Comprehensive Semen Evaluation for Male Fertility Assessment
Explore the detailed process of semen evaluation for male fertility, including gross examination, microscopic analysis, and biochemical assessment. Understand the factors affecting semen quality, such as volume, color, and motility, to assess reproductive health effectively.
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Presentation Transcript
SEMEN EVALUATION Evaluation of male fertility Evaluation the semen before artificial insemination Evaluation the effect of genital disease on male reproductive system Evaluation the effect of season on male reproduction Gross evaluation Microscopic evaluation Biochemical and biological evaluation
GROSS EXAMINATION Directly after semen collection Must be put in water bath in 37 C The color Volume Density Admixture (foreign bodies which may be mix with samples) For artificial vagina For male reproductive system
The color of semen The color of semen depend on the concentration of sperm Creamy or white creamy 0.75 - 1 million sperm / mm Milky 0.5 0.75 million sperm / mm Opaque or Cloudy 0.25 0.5 million sperm / mm Watery 0 0.25 million sperm / mm Abnormal color Yellowish orange presences of riboflavin (seminal plasma) Yellowish presences of urine Reddish presences of blood Greenish presences of feces Brownish presences of pus with necrotic tissues
The volume of semen ejaculation For bull 5-10 cm3 For ram 1-2 cm3 For buck 0.1-1 cm3 For stallion 50-100 cm3 The main factors which control the volume of semen ejaculation Species of animals The percentage of excitation Season The number of weekly ejaculation
Microscopic examination Sperm motility Mass motility Individual motility Sperm morphology Sperm concentration
Sperm motility Mass motility: the wave motion of sperms as a result of union many sperms in one direction which appear as a dark wave There are many grades for mass motility as follow: Grade 0 no any movement (0%) Grade 1 very few movement (<20%) Grade 2 mild movement (shaking movement) (20-50%) Grade 3 a movement with pale waves (50-70%) Grade 4 a movement with brownish waves (between pale and dark) (70-80%) Grade 5 a movement with dark waves (>80%)
Sperm motility Mass motility Diagnose directly after collecting Under light microscope under power X10 At first using warm slide or hot plate for slide
Sperm motility Individual motility: the movement of sporadic sperms in all directions of slide which give the real sperms movement inside the female reproductive system There are four types of individual movement: Progressive forward movement (important movement that shows the ability of sperms to reach the fertilization site( Progressive back down movement Non-progressive lateral movement Circular movement
Sperm motility Individual motility There are many grades for individual motility according to the number of sperms which move progressively forward: Grade 1 no any movement (0-5%) Grade 2 very few movement (20-40%) Grade 3 half of sperms move (40-60%) Grade 4 more than half of sperm move (60-80%) Grade 5 most sperms move (80-100%)
Sperm motility Individual motility It want semen dilution (1:20) or more Under microscope under power X40 Warm slide as well as cover slide
Determination the percentage of dead and live sperm Reflect the quality of semen and the ability of this sample for fertilization A special stain is important to determine the percentage of dead and live (the dead sperms absorb the stain due to the permeabilization of its membrane not like live sperm The acceptable ratio for dead sperm not more than 20% Special stain like eosin with blue aniline and nigrosine
Sperm morphology Sperm abnormalities ( 4-10%) and not more than 20% Primary abnormalities (occur during spermatogenesis) Genetics Diseases Environmental factors Nutritional factor Increasing in testis temperature Interfere with fertilization capacity It include : pyriform head, tapering head, narrow head, micro head, broken mid piece, enlarged mid piece, double head
Sperm morphology Sperm abnormalities Secondary abnormalities (occur outside testis especially during storage inside epididymis or ampulla) Problems in nutrition Interfere with fertilization capacity It include : cytoplasmic droplets
Sperm morphology Sperm abnormalities Tertiary abnormalities (occur during prepare slide or cold shock or osmotic shock (bent tail) or detach acrosome Doesn t reflect the ability of sperm for fertilization Include broken head, broken tail
Sperm morphology Sperm abnormalities For determination the sperm abnormalities Diluted semen (1:20) Special stain (eosin and nigrosine)
Sperm concentration Determine the fertility of male Determine the suitable concentrations of diluters or extender The effect of season The are many methods for this purpose: Direct sperm count Spectrophotometer Electronic Particle Counter
Sperm concentration Direct sperm count Hemocytometer Sperm dilution (1:200) (normal saline 0.9 NaCl) HgCl ( kill sperms to stop any movement) Ordinary method for counting like RBC counting Use 25 square ( if you used 5 squares should be X 5) No. sperm/ml = no. sperm in 0.1 mm3X 10 X dilution rate X 1,000
Sperm concentration Spectrophotometer (depending on Light transmittance) Calculate the intensity of light which reflected from the sample Electronic Particle Counter The instrument can be adjusted for particle size so that only the sperm cells in a sample will be counted
Biochemical and biological evaluation This type of evaluation depend on the measuring the activity rate of the sperms after ejaculation especially the metabolism of sperms This evaluation include: Oxygen take-up Fructolysis index PH Methylene blue reduction test
Biochemical and biological evaluation Oxygen take-up The amount of oxygen which are consumed by 100 million sperms per minute Especial instrument calculate that (microspirometer, Bercroft) If this ratio increase so the sample is ????
Biochemical and biological evaluation Fructolysis index The consuming of fructose is deal with the activity of sperms Determine directly after ejaculation After that incubate in 37 C for 1 hour Determine the amount of fructose which consumed The index for 1000 million sperms The normal value 1.4-2 mg/ hr
Biochemical and biological evaluation PH Any changing in PH is mean the activity of sperms and formation of lactic acid under oxygenated condition After ejaculation the amount of lactic acid is 63mg/cm3 which lead to form PH 6.6-6.9 After a time of incubation the PH will be decrease as a result of lactic acid formation
Biochemical and biological evaluation Methylene blue reduction test Reduction time depending on quality of semen The releasing of H ions from sperms reduce the stain from blue to colorless Good quality semen reduce the stain after 3-6 min. Poor quality semen reduce the stain after 9 min.
