Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias

16 t h International Congress on Animal Reproduction
Poster Abstracts 135
aimed at identifying the vaginal epithelial cells of maned sloths
(Bradypus torquatus) as a possible way to study the phases of the
estrous cycle of this animal.
Material and methods The samples for vaginal cytology were
obtained from four free ranging maned sloths living in a protected
area of coastal forest in the South of Bahia. The sterile gynecological
brush was inserted up to the necessary distance to reach the pelvic
channel. Two smears were immediately immersed in absolute alcohol
for ten seconds to produce cell fixation. Staining was performed using
rapid Panotic Kit (Laborclin®). The methodology for differential
evaluation of the cells was based on Schutte’s work (1967).
Results and discussion Maned sloths BT033, BT065, and BT042
presented, respectively, 32%, 33%, and 7% of parabasal epithelial
cells (PB); 58%, 22%, and 10% of small intermediate cells (SI); 9%,
18%, and 6% of large intermediate cells (LI); 4%, 13%, and 24% of
superficial epithelial cell with a nucleus (NS); 8%, 14%, and 53% of
anucleated superficial epithelial cell (AS). Two cell samples were
collected for maned sloth BT464 with a 15 months interval.
Cytological differences were observed between the two samples (1ª
and 2 ª): 7.5% and 17.5% of PB cells, 6.5% and 25% of SI cells, 12%
and 15.5 of LI cells, 9.5% and 19.5% of NS cells and 70% and 22.5%
of AS cells, respectively. It’s interesting to remark that the percentage
of vaginal epithelial cells varied among sloths and also for the same
animal. This result suggests that vaginal cytology of maned sloth can
be used as a tool to evaluate of estrous cycle.
References SCHUTTE AP. Canine vaginal cytology. I – Technique
and cytological morphology. J. Small Anim. Pract., v.8, p 301-306,
1967.
Keywords: cells, vaginal epithelia, estrous cycle.
P325
Effects of culture medium McCoy 5A Modified® (Sigma-
Aldrich M-9309) in semen motility post-thawing of
bottlenose dolphin (Tursiops truncatus, Tursiops
aduncus, Tursiops gilli)
Ugaz, C
Veterinary and research, Dolphinaris, Mexico
Due to the grate interest in captive marine mammals, in recent
decades works on reproduction biotechnology in these species
increased. One of the most charismatic species in captivity is the
bottlenose dolphin, in witch many different researches have been done
to improve procedures for their semen freezing (Robeck T. 2004;
Robeck T. 2001, Ugaz C. et al. 2006). Many researches in different
species have used the amino acids additions in freezing extender,
generally glutamine (50mM), proving good results on the motility
percentage after thawing (Khlifaoui M et al, 2005; Yahui L, et al,
2003; Trimeche A et al, 1999; Vidament M et al, 2001). In present job
semen of the three male of the three different species was use
(Tursiops truncatus, Tursiops aduncus, and Tursiops gilli); kept in
captivity under the same life conditions. At least 3 ejaculate each was
extracted (table 1). It was frozen using a refrigeration rate -0.18°
C/min. and freezing rate -8.5° C/min. The Triladyl® (Minitube)
extender was use, according to the manufacturer's instructions, with a
concentration of 800 million sperm/ml. Thawing was made at 36°C
for 60 seconds, McCoy 5A Modified ® (Sigma-Aldrich M-9309) at
36°C was added, in 1:1 (v/v) dilution. The main characteristics of this
culture medium are that it has a lot off amino acids, and it is enriched
with L-Glutamine (219.15 mg/L) and NaHCO3. All results indicated a
significant increase in motility post-thawing (P= 0.0004; R squared:
0.6889; T paired test) with McCoy 5A Modified ®, 49.57 % is higher
versus semen post-thawing without culture medium.
Poster 09 - Reproduction of Rabbit and Laboratory
Rodents
P326
Effect of reproductive rhythm on the oocyte quality and
fertility rate in primiparous does
Arias-Álvarez, M 1 *; García-García, RM 1 ; Revuelta, L 1 ; Rebollar, PG 2 ;
Lorenzo, PL 1
1Dpto. de Fisiología (Fisiología Animal). Facultad de Veterinaria-UCM.
Madrid. Spain; 2 Dpto. de Producción Animal. ETSIA-UPM. Madrid. Spain.
The fertility and prolificacy of the primiparous rabbit does are low
when they are inseminated (AI) in the short period after kindling.
Strong nutritional needs and a lactation status may have consequences
on the quality of their gametes. Oocyte maturation is the first step that
affects the successful fertilization and preimplantation embryo
development and finally influencing the economic profits. The aim of
this study was to compare the effect of different reproductive rhythms
(semi-extensive and extensive) on meiotic and cytoplasmatic
maturation measured as cortical granules migration (CG) of rabbit
oocytes matured in vitro. A total of 90 primiparous New Zealand x
California white rabbits, under semi-intensive (Group A: AI at 11
postpartum day (ppd), n=45) or extensive rhythm (Group B: AI at 32
ppd, post weaning, n=45), were synchronized by biostimulation 24
hours. Fifteen animals by group were euthanasized according to the
bioethics committee of the University. Cumulus oocyte complexes
(COC) were aspirated from ovarian follicles ≥ 1 mm in size of one
ovary and were matured in TCM-199 medium supplemented with
10% FCS, 10ng/ml EGF and 100ng/ml IGF. Afterwards, a total of
301 COC (n=188, group A and n=113, group B) were treated
progressively with 2mM hyaluronidase, 0.5% pronase, 4%
paraformaldehyde, 0.02% Triton X-100 and 7.5% BSA. Oocytes were
incubated with 100 μg/ml FITC-LCA for GC staining and with
10μg/ml Propidium Iodide for nuclear staining, and observed under a
confocal laser-scanning microscope. Chi-square test was performed to
analyse fertility, nuclear maturation and CG migration index. Fertility
rate was significantly higher in group B than group A (53.6% vs
100%, p