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      Ovulation induction in anovulatory southern white rhinoceros ( Ceratotherium simum simum) without altrenogest

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          Abstract

          Lack of ovulation is common in captive southern white rhino females and contributes to poor reproductive success. We show that ovulation can be induced efficiently with exogenous hormones using a single treatment protocol. This information can lead to improved genetic management of captive populations acting as insurance against extinction.

          Abstract

          All species in the extant Rhinocerotidae family are experiencing increased threats in the wild, making captive populations essential genetic reservoirs for species survival. However, managed species face distinct challenges in captivity, resulting in populations that are not self-sustaining. Captive southern white rhinoceros ( Ceratotherium simum simum) have low reproductive rates and presumed acyclicity is common among females. Although many females fail to ovulate, follicle growth may occur and ovulation can be hormonally induced. Female southern white rhino ( n = 6), housed as a bachelorette group, were determined to be ovulatory ( n = 1) or anovulatory ( n = 5) by serial ultrasound and fecal progestagen analysis. When follicles reached pre-ovulatory size (~35 mm), females ( n = 4) were induced to ovulate in 11 trials with a GnRH analog (4.5 mg, SucroMate™) via single intramuscular injection. Nine trials resulted in ovulation (81.8%), all between 36 and 48 hours post-treatment. Ovulations were confirmed by progestagen elevation above baseline coincident with visualization of a corpus luteum (CL). Luteal phases were characterized as short (<50 days) or long (≥50 days). Between short and long cycles, only the number of days of progestagen above baseline was significantly different ( P < 0.05), while days with visible luteal structures was not significant ( P = 0.11). Both cycle types were observed following both spontaneous and induced ovulations. Furthermore, we showed that longer cycle lengths do not necessarily indicate early pregnancy loss as none of the females were bred or inseminated during the study. While anovulation is common in the southern white rhino captive population, ovulation induction can be achieved efficiently and predictably for use in conjunction with artificial insemination or to facilitate natural breeding. This information will lead to more efficient use of assisted reproductive technologies to overcome reproductive challenges in this species and to generate genetically healthy captive populations as a hedge against extinction.

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          Relationships between patterns of Fecal corticoid excretion and behavior, reproduction, and environmental factors in captive black (Diceros bicornis) and white (Ceratotherium simum) rhinoceros

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            Angiogenesis in developing follicle and corpus luteum.

            Angiogenesis is a process of vascular growth that is mainly limited to the reproductive system in healthy adult animals. The development of new blood vessels in the ovary is essential to guarantee the necessary supply of nutrients and hormones to promote follicular growth and corpus luteum formation. In developing follicles, the pre-existing endothelial cells that form the vascular network in the theca layer markedly develop in response to the stimulus of several growth factors, mainly produced by granulosa cells, such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). The angiogenic factors also promote vessel permeability, thus favouring the antrum formation and the events inducing follicle rupture. After ovulation, newly formed blood vessels cross the basement membrane between theca and granulosa layers and continue a rapid growth to sustain corpus luteum development and function. The length of luteal vascular growth varies in cycling and pregnant animals and among species; both angiogenesis and subsequent angioregression are finely regulated by systemic and local factors. The control of angiogenic development in the ovary could be a useful tool to improve animal reproductive performances.
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              Blood flow: a key regulatory component of corpus luteum function in the cow.

              Prostaglandin F2alpha (PGF2alpha) is the primary luteolysin in the cow. During the early luteal phase, the corpus luteum (CL) is resistant to the luteolytic effect of PGF2alpha. Once mature, the CL becomes responsive to PGF2alpha and undergoes luteal regression. These actions of PGF2alpha coincide with changes in luteal blood flow (BF): PGF2alpha has no effect on BF in the early CL, but acutely increases BF in the peripheral vasculature of the mature CL within 30 min of PGF2alpha injection. During spontaneous luteolysis, luteal BF increases on Days 17-18 of the estrous cycle, prior to any decrease in plasma progesterone (P). The increase in luteal BF is synchronous with an increase in plasma PGFM levels, suggesting that pulsatile release of PGF2alpha from uterus stimulates the increase in luteal BF. Serial biopsies of these CL showed that mRNA expression for endothelial nitric oxide synthase (eNOS) together with endothelin-1 (ET-1) and angiotensin converting enzyme (ACE) increases on Days 17-18 when the luteal BF is elevated. On Day 19 when plasma P level firstly decreases, eNOS mRNA returns to the basal level whereas ET-1 and ACE mRNA remains elevated. Cyclooxygenase-2 (COX-2) mRNA expression increases on Day 19. In support of these data, an in vivo microdialysis study revealed that luteal ET-1 and angiotensin II (Ang II) secretion increases and precedes PGF2alpha secretion during spontaneous luteolysis. In conclusion, we show for the first time that an acute increase of BF occurs in the peripheral vasculature of the mature CL together with increases in eNOS expression and ET-1 and Ang II secretion in the CL during the early stages of luteolysis in the cow. We propose that the increase in luteal BF may be induced by NO from large arterioles surrounding the CL, and simultaneously uterine or exogenous PGF2alpha directly increases ET-1 and Ang II secretion from endothelial cells of microcapillary vessels within the CL, thereby suppressing P secretion by luteal cells. Taken together, our results indicate that an acute increase in luteal BF occurs as a first step of luteolysis in response to PGF2alpha. Therefore, local BF plays a key role to initiate luteal regression in the cow.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                Conserv Physiol
                Conserv Physiol
                conphys
                Conservation Physiology
                Oxford University Press
                2051-1434
                2019
                24 June 2019
                24 June 2019
                : 7
                : 1
                : coz033
                Affiliations
                [1 ]Reproductive Sciences, San Diego Zoo Institute for Conservation Research, Escondido CA, USA
                [2 ]Lead Keeper, San Diego Zoo Safari Park, Rhino Rescue Center, Escondido CA, USA
                Author notes
                Corresponding author: 15600 San Pasqual Valley Road, Escondido CA 92027, USA. Tel: 760-291-5489. Email: ppennington@ 123456sandiegozoo.org
                Article
                coz033
                10.1093/conphys/coz033
                6589769
                31249689
                bbb9cde7-e8c1-44cc-83ed-7b2fddf6c985
                © The Author(s) 2019. Published by Oxford University Press and the Society for Experimental Biology.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 6 March 2019
                : 10 May 2019
                : 14 May 2019
                : 16 May 2019
                Page count
                Pages: 11
                Funding
                Funded by: San Diego Zoo
                Award ID: 55352
                Categories
                Research Article

                anovulation,assisted reproductive technology (art),gnrh,white rhinoceros

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