الفهرس 
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Abstract
The current study was conducted at the International Live Stock Mangement Training Center (ILMTC), Sakha, belonging to the Animal Production Research Institute, Agricultural Research center, Ministry of Agriculture in cooperation with the dept. of Animal Production, Faculty of Agriculture, Mansoura University. The study aimed to compare different methods of evaluation of Holstein and buffalo semen through assessment of conventional methods for evaluation physical, chemical and morphological semen characteristics, along with Sphadex column filtration technique and osmotic shock at hypo and hyper osmotic levels. Semen was collected twice weekly from each of five healthy mature Holstein and five buffalo bulls as a rotein work of ILMTC. Immediately, after semen collection, one ejaculate was taken from each bull for 10 weeks (50 ejaculates for each species). The collected semen was transferred to a water bath at 37C(R@? (BSemen was evaluated for physical semen characteristics including, ejaculate volume, sperm cell concentration and percentages of gross motility, progressive motility, live, abnormal and intact acrosome spermatozoa. Grade of motility and sperm morphometric characteristics and biochemical characteristics of the seminal plasma were also determined. Sephadex filteration technique and osmotic test was also conducted on raw semen. Osmotic test designed to establish various levels of osmolarity and incubation times that would give the maximum percentage of curled spermatozoa at different hypoosmotic levels, (0, 50, ,100, 150, 200 and 300 mOsm) or to establish percentage of shrunk spermatozoa at hyperosmotic levels(400 and 600 mOsm) at 0, 15, 30, 45 and 60 minutes incubation times. Results:1. Physical characteristics of raw semen: (1).The present results indicated insignificant differences in physical characteristics of raw semen between Holstein and bufflo bulls including percentages of gross and progressive motility, grade of motility and percentages of live spermatozoa. However, ejaculate volume and percentage of sperm abnormality were significantly (P>0.001) higher by about 48% and 31.77%, respectively, in Holstein than buffalo bulls and average sperm cell concentration and spermatozoa with intact acrosome was significantly higher in buffalo than Holstein semen by about 24% and 4.48%, respectively. (2).Total count of spermatozoa and total out put of motile and live spermatozoa were significantly (p<0.05, P<0.01 and P<0.05, respectively) higher by about 17.18, 19.3 and 17.4% in Holstein than in buffalo buls. However, total out put of normal and intact acrosome spermatozoa did not differ significantly between both species. (3).All morphometric characteristics of spermatozoa including total length (67.35 vs. 65.17(So(Bm ) length (8.6 vs. 7.7 (So(Bm), breadth (4.3 vs. 3.9 (So(Bm) of head as well as lenth of middle (13.1 vs 12.0 (So(Bm) main (40.8 vs. 38.9 (So(Bm) and terminal (3.4 vs. 3.1 (So(Bm) pieces and head area (32.9 vs. 26.3 (So(Bm)were significantly higher in Holstein than buffalo spermatozoa. However, neck length and breadth/length of head did not differ significantly between both species (1.4 (So(Bm and 0.50 (So(Bm2, respectively). (4).Concentration of albumin showed the highest difference between both species, being significantly (p<0.001) higher in Holstein than buffalo seminal plasma by about 54%, while total proteins, globulin and phospholipids concentrations were significantly (p<0.001) higher in Holstein than buffalo seminal plasma by about 34, 20 and 38%, respectively. However, fructose concentration was significantly (P<0.001) higher by about 26% in buffalo than Holstein seminal plasma. (5).Activity of GOT, GPT and LDH was highly significant (P<0.001) in buffalo (52.6, 28.6 and 499.7 U/L) than Holstein (44.6, 24.3 and 390.9 U/L) seminal plasma . However, GOT/GPT ratio did not differ significantly between both species (1.87).