Oral Presentation at Rhodes College 2011

Kimber Jones, Andrew Kouba, and Erin Willis

Rhodes College Department of Biology

Memphis Zoo

Faculty Sponsor: Dr. Sarah Boyle

Snow Leopards (Panthera unica) 4,080-6,590 in wild (IUCN, 2010)Population decreased 20% in past 2 generations (IUCN, 2010)

Little known about reproductive physiology (Brown, 1994)Most felids reproduce poorly in captivity (Brown, 2006)

GOAL:To utilize a non-invasive technique to characterize and monitor

reproductive steroid hormone levels to build a comprehensive data set, draw comparisons between animals, and aid in conservation of the species as a whole

Fecal samples were collected every other day from leopards in zoos across the U.S.

Samples were dried at 60°C for 48-72 hours and ground into a fine powder

Testosterone was extracted from 0.25g of ground samples using a methanol extraction protocol optimized by our lab (Graham et al., 2010)

ELISA (Enzyme-linked immunosorbent assay ) quantified testosterone levels by utilizing a broad scale antibody for testosterone

Antibody R156/R157

Step 1-Add Antibody

Step 2- Add hormone of unknown amount

Step 3- Add labeled conjugate of known amount

Step 4- Add enzyme to react with conjugate

•Roosevelt Park Zoo•Young Adult (3 y.o)

minotdailynews.com

• Total Average- 171.06 ng/g

•Fall- 187.25

•Winter- 170.64

•Summer- 121.87

Roosevelt Park Zoo Male #2698

Date

Tes

tost

eron

e (n

g/g

fece

s)

0

100

200

300

400

500

•Memphis Zoo•Sub-Adult (2 y.o.)

Memphis Zoo Male # 22967

Date

8/2/

2010

8/16

/201

0

8/30

/201

0

9/13

/201

0

9/27

/201

0

10/11

/201

0

10/2

5/20

10

11/8

/201

0

11/2

2/20

10

12/6

/201

0

12/2

0/20

10

1/3/

2011

1/17

/201

1

1/31

/201

1

Tes

tost

eron

e (n

g/g

fece

s)

0

100

200

300

400

500

•Total Average- 110.22 ng/g

•Before 11/10- 79.27

•After 11/10- 141.18

•Brookfield Zoo•Adult (8 y.o)

Filip Stojković

Brookfield Zoo Male #2443

Date7/

20/2

009

8/31

/200

9

10/12

/2009

11/23

/2009

1/4/2

010

2/15

/201

0

3/29

/201

0

Tes

tost

eron

e (n

g/g

fece

s)

0

100

200

300

400

500

•Total Average- 92.52 ng/g

•Fall- 87.18

•Winter- 104.98

•Spring- 103.01

•Summer- 82.77

Established a non-invasive technique for monitoring hormone levels

Seasonal, regional, or age based trends Testosterone levels of below 100 ng/g

may be an indicator that the male is pre-pubertal

Testosterone levels were lowest in the summer months

Hormone levels can be an indicator or reproductive health

Testosterone levels of below 100 ng/g in an adult leopard could also be an indicator of infertility

These are preliminary results, but in the future we hope to compare our values to samples from wild animals

Timeoffun.com

Terry Whitaker

Topz10.com

•Research Team at the Memphis Zoo, Chelsea Peters, Dr. Sarah Boyle, Dr. Jon Davis, Nick Brydon, Stephen Leavelle, and Jordan Perchick, Emma Lendt, Michael Drake, Allison Graham, and James Artzer.

•CAN, Memphis Zoological Society, Roosevelt Park Zoo, Chicago Zoological Park, and C. Munro of UC Davis.

Brown J, Wasser S, Wildt D, Graham L. Comparative Aspects of Steroid Hormone Metabolism and Ovarian Activity in Felids, Measured Noninvasively in Feces. Biology of Reproduction. 1994;51:776-786.

Brown, J. Comparative endocrinology of domestic and nondomestic felids. Theriogenology. 2006;66:25-36.

Graham AW, Kouba AJ, Willis EL. 2010. Development of non-invasive reproductive monitoring techniques for endangered snow leopards and Amur leopards. Proc. of the annual meeting of the Society of Integrative and Comparative Biology.

Jackson, R., Mallon, D., McCarthy, T., Chundaway, R.A. & Habib, B. 2008. Panthera uncia.   In: IUCN 2010. IUCN Red List of Threatened Species. Version 2010.4.   <www.iucnredlist.org>. Downloaded on 14 December 2010.