Applications and Techniques for Fecal Biomarker Testing and Analysis
- Endocrine Analysis: Fecal hormone analysis has become an increasingly popular non-invasive method to monitor endocrine function in mammalian and avian species1–4. For example, fecal glucocorticoid levels represent a more meaningful and accurate assessment of stress than point-in-time blood glucocorticoid concentrations. Analytes of interest are easily measured using common laboratory techniques including chromatography and immunoassay (ELISA).
- Metabolic Profiling: Qualitative and quantitative analysis of low-molecular weight metabolites have important diagnostic and therapeutic applications. Metabolomics has become increasingly popular because of the ability to measure multiple metabolites directly from complex biologic systems and is also used to facilitate understanding of the causes of disease processes. Metabolites are typically analyzed via spectroscopy, chromatography, and immunoassay (ELISA).
- Genetic Analysis: Basic PCR techniques are used to assess paternity, diet, population genetics, community relationships, and spatial organization in various species. Mitochondrial DNA can used to determine species, and nuclear DNA is used differentiate individuals within a population, including gender determinations.
- Nutritional Analysis: Fecal samples may be used to determine dietary preferences and assess nutrient content in various species. Remains of prey in feces and predator DNA may be analyzed to study carnivore diets. Ecologic niche and differences in diet between and within species can be studied through stable isotope and elemental analysis.
- Disease Diagnosis: Noninvasively collected biologic samples are often analyzed to learn about disease transmission. Wildlife diseases may serve as indicators of pollution or degradation of ecosystems. Movements of wildlife prompted by environmental changes such as global warming may lead to movement of pathogens into susceptible host populations.
Utility of Fecal Biomarker Testing and Analysis in Wildlife Conservation Studies
Wildlife conservation scientists and anthropologists from the American Zoological Association (AZA), The International Society for Wildlife Endocrinology (ISWE), and the Society for Integrated & Comparative Biology (SICB) routinely use non-invasive protocols in their conservation and human-animal impact studies. Though a long-standing partnership with ISWE, Arbor Assays provides the specialized tools and resources for carrying out their important work.
Advantages of Using Fecal Samples for Biomarker Testing and Analysis
- Easy Collection: While blood sampling requires highly-trained personnel, fecal sample procurement can be done by anyone. Sample collection may be possible in the absence of the target animal. Efficient and cost-effective sampling make fecal biomarker testing suitable for long-term studies and repeated sampling.
- Simple transport and storage: Fecal samples require less manipulation than blood to procure, ship and store.
- Broad Applicability: Fecal samples contain a wide array of key physiologic and pathologic biomarkers.
Wildlife Studies Using Fecal Hormone Monitoring Include:
Brown, J. L., et al. (2020). Commonalities in management and husbandry factors important for health and welfare of captive elephants in North America and Thailand. Animals, 10(4), 737.
Brown, J. L., et al. (2019). Individual and environmental risk factors associated with fecal glucocorticoid metabolite concentrations in zoo-housed Asian and African elephants. PLoS ONE, 14(9)
Brown, J. L. & Graham, L. H. (1994). Comparative aspects of steroid hormone metabolism and ovarian activity in felids, measured non-invasively in feces. Biology of Reproduction, 51(4), 776-786.
Auer, K. E., et al. (2020). Measurement of fecal testosterone metabolites in mice: Replacement of invasive techniques. Animals 2020, 10(1), 165.
Pritchard, C. E., et al. (2020). Glucocorticoid and triiodothyronine concentrations do not correlate with behavior in vicuñas (Vicugna vicugna). General and Comparative Endocrinology, 286.
Handling Fecal Samples for Biomarker Analysis
- Processing and Storage: Samples should be collected into clean tubes and stored at 4°C short-term or frozen for long-term storage.
- Drying and Extraction: Samples are dried using passive drying or gentle heating (≤60ºC) prior to extraction or for long term storage. Biomarkers (e.g., hormones, metabolites & small molecules) must be extracted from fecal samples before analysis. Suitable protocols are described in detail here.
- Biomarker Analysis: Examples of fecal validated assays follow below.
Key Stress, Reproduction and Metabolic Biomarkers Found in Feces
- Cortisol and Corticosterone: Glucocorticoid hormones are used to study stress-related physiological phenomena in a wide range of animals. The primary function of glucocorticoids is to facilitate glucose release in response to an acute stressor, but have many pleiotropic effects related to the fight-or-flight response. Most large mammals secrete cortisol while corticosterone is primary found in birds and small mammals.
- Aldosterone: Aldosterone is one of the major mineralocorticoids in most species, its primary role being physiological regulation of electrolytes (sodium retention and potassium retention) and fluids (blood pressure) and coping of stress.
- Progesterone: Progesterone provides an important index of ovulation and pregnancy in mammals. It is also a crucial metabolic intermediate in the production of other endogenous steroids, including the sex hormones and the corticosteroids, and plays an important role in brain function as a neurosteroid.
- Testosterone: Testosterone and other androgen metabolites (e.g. Androstenedione) are used to assess reproductive state, sex, behavior, and social rank in many mammals.
- Thyroid Hormones: Thyroid hormone regulates metabolism, blood pressure, body temperature regulation and nutritional physiology Thyroid hormones (both thyroxine, T4 and triiodothyronine, T3) are particularly responsive to nutritional fluctuations, and are useful to characterize nutritional stress in metabolic studies.
We Can Measure Your Fecal Samples!
Send your samples to us and take advantage of our Sample Testing Services.
Our Fecal-Validated Assays
Arbor Assays ELISA kits have been validated for use with saliva for a variety of relevant biomarkers. Our assays are a cost-effective, high-quality alternative to other assay platforms and are available immediately.
|Product Name||Catalog #|
|17-Hydroxyprogesterone ELISA Kit||K053-H1 17-Hydroxyprogesterone ELISA Kit, 1 Plate|
|K053-H5 17-Hydroxyprogesterone ELISA Kit, 5 Plate|
|Aldosterone ELISA Kit||K052-H1 Aldosterone ELISA Kit, 1 Plate|
|K052-H5 Aldosterone ELISA Kit, 5 Plate|
|Allopregnanolone ELISA Kit||K061-H1 Allopregnanolone ELISA Kit, 1 Plate|
|K061-H5 Allopregnanolone ELISA Kit, 5 Plate|
|BCA Protein Dual Range Colorimetric Detection Kit||K041-H1 BCA Dual Range Protein Detection Kit, 2 Plate|
|Corticosterone Chemiluminescent ELISA Kit||K014-C1 Corticosterone Chemiluminescent ELISA Kit, 1 Plate|
|K014-C5 Corticosterone Chemiluminescent ELISA Kit, 5 Plate|
|Corticosterone ELISA Kit||K014-H1 Corticosterone ELISA Kit, 1 Plate|
|K014-H5 Corticosterone ELISA Kit, 5 Plate|
|Cortisol ELISA Kit||K003-H1 Cortisol ELISA Kit, 1 Strip-Well Plate|
|K003-H5 Cortisol ELISA Kit, 5 Strip-Well Plate|
|K003-H1W Cortisol ELISA Kit, 1 Solid Plate|
|K003-H5W Cortisol ELISA Kit, 5 Solid Plate|
|Cortisone Chemiluminescent ELISA Kit||K017-C1 Cortisone Chemiluminescent ELISA Kit, 1 Plate|
|K017-C5 Cortisone Chemiluminescent ELISA Kit, 5 Plate|
|Cortisone ELISA Kit||K017-H1 Cortisone ELISA Kit, 1 Plate|
|K017-H5 Cortisone ELISA Kit, 5 Plate|
|DHEA-S ELISA Kit||K054-H1 DHEA-S ELISA Kit, 1 Plate|
|K054-H5 DHEA-S ELISA Kit, 5 Plate|
|DNA Damage ELISA Kit||K059-H1 DNA Damage ELISA Kit, 1 Plate|
|K059-H5 DNA Damage ELISA Kit, 5 Plate|
|Epiandrosterone ELISA Kit||K063-H1 Epiandrosterone ELISA Kit, 1 Plate|
|K063-H5 Epiandrosterone ELISA Kit, 5 Plate|
|Estradiol ELISA Kit||K030-H1 Estradiol ELISA Kit, 1 Plate|
|K030-H5 Estradiol ELISA Kit, 5 Plate|
|Estriol ELISA Kit||K064-H1 Estriol ELISA Kit, 1 Plate|
|K064-H5 Estriol ELISA Kit, 5 Plate|
|Estrone ELISA Kit||K031-H1 Estrone ELISA Kit, 1 Plate|
|K031-H5 Estrone ELISA Kit, 5 Plate|
|Estrone-3-Glucuronide (E1G) ELISA Kit||K036-H1 Estrone-3-Glucuronide (E1G) ELISA Kit, 1 Plate|
|K036-H5 Estrone-3-Glucuronide (E1G) ELISA Kit, 5 Plate|
|Estrone-3-Sulfate (E1S) ELISA Kit||K038-H1 Estrone-3-Sulfate (E1S) ELISA Kit, 1 Plate|
|K038-H5 Estrone-3-Sulfate (E1S) ELISA Kit, 5 Plate|
|FRAP™ (Ferric Reducing Antioxidant Power) Detection Kit||K043-H1 FRAP™ (Ferric Reducing Antioxidant Power) Detection Kit, 2-plate|
|Levonorgestrel (LNG) ELISA Kit||K058-H1 Levonorgestrel (LNG) ELISA Kit, 1 Plate|
|K058-H5 Levonorgestrel (LNG) ELISA Kit, 5 Plate|
|Myeloperoxidase (MPO) Human ELISA Kit||K060-H1 Myeloperoxidase (MPO) ELISA Kit, 1 Plate|
|PGFM ELISA Kit||K022-H1 PGFM ELISA Kit, 1 Plate|
|K022-H5 PGFM ELISA Kit, 5 Plate|
|Pregnanediol-3-Glucuronide (PDG) ELISA Kit||K037-H1 Pregnanediol-3-Glucuronide (PDG) ELISA Kit, 1 Plate|
|K037-H5 Pregnanediol-3-Glucuronide (PDG) ELISA Kit, 5 Plate|
|Progesterone ELISA Kit||K025-H1 Progesterone ELISA Kit, 1 Plate|
|K025-H5 Progesterone ELISA Kit, 5 Plate|
|Progesterone Metabolites ELISA Kit||K068-H1 Progesterone Metabolites ELISA Kit, 1 Plate|
|K068-H5 Progesterone Metabolites ELISA Kit, 5 Plates|
|Testosterone ELISA Kit||K032-H1 Testosterone ELISA Kit, 1 Plate|
|K032-H5 Testosterone ELISA Kit, 5 Plate|
|Thyroxine (T4) ELISA Kit||K050-H1 Thyroxine (T4) ELISA Kit, 1 Plate|
|K050-H5 Thyroxine (T4) ELISA Kit, 5 Plate|
|Triiodothyronine (T3) ELISA Kit||K056-H1 Triiodothyronine (T3) ELISA Kit, 1 Plate|
|K056-H5 Triiodothyronine (T3) ELISA Kit, 5 Plate|
|Urea Nitrogen (BUN) Colorimetric Detection Kit||K024-H1 Urea Nitrogen (BUN) Colorimetric Detection Kit, 2 Plate|
|K024-H5 Urea Nitrogen (BUN) Colorimetric Detection Kit, 10 Plate|
- Fecal Extraction and Sample Handling Protocols
- Step-by-step instruction for steroid extraction from fecal samples
- Sample Processing Protocol Videos
- Tutorials on how to extract samples, and analyze preparations using immunoassay techniques
- Sample Testing Services
- Arbor Assays sample testing services lab will extract samples and measure fecal biomarkers for you. Send your samples to us and take advantage of our Sample Testing Services!
A History of Fecal Biomarker Testing
Fecal biomarker testing using assay methods has a long history. Evaluation of feces has been used to monitor the health and well-being of humans and animals for centuries. In modern times, fecal samples are often used to screen for disease, and to assess metabolic and nutritional status of humans and animal models5,6. Observations of the physical properties of fecal samples can indicate underlying pathology, and molecular biomarkers found in fecal samples are often used to screen for disease, and to monitor lineage and species identity.
- Brown, J. (2019). Individual and environmental risk factors associated with fecal glucocorticoid metabolite concentrations in zoo-housed Asian and African elephants. PLOS ONE 14(9), e0217326.
- Fauteux, D., et al. (2017). Assessing stress in arctic lemmings: Fecal metabolites reflect free corticosterone levels. Physiological and Biochemical Zoology, 90(3), 370-382.
- Lasley, B.L. & Kirkpatrick, J. F. (1991). Monitoring ovarian function in captive and free-ranging wildlife by means of urinary and fecal steroids. Journal of Zoo and Wildlife Medicine 22(1), 23-31
- Hunt, K.E. & Wasser, S. K. (2003). Effect of long‐term preservation methods on fecal glucocorticoid concentrations of grizzly bear and African elephant. Physiological and Biochemical Zoology 76(6), 918-928.
- Foell, D., et al. (2009). Monitoring disease activity by stool analyses: From occult blood to molecular markers of intestinal inflammation and damage. Gut, 58(6), 859-868.
- Desai, D., et al. (2007). Review article: Biological activity markers in inflammatory bowel disease. Alimentary Pharmacology & Therapeutics, 25(3), 247-255.