ABSTRACT BOOK

18TH JUNE 26-30TH

INTERNATIONAL 2016
CONGRESS LE VINCI IN TOURS
FRANCE
ON ANIMAL The cutting edge science…
in the garden of France.

REPRODUCTION
Reproductive physiology, pathology and
biotechnologies in domestic and wild
animal species.

ADMINISTRATIVE OFFICE

www.icar2016.org
PS117 - Binder of Sperm Proteins (BSP) and spermadhesins: immunodetection in the
reproductive fluids and binding patterns in sperm from Morada Nova rams
Authors and co-authors:
SOUSA Solange (1), TILBURG Maurício (1), SANTOS Fágner (1), VELHO Ana Luiza (1), MELO Révila (1), GUEDES Maria
Izabel (1), MOURA Arlindo (1)
(1) Federal University of Ceará, Animal Science, 60440-554 , Fortaleza, Brazil
Binder of Sperm Proteins (BSP) and spermadhesins are the major components of ruminant seminal plasma.
In bulls, BSP regulate capacitation by removing cholesterol and phospholipids from the sperm membrane.
In rams, however, BSPs functions have not been fully elucidated. Spermadhesins, in turn, are thought to
participate in gamete interaction. The present study was conducted to evaluate the presence of BSP and
spermadhesins in the reproductive fluids and their binding patterns to ram sperm. Our study used samples
from Morada Nova rams, a tropically adapted breed from the semi-arid region of Brazil (03°43'S; 38°32'W).
Semen was collected from six adult rams by artificial vagina and sperm and seminal plasma (SP) were
separated by centrifugation. After this first semen collection, rams were vasectomized and fluid from
accessory sex glands (FAG) was obtained by artificial vagina as well. Finally, animals were slaughtered and
fluids from the rete testis (RTF), cauda epididymidis (CEF), vesicular glands (VGF) and bulbourethral glands
(BGF) were obtained. Proteins of all fluids and extracted from epidymal sperm were separated by 1-D or 2-D
SDS-PAGE. Western blots using primary antibodies against BSPs and spermadhesins were carried out. Protein
binding to ejaculated sperm were evaluated by immunocytochemistry and confocal microscopy (LM710
– Zeiss), using the same antibodies. Proteins from RTF, FAG, BGF and CEF were identified using tandem
mass spectrometry. In silico protein interaction networks was obtained from STRING database (string-db.
org). Protein bands with 22 kDa and 14 kDa represented 22%, 32% and 28% of the total intensity of the all
bands detected in SP, FGA and VGF 1-D gels, respectively. Bands at 15 kDa had 15% of the total intensities
detected in 1-D gels from those three sample types. Based on Western blots, ram BSP and spermadhesins
were present in SP, FGA and VGF, but not in RTF, CEF and BGF. As determined by immunocytochemistry, BSP
were present on the midpiece and acrosome of ejaculated sperm, and spermadhesin was detected bound to
the acrosome region. Our previous results show that BSP and spermadhesin was not detected on epididymal
sperm. Based on molecular weight (kDa) and isoeletric point values, VGF 2-D map indicates the presence of
BSP and spermadhesins. Mass spectrometry confirmed their presence in FGA. In RTF, FBG and epididymal
sperm, these proteins were not detected. Protein network analysis shows that BSP can interact with
molecules involved in cellular responses, structural support, gamete interaction and fertilization. In addition,
spermadhesins can interact with proteins linked to fertilization. In conclusion, we describe a comprehensive
evaluation of BSP and spermadhesin expression in the ram reproductive fluids and their binding patterns to
sperm before and after ejaculation. In the ram, both BSP and spermadhesins likely play fundamental roles in
post-maturation events, from capacitation to sperm-egg interaction.

PS118 - Pasture allowance during pregnancy affects the histology of the testes in
newborn lambs
Authors and co-authors:
Raquel Raquel (1), Montaldo Sebastián (2), Genovese Patricia (2), Riaño Victoria (2), Bielli Alejandro (2)
(1) Facultad de Agronomía, Universidad de la República, Producción Animal y Pasturas, 11900, Montevideo, Uruguay
(2) Facultad de Veterinaria, Universidad de la República, Morfología y Desarrollo, 11600, Montevideo, Uruguay
Intrauterine subnutrition influences testicular development. The aim of the present work was to compare
the effect of 2 natural pasture allowances (PA) in Uruguay (32° SL) on testicular development in single
male newborn lambs. All procedures were conducted in compliance with recommendations of the Animal
Experiment Committee of the Universidad de la República. Multiparous Corriedale pregnant ewes (n=18),
carrying a single male fetus were used in a completely randomized block design (by topography; 3 blocks, 3
plots/treatment). Since 23 d before conception until 121-122 d of pregnancy, when they were shorn, 9 ewes
grazed on natural grassland at 2.9-3.8 kg dry matter/kg of live weight (LW) (group H) and 9 at 1.4-2.6 kg DM /
kg of LW (L). After shearing, all ewes grazed as a single mob on Festuca arundinacea (2.8 kg DM/kg LW until 2
days before lambing). Thereafter, all ewes grazed natural grassland ad libitum in small paddocks. All the ewes

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