Excreted steroids in primate feces over the menstrual cycle and pregnancy.

Department of Obstetrics and Gynecology, University of Washington Seattle, Seattle, Washington, United States
Biology of Reproduction (Impact Factor: 3.45). 12/1988; 39(4):862-72. DOI: 10.1095/biolreprod39.4.862
Source: PubMed

ABSTRACT Techniques were established for the extraction and measurement of 17 beta-estradiol (E2) and progestins (P4) from feces of Old World primates. Studies were conducted to show the sensitivity of these measures, means of preserving fecal samples in the field, effects of urinary contamination, and means to eliminate these effects. Our results show that excreted steroid measures can be used to distinguish between mid-follicular and luteal phases in the menstrual cycle, and to identify pregnancy by Day 20 of gestation; the steroid measures can also be used to identify ovulatory levels of E2 and to establish the length of the menstrual cycle. Urine was shown to contaminate the fecal sample and to confound the estimate of steroid levels in feces; prolonged storage (less than 6 h) was shown to change the steroid estimate. Both urinary contamination and storage-dependent changes were eliminated by the addition of ethanol to the sample. Preliminary results also suggest that effects of dietary fiber on steroid hormone levels are minimal when controlled quantitatively by adjusting for water content of the fecal sample. We conclude that these measurements of excreted steroids provide a valid, noninvasive measure of physiological state of the hypothalamic-pituitary-ovarian axis among free-ranging animals in the field.

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    ABSTRACT: By extracting steroid metabolites from feces, researchers can track endocrine activity noninvasively in free-ranging animals. Sample preservation is a critical component of such methods because steroid metabolites rapidly decompose. Here, we describe a method for preservation, field extraction, and radioimmunoassay of steroid metabolites (estradiol, progesterone, glucocorticoids, and testosterone) from the feces of wild female baboons (Papio spp.). This method is a modification of that developed by Stavisky [Socioendocrinology: noninvasive techniques for monitoring reproductive function in captive and free-ranging primates. PhD, Emory University, 1994.], which employs reversed-phase octadecylsilane cartridges to extract steroids from feces. In addition to providing physiological validation for this method, we examine variation in steroid concentration across different (1) collection times (morning vs. afternoon), (2) methanol extraction treatments (homogenized vs. hand-mixed), and (3) solid-phase extraction times (2 vs. 10 h after collection). We then examine the stability of sample storage at ambient and subzero temperatures to determine whether storage time significantly alters steroid concentrations. Our results show that hormone concentrations do not differ between morning and afternoon samples, homogenization yields significantly higher fecal steroid concentrations, and fecal steroids are stable in a methanol/acetone solution for up to 10 h. When stored at ambient temperatures, only glucocorticoid metabolites had some degradation over a period of up to 40 days. However, when stored at −10 °C, no significant steroid changes were observed for up to 400 days. This method is particularly suited for behavioral research because it permits delays between sample collection and sample processing, thus allowing behavioral observations to continue.
    Physiology & Behavior 05/2004; DOI:10.1016/S0031-9384(04)00103-9 · 3.03 Impact Factor
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    ABSTRACT: Arctic marine mammals are facing increasing levels of many anthropogenic stressors. Novel tools are needed for assessment of stress physiology and potential impacts of these stressors on health, reproduction and survival. We have investigated baleen as a possible novel tissue type for retrospective assessment of stress and reproductive hormones. We found that pulverized baleen powder from bowhead whales (Balaena mysticetus) contained immunoreactive cortisol and progesterone that were detectable with commercially available enzyme immunoassay kits. Both assays passed parallelism and accuracy validations using baleen extracts. We analysed cortisol and progesterone at the base of the baleen plate (most recently grown baleen) from 16 bowhead whales of both sexes. For a subset of 11 whales, we also analysed older baleen from 10, 20 and 30 cm distal to the base of the baleen plate. Immunoreactive cortisol and progesterone were detectable in all baleen samples tested. In base samples, females had significantly higher concentrations of cortisol and progesterone compared with males. Cortisol concentrations in older baleen (10, 20 and 30 cm locations) were significantly lower than at the base and did not exhibit correlations with age-class or sex. Progesterone concentrations were significantly higher in females than in males at all baleen locations tested and were significantly higher in pregnant females than in non-pregnant females. Four of five mature females showed dramatic variation in progesterone concentrations at different locations along the baleen plate that may be indicative of previous pregnancies or luteal phases. In contrast, all males and all immature females had uniformly low progesterone. Baleen hormone analysis is a novel approach that, with further methodological development, may be useful for deter-mining individual longitudinal profiles of reproductive cycles and stress responses. RM (2014) Baleen hormones: a novel tool for retrospective assessment of stress and reproduction in bowhead whales (Balaena mysticetus). Conserv Physiol 2: doi:10.1093/conphys/cou030.
    08/2014; 2(1):cou30. DOI:10.1093/conphys/cou030

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