Development of Artificial Insemination in the

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Development of Artificial Insemination in the
Greater One-Horned Rhinoceros (Rhinoceros
unicornis)
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Greater One-Horned Rhinoceros SSP Population

• Captive breeding program
• genetic management

• Natural breeding / AI
• ovulation near time of insemination

• Reproductive Physiology
• limited information

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Objectives

• Characterize the follicular, endocrine and
  behavioral
• dynamics of the GOH rhinoceros estrous cycle
• Determine if administration of exogenous
  hormone
• (GnRH) will induce ovulation
• Develop successful semen collection and
• cryopreservation techniques
• Develop a transcervical AI procedure

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Rectal Ultrasound -follicle selection/growth -ovul
ation
Hormone Analysis -estrogen -progesterone -LH
Behavior -whistling -urine spraying -increased
activity -decresed appetite
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Estrous Cycles 3 patterns
40 3.16 days
37.8 1.95 days
48 8.58 days
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single estrus
Ovulatory Estrous Cycles
multi-estrus
Preovulatory Follicle gt10cm
Ovulation
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Anovulatory Estrous Cycles
Hemorrhagic Follicle
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Can Exogenous Hormones be used to Induce
Ovulation?

• Hormone Cystorelin (GnRH) 500ug administered
  via intramuscular dart injection
• During 2 separate estrous cycles, Cystorelin was
  given on day of behavioral estrus, maximal
  follicle size (gt10cm) and peak estrogen
  concentrations
• Conclusion Cystorelin stimulates LH release that
  is measured in urine 2 hours post injection but
  does not cause ovulation

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Comprehensive Understanding

• Unique follicular growth dynamics
• Occurrence of anovulatory cycles
• Ovulatory estrous cycles of different length
• (36-61 days)
• Established biomarkers of ovarian function
• LH surge during estrus
• Progesterone rise prior to ovulation
• Ovulation 36-48 hr after behavioral estrus
• Exogenous GnRH stimulates LH release but does
  not
• advance ovulation in multi-estrus cycles and
  does not
• cause ovulation in a follicle destined to be
• anovulatory

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Sperm Collection
Electroejaculation
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Creative Solutions!
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• Semen cryopreservation study
• Sperm banking of genetically valuable male GOH
  rhinos

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Artificial Insemination (AI)
Goal transcervical semen deposition
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AI under standing sedation?
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Post-Ovulatory AI Attempts (n4)
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0
-1
-2
-3
-4
-5
-6
1
2
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Pre-Ovulatory AI Attempts (n2)
Uterine semen deposition
Custom Designed Tools uterine semen deposition
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• Comprehensive understanding of the estrous cycle
  of
• female GOH rhinoceros through the use of
• ultrasound, hormones and behavior

• Established technique to collect and
  cryopreserve semen
• from genetically valuable male GOH rhinoceros

• Nonsurgical inseminations accurately timed
  within
• 24 hours of ovulation

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Acknowledgements
White Oak Conservation Center

• Cincinnati Zoo Rhino Keepers and Veterinary Staff
• CREW Staff

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