In Vitro Fertilization and Embryo Development in Vitro and in Vivo in the Tiger (Panthera Tigris)1

Оплодотворение in vitro и развитие эмбрионов in vitro и in vivo у тигра (Panthera tigris)
Ann M. Donoghue, L. A. Johnston, U. S. Seal, Doug L. Armstrong, Ronald Tilson, Patricia Wolf, K. Petrini, Lee G. Simmons, T.S. Gross, David E. Wildt
1990-11-01

electroejaculationembryo culturein vitro fertilizationoocyte maturationtiger
A study was conducted to evaluate the adaptability to the tiger of an in vitro fertilization/embryo culture system previously developed in the domestic cat. In Trial I (July 1989), 10 female tigers were treated with either 2,500 (n = 5) or 5,000 (n = 5) IU eCG i.m. and with 2,000 IU hCG i.m. 84 h later. In Trial II (January 1990), 6 females (5 of which were treated in Trial I) were given 2,500 IU eCG i.m. and 2,000 IU hCG i.m. 84 h later. Twenty-four to twenty-six hours after hCG treatment, all tigers were subjected to laparoscopy, and oocytes were aspirated transabdominally. On the basis of follicular development (follicles greater than or equal to 2 mm in diameter), all females responded to exogenous gonadotropins (range, 6-52 follicles/female). Follicle number and oocyte recovery rate were unaffected (p greater than 0.05) by eCG dose or time of year. A total of 456 oocytes were collected from 468 follicles (97.4% recovery; mean, 28.5 +/- 3.4 oocytes/female). Of these, 378 (82.9%) qualified as mature, 48 (10.5%) as immature, and 30 (6.6%) as degenerate. During Trial I, 8 electroejaculates were collected from 7 male tigers, and in Trial II, 3 semen samples were collected from 3 males. Motile sperm were recovered on each occasion; the overall mean (+/- SEM) ejaculate volume was 7.5 +/- 0.7 ml, the number of motile sperm/ejaculate was 105.9 +/- 20.6 x 10(6), and the percentage of structurally normal sperm/ejaculate was 81.4 +/- 2.0%. After swim-up processing, 0.05 x 10(6) motile sperm were co-cultured with 10 or fewer tiger oocytes in a humidified atmosphere (38 degrees C) of 5% CO2 in air. Of the 358 mature oocytes inseminated, 227 (63.4%) were fertilized. Oocytes from 2 females became contaminated in culture and, therefore, were excluded from embryo cleavage calculations. Of the remaining 195 fertilized oocytes, 187 (95.9%) cleaved to the two-cell stage. No parthenogenetic cleavage was observed in noninseminated control oocytes (n = 20). Eighty-six good-to-excellent-quality two- to four-cell embryos were transferred surgically into the oviducts of 4 of the original oocyte donors in Trial I and 2 females in Trial II. A pregnancy occurred in 1 female in Trial II, and 3 live-born cubs were delivered by Caesarean section 107 days after embryo transfer. Of the 56 cleaved embryos cultured in vitro in Ham's F10 for 72 h, 14 (25.0%) were at the sixteen-cell stage, and 15 (26.8%) were morulae.(ABSTRACT TRUNCATED AT 400 WORDS)
1
Among recovered oocytes, 82.9% were mature, while 10.5% were immature and 6.6% were degenerate; eCG dose and season did not significantly affect yield.
2
Exogenous gonadotropins induced follicular development in all treated females, yielding 456 oocytes from 468 follicles and a 97.4% recovery rate.
3
In vitro fertilization occurred in 63.4% of inseminated mature oocytes, and 95.9% of evaluable fertilized oocytes cleaved to the two-cell stage without parthenogenetic cleavage in controls.
4
The domestic-cat in vitro fertilization and embryo-culture system was successfully adapted to tiger reproduction.
5
Tiger electroejaculates consistently contained motile sperm, averaging 105.9 × 10^6 motile sperm per ejaculate with 81.4% structurally normal sperm.

In vitro fertilization and embryo development of tiger (Panthera tigris) oocytes and embryos

Adaptability and developmental competence of tiger oocytes and embryos in a domestic-cat-derived in vitro fertilization and embryo culture system, including fertilization, cleavage, and in vivo development

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1990-11-01
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Ann M. Donoghue
L. A. Johnston
U. S. Seal
Doug L. Armstrong
Ronald Tilson
Patricia Wolf
K. Petrini
Lee G. Simmons
T.S. Gross
David E. Wildt
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