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Veterinary Ozone Protocol: Horses and Cattle
Systemic and local routes Madrid gives for horses and cows: autohemotherapy, rectal and vaginal insufflation, joint and spine injections, bagging, ozonated water, oil and mastitis.
Quick answer: Madrid gives autohemotherapy for horses and cows at 20–45 µg/mL, treating 2% of total blood volume. Treatment is 2 times a week for 10–20 treatments. The maximum is 45 µg/mL for blood, 40 for rectal and 35 for vaginal.
For veterinarians. Ozone use in animals should be done or directed by a licensed veterinarian. This sheet is a reference for them. It is not a guide for treating your own animal and not veterinary advice. Check your licensing board and local rules before using any route.
What are the basics of ozone therapy in horses and cattle?
- Madrid groups horses and cattle in one chapter and gives them the same dosing. Both have about 80 mL of blood per kg (0.08 L/kg), which is the number the blood doses are based on.
- Major autohemotherapy (MAH) treats about 2% of the animal's total blood volume. Madrid notes this is often hard to draw, and suggests rectal insufflation as a partner method when it is.
- Cattle have one extra route, intramammary infusion for mastitis, which Madrid describes as a way for dairy producers to cut antibiotic use.
- Condition-by-condition horse doses (hooves, joints, tendons, back, skin, foals, reproduction) are on the horse conditions sheet.
Sources: Madrid Declaration on Ozone Therapy, 4th edition (ISCO3, 2025), Addendum B (Veterinary Medicine), paraphrased, plus the published studies listed under More detail.
What dose and settings are used for ozone therapy in horses and cattle?
Systemic routes (horses and cows)
Concentrations in µg/mL (µg/NmL in Madrid).
| Route | Concentration | Volume | Frequency and course |
|---|---|---|---|
| Major autohemotherapy (MAH) | 20–45 | 2% of total blood volume. Total blood = 0.08 L × body weight in kg. | 2x/week, 10 to 20 treatments |
| Minor autohemotherapy (MiAH), 25 to 100 kg | 10–30 | 15 to 20 mL | 2x/week, 10 to 20 treatments |
| Minor autohemotherapy (MiAH), over 100 kg | 10–30 | 20 to 30 mL | 2x/week, 10 to 20 treatments |
| Rectal insufflation | 10–40 | 5 mL per kg | 2x/week up to once a day, 10 to 20 treatments |
| Vaginal insufflation | 10–35 | 8 mL per kg | 2x/week up to once a day, 20 treatments. Rinse with ozonated double-distilled water first. |
Worked example from Madrid's MAH formula: a 500 kg horse has 0.08 × 500 = 40 L of blood, and 2% of that is 0.8 L, or 800 mL, to treat. Madrid says the concentration depends on the animal's oxidative stress and the condition being treated. Madrid's text does not give an upper limit on the total gas dose, so a veterinarian sets it from the animal's size and response.
Local routes
| Route | Concentration | Volume and time | Frequency and course |
|---|---|---|---|
| Intramammary (cows, mastitis) | 35 | 1 L (35,000 µg total) | Once a day, 10 to 20 treatments |
| Bagging, to heal | 10–30 | Fill the bag; 20 to 30 min | 2 or 3 times a week, or daily, as the wound responds |
| Bagging, purulent infection | 50–60 | Fill the bag; 20 to 30 min | 2 or 3 times a week, or daily, as the wound responds |
| Intra-articular, degenerative | 10–12 | By joint size | 1 or 2 times a week, 10 to 20 treatments |
| Intra-articular, inflammatory | 15–20 | By joint size | 1 or 2 times a week, 10 to 20 treatments |
| Subcutaneous, degenerative | 5–10 | By area treated | 1 or 2 times a week, 10 to 20 treatments |
| Subcutaneous, inflammatory | 15–20 | By area treated | 1 or 2 times a week, 10 to 20 treatments |
| Paravertebral and muscle, 25 to 100 kg | 10–20 | 10 to 20 mL | 2 or 3 times a week for 9 treatments, weekly for 4, then every 15 days; 20 in all |
| Paravertebral and muscle, over 100 kg | 10–20 | 20 to 40 mL | 2x/week for 4, weekly for 8, then every 15 days; 20 in all |
| Fistula (fill the tract) | Infected 40; inflammatory 15–20 | Enough to fill the fistula | Daily or 3x/week, as the case responds |
| Ozonated water for cleaning wounds and cavities | 12–20 | Use right away; it loses about half its strength every 5 to 10 min | As needed |
| Ozonated oil on wounds and skin | PI 400 to 1,200 by stage (see the dogs and cats sheet) | Thin layer | Generally once a day |
Madrid's equine table (see the horse conditions sheet) lists horse joint doses of 3–8 mL at 20–35 µg/mL, higher than the general joint figures here (10–12 degenerative, 15–20 inflammatory). Start at the low end. Madrid also gives two time ranges for wound bagging: 20–30 min in this general section and 5–10 min in the equine wound table. Start with the shorter time and lower concentration.
Madrid describes the paravertebral schedule as twice a week for two weeks, then weekly for four to six weeks once the animal improves, then every 15 days until 20 sessions, and says the course can be shortened when signs resolve. Inject 0.5 to 2 cm to the side of the spine, on both sides, and check the muscles first for tender trigger points, which should be injected too. Distilled water for ozonated water: about 60 µg/mL can be reached at 14 °C in a sealed glass container in 10 to 15 min, but use 12 to 20 µg/mL on the animal.
What are the best tips when using ozone therapy in horses and cattle?
- Rectal insufflation is the simplest method, and works as a systemic route when MAH is not practical. Madrid also lists it for colitis, proctitis, and anal fistulas or fissures.
- Do the maths from body weight first. Madrid gives the MAH volume (2% of blood volume), rectal and vaginal volumes (5 and 8 mL per kg) and the concentration range, then the veterinarian picks the concentration for the condition.
- Madrid asks you to check the muscles for trigger points before paravertebral work, and to inject them as well.
- Close the bag around a wound only after the area is moist and cleaned, as Madrid describes.
What are the precautions for ozone therapy in horses and cattle, and who should avoid it?
- Fistulas: make sure the fistula does not connect with the airway. Gas can build up in a closed or cyst-like cavity and raise pressure sharply, which hurts and can be dangerous.
- Mastitis: Madrid's mastitis dose (35 µg/mL, 1 L) is 20 times the gas volume in the Argudo trial (50 mL at 35 µg/mL). The 2000 study used 1 to 5 L per quarter but at only about 6 µg/mL. Confirm the volume before using it.
- Rectal and vaginal volumes (5 and 8 mL per kg) are large for a big animal. Published horse rectal work used far smaller volumes (60 mL in one study of 10 horses). Confirm the volume for the animal's size.
- Sterility. Madrid asks for maximum sterility. Hair removal is mandatory, because hair carries germs into blood, joints and other tissue. Get signed informed consent from the owner.
- Never let anyone inhale the gas. Madrid says ozone must not be inhaled during any procedure, and calls nasal inhalation absolutely prohibited. Ventilate the room and use a destruct unit or suction where gas is vented.
- Materials. Madrid says to use materials made for each technique: latex-free, and PVC-free for some procedures. Use ozone-resistant (silicone) catheters and silicone-coated syringes.
- Routes Madrid says never to use: nasal inhalation, intra-arterial injection (air embolism), direct intravenous ozone gas, ozonated glucose or Lactated Ringer's by vein, and saline "ozonated" by beating a plastic bag with a syringe (phthalates, uncontrolled dose). It also does not recommend injecting ozonated distilled water into a joint or a vein.
- Go low and slow. For blood and rectal routes Madrid starts low and raises the concentration by 5 µg/mL every 2 to 3 sessions. In animals with chronic disease or high oxidative stress it suggests cofactor and antioxidant supplements and a low-dose lead-in. Inject gas slowly to avoid a vagal reflex, and consider sedation for painful methods.
- Contraindications. Madrid's veterinary addendum does not publish its own list of conditions that rule ozone out, and it gives no pregnancy rule. It states that the principles of the human Declaration apply, so the closest reference is Madrid's human list (section 2.1), shown below. The veterinarian decides animal by animal. Take extra care in a pregnant animal: for people, Madrid advises avoiding ozone in the first trimester and reserves later use for specialists, and the same caution is sensible before any blood, rectal, vaginal or uterine route in a pregnant animal.
- MediSkill does not advise ozonated saline. Madrid also describes ozonated saline (a drip, bladder and wound irrigation, uterine lavage). Those routes and doses are left off this sheet.
Closest reference: Madrid's human contraindications
From the Madrid Declaration on Ozone Therapy, 4th edition (ISCO3, 2025), section 2.1. This list is written for people and is shown as the nearest published reference, not as veterinary guidance.
- Direct inhalation of ozone gas. The fluid lining the lungs has no antioxidants to buffer it, so inhaled ozone is toxic.
- Glucose-6-phosphate dehydrogenase (G6PD) deficiency (favism, acute hemolytic anemia). Madrid lists it as a contraindication and recommends testing, but whether it is absolute is debated. MediSkill protocols start low and go slow; see the contraindications article.
- Toxic hyperthyroidism (severe Graves' disease)
- Thrombocytopenia (platelets under 50,000) and severe coagulation disorders
- Severe cardiovascular instability
- Acute alcohol intoxication
- Recent acute myocardial infarction
- Acute massive hemorrhage
- Convulsive states
- Hemochromatosis
- Patients receiving intravenous copper or iron therapy
More detail
Equipment
- Ozone generator with an ozone-resistant gas path (quartz glass, titanium, silicone) and a way to check the output concentration
- Medical oxygen tank and regulator (not an oxygen concentrator) for any gas given by rectum, vagina, injection or blood
- Latex-free, silicone-coated syringes, silicone catheters and ozone-resistant tubing; ozone destruct unit or good ventilation
- Large-bore sterile needles and tubing for large-animal blood draws; filter at the syringe tip
- Ozone-resistant bags sized for the limb or wound, with a vacuum pump to remove the gas afterward
What the studies used
Study doses are listed as printed in each paper. They are not Madrid dosing.
- In 54 Holstein cows with mild or moderate mastitis, recovery after three daily intramammary treatments of 50 mL ozone gas at 35 µg/mL was 78% (14 of 18), not different from an injected antibiotic (83%, 15 of 18). Ozonated saline did worse (33%) (Argudo 2017).
- In 19 cows with acute mastitis, 9 of 15 treated with intramammary ozone (1 to 5 L of gas per quarter) recovered without antibiotics, and 6 needed antibiotics after day 3. The paper reports no significant difference from antibiotics. Allocation was not stated (J Vet Med Sci 2000).
- In 400 Simmental cows, two postpartum infusions of 20 mL ozonated foam (within 6 hours of calving and 24 hours later) shortened days open (75.5 vs 98.0) and lowered inseminations per pregnancy (1.68 vs 2.42), both p < 0.05. The ozone concentration was not stated (Zobel 2014, Vet Arhiv).
- In 10 Thoroughbred geldings, one session of ozonated autohemotherapy (400 mL blood, 25 µg/mL) raised antioxidant capacity at 3 and 7 days compared with samples taken a month earlier (Tsuzuki 2016).
The evidence is thin: a few small trials and case series. The mastitis trials point to ozone gas working about as well as an antibiotic in mild to moderate cases, with small numbers.
Learn more
Background on the methods: autohemotherapy, rectal ozone, bagging and ozone oil. These articles are written for people, so use them for how the method works, not for animal doses.
Related protocols: Veterinary Ozone Protocol: Dogs and Cats, Veterinary Ozone Protocol: Horse Conditions.
Other veterinary protocols
For veterinarians. Educational reference only, not veterinary advice. Follow your scope of practice and local regulations.