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 />
140 Poster Abstracts<br />
P341<br />
The effect of various diluents on pigeon semen stored<br />
24h at 5 o C<br />
Klimowicz, M 1 *, Batkowski, F 2<br />
1Krakow Agricultural University, Faculty of Animal Breed<strong>in</strong>g and Biology,<br />
Department of Animal <strong>Reproduction</strong> and Anatomy, Al. Mickiewicza24/28, 30-<br />
059 Krakow, Poland; 2 Wroclaw University of Environmental and Live Science,<br />
Faculty of Veter<strong>in</strong>ary Medic<strong>in</strong>e, Stu<strong>de</strong>nt Scientific Association, ul. Norwida<br />
31, 50-375 Wroclaw, Poland<br />
The aim of the study was to i<strong>de</strong>ntify a suitable exten<strong>de</strong>r for pigeon<br />
semen preserved for 24h at 5 o C. Experiment was conducted 10 weeks.<br />
Semen was collected twice a week, from 40 fancy pigeons. After<br />
macroscopic analysis semen was pooled, diluted <strong>in</strong> Lake’s solution<br />
and BPSE exten<strong>de</strong>r, and divi<strong>de</strong>d <strong>in</strong> two equal parts. One part of semen<br />
was evaluated immediately and second one was stored at 5 o C and<br />
evaluated after 24h. Every sample of semen was sta<strong>in</strong>ed us<strong>in</strong>g 2<br />
different methods - conventional eos<strong>in</strong>-nigros<strong>in</strong> sta<strong>in</strong> to estimate<br />
morphology of spermatozoa and SYBR-14/PI to evaluate sperm<br />
viability by flow cytometry. Sperm motility and velocity parameters<br />
were estimated us<strong>in</strong>g computer-assisted semen analyser HTM IVOS<br />
12.2 (CASA). CASA analysis revealed that sperm motility (MOT),<br />
percentage of spermatozoa with a progressive motility (PMOT) were<br />
significantly higher <strong>in</strong> BPSE than <strong>in</strong> Lake’s solution after 24h storage<br />
(p≤0.05). Velocity parameters such as VAP, VSL, VCL, LIN, STR<br />
and the percentage of viable spermatozoa were not different between<br />
exten<strong>de</strong>rs. The percentage of morphologically normal spermatozoa<br />
was significantly higher at 0h <strong>in</strong> semen diluted <strong>in</strong> BPSE than <strong>in</strong><br />
Lake’s solution (85.48±4.83 and 75.12±3.45; p≤0.01), as well as after<br />
24h <strong>in</strong> vitro storage - 65.4±10.71 and 52.5±10.44 respectively<br />
(p≤0.05). The <strong>de</strong>formation and damage of spermatozoal acrosome<br />
was extremely higher <strong>in</strong> semen exten<strong>de</strong>d <strong>in</strong> Lake’s solution after <strong>in</strong><br />
vitro storage (p≤0.001). BPSE is a suitable semen exten<strong>de</strong>r for storage<br />
of pigeon ejaculates at 5 o C. This diluent has been capable of<br />
protection pigeon semen dur<strong>in</strong>g <strong>in</strong> vitro storage and could potentially<br />
improve methods of cryopreservation.<br />
P342<br />
The effect of short-term semen storage temperature and<br />
period on South African <strong>in</strong>digenous cock breeds<br />
Mphaphathi, ML*; Raito, MB; Mapeka, MH; Mantiziba, CW; Munyai, PH1;<br />
Boshoff, MP; Suzette, F and Nedambale, TL<br />
Agricultural Research Council-Livestock Bus<strong>in</strong>ess Division, Germplasm and<br />
Reproductive Biotechnologies, Private Bag X2, Irene, 0062, RSA<br />
The <strong>de</strong>velopment of short and long term storage of South African<br />
<strong>in</strong>digenous cock’s semen is nee<strong>de</strong>d to ensure a viable reserve of<br />
germplasm for artificial reproduction. The aim of this study was to<br />
exam<strong>in</strong>e the longevity of freshly collected semen of two different<br />
<strong>in</strong>digenous cock breeds at room temperature (25 o C) and low<br />
temperature (4 o C) for 4 h, 8 h, and 24 h. Abdom<strong>in</strong>al massag<strong>in</strong>g<br />
technique was used to collect cock’s semen of Naked neck (NN) and<br />
Venda (V) breed. Follow<strong>in</strong>g semen collection, spermatozoa was<br />
divi<strong>de</strong>d equally per treatment group; then microscopic characteristics<br />
(motility and survival rate) were evaluated un<strong>de</strong>r polariz<strong>in</strong>g BHTU<br />
microscope and the sperm concentration was measured by spermacue.<br />
Modified Brackett and Oliphant’s (BO) medium was used to dilute<br />
(1:2) <strong>in</strong>dividual ejaculates per treatment group. The ejaculates<br />
volume, concentration, and pH of NN and Venda breed were recor<strong>de</strong>d<br />
and evaluated for different time <strong>in</strong>tervals. Data was analyzed by<br />
ANOVA. Prelim<strong>in</strong>ary results <strong>de</strong>monstrated a higher concentration of<br />
sperm <strong>in</strong> NN (8.14 x 10 8 /ml) than <strong>in</strong> Venda (4.46 x 10 8 /ml) breed. In<br />
contrast, higher pH was recor<strong>de</strong>d <strong>in</strong> semen collected from Venda cock<br />
breed. However, there were no statistical differences <strong>in</strong> sperm motility<br />
and survival rate of semen stored at 25°C and 4°C between NN and<br />
Venda breed for all periods (Table 1). Regardless of time <strong>in</strong>tervals<br />
and cock breed, there was an <strong>in</strong>crease <strong>in</strong> <strong>de</strong>ad sperm percentage and<br />
pH over time for semen stored at 4 and 25 o C. In summary, semen<br />
collected from NN cock resulted <strong>in</strong> higher concentration of sperm.<br />
This study also <strong>in</strong>dicated changes <strong>in</strong>duced by storage temperature,<br />
breed and length of semen storage. Study on cryopreservation of NN<br />
and Venda cock semen is on progress.<br />
P343<br />
Changes <strong>in</strong> sperm quality of broiler bree<strong>de</strong>r males<br />
supplemented with organic selenium<br />
Végi, B*; Váradi, É; Ferencz<strong>in</strong>é Szőke, Zs; Barna, J<br />
Avian <strong>Reproduction</strong> Group, Research Institute for Animal Breed<strong>in</strong>g and<br />
Nutrition, Hungary<br />
The fertility <strong>de</strong>cl<strong>in</strong>e <strong>in</strong> the second half of reproduction cycle has been<br />
a permanent problem <strong>in</strong> broiler bree<strong>de</strong>rs’ production. The aim of the<br />
study was to f<strong>in</strong>d differences <strong>in</strong> the effect of <strong>in</strong>organic and organic<br />
selenium and the higher level of vitam<strong>in</strong> E on sperm parameters,<br />
ma<strong>in</strong>ly <strong>in</strong> the second half of the reproduction cycle <strong>in</strong> two types of<br />
broiler bree<strong>de</strong>rs. Selenium and the vitam<strong>in</strong> E play important role <strong>in</strong><br />
the antioxidant system of live organism and additionally, of the<br />
membrane of avian spermatozoa. Due to the membrane damages the<br />
sperm functions failure, which results <strong>in</strong> reduced fertility. Sperm<br />
parameters of 20-20 males from ROSS 308 and Hubbard broiler<br />
bree<strong>de</strong>rs were compared dur<strong>in</strong>g the whole reproduction cycle by<br />
weekly semen evaluations. The food of experimental groups was<br />
supplemented with 0.3 ppm Sel-Plex (Alltech) and 200 ppm vitam<strong>in</strong><br />
E, while the food of control groups conta<strong>in</strong>ed sodium selenite <strong>in</strong><br />
traces and 100 ppm vitam<strong>in</strong> E. Males of 26 weeks of age were placed<br />
<strong>in</strong> <strong>in</strong>dividual cages feed<strong>in</strong>g and keep<strong>in</strong>g accord<strong>in</strong>g to the management<br />
manual, until 61 weeks of age. Sperm collections were ma<strong>de</strong> twice a<br />
week by dorso-abdom<strong>in</strong>al massage accord<strong>in</strong>g to Burrows and Qu<strong>in</strong>n<br />
(1937). Concentrations were <strong>de</strong>term<strong>in</strong>ed by spectrophotometer<br />
(Accucell, IMV Technologies), motility by subjective scor<strong>in</strong>g from 0<br />
to 5, morphology of spermatozoa and live/<strong>de</strong>ad cell ratio <strong>in</strong> sta<strong>in</strong>ed<br />
smears by anil<strong>in</strong>e eos<strong>in</strong>. As a result, the 0.3 ppm organic selenium and<br />
200 ppm vitam<strong>in</strong> E affected differently <strong>in</strong> the two types of males.<br />
While <strong>in</strong> ROSS males they improved significantly the sperm motility,<br />
concentration and the ratio of live, morphologically normal<br />
spermatozoa <strong>in</strong> the second half of the production cycle, <strong>in</strong> the case of<br />
Hubbard males there were no any significant improvements <strong>in</strong> sperm<br />
traits. However, significant differences were found between the sperm<br />
qualities of the two types regard<strong>in</strong>g to the sperm volumes (.18 - .25<br />
ml/ejaculate), the concentrations (4.9 – 1.9 million/µL), and the <strong>de</strong>ad<br />
cells’ ratios (15 – 17.8 %) <strong>in</strong> ROSS vs. Hubbard control males,<br />
respectively. Thus, Hubbard males produced higher semen volume<br />
with significantly less sperm concentration, less abnormal sperm cells<br />
but significantly more <strong>de</strong>ad cells, than ROSS males. As a<br />
consequence, Sel-Plex with vitam<strong>in</strong> E could ma<strong>in</strong>ta<strong>in</strong> the <strong>in</strong>itial good<br />
sperm quality of ROSS males until the end of the cycle. However, <strong>in</strong><br />
the case of Hubbard males the spermatological performance seemed<br />
to be more stable dur<strong>in</strong>g the reproduction cycle and less accessible by<br />
exogenous factors.<br />
Poster 11 - <strong>Reproduction</strong> of Other Vertebrates (Fishes,<br />
Amphibians, Reptiles)<br />
P344<br />
Endogenous opioid system and sharpsnout seabream<br />
(Diplodus Puntazzo) milt: <strong>de</strong>tection of mu, <strong>de</strong>lta and<br />
kappa opiod receptors on sperm cells<br />
Aiudi, G*; De Sandro Salvati, A; Bucci, FA; Micera, E; Albrizio, M<br />
Department of Animal Production, University of Bari, Italy<br />
Sharpsnout seabream, (Diplodus puntazzo) is the third most farmed<br />
mar<strong>in</strong>e teleost species <strong>in</strong> Italy (Tad<strong>de</strong>i et al., Cryobiology 42:244,<br />
2001). Fish reproduction <strong>in</strong> aquaculture facilities is ma<strong>in</strong>ly obta<strong>in</strong>ed<br />
through environmental/pharmacological condition<strong>in</strong>g. These<br />
techniques may stress fish and <strong>de</strong>crease reproductive performances<br />
(Cleary et al., Aquac. Res. 33:829, 2002). Endogenous opioid pepti<strong>de</strong>s<br />
(EOPs) system is <strong>in</strong>volved <strong>in</strong> stress response (Arends et al., J.<br />
Endocr<strong>in</strong>ol., 163:149, 1999), and affects several physiological