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 />
Workshop Abstracts 9<br />
Materials and methods Semen collected from 3 adult Lacaune rams<br />
was diluted <strong>in</strong> milk at 1.10 9 sperm/ml and stored for 1 h (fresh semen)<br />
or 25 h (stored semen) at 15°C. Semen was then fluorescently sta<strong>in</strong>ed<br />
by addition of octa<strong>de</strong>cyl-rhodam<strong>in</strong>e B and MitoTrackerGreen FM,<br />
and 75.10 6 sperm were <strong>in</strong>sem<strong>in</strong>ated by endoscopy <strong>in</strong>to the base of<br />
each uter<strong>in</strong>e horn of estrus-<strong>in</strong>duced ewes (n = 6 ewes / treatment). In<br />
vivo imag<strong>in</strong>g of sperm was done un<strong>de</strong>r general anesthesia 4 h after<br />
<strong>in</strong>sem<strong>in</strong>ation. The uterus and oviducts were exteriorized through a<br />
mid-ventral <strong>in</strong>cision and the microprobe of the FCM was <strong>in</strong>serted <strong>in</strong>to<br />
different anatomical regions of the genital tract: the base, middle and<br />
tip of the uter<strong>in</strong>e horn, the uterotubal junction and the oviduct. Stilland<br />
vi<strong>de</strong>o-images were recor<strong>de</strong>d for each region of the genital tract,<br />
and the distribution of fluorescent motile sperm was <strong>de</strong>term<strong>in</strong>ed by<br />
count<strong>in</strong>g the number of motile sperm per field.<br />
Results The <strong>in</strong>fluence of uter<strong>in</strong>e contractions on sperm transport was<br />
clearly observed by <strong>in</strong> vivo FCM. In addition to sperm transport via<br />
massive uter<strong>in</strong>e contractions, <strong>in</strong>dividual motility of sperm could be<br />
imaged at the different regions of the genital tract. When fresh semen<br />
was <strong>in</strong>sem<strong>in</strong>ated, the concentration of motile sperm <strong>in</strong> the lumen 4 h<br />
after <strong>in</strong>sem<strong>in</strong>ation showed an <strong>in</strong>creas<strong>in</strong>g gradient along the uterus:<br />
10.0 ± 1.7, 15.3 ± 2.2, 22.6 ± 4.1 and 21.0 ± 2.9 sperm / field,<br />
respectively, for the base, middle and tip of the uter<strong>in</strong>e horn, and the<br />
uterotubal junction (p500µ (bias 0.0). However, the agreement was variable for<br />
CL (overall bias -3.72) ow<strong>in</strong>g to a less resolvable <strong>in</strong>terface between<br />
<strong>in</strong>dividual CL by ultrasonography. In conclusion, ultrasound<br />
biomicroscopy can be used to accurately assess ovarian follicular<br />
dynamics <strong>in</strong> vivo <strong>in</strong> the mouse.<br />
Research supported by the Natural Sciences and Eng<strong>in</strong>eer<strong>in</strong>g<br />
Research Council of Canada and the Saskatchewan Health Research<br />
Foundation.<br />
WS04-4<br />
Feed<strong>in</strong>g supplement improves spermatogenetic potential<br />
dur<strong>in</strong>g w<strong>in</strong>ter <strong>in</strong> young mer<strong>in</strong>o rams: Prelim<strong>in</strong>ary<br />
morphometric data<br />
Genovese, P 1 , Picabea, N 1 , V<strong>in</strong>oles, C 2 , Gil, J 3 , Bielli, A 1 *<br />
1Morphology and Development, Veter<strong>in</strong>ary Faculty, University of Uruguay,<br />
Uruguay; 2 Glencoe, INIA, Uruguay; 3 DILAVE Paysandú, MGAP, Uruguay<br />
Nutrition <strong>in</strong>fluences testicular <strong>de</strong>velopment before and after puberty.<br />
Quantitative histological techniques allow estimation of the actual and<br />
potential testicular spermatogenic capacity: greater sem<strong>in</strong>iferous<br />
epithelium volume reflects greater sperm produc<strong>in</strong>g capacity. The<br />
effect of nutritional supplementation on testicular <strong>de</strong>velopment was<br />
exam<strong>in</strong>ed <strong>in</strong> Mer<strong>in</strong>o rams (n=11) beg<strong>in</strong>n<strong>in</strong>g at 17 months of age. The<br />
rams divi<strong>de</strong>d <strong>in</strong>to 2 groups and fed differentially from March (late<br />
summer, breed<strong>in</strong>g season) until the time of castration <strong>in</strong> July (midw<strong>in</strong>ter,<br />
beg<strong>in</strong>n<strong>in</strong>g of the non-breed<strong>in</strong>g season) at the Glencoe INIA<br />
research station, Paysandú, Uruguay (Latitu<strong>de</strong>, 32 º south). The rams<br />
were given free access to native pasture at a stock<strong>in</strong>g rate of 4 rams<br />
per hectare. One group (n=5) was not given supplemental nutrition,<br />
while the other group (n=6) was supplemented (0.75% of live weight)<br />
with sorghum (70%) and soy meal (30%). Testes and epididymi<strong>de</strong>s<br />
were weighed immediately after castration. Testicular samples were<br />
processed for quantitative histological analysis. Paraff<strong>in</strong> sections (5<br />
µm thick) were sta<strong>in</strong>ed with hematoxyl<strong>in</strong>-eos<strong>in</strong>, and 30 images were<br />
taken (BX 50 Olympus microscope, CCD camera connected to pc) per<br />
ram. The diameter of the sem<strong>in</strong>iferous tubules and the fractional<br />
volume of the sem<strong>in</strong>iferous tubules, sem<strong>in</strong>iferous epithelium,<br />
sem<strong>in</strong>iferous tubule lum<strong>in</strong>a and testicular <strong>in</strong>terstitium were measured<br />
by means of computer assisted analysis with Image Pro Plus®<br />
software. Fractional volumes were estimated by count<strong>in</strong>g the<br />
percentage of po<strong>in</strong>ts <strong>in</strong> a po<strong>in</strong>t grid fall<strong>in</strong>g over the object of <strong>in</strong>terest.<br />
Absolute volumes of sem<strong>in</strong>iferous tubules, sem<strong>in</strong>iferous epithelium,<br />
sem<strong>in</strong>iferous tubule lum<strong>in</strong>a and testicular <strong>in</strong>terstitium were estimated<br />
by multiply<strong>in</strong>g fractional volumes by testicular volume (estimated<br />
from testicular weight). The nutritionally supplemented group of rams<br />
had a greater total testicular weight (g, 299.9 ± 24.0 vs 201.9 ± 42.8;<br />
P