1. Semen Analysis
Clinical Pathology

2. Semen Collection
Semen is often collected into an artificial vagina, usually while a teaser bitch is present.
An artificial vagina may be made of latex or disposable plastic.

3. May use electoejaculation in which a probe is attached to the pelvic nerves to stimulate ejaculation.

5. Semen (3 fractions) 1st- mainly prostatic fluid, few sperm
Released during the period of vigorous thrusting
2nd- Sperm rich portion of ejaculate
3rd- Mainly prostatic fluid, few sperm, majority of total volume of ejaculate.
Buck, bulls, tom, ram you should collect all three
Stallions, dogs, boars- collect 2nd and 3rd portions seperately

6. Semen Handling Techniques
Avoid marked temperature changes
Avoid exposure to water, disinfectants
Use clean, dry, warm equipment (37 C or 98 F)
Slides, coverslips, pipettes, stains.
Process soon after collection

7. Semen Evaluation
Color and consistency (normal is milky and moderately viscous)
Volume
Wave motion/sperm motility
Spermatozoa concentration
Morphology
Ratio live:dead
Presence of foreign cells/material

8. Volume of Ejaculate Measured in volumetric flask
Method of collection affects volume
Electroejaculation- volume is larger
Teasing with a female-volume is larger
Species variation
Dogs: ml
Stallion: 65 ml
Tom: 0.5 ml
Volume does not necessarily correlate with fertility

9. Sperm Motility Motility correlates with fertility
Improper handling can affect motility
Evaluate immediately after collection
Place a drop of semen on a warm slide, immediately cover with coverslip
Dilute with warm saline if high concentration of sperm

10. Classes of Sperm Motility
Can be classified as good, very good, fair, or poor.
Normal sperm should have greater than 70% motility
Examine under 100 x, may need to dilute concentrated samples
Poor is when there is less than 40% motility

11. Wave Motion Under low power 40x, look for swirling
Progressive motility- sperm are moving around all over slide
Non-progressive motility- sperm are only swimming in a similar pattern

13. Sperm Concentration Most important characteristic
Dilute a portion 1:100 with saline or red cell Unopette.
Using a hematocytometer, count total of sperm in the central grid
Multiply the number by 2 million
Boars/Stallions: 150 M/ml
Bulls: M/ml
Dogs: 300 M/ml
Cats: M/ml

15. Live:Dead Sperm Ratio
Place 1 drop eosin/nigrosin stain (make sure it is warm) and mix gently with a drop of semen on a warm slide.
After several seconds, smear like blood smear.
Live sperm resist staining-appear white against a blue-black background
Dead sperm take up the eosin and stain pinkish red
Examine and observe 200 cells

17. Sperm Morphology Can examine on eosin/nigrosin stained smear
Other stains: india ink, H&E, Wrights
Observe cells
Record % of abnormal cells
Divide problems into head, neck, midpiece, and tail problems.
Primary abnormalities occur during sperm production.
Secondary occur from storage in the epididymis until the smear is made

19. Double headed Sperm

20. Misshapen Head

21. Elongated Head

22. Pear shaped head and bent midpiece

23. Proximal Droplet

24. Distal Droplet

25. Detached Head

26. Bent Tail or Midpiece

27. Coiled Tail

29. http://www. vivo. colostate. edu/hbooks/pathphys/reprod/semeneval/conc