Sperm Evaluation

Question 1

Appearance and volume

Answer 1

= milky white single cream

• watery/grey = low conc of sperm
• seminal vesiculitis = clumped, blood and consist of discharge (inflamtion of seminal vesicles
• abnormalities ~ blood = haemospermia, urine = urospermia
• volume 30-250ml
• gel fraction = 20-40%

Question 2

Motility

Answer 2

• semen extended before assessments, otherwise clumps

- % progressive forward motility = >60% good

Question 3

Concentration

• ideal
• how to measure

Answer 3

30-600 x 10^6 sperm/ml
for fertility need at least 100x10^6 (AI = 100-200x10^6

• measure on haemocytometer invented by Louis-Charles Malassez
• thick glass microscope slide with 2 chambers engraved with perpendicular lines
• diluted sperm sample 1:100 loaded by capillary action into chamber and count sperm on grid
• known depth/volume/size (translate to original sample)
• A: inexpensive, discolouration of sample not affect acuracy of count
• counting takes a long time, variability from dilution/loading errors common

Question 4

Morphology

• Nigrosin
• primary, secondary and tertiary abnormalities
• normal % for AI

Answer 4

• live:dead ~ Nigrosin = eosin vital stain indicates % live
• can visualise abnormalities
• primary abnormalities = indicate problems with spermatogenesis/ sperm maturation (e.g. extra/no tails, cytoplasmic droplets
• secondary abnormalities = indicate sperm damage at ejaculation (bent tails, detatched heads/tails)
• tertiary = problem with semen collection/handling
• 65%

Question 5

Semen collection - what must the equipment be like

Answer 5

• sterile
• warmed to 37 degrees C
  = not cold shock sperm or contaminate semen quality
• keep graduated cylinders in incubator

Question 6

Computer assisted sperm analysis

Answer 6

• developed in 1980s
• greater repeatability than visual estimates is (closeness of the agreement between results of successive measurements of the same measure)
• capture sperm frame by frame playback video

Question 7

pH

• normal range for sperm
• influenced by
• what is high ph associated with

Answer 7

• pH meter
• normal = 7.2-7.7
• influenced by: season, freq of ejaculation, sperm concentration
• negative correlation between seminal volume and pH/ between no. spermatozoa in ejaculate and pH
• ( increase ph = decrease spermatozoa)
• high pH associated with urospermia or infection in genital tract

Question 8

Microbiology and cytology

- which bacteria can cause infection to mare

Answer 8

• pseudomonas aeruginosa
• Klebsiella pneumoniae
• both cause pneumonia and blood infections
• wash penis or use antibiotics on infected semen for AI

Question 9

Longevity of sperm motility

Answer 9

• longevity tests determined at 5 degrees C

- cri

Question 10

Longevity of sperm motility

• temperature used now
• indicates
• ideal sperm motility after 24 hours being cool
• influenced by

Answer 10

• longevity tests determined at 5 degrees C
• critical for cool shipped semen
• indicates how well sperm tolerate stress in environment = relate to lifespan in female repro tract
• knowing longevity determines optimal time period for shipping and insemination
• idealy retain >30% motility after 24 hours
• influenced by: dilution factor, final concentration of extended semen