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Invasive and Non-invasive Methods of Hormone Monitoring in Captive Wildlife

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Invasive and Non-invasive Methods of Hormone Monitoring in Captive Wildlife Bruce W. Christensen, DVM Reproductive Endocrinology Cyclicity Gender, age, sociosexual ... – PowerPoint PPT presentation

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Title: Invasive and Non-invasive Methods of Hormone Monitoring in Captive Wildlife


1
Invasive and Non-invasive Methods of Hormone
Monitoring in Captive Wildlife
  • Bruce W. Christensen, DVM

2
Why do we care about hormones?
1. Reproductive Endocrinology
2. Stress (cortisol)
3
Reproductive Endocrinology
  • Cyclicity
  • Gender, age, sociosexual, seasonality effects
  • Spermatogenesis/ovulation timing
  • Ovulation type (spontaneous v. induced)
  • Clues to overcome infertility
  • Timing of AI
  • Pregnancy diagnosis
  • Parturition date prediction

4
Stress
  • Impaired immune system
  • Increased disease
  • Decreased androgen production
  • Decreased spermatogenesis
  • Decreased libido
  • Disrupts reproductive cycles

5
How do we monitor hormone concentrations?
  • Invasive
  • venipuncture
  • Non-invasive
  • Feces, urine, saliva

6
Monitoring hormones Invasive
7
Monitoring Hormones Non-invasive
NON-INVASIVE MONITORING OF STRESS IN NON-PREGNANT
AND PREGNANT OKAPI (Okapia johnstoni)
Bruce Christensen, Marcie Oliva, Karin Andra,
Linda Penfold
8
Fecal/urinary hormone monitoring
  • Advantages
  • Sample collection
  • Low stress
  • Less noise
  • Disadvantages
  • Cross-reaction of metabolites
  • Labor intensive
  • Protein hormones
  • Cannot measure diurnal variation

9
Direct ELISA
10
Solid Phase
11
Add sample (antigen)
12
Adsorption
13
Wash
14
Add antibody/enzyme complex
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
15
Incubate
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
16
Wash
Y
Y
Y
Y
Y
Y
17
Add color reagent
Y
Y
Y
Y
Y
Y
18
Add color reagent
Y
Y
Y
Y
Y
Y
19
Read
Y
Y
Y
Y
Y
Y
20
Competitive ELISA
21
Solid Phase
22
Add Antibody/adsorption
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
23
Wash
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
24
Add Sample
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
25
Add competitive marker
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
26
Incubate
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
27
Wash
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
28
Add color reagent
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
29
Incubate
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
30
Read
Y
Y
Y
Y
Y
Y
Y
Y
Y
Y
31
Endangered Species
1980
32
Cheetah-specific research
  • Reproductive endocrinology
  • Timing and type of ovulation
  • Ovulation induction
  • Sperm collection and processing

33
Cheetah AI
  • 10/24 AI successful litters (41)
  • 3 from African cheetahs

Wildt et al. 1997, Howard unpublished
Photos by Jessie Cohen, National Zoological Park
34
Rhinoceros Endocrinology
?
35
Acyclic flatliners
Schwarzenberger et al. 1998 Brown et al 2001
36
Rhino Ovarian Cycle
  • Fecal P4 metabolites
  • 4 m. postpartum inactivity
  • 4-7 m. cyclicity
  • Long-day breeders
  • Rainy season

Garnier et al 2002
37
Sumatran rhinoceros
  • Serum P4 LH
  • Fecal P4 metabolites
  • Induced ovulators

Roth et al. 2001
http//www.cincyzoo.org.html
38
Indian rhinoceros
  • Ovarian cycle
  • Follicular phase
  • Luteal phase
  • Pregnancy

Schwarzenberger et al. 2000
http//www.miamimetrozoo.com
http//www.giconline.org/home.htm
39
Pregnancy and cyclicity
  • Fecal urinary metabolites
  • Hindl et al 1992
  • Schwarzenberger et al 1993, 1998
  • Garnier et al 1998
  • Patton et al 1998
  • Stoops et al (in prep)

http//www.giconline.org/home.htm
40
Male White Rhinos
  • Fecal androgen metabolites
  • Territorial males seasonal
  • Positive effect of female presence

Kretzschmar et al 2003
41
STRESS
  • Fecal cortisol metabolites
  • Black and white rhinoceros
  • Captivity

Turner et al 2002
42
Translocation
  • Wild black and white rhinos
  • Fecal cortisol and P4 metabolites
  • Current management decisions

Czekala et al, in preparation
43
Stress in Captivity
  • Captive black rhinoceros
  • Fecal cortisol metabolites
  • Exposure to public
  • Exposure to conspecifics

Carlstead and Brown, in preparation
http//www.honoluluzoo.org/black_rhinoceros.htm
44
  • gt 100,000 circa 1900
  • lt 2,300 circa 1992

www.rhinos-irf.org
45
Black Rhinoceros
  • Currently 3,610
  • 185 captive
  • Limited success captive breeding
  • 2 births in last 6 months

Kibo and her calf, Jello, photo courtesy of
Miami Metrozoo
46
  • 50-200 low
  • Mostly now in S. Africa.

www.rhinos-irf.org
47
  • lt 200
  • gt2500

www.rhinos-irf.org
48
Rhino Repro Studies
  • Studies in female reproductive physiology
  • Ultrasound of ovarian cyclicity fetal growth
  • Progesterone, estrogen, LH cyclicity
  • Pregnancy diagnosis
  • Uterine biopsy
  • Male reproductive physiology???
  • Testosterone concentrations in free-ranging white
    rhinoceros

Photo courtesy of Cleveland Metroparks Zoo
Photo courtesy of Kansas City Zoo
49
Hypothesis
  • Serum testosterone in male rhinoceros will show
    variation with
  • Age
  • Time of year
  • Stress
  • Female presence

Masshaki, photo courtesy of San Francisco Zoo
50
Methods
  • 363 serum samples from 18 male black rhinoceros

51
Methods
  • Testosterone concentration by EIA
  • Grouped by
  • Age
  • Season
  • Proximity to and of females
  • Stats
  • Mann-U-Whitney test, Wilcoxon sign rank test
  • Student paired t test, linear regression
  • P 0.05 were considered significant

52
Results Age

P lt 0.05
P gt 0.05
53
Results Age

54
Results Season
P gt 0.05
55
Results Social
P gt 0.05
Adjacent to female
Directly with female
56
Results Social

n 4
n 4
P lt 0.05
57
Discussion Age
Photos courtesy of Brookfield Zoo and Fossil Rim
Wildlife Center
58
Discussion Season
www.rhinos-irf.org
59
Busch Gardens, Tampa Florida
60
Discussion Female Proximity
Photo courtesy of Virginia Zoo
61
Discussion Female Number
Photos courtesy of Miami Metrozoo and White Oak
Conservation Center
62
Conclusions
  • Captive, male black rhinoceros do not show a
    fluctuation in their serum testosterone
    concentration according to time of year.
  • Testosterone concentrations rise during the first
    eight years of life, and then no longer show a
    correlation with age.
  • Serum concentrations were significantly higher in
    adults ( 8 y) compared to juveniles.
  • Close versus immediate proximity to females was
    not important to male serum testosterone
    concentrations.
  • Number of females or the introduction of a new
    female may play a role in serum testosterone
    concentration in captivity.

63
White Rhino Age
64
White Rhino Age
65
White Rhino Female Presence
www.hoglezoo.org
66
Acknowledgements
  • Albuquerque Biological Park
  • Baltimore Zoo
  • Brookfield Zoo
  • Buffalo Zoo
  • Busch Gardens
  • Caldwell Zoo
  • Chicago Zoological Park
  • Cleveland Metroparks Zoo
  • Dallas Zoo
  • Denver Zoological Gardens
  • Disneys Animal Kingdom
  • Fort Worth Zoological Park
  • Fossil Rim Wildlife Center
  • Gladys Porter Zoo
  • Henry Vilas Zoo
  • Houston Zoo
  • Indianapolis Zoo
  • Knoxville Zoological Garden
  • Kansas City Zoo
  • UF CVM Resident Research Grant
  • White Oak Conservation Park

Jello photo courtesy of Miami Metrozoo
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