 'Marine mix' and down-regulation of ERbeta by Mjosa mix. Up-regulation of CYP1A1 expression and activity were seen following treatment with Marine mix, but not Mjosa mix. Increased CYP2B1 activity was noted after treatment with both 'Marine mix' and Mjosa mix. Both mixtures increased luteinizing hormone (LH)-stimulated progesterone and testosterone secretion, follicle-stimulating hormone (FSH)-stimulated oestradiol secretion, and CYP19 activity. These results suggest that: (1) 'Marine mix' is a mixed-type CYP inducer; (2) 'Mjosa mix' is an inducer of ERbeta and CYP2B; and (3) both 'Marine mix' and 'Mjosa mix' stimulate aromatase activity as a consequence of oestradiol secretion through activation of CYP19.

Persistent organic pollutants (POP) occur as mixtures in nature and it is difficult to predict the toxicity of such mixtures based on knowledge about toxicity and mechanisms of action for single compounds. The present knowledge on the combined toxic effects and modes of actions of exposure to mixtures is limited. Thus, the scientifically based hazard and risk assessment of POP requires analytical and toxicological data from studies with environmental mixtures of POP. The application of genome wide transcription profiling in toxicology, in combination with classical endpoints, will improve the current understanding of the mechanisms of toxic processes. Furthermore, gene expression data may be useful in establishing new hypothesis and discovering new biomarkers for known toxicity as well as not yet recognized toxicity endpoints. In the present study, developmental and reproductive effects of lifelong exposure to environmental relevant concentrations of two natural mixtures of POP were investigated using classical and molecular methods in a controlled zebrafish model. The mixtures used were extracted from burbot (Lota lota) liver originating from freshwater systems in Norway: one mixture with high levels and one mixture with background levels of polybrominated diphenyl ethers (PBD), polychlorinated biphenyls (PCB), and DDT. The concentration of POP in the zebrafish ranged from levels detected in wild fish from Lake Mjosa, to concentrations reported in human and wildlife populations. Phenotypic effects observed in both exposure groups included (1) reduced survival, (2) earlier onset of puberty, (3) increased male/female sex ratio, and (4) differences in body weight at 5 mo of age. Interestingly, genome-wide transcription profiling showed changes in regulation of genes involved in endocrine signaling and growth. The transcriptomics changes included (1) key regulator genes for steroid and thyroid hormone functions (cga, ncoa3), (2) insulin signaling and metabolic homeostasis (pik3r1, pfkfb3, ptb1), and (3) p53 activation (mdm4). The effects observed in the experimental zebrafish model raise the question of whether chemical pollution represents a risk to the reproductive health of wild fish inhabiting the freshwater system.

The purposes of this review are to (1) evaluate human and experimental evidence for adverse effects on reproduction and development in humans, produced by exposure to phthalates, and (2) identify knowledge gaps as for future studies. The widespread use of phthalates in consumer products leads to ubiquitous and constant exposure of humans to these chemicals. Phthalates were postulated to produce endocrine-disrupting effects in rodents, where fetal exposure to these compounds was found to induce developmental and reproductive toxicity. The adverse effects observed in rodent models raised concerns as to whether exposure to phthalates represents a potential health risk to humans. At present, di(2-ethylhexyl) phthalate (DEHP), di-n-butyl phthalate (DBP), and butyl benzyl phthalate (BBP) have been demonstrated to produce reproductive and developmental toxicity; thus, this review focuses on these chemicals. For the general population, DEHP exposure is predominantly via food. The average concentrations of phthalates are highest in children and decrease with age. At present, DEHP exposures in the general population appear to be close to the tolerable daily intake (TDI), suggesting that at least some individuals exceed the TDI. In addition, specific high-risk groups exist with internal levels that are several orders of magnitude above average. Urinary metabolites used as biomarkers for the internal levels provide additional means to determine more specifically phthalate exposure levels in both general and high-risk populations. However, exposure data are not consistent and there are indications that secondary metabolites may be more accurate indicators of the internal exposure compared to primary metabolites. The present human toxicity data are not sufficient for evaluating the occurrence of reproductive effects following phthalate exposure in humans, based on existing relevant animal data. This is especially the case for data on female reproductive toxicity, which are scarce. Therefore, future research needs to focus on developmental and reproductive endpoints in humans. It should be noted that phthalates occur in mixtures but most toxicological information is based on single compounds. Thus, it is concluded that it is important to improve the knowledge of toxic interactions among the different chemicals and to develop measures for combined exposure to various groups of phthalates.

Contents The effects of human chorionic gonadotropin (hCG) stimulation on fat skatole concentrations and hepatic activities of cytochromes P4502E1 (CYP2E1) and P4502A (CYP2A) were studied in Landrace and Duroc breeds of entire male pigs. Pigs were divided into four groups: two control groups of each breed, without hCG stimulation (n = 20 for each breed), and two experimental groups of each breed, with hCG stimulation (n = 18 for each breed). Pigs were slaughtered 3 days after hCG stimulation and activities of CYP2E1 and CYP2A were measured in liver homogenate. Activities of both CYP2E1 and CYP2A were lower in hCG-stimulated pigs than control pigs for both Landrace (p = 0.005 for CYP2E1, p = 0.016 for CYP2A) and Duroc breeds (p = 0.003 for CYP2E1, p = 0.001 for CYP2A), and skatole concentrations in fat were higher in the hCG-stimulated pigs of both breeds (p

SummaryPurposeSeveral antiepileptic drugs (AEDs) induce changes in endocrine function in women with epilepsy. Levetiracetam (LEV) is one of the newer AEDs, and to date no endocrine side-effects have been reported in humans. However, a recent study on ovarian follicular cells from prepubertal pigs showed that LEV affected basal steroid hormone secretion. The aim of the present study was to investigate possible effects of the drug on endocrine function and ovarian morphology in non-epileptic rats.MethodsThirty female Wistar rats were fed per-orally with either 50mg/kg LEV (n=15) or 150mg/kg LEV (n=15) twice daily for 90–95 days. Twenty rats received a control solution. The rats were killed in the dioestrus phase of the oestrous cycle. Serum concentrations of testosterone, 17β-oestradiol, progesterone, follicle stimulating hormone (FSH), luteinizing hormone (LH) and LEV were measured, and the ovaries examined histologically.ResultsMean ovarian weight showed a significant, dose-dependent increase after LEV treatment. Mean numbers of ovarian follicular cysts were not changed, but the numbers of corpora lutea and secondary follicles were significantly higher in the treated animals. Serum testosterone was significantly increased in treated animals (0.50nmol/l versus 0.16nmol/l in controls, p

The aim of the present study was to study the effects of social instability (regrouping) during the second trimester (7 weeks) of pregnancy on aggression, cortisol concentrations and growth in goats and its consequences for survival, growth and some aspects of behavioural development in the kids. Six weeks after mating, 32 goats were distributed into eight groups. In four of the groups, randomly chosen pairs of goats were rotated between groups every Monday morning (around 08:30 h) for 7 weeks (unstable groups), starting 6 weeks into gestation (second trimester). The remaining four groups were kept stable throughout the entire pregnancy (stable groups). The adult goats were video recorded for 6 h twice a week for the first, second, fourth and seventh regrouping and for the 2 last weeks before expected birth. Blood samples of the adult goats were collected in a period from I week before the start of the first regrouping and until I week before expected birth. Blood samples from a maximum of two kids from each litter were collected at 3 weeks of age. Two kids from each litter were subjected to two types of behavioural tests: a 'social test' at the age of 1 and 7 weeks and a 'novel object test' at the age of 5 weeks. Except for the higher aggression level in the unstable groups, social instability did not have any other significant effects on factors such as growth, cortisol level or kid production in the goats. There were only minor effects on the behavioural development in the goat kids. However, kids from the unstable groups showed more escape attempts in the first trial of a,social test', spent more time in contact with a novel object and unfamiliar kids, and showed less fear (escape attempts) after they were given one exposure to the social test situation. Furthermore, kids from unstable groups had a lower basal cortisol level than kids from stable groups. (C) 2008 Elsevier B.V. All rights reserved.

Milk progesterone measurements were used to study post-partum ovarian activity and risk factors for ovarian dysfunction in Zebu cows reared under a traditional pastoral management system in three villages in Morogoro region, Tanzania. Milk samples for progesterone analysis were collected twice per week from a total of 98 lactations. The most prevalent ovarian dysfunction detected was delayed onset of post-partum ovarian activity and cessation of cyclical ovarian activity after a period of normal ovarian function. Prolonged luteal phases were not observed. Resumption of post-partum ovarian activity was observed in 61 (62.2%) cows. Pregnancy was diagnosed in 43 (43.8%) out of 98 cows and was terminated by abortion in 7 (16.3%) cows. The median number of days from calving to resumption of ovarian activity was lower in cows that calved during the dry season (167 days) as compared with those that calved in the rainy season (200 days, P=0.03), probably because ovarian function was stimulated in cows entering the rainy season first after the puerperal phase had ended. Cyclical ovarian activity ceased in seven (11.5%) cows, and all but one case of cessation of ovarian activity took place during the dry season. Cows in their first and second parities experienced prolonged intervals (244 days) between calving and onset of ovarian activity compared with cows in their third or later parity (172 days, P=0.03). Cows with marked losses in body condition experienced prolonged intervals between calving and onset of ovarian activity (229 days) as compared with cows that lost1 body condition unit (172 days, P=0.03). The time interval from calving to resumption of ovarian activity varied between the three villages of the study (142, 192 and 229 days, P=0.05), indicating local differences in management practices.

Samples (blood or tissue fluid) from 594 arctic foxes (Alopex lagopus), 390 Svalbard reindeer (Rangifer tarandus platyrhynchus), 361 sibling voles (Microtus rossiaemeridionalis), 17 walruses (Odobenus rosmarus), 149 barnacle geese (Branta leucopsis), 58 kittiwakes (Rissa tridactyla), and 27 glaucous gulls (Larus hyperboreus) from Svalbard and nearby waters were assayed for antibodies against Toxoplasma gondii using a direct agglutination test. The proportion of seropositive animals was 43% in arctic foxes, 7% in barnacle geese, and 6% (1 of 17) in walruses. There were no seropositive Svalbard reindeer, sibling voles, glaucous gulls, or kittiwakes. The prevalence in the arctic fox was relatively high compared to previous reports from canid populations. There are no wild felids in Svalbard and domestic cats are prohibited, and the absence of antibodies against T. gondii among the herbivorous Svalbard reindeer and voles indicates that transmission of the parasite by oocysts is not likely to be an important mechanism in the Svalbard ecosystem. Our results suggest that migratory birds, such as the barnacle goose, may be the most important vectors bringing the parasite to Svalbard. In addition to transmission through infected prey and carrion, the age-seroprevalence profile in the fox population suggests that their infection levels are enhanced by vertical transmission.

Androstenone is a 16-androstene steroid pheromone produced in the Leydig cells in the testis, and considered to be one of the major compounds responsible for boar taint. In entire male pigs, progress of sexual maturation has been related to an increase in androstenone levels in fat. Onset of puberty and subsequent reproductive function involves genetic factors affected by the internal and external environment. In this study entire male cross-bred pigs were housed under two different light regimens in order to manipulate the onset of puberty. DNA flow cytometry (FCM) was used to study spermatogenesis and monitor the proportions of haploid (1 n), diploid (2 n), and tetraploid (4 n) testicular cells, with conventional histological evaluation used as the reference technique. Agreement between these two methods was found to be good. The best fit model explained 34% of the variation in the androstenone concentrations. Sexual maturation in boars of 125–146 days of age, as assessed by DNA FCM, was not significantly associated with the variation in androstenone concentrations in adipose tissue when various independent variables (breed, age, light strategy, skatole concentrations in fat, and length of the bulbourethralis glands) were included in this model. These findings support the suggestion that selection against androstenone may be an option in the breeding of entire male pigs.

The aim of this study was to investigate if environmentally relevant doses of the putative estrogenic non dioxin-like PCB 153 and the dioxin-like PCB 126 caused changes in bone tissue in female goat offspring following perinatal exposure. Goat dams were orally dosed with PCB 153 in corn oil (98 microg/kg body wt/day) or PCB 126 (49 ng/kg body wt/day) from day 60 of gestation until delivery. The offspring were exposed to PCB in utero and through mother's milk. The suckling period lasted for 6 weeks. Offspring metacarpal bones were analysed using peripheral quantitative computed tomography (pQCT) after euthanisation at 9 months of age. The diaphyseal bone was analysed at a distance of 18% and 50% of the total bone length, and the metaphyseal bone at a distance of 9%. Also, biomechanical three-point bending of the bones was conducted, with the load being applied to the mid-diaphyseal pQCT measure point (50%). PCB 153 exposure significantly decreased the total cross-sectional area (125 mm(2)+/-4) versus non-exposed (142 mm(2)+/-5), decreased the marrow cavity (38 mm(2)+/-4) versus non-exposed (50 mm(2)+/-3) and decreased the moment of resistance (318 mm(3)+/-10) versus non-exposed (371 mm(3)+/-20) at the diaphyseal 18% measure point. At the metaphyseal measure point, the trabecular bone mineral density (121 mg/cm(3)+/-5) was increased versus non-exposed (111 mg/cm(3)+/-3). PCB 126 exposure did not produce any observable changes in bone tissue. The biomechanical testing of the bones did not show any significant changes in bone strength after PCB 153 or PCB 126 exposure. In conclusion, perinatal exposure to PCB 153, but not PCB 126, resulted in altered bone composition in female goat offspring.

Pregnant does (10 goats/group) were dosed orally either with polychlorinated biphenyl (PCB) 153 (98 microg/kg body weight/d) or PCB 126 (ng/kg body weight/d) dissolved in corn oil or with corn oil only (control group) from gestation day (GD) 60 until delivery. An additional group (n = 5) of pregnant does received the synthetic estrogen diethylstilbestrol (DES; 0.4 microg/kg body weight/d) by intramuscular injection using the same treatment schedule as for the PCB groups. Blood samples for immune analysis were collected at wk 0, 1, 2, 4, 6, and 8 of age. The effects of perinatal PCB exposure on postnatal humoral immune responses were examined by assessing the levels of total immunoglobulin G (IgG) and immunoglobulins to specific microbes at wk 0, 1, 2, 4, 6, and 8 of age, and immune responses following immunization of kids at 2 wk of age. PCB 153 exposure suppressed maternal and neonatal immunity, as demonstrated by reduced transfer of maternal IgG and specific antibodies to the environmental microbes Arcanobacterium pyogenes, Mannheimia haemolytica, and reovirus (REO-1). Furthermore, PCB 153 reduced the level of maternal antibodies to Mycobacterium avium paratuberculosis and equine influenza virus (EIV-1) in the newborn kids. The antibody response against EIV-1 was significantly higher in PCB 153-exposed kids 2 wk following immunization. PCB 126 exposure reduced the levels of maternal antibodies to REO-1. In contrast, gestational exposure to PCB 126 increased the concentrations of maternal antibodies to tetanus toxoid. No differences from controls in plasma total IgG levels at birth or colostrum IgG concentrations were observed in the PCB 126-treated does. However, a significant reduction in IgG levels from GD 60 until delivery was found in this group. Gestational exposure to DES reduced the concentrations of maternal antibodies against A. pyogenes, M. haemolytica, M. avium Paratuberculosis, and REO-1. These results suggest that perinatal exposure to low doses of PCB 126 and PCB 153 affects the maternal immunity in kids. The difference in responses between PCB 126 and PCB 153 treatment groups may strengthen the hypothesis that PCBs mediate immunotoxic effects through both AhR-dependent and -independent mechanisms. The observation that the effects produced by PCB 153 resembled those produced by DES raises the question of whether this congener may modulate immunity by estrogenic mechanisms.

Plasma total protein (TP), albumin (ALB) and globulin (GLOB) concentrations and albumin/globulin ratio (A/G) were analysed from blood samples collected from free-ranging reindeer (Rangifer tarandus tarandus) herds at varying times of year. The same parameters were followed in nine captive reindeer with varying protein and energy intake. Variation in the blood constituents of free-ranging reindeer was analysed in relation to different extrinsic and intrinsic factors and compared to findings from captive animals, allowing the analysis of effects of protein and energy intake. There was large overall variation in TP, ALB, GLOB and A/G ratio in the free-ranging animals, ranging between 36-110 g/L, 18-59 g/L, 17-59 g/L and 0.5-2.1. The variation between months and years was significant for all variables except the A/G ratio, where no year effect was noted. Increase in live body mass was associated with a small significant increase and pregnancy with a small significant decrease in all dependent variables, except for the A/G ratio. Age did not have a significant effect on any of the blood constituents when body mass was included in the same model. In captive animals, feeding lichens with low protein content was related to a significant decline in TP, ALB and GLOB, but not in the A/G ratio, whereas feeding commercial ration increased plasma TP, ALB and GLOB significantly. Extrinsic factors such as season and year explained majority of variation in the blood constituents of free-ranging reindeer, whereas body mass, pregnancy and age had only a minor influence. It is concluded that plasma TP and ALB, and to a lesser extent GLOB and A/G ratio may serve as nutritional biomarkers of reindeer.

Since the polar bear (Ursus maritimus) is among the most highly organochlorine-contaminated species of the Arctic mammals, there is growing concern that in addition to the natural stressors in the polar bear's environment, several organochlorines (OCs) may be able to change basic endocrine pathways. Alterations in the hypothalamic-pituitary-adrenal (HPA) axis may affect plasma cortisol concentrations and inhibit physiological processes involved in the maintenance of homeostasis in a way that may endanger the animal's health. Between 1995 and 1998, samples were collected from 121 male and 130 female free-ranging polar bears from the Svalbard area. The aim of the study was to investigate relationships between plasma cortisol concentrations, biological factors, and OCs. The variation in plasma cortisol concentrations was determined for the total sample. Axillary girth and body mass together with their interactions explained more than 50% of the variation in the plasma cortisol concentration. The sum of pesticides (Sigma pesticides) combined with the sum of polychlorinated biphenyls (Sigma PCBs) and their interactions explained over 25% of the variation in the cortisol concentration. Although Sigma pesticides contributed negatively and Sigma PCBs contributed positively to the variation in the plasma cortisol, the over-all contribution of the OCs to the plasma cortisol variation was negative. Despite the complexity on stress responses and the interactions with environmental factors, this study demonstrated that high concentrations of OCs in polar bears might alter plasma cortisol concentrations.

Pregnant does (10 goats/group) were dosed orally with either PCB 153 or PCB 126 dissolved in corn oil or only corn oil (control group) from day 60 of gestation until delivery. Effects on in vitro mitogen-induced lymphocyte proliferation and blood cell counts in their goat kids exposed to low levels of PCB 153 and PCB 126 during gestation and lactation were assessed. The concentrations of PCB 153 and PCB 126 in adipose tissue in the goat kids 9 mo postpartum were 5800 ng/g (fat weight) and 0.49 ng/g (fat weight), respectively. Kids exposed to PCB 153 had a significantly higher number of white blood cells, neutrophils, and lymphocytes at 2 wk of age compared to controls. In the kids exposed to PCB 126 there was a significantly lower concentration of monocytes at 2, 4, and 8 wk of age. The mean lymphocyte response to phytohemagglutinin (PHA) and to concanavalin A (Con A) was significant lower in the PCB 153 compared to the control group at wk 2, 4, and 8 postnatally. The results of the present study support previous reports on immunotoxic effects of PCB exposure in animals. However, this is the first report to demonstrate immunotoxicity in animals by using low doses of PCB 153. The difference in results between PCB 126 and PCB 153 treatment groups may strengthen the hypothesis that PCBs mediate immunotoxic effects through both AhR-dependent and -independent mechanisms.

The aim of the present study was to investigate if environmental doses of PCB 153 and PCB 126 could produce effects in a controlled animal model. Possible adverse effects on the hypothalamic-pitutitary-gonadal axis were examined by measuring gonadotrophins and gonadal steroid hormone concentrations in goat kids exposed during gestation and lactation. The concentrations of PCB 153 and PCB 126 in adipose tissue in the goat kids 9 months post-partum were 5800 ng/g (fat-weight, range; 2900-12700 ng/g) and 0.49 ng/g (fat-weight, range; 0.28-0.80 ng/g), respectively. The pre- and post-pubertal plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (Prl) and progesterone (P4) were analysed. LH, FSH, Prl, and P4 were also measured during an induced oestrus cycle. The prepubertal LH concentration was significantly lower, the puberty was delayed and the P4 level during the luteal phase of an estrous cycle was higher in the group exposed to PCB 153. No significant effect of PCB 153 exposure was found on Prl and FSH. PCB 126 did not produce any effects at the exposure level tested in this study. In conclusion, perinatal exposure to PCB 153 affected the reproductive function and the puberty maturation in goats.

The aim of the present study was to gain knowledge about the disposition of the PCB congeners 126 and 153 in a goat model where pregnant does were given oral doses of PCB from day 60 of gestation until delivery. The goat kids were thus exposed to PCB during gestation and lactation. The doses of PCB 153 and PCB 126 were 98 and 49 ng/kg body wt/day, respectively. PCB levels were measured in plasma from the does at day 90 of gestation and at delivery, and in plasma from the kids at birth and four weeks post partum. Concentrations of PCB were analysed in brain, liver and fat tissue from the does six weeks after delivery and in liver and fat tissue from the kids at nine months of age. The ratio of estimated body burden to ingested dose in the does, was 0.95 +/- 0.07 for PCB 153 and 0.41 +/- 0.03 for PCB 126. Approximately 9% of PCB 153 and 6% of PCB 126 was transferred from the mothers to their kids during gestation and lactation. Prenatal exposure contributed to a much lower fraction of the body burden than postnatal PCB intake via milk, due to the fact that almost 100% of the PCB 153 transferred from the does to kids was transferred via milk, and the PCB 126 intake via milk was threefold higher than the calculated body burden. The hepatic PCB 126 concentration in both does and kids was significantly higher than the concentration in fat, in contrast to PCB 153, where the highest concentrations were found in fat. A significant difference in body burden between the does at delivery, the newborn kids, and the kids at four weeks of age, did not influence the plasma concentration of PCB 153 on a fat weight basis, which showed no difference with sampling time. Our results suggest that PCB 126 and PCB 153 have different pharmacokinetic properties. The higher levels of PCB 126 in liver tissue compared to fat tissue confirm the concept of hepatic sequestration. The similar blood concentration of PCB 153 in the does at delivery and their newborn kids despite the considerable difference in body burden, suggests a high degree of placental transfer. This supports previous observations that low molecular weight, lipid-soluble, non-polar chemicals reach the fetus to the greatest possible extent. For PCB 153, the body burden may provide the appropriate dose metric at steady state, but may give a minor underestimation of PCB 126 at low environmental exposure levels due to hepatic sequestration.

Even though theoretical models show that parasites may regulate host population densities, few empirical studies have given support to this hypothesis. We present experimental and observational evidence for a host-parasite interaction where the parasite has sufficient impact on host population dynamics for regulation to occur. During a six year study of the Svalbard reindeer and its parasitic gastrointestinal nematode Ostertagia gruehneri we found that anthelminthic treatment in April-May increased the probability of a reindeer having a calf in the next year, compared with untreated controls. However, treatment did not influence the over-winter survival of the reindeer. The annual variation in the degree to which parasites depressed fecundity was positively related to the abundance of O. gruehneri infection the previous October, which in turn was related to host density two years earlier. In addition to the treatment effect, there was a strong negative effect of winter precipitation on the probability of female reindeer having a calf. A simple matrix model was parameterized using estimates from our experimental and observational data. This model shows that the parasite-mediated effect on fecundity was sufficient to regulate reindeer densities around observed host densities.

The aim of the present study was to investigate whether exposure of porcine ovarian follicular cells to clinically relevant concentrations of valproate affected steroid production in vitro and to which extent these effects were reversed by removal of the drug from the culture medium. Small and medium follicles were obtained from ovaries collected, respectively, at days 8-10 and 14-16 of the estrous cycle. Theca and granulosa cells were collected from follicles and co-cultured in one well. To show whether the effect of valproate was reversible, cells were cultured for 24, 48, or 72 h with valproate 100 or 250 microg/ml. The medium was then changed to fresh medium without drugs for an additional 24, 48, or 72 h. Valproate added to the culture medium caused a significant reduction of estradiol secretion with concomitant increase in both testosterone and progesterone secretion by small follicles. In medium-sized follicles, 100 microg/ml valproate was without effect on estradiol secretion while 250 microg/ml caused a small, but statistically significant decrease. The effects of valproate on steroid secretion patterns were irreversibly independent of concentration, exposure time, and time of restoration after drug exposure up to 72 h in both small and medium follicles. In conclusion, the present study demonstrated that even short-term valproate treatment disrupted follicular steroidogenesis in isolated ovarian follicular cells resulting in increased testosterone and progesterone and decreased estradiol secretion. It was not possible to reverse the steroidogenic effects of valproate by removing the drug from the cell cultures.

Research on Alzheimer's disease (AD) has indicated an association between hormones of the hypothalamic-pituitary-gonadal (HPG) axis and cognitive senescence, indicating that post meno-/andropausal changes in HPG axis hormones are implicated in the neuropathology of AD. Studies of transgenic mice with AD pathologies have led to improved understanding of the pathophysiological processes underlying AD. The aims of this study were to explore whether mRNA-levels of gonadotropin-releasing hormone (Gnrh) and its receptor (Gnrhr) were changed in plaque-bearing Alzheimer's disease transgenic mice and to investigate whether these levels and amyloid plaque deposition were downregulated by treatment with a gonadotropin-releasing hormone analog (Gnrh-a; Leuprorelin acetate). The study was performed on mice carrying the Arctic and Swedish amyloid-β precursor protein (AβPP) mutations (tgArcSwe). At 12 months of age, female tgArcSwe mice showed a twofold higher level of Gnrh mRNA and more than 1.5 higher level of Gnrhr mRNA than age matched controls. Male tgArcSwe mice showed the same pattern of changes, albeit more pronounced. In both sexes, Gnrh-a treatment caused significant down-regulation of Gnrh and Gnrhr mRNA expression. Immunohistochemistry combined with quantitative image analysis revealed no significant changes in the plaque load after Gnrh-a treatment in hippocampus and thalamus. However, plaque load in the cerebral cortex of treated females tended to be lower than in female vehicle-treated mice. The present study points to the involvement of hormonal changes in AD mice models and demonstrates that these changes can be effectively counteracted by pharmacological treatment. Although known to increase in normal aging, our study shows that Gnrh/Gnrhr mRNA expression increases much more dramatically in tgArcSwe mice. Treatment with Leuprorelin acetate successfully abolished the transgene specific effects on Gnrh/Gnrhr mRNA expression. The present experimental approach should serve as a platform for further studies on the usefulness of Gnrh-a treatment in suppressing plaque development in AD.

Norwegian School of Veterinary Science, PB 8146 Dep, 0033 Oslo, Norway Department of Neuropsychiatry and Psychosomatic Medicine, Division of Surgery and Clinical Neuroscience, Oslo University Hospital –— Rikshospitalet, Sognsvannsveien 20, 0027 Oslo, Norway c Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, Bearsden Road, Glasgow G61 1QH, Scotland, UK Center for the Study of Human Cognition, Department of Psychology, University of Oslo, PB 1094 Blindern, 0317 Oslo, Norway KG Jebsen Centre for Psychosis Research, Division of Mental Health and Addiction, Oslo University Hospital Ulleval, Kirkeveien 166, PO Box 4956 Nydalen, 0424 Oslo, Norway Department of Psychology, University of California, Berkeley, CA, USA

The cellular composition of the silver fox testis assessed by DNA flow cytometry and histological analysis exhibited marked circannual alterations. The proportion of haploid cells increased from late October to the breeding season in February, while that of diploid cells decreased and that of tetraploid cells fluctuated during the same period. Towards the end of March these changes were reversed. The seasonal variations in testicular histology paralleled the changes in distribution of cells from the different DNA populations. In August, 69% of the tubules contained spermatogonia as the only type of germ cell, while the remaining 31% also contained a few primary spermatocytes. In late October more than 50% of the tubules contained spermatocytes, and during the period of further activation from early December-February the seminiferous epithelium included round and/or elongated spermatids as well. In February, all tubules contained complete associations of germ cells, whereas in late March tubules with spermatogonia only and spermatogonia together with a few spermatocytes reappeared. In May, only such tubules could be found indicating total regression. Plasma concentrations of FSH and LH increased from early November, both gonadotrophins reaching maximum levels in December or early January, and then both declined during the second part of January, immediately prior to the actual breeding season. LH values showed a few smaller peaks in the beginning of June, whereas FSH levels were generally low until the next period of testicular reactivation. Testosterone concentrations were also low during most of the year but rose in November and December to reach a peak in January and a second peak in June. In animals immunized against inhibin the distribution of haploid, diploid and tetraploid cells did not deviate to any great extent from that in the controls, except in March when the immunized males had a markedly lower proportion of tetraploid cells, and in May, when they had a distinctly higher proportion of haploid cells. These findings were partly reflected by the histology. In the immunized animals, plasma FSH levels started to increase at approximately the same time but peaked higher and remained elevated almost 1 month longer than in the controls, whereas both the rise and decline in LH levels generally coincided with the variations in these animals, but the values were mostly higher. The testosterone profiles were similar to those in the controls except that the maximum values were also usually higher.

The effects of different levels of concentrate, milk yield, and energy balance (EB) on postpartum ovarian activity were studied in 164 lactations from 108 moderate yielding, dual-purpose cows. All cows were of the breed Norwegian Cattle. The cows were assigned to three different concentrate allocations and had free access to roughage. Cows in the high energy group were fed as suggested by Norwegian standards, whereas cows in the low energy group were supplied with approximately 40% less concentrate than recommended. Cows in the medium energy group were fed as the low energy group prepartum and as the high energy group during the first 90 d in milk (DIM). Energy balance was estimated by subtracting energy required for maintenance and lactation from energy intake. Ovarian activity was expressed in categories (early, mid and late responders) of days to first luteal response (milk progesterone concentration >5 ng/ml) and days to first regular cycle. Energy balance and milk yield throughout the first 100 DIM were compared between categories of concentrate allocation and ovarian activity using repeated measures mixed models. Decreasing level of concentrate was related to negative EB and declining milk yield. Both negative EB and high milk yield were associated with delayed postpartum ovarian activity. Concentrate allocation did not, however, alter days to first luteal response or days to first cycle. This study demonstrated that although negative EB was associated with delayed ovulation, Norwegian Cattle adapted to less energy-dense rations by reducing milk yield rather than delaying onset of ovarian activity.

Relationships between light exposure, milk yield, and reproductive traits in Norwegian Red cattle were studied using data from 1538 farms in southern Norway during the darkest part of the year (November 1 to February 28). Production level was expressed as milk yield at first artificial insemination (AI) and 305-d milk yield. Reproductive performance was calculated using the following dependent variables: age at first AI, age at first calving, days open, calving interval, number of AI per cow, and nonreturn rate at 60 d. Days open and calving interval were both 4.0 d shorter, number of AI per cow was 0.07 less, nonreturn rate was 3.1% greater, and daily milk yield at first AI was 0.5 kg greater in herds that used dim illumination at night compared with herds that used no illumination at night. Milk yield at first AI was 0.5 kg greater in herds with photoperiods >12 h. Photoperiods >12 h were associated with a reduction in age at first AI by 4.8 d and age at first calving by 6.6 d. We concluded that dairy cattle kept at high latitudes during winter should be exposed to dim illumination at night and a minimum photoperiod of 12 h to stimulate reproductive performance and milk yield.

Two qualitative on-farm milk progesterone test kits were used for early pregnancy diagnosis in goats. One kit was based on latex agglutination (LA) and the other on enzyme-linked immunosorbent assay (ELISA). The accuracy of early pregnancy diagnosis by these two methods was compared with the accuracy of oestrous observation (OeO) and the level of plasma progesterone measured by radioimmunoassay (RIA). Dairy goats (n = 73) from a single herd were used for collection of milk and blood samples 20 days after breeding. Ultrasound examination 50 days after mating found that 49 (67%) goats were pregnant, and 24 (33%) were not pregnant. Using ultrasound as a reference method, the accuracy of early pregnancy diagnosis by RIA and OeO was 92% and 86%, respectively, and for non-pregnancy 100% for both RIA and OeO. The accuracy of early pregnancy diagnosis by ELISA and LA was 82% and 79% and for non-pregnancy, 88% and 100%, respectively. The kappa-statistic for RIA, OeO, ELISA and LA was 0.93, 0.84, 0.77 and 0.73, respectively. It was concluded that LA and ELISA tests can be used for early pregnancy diagnosis in dairy goats. However, in the herd studied, early pregnancy diagnosis by OeO was as good as that achieved with progesterone determination using the kits.

The effect of Listeria monocytogenes infection on hormone levels in pregnant goats was studied. Four goats (Group I) received an intravenous inoculation of a bacterial culture (Type 1) on Days 69-77 and another four goats (Group II) received a similar inoculation on Days 105-106 of gestation. Five non-inoculated goats were used as controls. Plasma was analysed for progesterone, oestrone sulphate and 15-ketodihydro-PGF2 alpha. The status of the foetus was followed using real-time ultrasonography. Three of the four goats in Group I aborted 8-10 days after inoculation. The fourth goat gave birth to a normal live kid at term. The three goats which aborted showed clinical signs of disease in connection with abortion. In Group II, all goats aborted after 9-11 days. All the goats showed clinical symptoms of disease from a few days after inoculation and the symptoms continued until abortion. The clinical symptoms of disease were more pronounced in Group II than in Group I. L. monocytogenes was isolated from all aborted foetuses. None of the control goats aborted. Ultrasound examination revealed foetal death either immediately before or up to 2 days before abortion. Mummification had begun in the foetus that had been dead for 2 days before expulsion. In comparison with pre-inoculation plasma levels in Group I, a significant decrease (P0.05) in progesterone levels and an increase in 15-ketodihydro-PGF2 alpha levels were observed from Days 4 and 6 after inoculation, respectively. In Group II, a significant decrease (P0.05) in progesterone levels and an increase in 15-ketodihydro-PGF2 alpha levels in plasma were observed from Days 8 and 6, respectively. The oestrone sulphate levels decreased slightly in the inoculated goats a few days before abortion. The pattern of changes in levels around abortion was similar to the pattern present in the control animals around parturition. However, oestrone sulphate levels did not increase in the inoculated groups before abortion in contrast to goats which delivered healthy kids. The changes in levels of 15-ketodihydro-PGF2 alpha in goats that aborted indicated that the endocrine foetal-placental function was disturbed, which was most likely due to the establishment and development of L. monocytogenes in the placenta and foetus.

Blood samples were collected from eleven 1.5 year old female reindeer three times a week from September 1992 until February 1993 and daily in October and November 1992. Blood samples were collected every third hour for a period of three weeks from two females that showed regular oestrous cycles. Plasma progesterone, oestradiol, LH and 15-ketodihydro-PGF2 alpha were analysed to characterize variations in ovarian function. Reindeer are seasonally polyoestrous. Average duration of oestrous cycles was 19.4 days (range: 13-33 days). Short periods of high progesterone concentrations (4-8 days duration) occurred in five animals before the onset of regular oestrous cycles. The first regular cycle had a significantly longer luteal phase than did the following cycles. During luteolysis 15-ketodihydro-PGF2 alpha was released in a pulsatile pattern. Maximal oestradiol concentrations preceded preovulatory LH peaks by about 3 h in the two frequently sampled animals. The duration of LH surges was 12 and 15 h, respectively.

A major response of the body to stress is the secretion of adrenocorticotrophic hormone (ACTH) by the adenohypophysis resulting in increased blood Cortisol levels. Corticotropin releasing factor (CRF) controls the release of ACTH, mediating a direct effect at the level of the pituitary (Guillame et al. 1992). An animal's perception of a stressful event and the resulting increase in Cortisol production can be influenced by previous experience, season, time of day, and the animal's sex, age, and condition (Dantzer & Mormede 1983, Moberg 1985).

Kjxstad, H., E. Ropstad and K. Andersen Berg: Evaluation of spermatological parameters used to predict the fertility of frozen bull semen. Acta vet. scand. 1993,34,299-303.– Post-thaw motility, velocity and acrosome integrity of frozen semen were determined in 18 bulls with varying fertility (average non-return rates: 71.3 (± 2.8) - range: 65.2-75.7). Five semen straws were investigated from each bull. The average values for sperm motility (percentage motile spermatozoa), sperm velocity (graded from 0-3) and acrosome integrity (proportion of spermatozoa with intact acrosome) were 67.5%, 2.5 and 79.3%, respectively. Significant correlations were found between sperm motility and velocity, but not between sperm motility and acrosome integrity. Both sperm motility and velocity were significantly related to bull fertility. It was concluded that of the post-thaw semen characteristics investigated in this study these 2 parameters provided a reliable basis for prediction of bull fertility.

Organohalogen contaminants detected globally in avian wildlife, including populations from the Arctic, have been related to various reproductive hormone potencies, and altered hormonal balance and functions. Besides legacy organochlorine (OC) substances, that is, polychlorinated biphenyls (PCBs) and OC pesticides and by-products, endocrine-disruptive properties have been defined for chemicals of new and emerging environmental concern, such as polybrominated diphenyl ethers (PBDEs) and metabolically derived products like methylsulfonyl (MeSO2)- and hydroxyl (OH)-PCBs. We investigated the relationships between plasma concentrations of selected legacy OCs, PBDEs, and MeSO2- and OH-PCB metabolites and the circulating reproductive hormones testosterone (T), 17beta-estradiol (E2), and progesterone (P4) in incubating male and female glaucous gulls (Larus hyperboreus) from the Norwegian Arctic. Principal component and regression analyses demonstrated that P4 levels in male glaucous gulls were associated positively with variations of sum (Y) PCB, dichloro-diphenyl-trichloroethane (sigmaDDT), chlordane (sigmaCHL), and sigmaPBDE concentrations, which were the most recalcitrant organohalogens determined in glaucous gulls. No such relationship was found for female glaucous gulls as well as between concentrations of any of the selected organohalogens and levels of T for both sexes. The E2 was not detected in any plasma samples. Present results were highly suggestive that exposure to high organohalogen concentrations in glaucous gulls, particularly the most persistent compound classes, may have the potential to interfere with steroidogenesis and impinge on circulating P4 homeostasis. Because significant effects were found in males exclusively, it cannot be completely ruled out that male glaucous gulls are more sensitive than females to organohalogen-mediated alteration of P4 synthesis and breakdown.

Blood selenium levels were found to fluctuate throughout the year, being highest during the indoor season when the greatest amounts of compound concentrates were fed. From October to January the average blood selenium levels increased from 0.10 μg/ml to 0.18 μg/ml (15 cows). Subcutaneous injections of barium selenate (500 mg selenium) increased the blood selenium levels significantly. The treated group (15 cows) reached average levels of about 0.21 μg/ml blood during the indoor season. The effect of oral supplementation of sodium selenite (for 2 months) on the levels of plasma selenium, blood selenium and glutathione peroxidase activity was investigated. Plasma selenium was found to give an immediate reflection of the daily selenium intake. Maximum activity of glutathione peroxidase was reached 1 month after the end of the supplementation period. It is concluded that if selenium enriched concentrates are used in a normal feeding regimen, further supplementation with selenium does not seem to be necessary.

The study was undertaken to establish the relationships between milk and plasma urea levels and protein supply, using traditional and new measures of feed protein value. Samples of milk and plasma were taken before morning feeding twice weekly for the first 3 months post partum from 21 multiparous cows and 7 heifers. Samples of rumen fluid were collected every second week. The cows were distributed into 4 different feeding groups. Feeding regimens were factorial with respect to protein content of the concentrates (i.e. Low protein (Lp): 12.5% digestible crude protein (DCP) vs. High protein (Hp): 17.5% DCP) and concentrate allowances (i.e. Low energy (Le): substandard vs. High energy (He): standard). Silage was offered ad libitum. The within-animal coefficients of correlation between milk and plasma urea levels, and between milk urea levels and the levels of ammonia in the rumen fluid wer r = 0.88 (p less than 0.001) and r = 0.75 (p less than 0.001), respectively. Analysis of variance showed the following sources of urea variation to be significant: Protein balance in the rumen (PBV), intake of DCP, intake of amino acids absorbed in the intestine (AAT), intake of fattening feed units, and the individual animal factor. A similar tendency was found for the variation of ammonia in rumen fluid. The coefficients of determination (R2 model) were not increased by introducing protein balance (BADCP), the amount of PBV and AAT in the dry matter ingested, or the protein/energy ratio as alternative parameters of protein intake in the models. The levels of protein or energy intake did not significantly influence the levels of plasma albumin. The correlations between urea levels and other plasma indicators of metabolic status and liver function were generally low.

Selenium levels were assayed and related to reproductive and health performance in two different materials, comprising 17 dairy herds without reproductive problems (Material I) and 47 herds with reproductive problems (Material II). A high correlation between plasma and blood selenium levels was found (r=0.79). A majority of dry cows and pregnant heifers showed marginal or deficient selenium levels. Among herds with reproductive problems, 43% of the herds had a low selenium status (

Two feeding experiments were carried out in 2 successive years with 28 cows of the Norwegian Red Cattle (NRF) in each experiment. The cows were randomly distributed into 4 groups and subjected to different feeding regimens from 1 month prior to calving until 12 weeks after calving. The experimental design was factorial (2 x 2) with respect to protein content of the concentrate (17.5% digestible crude protein (DCP) v.s. 12.5% DCP) and concentrate allowances (standard v.s. substandard allowances after calving). Silage was offered ad libitum. Samples for estimation of serum immunoglobulin-G, white blood cells and lymphocyte responses to the mitogens phytohemagglutinin, concanavalin A and pokeweed mitogen were collected 4 weeks prior to expected calving, and 2, 4 and 8 weeks after calving. The levels of milk immunoglobulin-G were estimated at calving and 2, 4 and 8 weeks after calving. A significant positive relationship was found between the estimated energy balance and the lymphocyte response to mitogens. Little evidence was found for the existence of a significant relationship between the immunologic parameters and plasma indicators of metabolic status. The lymphocyte response to phytohemagglutinin and levels of serum immunoglobulin-G increased, while levels of milk immunoglobulin-G decreased during the period from calving to 8 weeks after calving. Increased milk somatic cell counts were associated with a significant decrease in the lymphocyte responses to mitogens.

Bulk milk samples were collected from 256 dairy herds from 4 districts in Southern Norway: Gudbrandsdal (n = 35), Bergen (n = 15), Hedmark (n = 117) and Hardanger (n = 89). A total of 6–8 samples were collected in each herd during the indoor season. Data concerning reproductive performance, milk production and nutrition were registered for a period of 15 months starting 7 months before the first milk sample was collected. The urea levels obtained during the sampling period were averaged generating a mean urea level of each herd. The mean urea levels were related to reproductive performance. The urea levels within districts showed only minor variations during the sampling period. The mean urea level in Hardanger was significantly higher (p < 0.05) than in the other distrikts. This district also had the lowest fertility. Significant correlations between mean urea level and fertility were observed only in Hedmark. Fertility status (FS), an integrated index, and number of inseminations per animal inseminated were significantly affected by the interaction between mean urea level and district. Herds in Hardanger with cows treated for ovarian cysts had a significantly higher mean urea level (p = 0.02) than herds with non-treated cows. The results appear to support the notion that a negative relationship may exist between bulk milk urea levels and reproductive performance. The great district variations observed indicate, however, that the practical value of such measurements is limited.

Blood samples were collected every 2 h during a 24 h period from 6 cows of one herd and 10 cows of another herd. In a third herd 9 cows were sampled every 2 h from 6 a.m. to 8 p.m. Concentrations of total bile acids, acetoacetate, glucose and free fatty acids were determined in blood plasma. A marked difference in individual bile acid concentrations and patterns of diurnal variation was found. For most cows the highest bile acid values were observed between 2 and 6 a.m. (overall mean (+/- SD) at 6 a.m.: 104 +/- 84 mumol/l, range: 20-307 mumol/l). Fourteen cows with a bile acid value greater than 90 mumol/l at 6 a.m. ("high BA") were characterized as a group by showing a pronounced decrease in the mean bile acid concentration after morning feeding. In the group of 11 cows with a 6 a.m. bile acid value less than 90 mumol/l ("low BA") the time of day did not contribute significantly to the bile acid variation. For the "high BA" group a nearly synchronous variation between the mean values of the 3 feeding dependent parameters (acetoacetate, glucose and free fatty acids) and the mean values of bile acids was found. The within animal coefficients of correlation between bile acids and the feeding dependent parameters were significantly higher in the "high BA" group than in the "low BA" group. No direct connection was found between bile acid levels and the quantity of concentrates fed or the individual milk yield.

Relationships among body condition score (BCS), milk constituents, and resumption of postpartum luteal function were studied in 162 lactations of first- and second-parity Norwegian dairy cows. Milk components included acetone, lactose, fat, protein, urea, and ratios of fat to protein and fat to lactose. Milk progesterone concentrations were used to determine intervals from calving to first luteal response (> 5 ng/ml). Intervals to first luteal response were divided into categories of early (≤ 24 d) or late (> 24 d) responses. Higher BCS were observed during wk 4 and 5 postpartum among both first- and second-parity cows with early compared with delayed luteal responses. Second-parity cows with early onset of luteal function also had higher BCS from wk 6 through 12, whereas first-parity cows with early onset of luteal function had higher BCS from wk 13 through 15. Higher acetone levels from wk 2 through 4 postpartum were associated with late luteal response in second-parity cows. Greater milk lactose content during wk 1, 2, 3, 6, 7, and 8 postpartum and higher fat fractions during wk 4 postpartum were related to early luteal response in second-parity cows. Relationships between milk constituents and onset of luteal function were less evident and occurred later postpartum among first-parity cows than among second-parity cows. Measures of weekly milk composites obtained during the early postpartum period and BCS were closely associated with postpartum resumption of luteal function. Acetone and lactose values in milk from the first 4 wk postpartum predicted postpartum luteal function in second-parity cows at a sensitivity of 0.84 and specificity of 0.86.

Embryonic and larval zebrafish (Danio rerio) behavior is commonly used to identify neurotoxic compounds. Here, we investigated whether sub-lethal exposures to the common solvents dimethyl sulfoxide (DMSO, 0.01-1%) and methanol (MeOH, 0.01-1%), or the anti-fungal agent methylene blue (MB, 0.0001-0.0005%), can influence larval behavior in a simple light/dark paradigm conducted in 96-well plates. In addition, we tested whether the media volume within the behavioral arena or the zebrafish strain, AB wild type, AB Tübingen (AB/TU), or Tüpfel long-fin (TL), could also influence larval behavior. Following the single exposures, we co-exposed larvae to DMSO and either MB or two other compounds with known behavioral effects in larval zebrafish, flutamide and perfluorooctanesulfonic acid (PFOS). We found ≥0.55% DMSO and 0.0005% MB significantly affected larval behavior, but there was no effect of MeOH. Similarly, TL showed less movement compared to AB and AB/TU strains, whereas lower media volumes also significantly reduced larval movement. However, all strains responded similarly to DMSO and MB. In the co-exposure studies, we found either additive or interaction effects between DMSO and either MB, flutamide, or PFOS, depending on the behavioral endpoint measured. In addition, media volume had no effect on the DMSO concentration response curve, but again we observed additive effects on behavior. In conclusion, methodology can lead to alterations in baseline locomotor activity and compounds can have additive or interaction effects on behavioral endpoints. However, we found no evidence that strain effects should be a concern when deciding on solvents for a simple light/dark behavioral test in larval zebrafish.

Background The EU 6th Framework Program (FP)-funded Health and Environment Network (HENVINET) aimed to support informed policy making by facilitating the availability of relevant knowledge on different environmental health issues. An approach was developed by which scientific agreement, disagreement, and knowledge gaps could be efficiently identified, and expert advice prepared in a way that is usable for policy makers. There were two aims of the project: 1) to apply the tool to a relevant issue; the potential health impacts of the widely used plasticizers, phthalates, and 2) to evaluate the method and the tool by asking both scientific experts and the target audience, namely policy makers and stakeholders, for their opinions. Methods The tool consisted of an expert consultation in several steps on the issue of phthalates in environmental health. A diagram depicting the cause-effect chain, from the production and use of phthalates to potential health impacts, was prepared based on existing reviews. This was used as a basis for an online questionnaire, through which experts in the field were consulted. The results of this first round of consultation laid the foundation for a new questionnaire answered by an expert panel that, subsequently, also discussed approaches and results in a workshop. One major task of the expert panel was to pinpoint priorities from the cause-effect chain according to their impact on the extent of potential health risks and their relevance for reducing uncertainty. The results were condensed into a policy brief that was sent to policy makers and stakeholders for their evaluation. Results The experts agreed about the substantial knowledge gaps within the field of phthalates. The top three priorities for further research and policy action were: 1) intrauterine exposure, 2) reproductive toxicology, and 3) exposure from medical devices. Although not all relevant information from the cause-effect chain is known for phthalates, most experts thought that there are enough indications to justify a precautionary approach and to restrict their general use. Although some of the experts expressed some scepticism about such a tool, most felt that important issues were highlighted. Conclusions The approach used was an efficient way at summarising priority knowledge gaps as a starting point for health risk assessment of compounds, based on their relevance for the risk assessment outcome. We conclude that this approach is useful for supporting policy makers with state-of-the-art scientific knowledge weighed by experts. The method can assist future evidence-based policy making.

Aim: Apply a recently developed expert elicitation procedure to evaluate the state of the current knowledge of the two brominated flame retardants (BFRs) most commonly used today; decabromo-diphenyl ether (decaBDE) and hexabromocyclododecane (HBCD) and their potential impact on human health in order to support policy considerations. This expert elicitation was organized by the HENVINET (Health and Environment Network) Consortium. Method: The HENVINET expert elicitation procedure that was used in the evaluations of decaBDE and HBCD is a rapid assessment tool aimed at highlighting areas of agreement and areas of disagreement on knowledge-related key issues for environment and health policy decision making. Results: The outcome of the expert consultation on BFRs was concrete expert advice for policy makers with specific priorities for further action made clear for both stakeholders and policy makers. The experts were not in agreement whether or not the knowledge currently available on decaBDE or HBCD is sufficient to justify policy actions, but most experts considered that enough data already exists to support a ban or restriction on the use of these compounds. All experts agreed on the necessity of more research on the compounds. Priority issues for further research were, among others: more studies on the extent of human exposure to the compounds. more studies on the fate and concentration in the human body of the compounds.