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Biomedical subjects

E Möstl

Publications and source records attributed to E Möstl.

At least 19 recordsLinked to original sources
PubMed2005

Ligula intestinalis infection as a potential source of bias in the bioindication of endocrine disruption in the European chub Leuciscus cephalus.

European chub Leuciscus cephalus collected from five localities in the lowland and subalpine regions of Austria were analysed for oestrogenic effects of endocrine-disrupting chemicals and the presence of the plerocercoid of the tapeworm Ligula intestinalis. Of 1494 chub analysed, only seven (six males, one female) were found to be infected with single, but large plerocercoids up to 15 cm in length. Ligula-infected fish showed comparatively immature gonads, as demonstrated by the gonadosomatic index and gamete developmental stages. Plasma levels of the egg precursor protein vitellogenin also showed concentrations ranging below the detection limit. The present results indicate that chub infected with L. intestinalis and exposed to exogenous oestrogenic compounds can result in reduced gonadal maturation and produce false oestrogen-positive diagnoses in male fish. For plasma vitellogenin levels, L. intestinalis infections can result in false oestrogen-negative diagnoses in male and female fish.

Animals↗
PubMed2005

Stress hormones in mammals and birds: comparative aspects regarding metabolism, excretion, and noninvasive measurement in fecal samples.

A multitude of endocrine mechanisms are involved in coping with challenges. Front-line hormones to overcome stressful situations are glucocorticoids (GCs) and catecholamines (CAs). These hormones are usually determined in plasma samples as parameters of adrenal activity and thus of disturbance. GCs (and CAs) are extensively metabolized and excreted afterwards. Therefore, the concentration of GCs (or their metabolites) can be measured in various body fluids or excreta. Above all, fecal samples offer the advantages of easy collection and a feedback-free sampling procedure. However, large differences exist among species regarding the route and time course of excretion, as well as the types of metabolites formed. Based on information gained from radiometabolism studies (reviewed in this paper), we recently developed and successfully validated different enzyme immunoassays that enable the noninvasive measurement of groups of cortisol or corticosterone metabolites in animal feces. The determination of these metabolites in fecal samples can be used as a powerful tool to monitor GC production in various species of domestic, wildlife, and laboratory animals.

Adrenal Glands↗
PubMed2005

Corticosterone in chicken eggs.

Birds are discussed as models for prenatal stress. In this study, several experiments were conducted to gain basic knowledge of if, how, and when maternal adrenocortical activity is reflected by corticosterone concentrations in the egg. Radiolabeled corticosterone was administered to 10 laying hens to investigate the uptake into as well as the distribution within the eggs. The yolk was dissected in concentric layers and analyzed. Less than 1% of the administered radioactivity entered the egg but was, however, not evenly distributed. On the day after injection, highest radioactivity (Bq/g) was detected in the albumen and the outmost layer, whereas concentration peaked 4-7 days later in the inner layers. In two other experiments, increased plasma levels of corticosterone were induced by injection of adrenocorticotropic hormone (ACTH) or feeding of corticosterone. Again, yolk disks were cut in layers and analyzed with a corticosterone enzyme immunoassay. No effect of the ACTH administration was detected, whereas feeding of corticosterone resulted in increased immunoreactive corticosterone concentrations in the yolk. Straight-phase high-performance liquid chromatographic (HPLC) separations were also performed to characterize immunoreactive steroids in the yolk. Two close-eluting peaks at the approximate elution position of corticosterone could be observed after the feeding experiment, whereas in untreated control eggs they were absent. It was concluded that transfer from plasma to egg is low for corticosterone and that further investigations concerning the transport mechanisms and the exact nature of yolk steroids are necessary.

Adrenocorticotropic Hormone↗
PubMed2004

Measurement of corticosterone metabolites in chicken droppings.

(1) A non-invasive technique for stress assessment is needed. Therefore, an enzyme immunoassay (EIA) for measurement of glucocorticoid metabolites in chicken droppings was established and validated. (2) Radiolabelled corticosterone was administered intravenously to detect the time course of excreted metabolites. The metabolites were then characterised by chemical and immunological methods to find a suitable antibody. (3) Reversed-phase high-performance liquid chromatography (RP-HPLC) separations of the peak concentration samples revealed that corticosterone was extensively metabolised, mainly to more polar substances. (4) HPLC fractions were tested in several EIAs for glucocorticoid metabolites, where the highest quantities were detected by a newly established cortisone assay, measuring metabolites with a 3,11-dione structure. (5) The biological relevance of this cortisone EIA was confirmed by stimulation of adrenocortical activity by adrenocorticotropic hormone (ACTH). (6) With this newly developed EIA it should be possible to measure adrenocortical activity non-invasively in chickens and other galliformes, thus providing a tool for a variety of research fields, such as poultry production, ethology and behavioural ecology.

Adrenocorticotropic Hormone↗
PubMed2004

Coping capacity of dairy cows during the change from conventional to automatic milking.

In conventional milking systems, dairy cows are driven to the milking stall twice or thrice daily, whereas in automatic milking systems (AMS), the cows enter the milking stall voluntarily. In this study, noninvasive methods were used to analyze the physiological reaction of 17 cows toward the changeover from conventional to automatic milking. Milk yield and composition were analyzed. Heart rate was recorded continuously, and feces was sampled twice daily to determine cortisol metabolites (11, 17-dioxoandrostanes) for a period of 2 wk. During the first visit to the AMS (without milking), heart rate was elevated compared with parlor milking by 35 +/- 3 beats per minute (bpm) above basal heart rate (P < 0.05). Heart rate during the first milking in AMS (eighth visit) was already similar to the heart rate previously measured during milking in the parlor (18.1 +/- 2.2 bpm above basal level). Concentration of fecal cortisol metabolites was unchanged during the change-over compared with parlor milking. A decreased (P < 0.05) milk yield of 68 +/- 7% relative to previous parlor yield during the first AMS milking indicated a disturbance of milk ejection in most cows. Individual yields ranged from 8 to 96% of the previous parlor yield. To examine the relationship between adrenal cortex sensitivity and the coping process, an ACTH challenge experiment was performed after the changeover period. Cows that released more cortisol after ACTH injection, indicating a higher adrenal cortex sensitivity, had a less enhanced heart rate and a near normal milk ejection during the first AMS milkings (P < 0.05). In conclusion, the reactions toward the changeover to AMS milking varied widely within cows. Adaption to the AMS was easier in animals with a higher adrenal cortex sensitivity to ACTH.

Adaptation, Physiological↗
PubMed2003

Distribution and origin of steroid hormones in the yolk of Japanese quail eggs (Coturnix coturnix japonica).

The yolk of avian eggs contains steroid hormones, which may influence the development and behaviour of hatched birds. The aim of the present study was to investigate the concentration as well as the distribution of various gonadal steroids in the yolk spheres of quail eggs. Steroid concentrations of dissected yolk layers were analysed after alcoholic extraction using enzyme immunoassays (EIAs) for progesterone, androstenedione and testosterone. To monitor the uptake of testosterone into the yolk, radioactive testosterone was injected i.m. into six female quails. The radioactivity of yolk layers of subsequently laid eggs was measured by liquid scintillation counting. Progesterone concentrations were highest in the outer layer (median: 2265 nmol/kg). Androstenedione (median: 453 nmol/kg), as the major androgen, and testosterone (median: 99 nmol/kg) reached their highest concentrations in interior layers, whereas in the centre the concentration of all three hormones was low. No significant variation of steroid levels in yolk layers of subsequently laid eggs was found. The highest radioactivity was detected in the outer yolk layer in those eggs laid 1 day after injection and in subsequently laid eggs was measured nearer to the centre. These results indicated local origin of the steroid hormones especially because of the result that only 0.1% of the radioactivity entered the yolk. We conclude that steroid concentrations in the yolk layers reflected progesterone and androgen production of the cells of the follicular wall at the time.

Androstenedione↗
PubMed2003

Fecal corticosteroids in a territorial bird selected for different personalities: daily rhythm and the response to social stress.

In this study we tested the hypothesis that in a passerine bird (great tit, Parus major) individuals differing for coping strategies differ in the magnitude of the adrenocortical response to social stress as well. Furthermore, we aimed at characterizing daily rhythms in corticosteroid release before and after social stress. We used 16 males from either of two lines bidirectionally selected for different coping strategies (fast and slow explorers). Social stress was induced by confrontation with an aggressive resident male. Corticosteroid metabolites were analyzed in feces collected at 90-min intervals from 900 to 1630 h on a baseline day, on the day of the social conflict, and on the following day. In both days and in both lines levels varied with time of day in a robust rhythm with a peak in the first sample of the morning and a trough at the end of the light phase. This rhythm correlates with activity (perch hopping). An overall increase in levels relative to baseline day was observed between 30 and 140 min after the challenge. Birds of the less aggressive and more cautious line (slow explorers) showed a trend for a higher response compared to birds of the more aggressive and bolder line (fast explorers), which showed almost no response. On the day after the challenge the birds of the slow line exhibited significantly reduced corticosteroid secretion, probably due to an increased negative feedback. The results provide evidence for a physiological basis of different coping strategies in birds, emerging in response to social stress and with a pattern similar to that in other vertebrates.

Adaptation, Psychological↗
PubMed2003

Glucocorticoid metabolites inhibit the metabolism of androstenedione in red blood cells of ruminants.

The present work aimed to confirm that erythrocytes of ruminants, in general, are capable of converting 17-oxo to 17-hydroxysteroids. Special attention was given to 11-oxoaetiocholanolone (a cortisol metabolite) and its possible interaction with androstenedione as substrates of 17-hydroxysteroid dehydrogenases (17-OH SDH). Blood samples were taken from cattle, sheep and goats (n = 3). Aliquots (100 or 300 microl) of washed red blood cell (RBC) suspensions were incubated in triplicates with Ringer's/glucose solution (1 ml) containing either androstenedione (10 ng) or 11-oxoaetiocholanolone (100 ng) or a mixture of 10 ng of each. Incubations were performed on a shaker at 38 degrees C for 10, 20, 40 or 80 min, respectively. After centrifugation the supernatants were stored at -24 degrees C until analysis. Concentrations of added steroids were measured with enzyme-immunoassays to monitor their decrease. The 17-OH SDH activity of RBC was highest in cattle followed by goats and sheep, and 11-oxoaetiocholanolone was a better substrate than androstenedione. Concentrations of the latter decreased more pronounced, if incubated alone. High performance liquid chromatography separations of the metabolites of 17-oxosteroids revealed the presence of both, a 17beta- and 17alpha-hydroxylated product formed by erythrocytes of sheep and goats, but only the latter in cattle. The results demonstrated that 11-oxoaetiocholanolone was also a substrate of RBC 17-OH SDH and inhibited the metabolism of androstenedione. Therefore, in ruminants, there might be an interaction between cortisol metabolites and gonadal steroids on the level of peripheral steroid metabolism.

Androstenedione↗
PubMed2003

Determination of autoantibodies to thyroglobulin, thyroxine and triiodothyronine in canine serum.

Enzyme immunoassays (EIAs) for the determination of autoanti-bodies (AA) to thyroid antigens in canine serum were developed. Streptavidin (SA) was immobilized as capture molecule on microtitreplates (MTP). Thyroglobulin (Tg) purified from canine thyroids and the thyroid hormones thyroxine and triiodothyronine (T3 and T4) were conjugated to biotin labelling reagents and attached to the MTP over the SA-biotin bridge. Bound AA were detected with anti-dog-immunoglobulin G (IgG) labelled with horseradish peroxidase. Serum samples from dogs which were allotted to four groups were analysed: A (n = 31), biochemical evidence of hypothyroidism; B (n = 76), clinical signs of hypothyroidism; C (n = 47), euthyroid with non-thyroidal disease; D (n = 186), clinically healthy. The validity of the assays was tested with two different methods. After thiophilic absorption chromatography of positive sera, a positive reaction in the EIA was only detected in those fractions which coeluted with the canine IgG standard. Furthermore, the positive reaction was blocked by the addition of the corresponding antigen. In 55% of the hypothyroid dogs AA to Tg and/or T3 and T4, respectively, were found (up to a titre of 1 : 1600). In group B 34% of the dogs were diagnosed positive, but the titre was lower (up to 1 : 400). In the groups C and D the number of dogs with AA and their titre was significantly lower. Two different methods for distinguishing positive and negative test results were compared in order to increase the specificity of the tests without decreasing the sensitivity. The EIAs are precise and based on high agreement with previous reported assays able to discriminate dogs with thyroiditis from healthy ones. These assays represent a good alternative to the isotope assays generally used for the analysis of AA to T4 and T3.

Animals↗
PubMed2002

Hormones as indicators of stress.

Animal welfare is of increasing importance and absence of chronic stress is one of its prerequisites. During stress, various endocrine responses are involved to improve the fitness of the individual. The front-line hormones to overcome stressful situations are the glucocorticoids and catecholamines. These hormones are determined as a parameter of adrenal activity and thus of disturbance. The concentration of glucocorticoids (or their metabolites) can be measured in various body fluids or excreta. Above all, fecal samples offer the advantage that they can be easily collected and this procedure is feedback free. Recently, enzyme immunoassays (EIA) have been developed and successfully tested, to enable the measurement of groups of cortisol metabolites in animal feces. The determination of these metabolites in fecal samples is a practical method to monitor glucocorticoid production.

Animal Welfare↗
PubMed2002

Measurement of cortisol metabolites in faeces of ruminants.

Twenty-one metabolites were detected in faecal samples collected after infusion of (14C)cortisol into the jugular vein of sheep. Using high-performance liquid chromatography/radiometric analysis plus mass spectrometry. One group of metabolites had molecular weights of between 302 and 308, and another group of 350, which indicates that the substances have a C19O3 or a C21O4 structure. Therefore, an enzyme immunoassay against 5beta-androstane-3alpha-ol-11,17-dione-17-CMO:BSA was established. Faecal samples were collected from 10 cows immediately after transport and then during a course in which non-invasive diagnostic procedures were being taught (course 1). For comparison, faeces were sampled from another 5 cows that were being used for teaching invasive procedures (course 2). Six cows from a university farm served as controls. In the animals used in course 1, the highest concentrations of cortisol metabolites were measured immediately after transport to the university (median value: 2.2 micromol/kg faeces). During the first 5 days at the university, the concentrations decreased to 0.52 micromol/kg (median) and remained at this level during the rest of the course. The median concentration in the samples that were taken during coursc 2 (collected about 2 months after transport) was 0.48 micromol/kg. There was no significant difference in the excretion of cortisol metabolites between these cows and the controls. We conclude from these data that, using the enzyme immunoassay against 5beta-androstane-3alpha-ol-11,17-dione-17-CMO, we were able to detect transport/novel environment stress but not the potential disturbance that cows experience during diagnostic procedures.

Animals↗
PubMed2002

Immunohistochemical distribution of oestrogen and progesterone receptors and tissue concentrations of oestrogens in the cervix of non-pregnant cows.

An immunohistochemical study of the expression of oestrogen (ER) and progesterone receptors (PR) in different regions along the longitudinal and vertical axes of the cervix of non-pregnant cows was performed. Animals were separated into two groups depending on the presence or absence of a functional corpus luteum in their ovaries, as indicated by blood progesterone concentrations. The high progesterone group (HP4) had serum progesterone concentrations > 2.0 ng mL(-1) (n = 6) and the low progesterone group (LP4) had serum progesterone concentrations < or = 0.5 ng mL(-1) (n = 4). Significantly higher concentrations of oestrogen were found in the cervical tissue of animals in the LP4 group than those in the HP4 group (473 +/- 53 v.149 +/- 46 pg g(-1) wet weight; P < 0.01). Furthermore, there was a significant effect of tissue layer (epithelium to deep stroma) on the number of ER (P < 0.01) and PR (P < 0.05) immunoreactive nuclei per 1000 cells. For both ER and PR the proportion of cells expressing the receptor increased from epithelium to subepithelial stroma (P < 0.01) and from subepithelium to deep stroma (ER P < 0.05; PR P =0.061). When the number of receptor-positive cells were expressed per mm2 tissue, differences between the subepithelial stroma and the deep stroma became even more marked. In addition, the vaginal part of the cervix had significantly more (P < 0.01) ER and PR immunoreactive nuclei per 1000 cells than the uterine part, but these differences were no longer apparent when a correction was made for cell density. There was no relationship between progesterone status of the animals, nor local tissue oestrogen concentrations and ER or PR immunoreactivity in the cervix of these non-pregnant cows. Instead, a strong relationship between both longitudinal and vertical positioning of tissue in the cervix and expression of both receptor types was shown. In addition, a strong correlation between ER and PR expression in the subepithelial stroma (R = 0.85, P < 0.01) and the deep stroma (R = 0.83 P < 0.01) was evident. In conclusion, these results demonstrate that in studies of steroid hormone receptor expression in the cervix, careful description of sampling site and depth are necessary if the results are to be interpreted meaningfully.

Animals↗
PubMed2001

Social, state-dependent and environmental modulation of faecal corticosteroid levels in free-ranging female spotted hyenas.

Little is known about to what extent the sensitivity of the hypothalamic-pituitary-adrenal (HPA) axis may be state dependent and vary in the same species between environments. Here we tested whether the faecal corticosteroid concentrations of matrilineal adult female spotted hyenas are influenced by social and reproductive status in adjacent ecosystems and whether they vary between periods with and without social stress. Females in the Serengeti National Park frequently become socially subordinate intruders in other hyena territories by undertaking long-distance foraging trips to migratory herds, whereas in the Ngorongoro Crater they usually forage inside their own small territories on resident prey. The faecal corticosteroid concentrations in Serengeti females were significantly higher than in Ngorongoro females. Energy expenditure by lactation is exceptionally high in spotted hyenas and this may be reflected in their corticosteroid levels. The faecal corticosteroid levels in both populations were higher in lactating than in non-lactating females. During periods of social stability, faecal corticosteroid concentrations increased in non-lactating females but not in lactating females as social status declined. Lactating Serengeti females had significantly higher faecal corticosteroid concentrations during periods with acute severe social stress than during periods without, indicating that the HPA axis is sensitive to social stimuli even in lactating females. So far few studies have used non-invasive monitoring methods for assessing social stress in freeranging animals. This study demonstrates for the first time, to the authors' knowledge, that corticosteroid concentrations may differ between periods with and without social stress for a free-ranging female mammal and that the modulating effect of social status may depend on reproductive status.

Adrenal Cortex Hormones↗
PubMed2001

Faecal oestrogens and progesterone metabolites in mares of different breeds during the last trimester of pregnancy.

Non-invasive pregnancy diagnosis in mares by measuring faecal oestrogens has been performed over years with great accuracy. However, results have indicated breed-related differences in the amount of excreted steroids during late pregnancy. Therefore faecal samples were collected during the last 4 months of pregnancy of Thoroughbred (n = 10), New Forest pony (n = 9), Shetland pony (n = 10) and Iceland pony mares (n = 11). Concentrations of oestrogens, 20alpha-hydroxy- and 20-oxopregnanes were measured using enzyme immunoassays. Breed differences concerning both levels (though significant only in case of oestrogens) and time course of measured steroids were observed. There was a highly significant time effect (p < 0.00001) and an interaction between time and breeds (p < 0.02) for all steroids measured, suggesting that the time effect differs for different breeds. Oestrogen concentrations showed a decrease towards parturition, whereas in 20alpha-hydroxy- and 20-oxopregnane levels a pronounced increase was found 2 and I months, respectively, before parturition. A breed effect was only significant (p = 0.001) when comparing oestrogen concentrations and was mainly due to Iceland ponies, which had the lowest concentrations especially during the last 2 months of pregnancy. An almost significant (p = 0.06) breed effect was found for 20-oxopregnanes. In Iceland mares an additional increase in faecal pregnane content was already observed earlier, reaching maximum levels before the 60th day ante-partum (a.p.), followed by a decrease until the 30th day a.p. The ratio of 20-oxopregnanes to oestrogens in the samples was significantly higher (p < 0.006) in Iceland ponies in comparison with any other breed throughout all months before parturition. The breed differences observed in the amounts of oestrogens and/or progestagens present during late pregnancy may demonstrate micro-evolutionary changes in the endocrine system of a species.

Animals↗
PubMed2001

[Effect of vaccination on fecal cortisol metabolites in cats and dogs].

The aim of this study was to apply a cortisol metabolite determination in the faeces of cats and dogs for monitoring disturbances. In this experiment faeces from every spontaneous defecation of 10 cats and 10 dogs (5 of each sex) were collected starting from one day before until two days after the yearly vaccination. Concentrations of 11,17-dioxoandrostanes (cat) and cortisol equivalents (dog) were determined by enzyme immunoassay (EIA). Faecal cortisol metabolites increased and reached peak concentrations (median: 412% in cats and 417% in dogs, respectively above baseline values) in one of the next two samples following the vaccination. This indicated an activation of the adrenocortex, the degree to which the different parts (physical and psychological components) of the whole vaccination procedure contributed to it was not evaluated. From this experiment we conclude that measuring cortisol metabolites in the faeces is a non-invasive method to monitor stressful conditions in cats and dogs and thus is a valuable tool for evaluating animal welfare.

Animals↗
PubMed2001

Oestrogen concentrations in the faeces of bitches.

Few data are available concerning excretion of oestrogen from bitches. During anoestrus, oestrogen concentrations are low in the blood of bitches and, therefore, a sensitive method for measuring concentrations of these hormones by analysis of faeces was developed. Daily faecal samples from bitches (n = 25) of various breeds were analysed (collected for 20 days after the start of pro-oestrous vulvar sanguinous discharge), as well as single samples from bitches in anoestrus (n = 30; samples from 16 of these bitches were also analysed during pro-oestrus and oestrus) and ovariohysterectomized bitches (n = 18). Samples (0.5 g mixed with 0.5 ml water) were extracted and after the samples were cleaned the unconjugated oestrogen content was measured, using a group specific enzyme immunoassay. Oestrone and oestradiol were identified as the predominant immunoreactive oestrogens by HPLC. In samples collected on the first day of pro-oestrous vulvar sanguinous discharge, oestrogen content was 9.3-231.1 nmol kg-1 faeces (median 45.5 nmol oestrogen kg-1 faeces). On day 8 of discharge, oestrogen content was 21.8-539.5 nmol kg-1 faeces (median 114.1 nmol oestrogen kg-1 faeces). Oestrogen content in bitches in anoestrus was 2.2-65.2 nmol kg-1 faeces (median 18.5 nmol oestrogen kg-1 faeces), whereas ovariohysterectomized bitches excreted lower amounts of oestrogens (range 0.7-18.1 nmol kg-1; median 4.4 nmol kg-1). The differences between oestrogen content in ovariohysterectomized bitches and bitches during anoestrus and oestrus were significant (P < 0.001). It is concluded that the method developed in the present study is suitable for monitoring the low production of oestrogens during anoestrus.

Anestrus↗
PubMed2000

Seasonal relationships between plasma and fecal testosterone in response to GnRH in domestic ganders.

In two groups (eight individuals each) of socially acquainted, outdoor-kept, domestic ganders (male Hungarian white: Anser domesticus), basal and GnRH-stimulated plasma testosterone (T) concentrations were compared with fecal testosterone metabolites (TM) in and between three seasons, spring peak of reproductive activity, summer photorefractoriness, and fall sexual reactivation. Plasma was sampled 90 min following the challenge and T was analyzed by radioimmunoassay following the GnRH challenge. Fecal TM were measured by enzyme immunoassay using two group-specific antibodies against 17beta-OH-androgens or a novel antibody against 17-oxo groups, which was found to react with major testosterone metabolites without prior hydrolytic deconjugation. Baseline plasma T and systemic levels were high in spring and fall but low in summer. Plasma T increases in response to GnRH were followed by significantly elevated fecal TM levels 2 to 6 h following the challenge in spring and fall. In fall, at high plasma T levels, fecal TM levels were disproportionally lower than in spring. Variability of TM levels was two to five times higher in feces than in plasma, which explains why correlations between individual plasma T and fecal TM levels generally remained nonsignificant. This points to a low-level short-term relationship between the excreted TM and the plasma T levels. However, the reliability of the method was demonstrated by standard inter- and intraassay variabilities and by a high correspondence between results obtained by the two assays. It is suggested that, with appropriate sample size, fecal TM reflects plasma T increase. However, fecal TM was more variable than the plasma T, and fecal TM responses to GnRH did not always parallel the plasma T response. In addition, seasonal changes in androgen excretion regimes must be taken into account.

Animals↗