Semen Collection and Evaluation Flashcards

1
Q

when do we do fertility evaluation?

A

 PREPURCHASE
 PREBREEDING
 QUESTIONABLE FERTILITY

Routinely in management of valuable stallions and bulls

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2
Q

where and how do we collect semen?

A

GOOD FACILITY – safety
* GOOD EQUIPMENT *TRAINED PERSONNEL
* METHODS:
 artificial vagina (AV) – stallions, bulls, small ruminants, camelids, rabbits, toms
 electro-ejaculator – bulls, small ruminants, toms & zoo animals
 manual – dogs, boars, poultry

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3
Q
  • Factors Affecting Semen Quality:
A

 Health of the animal
 Season of the year
 Testicular size
 Frequency of collection
 Intensity of teasing
 Method of collection
 Method of handling
 Environmental temp

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4
Q

Semen Handling principles - after collection

A
  • Pre-warm all slides, tubes etc.
  • Warm microscope stage
  • Keep semen warm
  • Evaluate quickly
  • Avoid contamination with lube, soap, dirt etc.
  • All-plastic syringes
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5
Q

How do we “get the goods”?

A
  • Species dependent
  • Bull,Buck,Ram– Electroejaculator or trained to artificial vagina (AV)
  • Stallion – AV (NO EEJ)
  • Boar and Dog– manually into an AV
  • Best to have a female in estrus present
  • Stallion – AV (NO EEJ)
  • Boar and Dog– manually into an AV (exception bulls for EEJ)
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6
Q

SEMEN EVALUATION metrics

A
  • COLOUR
  • VOLUME
  • pH
  • MOTILITY
  • CONCENTRATION
  • MORPHOLOGY
  • CYTOLOGY
  • SEMEN CULTURE
  • Advanced tests – eg. CASA, Fluorescent stains – acrosome, membrane integrity, DNA
    *Oocyte penetration assay
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7
Q

how do we evaluate semen colour? what factors affect this

A

CREAMY, MILKY, SKIM MILK, WATERY

ABNORMAL: Yellow (Pus, Urine) Red/Brown (Pus, Blood)

Factors affecting:
- Contamination
- Frequency and method of collection (determines concentration in sample)

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8
Q

if semen has a red colour, what does this point to?

A
  • Blood–red-brown or red

Stallions:
* Urethral lacerations
* Urolithiasis
* Penile lesions
– Trauma
– Coital exanthema
– Tumours

Dogs: prostatitis, penile lesions

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9
Q

what creates variation in semen volume for collection?

A
  • Varies between species
    – Eg. 2-4 mls for bull, 60-100 mls for stallion
  • Method of collection
     AV vs EE
     Mixing fractions
  • Frequency of collection
  • Teasing intensity – stallion
    *Season – seasonal breeders
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10
Q

how do we measure semen motility?

A
  • Gross Motility – place a drop on a slide - - waves and eddies – only in species with highly concentrated sperm (bull, ram, buck)

Individual Motility – with coverslip:
* Total Motility - % of cells that are moving
* Progressive Motility - % of cells moving forward in a progressive manner

  • Velocity
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11
Q

how do we score gross motility for semen? What species?

A
  • Swirling, Waves, Eddies seen under low power without a coverslip

Bull, Ram, Buck
Good gross motility is a factor of high concentration, a large percentage of progressively motile cells, and high velocity of movement

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12
Q

individual sperm motility is used as a metric for what species? what are the subcategories?

A

Stallion, dog, boar, bull, ram, buck

  • Total Motility - % of all cells that are moving in any direction
  • Progressive Motility - % of cells moving forward in a progressive manner
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13
Q
  • Factors Affecting sperm motility:
A

 Morphology
 Temperature – cold/heat shock
- use prewarmed, clean slide, warm stage
 Time between collection & evaluation
 Detergents, lubricants, latex, water
 Contamination with dirt, epithelial cells

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14
Q

what does sperm concentration allow us to calculate?

A
  • Allows calculation of total number of sperm in ejaculate - expected range or not
    > Calculation of breeding doses
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15
Q

methods of determining sperm concentration?

A
  • Hemocytometer
  • Densimeter
  • Spectrophotometer (Spermacue)
  • Nucleocounter (also does membrane integrity)
  • CASA
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16
Q

Factors Affecting sperm Concentration

A
  • Season – esp. seasonal species (stallion, ram)
  • Amount of teasing (increased teasing results in increased seminal plasma volume — lowers concentration but overall usually increases total numbers)
  • Dog - Collection of all fractions together versus separating them out
  • Bull – Method of collection - EEJ vs. AV
17
Q

how do we look at sperm to assess morphology?

A

*Ideal – wet mount under Oil Immersion using DIC microscopy
*Eosin – Nigrosin stain
* Oil immersion - 100X magnification
* Count minimum of 100 cells
* % of normal and abnormal cells

18
Q

types of sperm morphological defects we look for?

A

Defects – Head, Mid-piece, Tail, Proximal Droplets, Distal Droplets, Loose Heads

19
Q

what are ‘droplets’ on sperm?

A

Droplets are the remnants of spermatid residual cytoplasm that remain attached

20
Q

what can we see on sperm cytology and what can this tell us? how do we do it?

A

*Diff-Quick WBC stain on feathered edge

  • Cytology Scores (Cells/hpf):
    0= <1 ; 1+ = 1-3 ; 2+ = 4-6 ; 3+ = 7-10
    4+ = >10

Neutrophils > Culture

Round cells (immature spermatogonia) – associated with testicular degeneration