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Preservation of Liquid Semen and Arti?cial Insemination by Liquid and Frozen Semen in Native Sheep

Dr. Azharul Islam Talukder
CSO
Goat and Sheep production research division, BLRI, Savar, Dhaka.

Md. Nazmul Hasan,
SO
Conservation and improvement of native sheep through community and commercial farming project, BLRI, Savar, Dhaka.

Md. Rakibul Hasan
SSO
Poultry Production Research Division, BLRI, Savar, Dhaka

Shahin Pervage

Abstract/ Executive Summary:

An experiment was undertaken to determine the conception rate of native sheep by using Artificial Insemination with liquid ram semen. The semen was collected from ram using artificial vagina and was stored in a refrigerator (4⁰C) for three days. The volume of semen was extended with egg yolk citrate diluter. A total of 63 ewes were inseminated with stored liquid semen collected from 15 rams by AV method. The total number of spermatozoa, live-dead, normal-abnormal, sperm motility and the pH was observed regularly. The average semen volume per ejaculate was 0.76-1.00ml and the sperm concentration was 2.37x109 – 4.30x109 per ejaculate. The number of normal spermatozoa and the pH was almost similar irrespective of days of storage. Number of live spermatozoa and the sperm motility were reduced with the increasing age of semen. The average conception rate (%) was obtained as 63.61, 61.90, 52.38 and 47.61 with sperm in zero, 1st, 2nd and 3rd day storage respectively.

Keywords: Native sheep, Artificial insemination, Liquid semen, Frozen semen

Location: Goat and Sheep Production Research Division, BLRI, Savar, Dhaka,

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Sheep

Objectives:

a) To know the conception rate of native sheep by using AI technique

b) For reducing the inbreeding problem and increasing the purity of breed

c) For proper use of outstanding ram

d) For storage of ram semen

Methodology:

A total of 15 rams and 63 ewes were used in this experiment where the ewes were inseminated with diluted liquid (4⁰C) semen of either zero (0), 1st, 2nd and 3rd day storage. Concentrate was fed twice daily @ 200g per head with 7-8 hours grazing per day. Rams were always kept separately from the ewes. Semen was collected from each ram twice a week. The volume of semen was measured with a calibrated collection vial. Sperm concentration was measured by haemocytometer method. The pH of semen was measured directly with the aid of nitragene paper while the individual motility of sperm was observed under microscope. Then the semen was mixed with diluter (Egg-yolk-citrate), fixed the sperm concentration at 300x106/ml and finally kept in a refrigerator (4⁰C). The semen was further evaluated for its quality in zero (0), 1st, 2nd and 3rd days in terms of motility, pH; presence of live, dead and normal, abnormal sperm. For counting the presence of live-dead and normal-abnormal sperm in the semen, Eosin-nigrosin and Rose Bengal stain were used. After completion of microscopic evaluation, semen sample was diluted with egg-yolk citrate (each 100ml diluter contained 20ml egg-yolk, 80ml 2.94% sodium-citrate buffer, penicillin @1000IU/ml, streptomycin @1000mg/ml, 6mg/ml sulphanilamide powder), fixed the sperm concentration at 300x106/ml and kept in refrigerator (4⁰C) up to three days. The diluted fresh semen at 1st, 2nd and 3rd days of preservation were evaluated for sperm motility, pH and presence of live, dead and normal, abnormal sperm using the same procedure. The estrus of the ewes was observed twice everyday with the aid of a vesectomized ram and inseminated after 12 hours from the onset of heat. The ewes that were in heat kept in pan separately. Cervical insemination was performed after 12 hours of showing the heat. Insemination with diluted semen was performed using pipette attached with a syringe without needle. Before the semen was expelled out of the insemination pipette, the speculum was removed to avoid pneumovagina and subsequent discharge of semen. Assumed conception rate was calculated on the basis of the ewes not coming into heat again within forty days. Data were treated and statistically analyzed by using Statistical Package for Social Science (SPSS, version 11.5). Mean comparisons were made using Duncan multiple range test (DMRT).

Source of Information: BLRI. Annual Research Review Workshop 2009-10. BLRI, Savar, Dhaka

Article Link (Online Journal):

Funding Source:

1. Government

Budget: 250000

Total Budget (Tk.) : 250000

Achievement/Findings:

The semen volume, sperm concentration and no. of spermatozoa per ejaculate were 0.5 to 1.20 ml/ejaculate, l.9x10⁹ to 2.2x10⁹ sp/ml and 0.96x10⁹ to 3.30x10⁹ sp/ml, respectively. The sperm concentrations of raw semen were l.9x10⁹ to 2.2x10⁹ sp/ml. The average volume of semen per ejaculate was 0.85 ml. Conception rates to Al of cervical fresh and double cervical frozen methods of arti?cial insemination. The conception rate obtained using fresh diluted semen was comparatively higher (52.38%) than the cervical frozen semen (25.00%). The acceptable conception rates (52.38%) of Al were achieved with cervical fresh group while the conception rate (25.00%) for cervical frozen semen was extremely poor. Low conception rates following cervical insemination of frozen semen in ewes is attributable to the combined influences of poor sperm transport through the cervix and lower than optimum numbers of motile sperm in the inseminate. Conception rates with fresh or chilled semen are good (65 to 75%) but unacceptably low (10 to 30%).

Knowledge Management: Report/Proceedings