Step-by-Step Guide to Fresh Semen Artificial Insemination in Dogs

July 26, 2025

How to Perform Fresh Semen AI in Dogs

A practical step-by-step guide for breeders on using fresh semen artificial insemination safely and effectively.

Artificial insemination (AI) using fresh semen is a safe, simple, and effective way to breed dogs — especially when natural mating isn’t ideal. Done correctly, this method maintains semen quality, reduces stress on the dogs, and increases success rates with the right timing.

Here’s a clear, step-by-step guide to help you confidently perform fresh semen AI at home or in a clinic setting.

What You’ll Need

Before you begin, gather all equipment to ensure a smooth and hygienic process:

Semen collection cone and sterile container
Semen safe syringe (ideally 10–20 mL)
Insemination tube (pipette)
Sterile semen safe lubricant
Gloves
F10 disinfectant for surfaces
Paper towel or wipes
Tail wrap (optional) - Cohesive bandage
Helper/handler (recommended)

Step 1: Prepare the Area and Equipment

Choose a quiet, clean, stress-free area for both dogs. Disinfect all surfaces with F10. Lay out all equipment.

Wear gloves, and ensure everything is within reach.

Step 2: Collect the Semen

Ensure the stud is excited by allowing him to approach the females rear.

Use a collection cone/sleeve to collect semen manually. Do this by gently sliding the skin covering the penis back to expose the bulb of the penis. Slide the collection sleeve onto the dogs penis and hold gentle steady pressure above the bulb of the penis to encourage an erection. Allow him to thrust or if he needs encouragement, stimulate above the bulb of the penis.

Only the second fraction (sperm-rich) is needed — stop collection before the third watery fraction begins. Once this is collected, remove the collection sleeve.

Keep the sample warm (body temperature) and protected from light and cold air.

Transfer to the sterile syringe immediately if inseminating within minutes.

Note: If inexperienced, ask a vet or mentor to assist until you’re confident.

Step 3: Restrain and Position the Female

Have a second person gently hold the female. The female should be stood with her back legs behind her pelvis much like a show stack. This flattens the vaginal vault, making insertion of the pipette to the cervix easier.

Step 4: Load the Semen

Attach the insemination pipette to the syringe preloaded with semen

Apply a small amount of sterile lubricant to the tip of the pipette

Step 5: Insert the Pipette

Gently guide the pipette (tube) into the top of the vulva to avoid urethral entrance. Be gentle and travel up, then flat, then slightly down.

Step 6: Inseminate the Semen

Slowly depress the syringe plunger to deposit the semen. Then fill the syringe with air or semen extender and push any remaining semen through the pipette into the vulva.

Once done, remove the pipette gently

Hold the female's rear up for 5–10 minutes and massage the vulva upward to help semen stay in place

Keep the female standing quietly for 10–15 minutes without toileting or sitting.

Timing Tips

Fresh semen should ideally be inseminated

Within 15–30 minutes of collection
During the female’s fertile window (based on progesterone or LH testing)

Repeat the AI in 48 hours for best results (depending on ovulation timing).

Final Thoughts

With the right technique, timing, and hygiene, fresh semen artificial insemination is an excellent breeding method for both novice and experienced breeders. It’s safe, efficient, and maximises conception without the stress of natural mating.