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 />
110 Poster Abstracts<br />
was diluted 1:1 with Botu-Semen ® (skim-milk based exten<strong>de</strong>r), split<br />
<strong>in</strong> two aliquots and centrifuged at 600xg for 10 m<strong>in</strong>utes. Pellets were<br />
ressuspen<strong>de</strong>d us<strong>in</strong>g Botu-Crio ® (egg-yolk based exten<strong>de</strong>r with<br />
am<strong>in</strong>oacids, 1% glycerol and 4% ami<strong>de</strong>s - Biotech Botucatu ® , Brazil),<br />
or Botu-Crio ® Egg Free ® exten<strong>de</strong>r (same basis without egg-yolk -<br />
Biotech Botucatu ® , Brazil), to a f<strong>in</strong>al concentration of<br />
100x10 6 sperms/mL. Samples were prepared at room temperature.<br />
After ressuspension, samples were loa<strong>de</strong>d <strong>in</strong>to 0.5mL straws,<br />
stabilized for 20m<strong>in</strong> at 5°C <strong>in</strong> refrigerator then placed 6 cm above<br />
nitrogen level <strong>in</strong> isopor box (42L) filled with 3 cm of liquid nitrogen<br />
for 20 m<strong>in</strong> and f<strong>in</strong>ally immersed. Samples were thawed at 46°C for<br />
20s and ma<strong>in</strong>ta<strong>in</strong>ed <strong>in</strong> 1.5mL plastic tubes <strong>in</strong> dryblock at 37°C and<br />
evaluated for sperm motility by CASA (IVOS 12) and plasma<br />
membrane <strong>in</strong>tegrity with fluorescent probes (PI and CFDA). ANOVA<br />
and Tukey test were performed by GraphPad InStat program. Results<br />
obta<strong>in</strong>ed for Botu-Crio ® and Botu-Crio Egg Free ® exten<strong>de</strong>rs were:<br />
total motility (61.4±16.2 x 61.4±15.9%), progressive motility<br />
(27.0±9.8 x 26.7±10.6%), VAP (path velocity) (92.6±13.3 x<br />
97.1±13.8µm/s), VSL (straight l<strong>in</strong>e velocity) (73.0±8.8 x<br />
75.0±10.3µm/s), VCL (curvil<strong>in</strong>ear velocity) (167.5±21.7 x<br />
176.3±22.9µm/s), ALH (amplitu<strong>de</strong> of lateral head displacement)<br />
(6.7±0.5 x 7.0±0.4µm), STR (straightness) (78.1±5.2 x 77.0±2.4%),<br />
LIN (l<strong>in</strong>earity) (46.2±5.1 x 43.3±1.9%) and plasma membrane<br />
<strong>in</strong>tegrity (53.4±11.5 x 57.4±7.0%), respectively. No difference<br />
(P>0.05) was observed between exten<strong>de</strong>rs. In conclusion, as Botu-<br />
Crio Egg Free ® , which is a completely chemically <strong>de</strong>f<strong>in</strong>ed media, has<br />
shown similar results when compared to Botu-Crio ® , it may be an<br />
alternative for freez<strong>in</strong>g equ<strong>in</strong>e semen. Further researches are<br />
necessary to evaluate <strong>in</strong> vivo fertility when us<strong>in</strong>g this exten<strong>de</strong>r.<br />
P248<br />
Vag<strong>in</strong>al biotic flora and its antibiotic resistance profile <strong>in</strong><br />
terras <strong>de</strong> miranda jennets<br />
Payan-Carreira, RM 1 *, Mota, VR 2 , Barroso, AT 2 , Quaresma, M 3 , Saavedra,<br />
MJ 4<br />
1Zootecnia Dept., University of Trás-os-Montes e Alto Douro, Portugal;<br />
2University of Trás-os-Montes e Alto Douro, Portugal; 3 Veter<strong>in</strong>ary Teach<strong>in</strong>g<br />
Hospital, University of Trás-os-Montes e Alto Douro, Portugal; 4 Veter<strong>in</strong>ary<br />
Cl<strong>in</strong>ics Dept., University of Trás-os-Montes e Alto Douro, Portugal<br />
In this work authors report the prelim<strong>in</strong>ary results on the<br />
characterization of the biotic flora found <strong>in</strong> the vag<strong>in</strong>a of Terras <strong>de</strong><br />
Miranda jennets. Vag<strong>in</strong>al swab samples were obta<strong>in</strong>ed from the<br />
vestibule and transported <strong>in</strong> transport medium for <strong>de</strong>livery to the<br />
Microbiology Lab. This work was carried <strong>in</strong> a group of 21 jennets,<br />
aged between 4 to 20 years old and the samples were obta<strong>in</strong>ed <strong>in</strong> two<br />
retractions (15 animals + 6 animals). For the bacteriological analysis<br />
of the vag<strong>in</strong>al samples different and selective culture media were<br />
used, aim<strong>in</strong>g a more significant bacterial growth. For the prelim<strong>in</strong>ary<br />
i<strong>de</strong>ntification of the isolated ones a set of biochemical tests was<br />
carried through (oxidase, catalase, <strong>in</strong>dol production, use of the citrate,<br />
metil red, vogues proskauer and lactose fermentation). Antimicrobial<br />
susceptibility test<strong>in</strong>g was performed by disc-diffusion methods to<br />
twenty seven antimicrobials (by CLSI recommendations), <strong>in</strong>clud<strong>in</strong>g<br />
different beta-lactams, qu<strong>in</strong>olone and am<strong>in</strong>oglicosi<strong>de</strong> antibiotics.<br />
From the vag<strong>in</strong>al samples, bacteria’s perta<strong>in</strong><strong>in</strong>g to the different<br />
generic groups were found: Gram-positive bacteria, such as<br />
Staphylococcus spp. and Enterococcus spp.; and Gram-negative<br />
bacteria, such as Enterobactereaceae (E. coli, Klebsiella and<br />
Enterobacter) and not Enterobactereaceae (Pseudomonas spp.).<br />
Regard<strong>in</strong>g to the resistance profile to antibiotics, the most frequently<br />
observed resistance was to amoxicill<strong>in</strong> and ticarcill<strong>in</strong> (penicill<strong>in</strong>s),<br />
even when associated to the beta-lactamases <strong>in</strong>hibitor (acid<br />
clavulanic), as well as the cephalospor<strong>in</strong> of 1st generation studied<br />
(cephalot<strong>in</strong>). Resistance to aztreonam (antibiotic beta-lactam of the<br />
group of the monobactams) was also observed <strong>in</strong> some cases.<br />
P249<br />
Does the microbial flora <strong>in</strong> the ejaculate affect the<br />
freezeability of stallion sperm<br />
Pena, FJ 1 *, Ortega Ferrusola, C 2 , Gonzalez Fernan<strong>de</strong>z, L 2 , Macias Garcia,<br />
B 2 , Rodriguez-Mart<strong>in</strong>ez, H 2 , Tapia, JA 2 , Alonso, JM 2<br />
1<strong>Reproduction</strong>, University of Extremadura, Spa<strong>in</strong>; 2 Spa<strong>in</strong><br />
In an attempt to evaluate the possible relationship between the<br />
microbial flora <strong>in</strong> the stallion ejaculate and its ability to freeze, 3<br />
ejaculates from 5 stallions were frozen us<strong>in</strong>g a standard protocol.<br />
Before freez<strong>in</strong>g, an aliquot was removed for bacteriological analysis.<br />
Bacterial growth was observed <strong>in</strong> all the ejaculates studied. The nonparametric<br />
Mann–Whitney U-test was used to directly compare pairs<br />
of values. The Spearman non-parametric test was used to study<br />
correlations between microbiological f<strong>in</strong>d<strong>in</strong>gs (presence and quantity,<br />
as number of CFU) and sperm quality post-thaw. The isolated<br />
microorganisms were: Staphylococcus spp and Micrococcus spp (<strong>in</strong><br />
all the stallions), β-haemolytic Streptococcus (<strong>in</strong> stallions 3 and 4),<br />
Corynebacterium spp (<strong>in</strong> stallions 1, 3-5), Rhodococcus spp (<strong>in</strong><br />
stallion number 2), Pseudomonas spp (<strong>in</strong> stallion number 1) and<br />
Klebsiella spp <strong>in</strong> stallions 1, 3 and 5. The presence and richness of<br />
Klebsiella and β-haemolytic Streptococcus <strong>in</strong> the ejaculate were<br />
related to two sperm variables post-thaw, namely the proportion of<br />
<strong>de</strong>ad spermatozoa (EH+ cells; r=0.55, P