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Elephant Progesterone Assay Protocol


African Elephant Progesterone Assay Submission Protocol


The Indianapolis Zoo offers solid-phase radioimmunoassay for the direct, quantitative measurement of serum progesterone in African Elephants.



The progesterone assay may be run on serum or plasma (collected in either EDTA or heparinized blood tubes).  While the assay manufacturer found virtually no measurable difference in hormone detection between serum and plasma, most institutions submit serum for testing.  Manufacturer testing has also shown results to be unaffected by bilirubinemia or hemolysis.  If possible, it is recommended that the serum or plasma be separated from red cells within 30 to 40 minutes of collection.


NOTE:  Some types of centrifuges tend to heat the blood tubes as they spin.  It has not been determined if this heating of the serum or plasma damages the proteins of the different hormones, but it could possibly affect the measured results.  In cases where this is a concern, some institutions have chosen to let serum tubes sit overnight (refrigerated) before spinning.  This allows the blood to fully clot, red cells to settle out from the serum and thereby requiring only three or four minutes to centrifuge the next day.  While this technique may be helpful for various types of hormone assays, it may render the serum unsuitable for standard chemistry analysis.  Regardless of the processing technique used; if serial hormone levels are to be monitored, maintaining a consistent protocol is of primary importance.



Serum or plasma should be stored and shipped in polypropalene cryovials which can withstand ultra-low freezing and are less likely to break during shipping.  Glass tubes or vials are not suitable for test submission.  Prior to submission, specimens may be kept refrigerated at 2-8 degrees celsius for up to seven days, frozen at -20 degrees C for two months or at -80 degrees C in an ultra-low freezer indefinitely.  Attempts should be made to avoid repeated thawing and freezing of the sample.  For test submission, each vial needs to be labeled with the animal's ID and date collected.  A list should accompany the vials itemizing the number of vials shipped, animal ID number(s), and dates of collection.


All sample submissions need to be prearranged with the Indianapolis Zoo's Veterinary Technician prior to shipping.  Samples should be packed on standard ice packs and shipped for next morning delivery.  Shipping on dry ice is acceptable, but not necessary.


Please do not ship for Saturday or Sunday delivery.



Requested submission volume for progesterone assay - 1.0 ml per sample  (minimum testable volume - 0.3 ml)


The extra serum or plasma is requested for submission in the case that repeat analysis is necessary for verification.  Unused sample volumes will be kept for a period of two weeks after results have been reported to the submitting institution to allow for additional assay requests.  If no further analysis is requested, leftover samples will be disposed of.  No returns of samples will be possible.


Maximum turnaround time for assay results is 14 days.  Most results will be available 7 days after receipt of samples.  Results will be reported by fax with a hard copy being mailed (if desired).  Phone results may be arranged by special request.


All collection information and hormone assay results remain the sole property of the submitting institution and will not be used by or shared with any other facility (including the Indianapolis Zoo) without the expressed written consent of the submitting institution.


 Ship to:  The blood serum samples should be sent to the Indianapolis Zoo per directions of the Indianapolis Zoo veterinary technicians.    Samples will be assayed by the Indiana University Analye Core Lab


Contact Person:  Ellen Clark, C.V.T, Jennifer Niederlander, C.V.T.        

                             Phone:  (317) 630-5163


COST: The current cost of  running the progesterone assay is approximately $5.25 per  sample.


PAYMENT: Indiana University will invoice the zoo sending the samples. Payment instructions will be included in the invoice.



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