TITLE: Sample Collection: Primate Feces for DNA/RNA
AUTHORS: Alicia Rich

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">All of the methods and protocols for this project have been reviewed and approved by the Otterbein University Institutional Animal Care and Use Committee, Otterbein University's Environmental Health and Safety Officer, the AZA Prosimian Taxonomic Advisory Group, and the individual collaborating AZA Institutions.</div><div class = "text-block">The pygmy slow loris (Nycticebus pygmaeus) captive population is declining at a rate that jeopardizes their future in the Species Survival Plan (SSP).  Experts agree that diet is somehow compromising reproduction, and this study will investigate dietary estrogenicity. Many plants produce chemicals known as phytoestrogens that activate estrogen receptors. The phytoestrogen-fertility relationship is well-documented in livestock, rhinoceroses, cheetahs, and pandas, and this study will characterize the pattern in slow lorises. We will adapt existing biomedical methods to investigate dietary estrogens and reproductive outcomes in captive pygmy slow lorises. Gene expression profiles will serve as proxies for in vivo estrogen receptor activity, and our heterologous cell-based receptor reporter assay will quantify in vitro estrogen receptor activation. Generalized linear mixed-effects models will integrate these parameters with reproductive outcomes to test the effect of gene expression profiles and diet estrogenicity on fertility. This long-term project will ultimately provide one of the first deep physiological examinations of phytoestrogen’s proximate and ultimate influences on primate reproduction.</div></div>

[STEPS]
?. [Sample Collection]
Gather fresh feces (
If it is not possible to gather fresh feces indirectly from a reliably identified individual, then follow the rectal swab directions in the direct sample collection procedure.
Always wear a facial covering and nitrile examination gloves while collecting samples.
?. [Sample Collection]
Label one of the sterile tubes provided (pre-filled with Zymo® DNA/RNA Shield) with your initials, the date (YYYYMMDD), the time (HH:MM:SS), animal code (PSL for pygmy slow loris), sample type (feces), and animal name or ID.  Open the tube but take care not to touch the inside of the tube, lid, or the outside of the attached scoop to your glove or any other surface. Use the scoop to gather ~0.5-1 ml of feces and deposit it into the tube.
?. [Sample Collection]
Make sure there is just enough   to submerge the entire sample in the tube. If there is too much sample for one tube and you have an extra, then you can fill a secondary tube and label it as such. We will use matched tubes like this for validation in the lab. Close the lid as tightly as possible and use parafilm (included) to reinforce the outside of the closure.
?. [Sample Storage]
Place this tube inside of a clearly labeled sample bag (also provided), and put that into the envelope with postage, already addressed to Alicia Rich at Otterbein University. Please drop this envelope in the mail as soon as possible (within 24-48 hours of sample collection).
Remove your gloves inside out and safely dispose of them in the proper area. Always wash your hands thoroughly after collecting a specimen.