Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias
Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias
Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias
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16 t h International Congress on Animal <strong>Reproduction</strong><br />
Poster Abstracts 167<br />
un<strong>de</strong>r extensive rear<strong>in</strong>g with natural mat<strong>in</strong>g and are characterized by<br />
lack of selection, poor record keep<strong>in</strong>g and <strong>in</strong>frequent veter<strong>in</strong>ary<br />
assistance. Besi<strong>de</strong>s, breed<strong>in</strong>g bulls are selected based ma<strong>in</strong>ly on<br />
phenotype, neglect<strong>in</strong>g the importance of the breed<strong>in</strong>g soundness<br />
evaluation (BSE) (1).<br />
Materials and Methods A s<strong>in</strong>gle BSE was performed <strong>in</strong> 38 sires<br />
placed <strong>in</strong> 26 beef cattle farms managed un<strong>de</strong>r extensive rear<strong>in</strong>g and<br />
cont<strong>in</strong>uous mat<strong>in</strong>g from the south area of Costa Rica. Bulls belonged<br />
to Brahman (n=8), Simmental x Brahman crosses (n=17), Simmental<br />
(n=6), Brown Swiss x Brahman (n=3) and other crosses (n=4). These<br />
bulls were un<strong>de</strong>r s<strong>in</strong>gle sired mat<strong>in</strong>g breed<strong>in</strong>g with 24.0±13.6 (5-73)<br />
adult cows. After the BSE, bulls were classified as Sound: Bulls that<br />
fitted the physical exam, with a scrotal circumference (SC) accord<strong>in</strong>g<br />
to breed and age standards (2), healthy reproductive organs and<br />
without <strong>de</strong>viations <strong>in</strong> the spermiogramme. Deferred: Sires who did<br />
not meet the above requirements but with a favorable prognosis for<br />
recover<strong>in</strong>g. Unsound: Serious cl<strong>in</strong>ical and spermiogramme<br />
abnormalities compromis<strong>in</strong>g gravely their potential breed<strong>in</strong>g<br />
efficiency. Simultaneously, the conception rate (CR) was evaluated <strong>in</strong><br />
the herds by rectal palpation and ultrasound. Thereafter, the cows<br />
were ranked as pregnant or not, be<strong>in</strong>g the cycl<strong>in</strong>g activity <strong>de</strong>term<strong>in</strong>ed<br />
by the presence of a corpus luteum.<br />
Results and Discussion Mean SC (cm) and age (yrs) for sound<br />
(n=23), <strong>de</strong>ferred (n=5) and unsound (n=10) sires were 37.4±3.3 and<br />
4.5±2.4, 39.0±4.4 and 4.7±2, 37.6±5.2 and 4.2±2 respectively. In the<br />
unsound group, one bull had low SC and the remnant had cl<strong>in</strong>ical and<br />
spermiogramme <strong>de</strong>viations typical of testicular <strong>de</strong>generation. The CR<br />
calculated solely upon the cycl<strong>in</strong>g cows was 82.5%±12.1 (64-100) for<br />
sound, 77.4%±27.2 (36-100) for <strong>de</strong>ferred and 28.4%±30.5 (0-79) for<br />
the unsound sires. The CR was p0.05 when<br />
compar<strong>in</strong>g the sound vs. the <strong>de</strong>ferred group. Given the conditions of<br />
cattle rear<strong>in</strong>g prevail<strong>in</strong>g <strong>in</strong> the tropics, the BSE is an efficient tool <strong>in</strong><br />
or<strong>de</strong>r to i<strong>de</strong>ntify those bulls with an unsound andrological status. The<br />
presence of those sires <strong>in</strong> the herd, contributes to impair seriously the<br />
productive proficiency of the beef cattle system.<br />
P424<br />
Effect of epididymis storage temperature and<br />
cryopreservation on mitochondrial potential, membrane<br />
<strong>in</strong>tegrity and motility of bov<strong>in</strong>e epididymal sperm<br />
Nichi, M 1,2 *, Rijsselaere, T 3 , Goovaerts, IGF 2 , Van Soom, A 3 , Barnabe, VH 1 ,<br />
De Clercq, JPB 2 , Bols, PEJ 2<br />
1Department of Animal <strong>Reproduction</strong> (VRA), Faculty of Veter<strong>in</strong>ary Medic<strong>in</strong>e<br />
and Zootechny (FMVZ), University of São Paulo (USP), Brazil; 2 Laboratory of<br />
Veter<strong>in</strong>ary Physiology, Faculty of Pharmaceutical, Biomedical and Veter<strong>in</strong>ary<br />
Sciences, University of Antwerp, Belgium; 3 Department of <strong>Reproduction</strong>,<br />
Obstetrics and Herd Health, Faculty of Veter<strong>in</strong>ary Medic<strong>in</strong>e, Ghent University,<br />
Belgium<br />
Introduction Cryopreservation of sperm recovered from<br />
epididymi<strong>de</strong>s is potentially a useful tool <strong>in</strong> case of unexpected <strong>de</strong>ath<br />
of animals with high genetic value, species endangered of ext<strong>in</strong>ction,<br />
or when the collection of sperm by other means is not possible.<br />
Studies <strong>in</strong>dicate that ma<strong>in</strong>ta<strong>in</strong><strong>in</strong>g the epididymis at a lower<br />
temperature dur<strong>in</strong>g storage and transport improves the quality of the<br />
retrieved sperm. The objective of this experiment was to study<br />
whether the storage temperature of the epididymis follow<strong>in</strong>g slaughter<br />
<strong>in</strong>fluences sperm membrane resistance aga<strong>in</strong>st lipid peroxidation,<br />
resistance to cryopreservation, and fertiliz<strong>in</strong>g capacity, and if the<br />
percentage of cytoplasmic droplets would play a role <strong>in</strong> this event.<br />
Methods Thirty two epididymi<strong>de</strong>s (16 bulls) were collected after<br />
slaughter and divi<strong>de</strong>d <strong>in</strong>to two groups, stored at 4 and 34ºC for 2-3<br />
hours, after which semen was collected from the caudae<br />
epididymi<strong>de</strong>s. The epididymal sperm was then evaluated us<strong>in</strong>g<br />
CASA, and an aliquot was sta<strong>in</strong>ed with SYBR14/PI/JC1 to evaluate<br />
membrane <strong>in</strong>tegrity and mitochondrial membrane potential. Sperm<br />
was then frozen us<strong>in</strong>g an automatic <strong>de</strong>vice. Post thaw sperm was also<br />
analyzed us<strong>in</strong>g CASA and SYBR14/PI/JC1.<br />
Results A marked negative effect of cryopreservation on sperm<br />
motility was observed <strong>in</strong> the samples collected from epididymi<strong>de</strong>s<br />
stored at 4ºC. Significant effects of temperature of epididymal storage<br />
and of cryopreservation were observed on motility parameters as well<br />
as on cell viability and mitochondrial activity. Pre-cryopreservation<br />
sperm motility, progressive motility, and velocity were higher <strong>in</strong><br />
samples from epididymi<strong>de</strong>s stored at 4 o C, while post-thaw sperm<br />
motility, progressive motility, and velocity did not differ between<br />
samples from epididymi<strong>de</strong>s stored at 4 or at 34 o C. Although the drop<br />
on motility was not evi<strong>de</strong>nt on sperm samples collected from<br />
epididymi<strong>de</strong>s stored at 34ºC, the effect of temperature before<br />
cryopreservation may have <strong>in</strong>fluenced those results. Strong<br />
correlations (r>0.6) were found for samples collected from<br />
epididymi<strong>de</strong>s stored at 34ºC between pre-freeze sperm motility<br />
<strong>in</strong><strong>de</strong>xes and mitochondrial potential and between post-thaw sperm<br />
motility and membrane <strong>in</strong>tegrity.<br />
Conclusions Results <strong>in</strong>dicate that the conditions <strong>in</strong> which<br />
epididymi<strong>de</strong>s are handled after cryopreservation may <strong>in</strong>fluence postthaw<br />
sperm quality, especially due to an impaired mitochondrial<br />
potential <strong>in</strong> sperm samples collected from epididymi<strong>de</strong>s stored at<br />
higher temperatures. This effect may have caused a higher<br />
susceptibility to membrane damages due to cryopreservation.<br />
P425<br />
Assessment of spermatozoal characteristics <strong>in</strong> exten<strong>de</strong>d<br />
boar semen <strong>in</strong>cubated at 18 o C us<strong>in</strong>g flow cytometry<br />
Niżański, W 1 *; Partyka, A 2 ; Dubiel, A 1 ; Łukaszewicz, E 2<br />
1Department of <strong>Reproduction</strong>, University of Environmental and Life Sciences<br />
<strong>in</strong> Wrocław, Poland; 2 Department of Poultry Breed<strong>in</strong>g, University of<br />
Environmental and Life Sciences <strong>in</strong> Wrocław, Poland<br />
Exten<strong>de</strong>d liquid boar semen is commonly used for <strong>in</strong>sem<strong>in</strong>ation for up<br />
to 5 days after collection. A litter size and farrow<strong>in</strong>g rates are reduced<br />
when us<strong>in</strong>g semen stored > 3 days. The aim of our study was to<br />
estimate changes of spermatozoal characteristics <strong>in</strong> exten<strong>de</strong>d boar<br />
semen <strong>in</strong>cubated at 18oC for 240 hrs us<strong>in</strong>g fluorescent sta<strong>in</strong><strong>in</strong>g and<br />
flow cytometry. The study was carried out on 35 ejaculates collected<br />
from 7 boars. Semen samples were exten<strong>de</strong>d <strong>in</strong> Safe Cell+ diluent<br />
(IMV Technologies). Spermatozoal characteristics were evaluated at<br />
24, 48, 96, 168 and 240 hrs after collection. Spermatozoal viability<br />
was <strong>de</strong>term<strong>in</strong>ed by SYBR-14/PI fluorescent sta<strong>in</strong><strong>in</strong>g. The DNA status<br />
of spermatozoa was assessed us<strong>in</strong>g the metachromatic properties of<br />
acrid<strong>in</strong>e orange (AO) <strong>in</strong> the sperm chromat<strong>in</strong> structure assay (SCSA).<br />
DNA fragmentation <strong>in</strong><strong>de</strong>x (DFI) and high DNA sta<strong>in</strong>ability (HDS)<br />
were evaluated. Acrosomal <strong>in</strong>tegrity was measured by Pisum sativum<br />
(PSA) agglut<strong>in</strong><strong>in</strong> labeled by a fluorescent probe that sta<strong>in</strong>s acrosomereacted-<br />
or damaged spermatozoa only. The mitochondrial function of<br />
spermatozoa was evaluated with Rhodam<strong>in</strong>e 123 (R123). Samples<br />
were analysed <strong>in</strong> a FACSCalibur flow cytometer (Becton Dick<strong>in</strong>son,<br />
San Jose, CA). The percentage of live spermatozoa did not change<br />
significantly dur<strong>in</strong>g the 168 hrs of storage. A significant <strong>in</strong>crease of<br />
the <strong>de</strong>ad spermatozoa percentage (p