Ecreted 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: 3.32). 12/1988; 39(4):862-72. DOI: 10.1095/biolreprod39.4.862
Source: PubMed


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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    • "Several methods have been used, including storage of faecal material in alcohol (Wasser et al., 1988; Lynch et al., 2003; Galama et al., 2004), the drying of either raw faecal material (Brockman and Whitten, 1996; Galama et al., 2004; Gould et al., 2005) or faecal extract in the field (Santymire and Armstrong, 2010) and storage of faecal extract on filter paper (Shideler et al., 1995); however, each method has potential constraints. "
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    ABSTRACT: Non-invasive hormone analysis is a vital tool in assessing an animal's adrenal and reproductive status, which can be beneficial to in situ and ex situ conservation. However, it can be difficult to employ these techniques when monitoring in situ populations away from controlled laboratory conditions, when electricity is not readily available. A practical method for processing faecal samples in the field, which enables samples to be extracted soon after defaecation and stored in field conditions for prolonged periods prior to hormone analysis, is therefore warranted. This study describes the development of an optimal field extraction method, which includes hand-shaking faecal material in 90% methanol, before loading this extract in a 40% solvent onto HyperSep™ C8 solid-phase extraction cartridges, stored at ambient temperatures. This method was successfully validated for measurement of adrenal and reproductive hormone metabolites in faeces of male and female black rhinoceros (Diceros bicornis) and was rigorously tested in controlled laboratory and simulated field conditions. All the hormones tested demonstrated between 83 and 94% and between 42 and 89% recovery of synthetic and endogenous hormone metabolites, respectively, with high precision of replication. Furthermore, results obtained following the developed optimal field extraction method were highly correlated with the control laboratory method. Cartridges can be stored at ambient (cool, dry or warm, humid) conditions for periods of up to 6 months without degradation, before re-extraction of hormone metabolites for analysis by enzyme immunoassay. The described method has great potential to be applied to monitor faecal reproductive and adrenal hormone metabolites in a wide variety of species and allows samples to be stored in the field for up to 6 months prior to analysis. This provides the opportunity to investigate hormone relationships within in situ populations, where equipment and facilities may previously have been limiting.
    Conservation Physiology 02/2014; 2(1):cot037. DOI:10.1093/conphys/cot037
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    • "Although evidence of effects of phytoestrogen consumption similar to those of laboratory studies are difficult to obtain in field studies, indirect measures can provide evidence of changes in either the HPG or HPA axes, indicating disruption of reproductive capabilities. These hormonal effects can be examined noninvasively using the measurement of excreted estradiol and cortisol metabolites in the feces (Heistermann et al., 1993; Touma and Palme, 2005; Wasser et al., 1988; Whitten et al., 1998; Ziegler and Wittwer, 2005). Thus, fecal samples were collected immediately upon defecation from ten known adult males (out of 13 in the group). "
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    ABSTRACT: Numerous studies have examined the effects of anthropogenic endocrine disrupting compounds; however, very little is known about the effects of naturally occurring plant-produced estrogenic compounds (i.e., phytoestrogens) on vertebrates. To examine the seasonal pattern of phytoestrogen consumption and its relationship to hormone levels (407 fecal samples analyzed for estradiol and cortisol) and social behavior (aggression, mating, and grooming) in a primate, we conducted an 11-month field study of red colobus (Procolobus rufomitratus) in Kibale National Park, Uganda. The percent of diet from estrogenic plants averaged 10.7% (n=45weeks; range: 0.7-32.4%). Red colobus fed more heavily on estrogenic Millettia dura young leaves during weeks of higher rainfall, and the consumption of this estrogenic item was positively correlated to both their fecal estradiol and cortisol levels. Social behaviors were related to estradiol and cortisol levels, as well as the consumption of estrogenic plants and rainfall. The more the red colobus consumed estrogenic plants the higher their rates of aggression and copulation and the lower their time spent grooming. Our results suggest that the consumption of estrogenic plants has important implications for primate health and fitness through interactions with the endocrine system and changes in hormone levels and social behaviors.
    Hormones and Behavior 09/2012; 62(5). DOI:10.1016/j.yhbeh.2012.09.005 · 4.63 Impact Factor
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    • "pigra) have been extracted, and the efficacy of the extraction of steroid metabolites radio marked with tritium have been evaluated (Van Belle et al., 2009). However, due to factors which may or may not modify the measurement of efficiency of hormone extraction in feces from the species (Wasser et al., 1988; Brown et al., 1994; Ziegler et al., 1996; Terio et al., 2002; Palme, 2005), we consider that is necessary to add radio marked hormones to the fecal samples in order to measure the quantity of said tracer at the time of implementing a new method of extraction in such a way that the recovered percentage of the tracer makes it possible to evaluate the recovery efficacy in the biological sample. The objectives of this study were: (1) to determine whether the physical condition of the sample (moist or lyophilized feces) and the type of solvent may influence the extraction efficacy of the hormones 125 Iestradiol and 125 I-progesterone added to the stratum (feces); (2) to compare the protocol for extraction in this study, agitation vs. ebullition, in order to obtain and value the percentage of increased extraction efficiency of the radio marked hormone added to moist and lyophilized feces of howler monkeys; and (3) to validate the hormonal extraction technique in the lyophilized feces of A. pigra. "
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    ABSTRACT: Several fecal steroid extraction techniques have been developed to measure the ovary function in different species of mammals. However, regardless of the method of extraction and the sample type chosen, it has been observed that they can yield results with different percentages of recuperation. The objective of this study was to determine whether the type of substratum, solvent and extraction method used have any influence on the extraction efficiency in the feces of Alouatta pigra (black howler monkey). For this purpose we used two methods: agitation and ebullition. With each method, we utilized moist and lyophilized feces. The validation of radioimmunoassay method was accurate and precise for quantify estradiol and progesterone in lyophilized feces of A. pigra. To both of which ethanol and methanol, absolute and at 80%, were added, besides the hormones (125)I-Estradiol and (125)I-Progesterone. The extraction efficiency for (125)I-Estradiol was from 87.72 ± 3.97 to 41.24 ± 2.67%, and for (125)I-Progesterone from 71.15 ± 4.24 to 42.30 ± 1.19% when we used the agitation method. Whereas with the ebullition method, the extraction efficiency for (125)I-Estradiol ranged from 86.89 ± 2.66 to 71.68 ± 3.02% and for (125)I-Progesterone from 98.31 ± 1.26 to 85.40 ± 1.98%. Due to the differences found in these assays, which depend on the method used, the type of feces employed and the type of solvent added to them, we recommend the ebullition method and the lyophilized feces of A. pigra for extracting the hormones, since in moist feces there may exist variables which might interfere in the quantification of (125)I-Estradiol and (125)I-Progesterone.
    Frontiers in Physiology 12/2011; 2:97. DOI:10.3389/fphys.2011.00097 · 3.53 Impact Factor
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