This paper was presented in the last World Equine Veterinary Association Mondial Congress on September 1997

Ozone treatment and blood lactate variation after thoroughbred races horses

Scrollavezza P, Ferrari F*, Martini FM, Isola M.

Surgery Institute, Parma University, Italy; * DVM, Innsbruck, Austria.


The post-exercise lactate blood concentration rise in athlete animals, is considered an anaerobic metabolism indicator (Jones & Campbell, 1982). Particularly in horses, a concentration of 4 mM/l is adopted as threshold above which anaerobic metabolism is predominant (Landry & Orban, 1978; Hagberg, 1984).

An inappropriate training, poor athletic condition and fatigue are strictly related to a significant rise of post-exercise lactate concentration (Persson, 1983).

Many attempts have been made to delay the beginning of aerobic metabolism, therefore improve horse performance. It’s been tried, with conflicting outcomes, the administration of substances, such as readily digestible sugar, amino acid and l-carnitina which increase anaerobic metabolism (Frape, 1988; Rose et al., 1989; Iben et al., 1992; Younglove et al., 1994). Others weighed the effects of the administration of alkalising substances in order to decrease the effects of metabolic acidosis that follows anaerobic metabolism (Lawrence et al., 1990; Harkins et Kamerling, 1992).

We have tested the effectiveness of ozonized autohemotherapy in horse to examine the effects on aerobic/anaerobic energetic metabolism in maximal effort, as it’s been described in human sport medicine. There are not papers about the specific use of ozone in human being sport medicine, but it is well-know that athletes, especially in Eastern Europe, use this method to improve energetic metabolism and to increase the catabolism of the energetic pathway substances after a maximal effort.


Ten thoroughbreds (5 males and 5 females) aged 4±1 years and weighing 414±20 kg have been made race, 2 at a time, for 800 meters.

Fifteen minutes before the race and 2, 5, 10, 30 and 60 minutes after the end of the race, a blood sample from jugular vein has been collected with Vacutainer® and immediately analyzed to test blood lactate concentration with an autocalibrating lactate analyser.

?After the race, the horse have been walked for 15’ then subjected to shower and finally put in the stable, fasted till the end of the experiment.

?Because sprinter quickly accumulate lactate during a maximal effort, our trial was conducted in a 800 meters gallop race (Svedenagh e Sjodin, 1984).

After a one month treatment with ozone autohemotherapy, the same samples were collected on the same horses before and after the same race on the same track. In particular, the first part of the trial dates October 27 and the second part, after ozone treatments, November 24, 1996. Weather and track conditions were practically the same on both dates, while temperature and humidity were respectively 8°C- 48% and 5°C - 35%.

For each horse, ozone treatment has been repeated 9 times, one every three days, using an amount of 2000 ml of heparinized blood ozonized at 30 mg/ml.

Horse were ridden by 2 professional jockey weighing 62,8 and 67,3 kg.

Data was analysed with the coupled t-test of Student. Statistical significance was set at P < 0,05. 


In order to exclude the variables that usually arises in this kind of experiment and that affect analysis results (samples taken in aerobiosis, incorrect centrifugation, sample prolonged stay at room temperature before and/or after storage) we have taken the samples with the vacuum technique in anaerobiosis, with 2 samples at a time immediately analysed on the field.

The horses raced 2 at a time at top speed both for the method of analysis chosen and for their psychological (then hormonal) stress would be the same as those in normal races. For the same reason, the horse received the same treatment they usually receive after a race, relaxing walk and cold shower before stabling.

Before the race, blood lactate values in horse treated with ozone don’t differ significantly from same horses not treated (Fig. 1).

Two, 5, 10, 30 and 60 minutes after the end of the race, blood lactate values in horse treated with ozone were significantly lower compared to the same horses not treated (Fig. 2-6).

General mean of blood lactate values in horse treated or not with ozone is represented in figure 7.

As earlier described by other Authors (Krzywanek et al., 1970; Auvinet and Demonceau, 1992), we observed the blood lactate concentration peak in not treated horses at 10 minutes after the race, while lactate concentration peak in the same horses treated with ozonized autohemotherapy was pointed out after only 5 minutes from the end of the race.

Blood lactate concentration observed one hour after the end of the race in horses treated with ozonized autohemotherapy has been even lower than the value indicated as threshold of aerobic-anaerobic metabolism, while in horses not treated was still above the threshold (3,6 vs. 7,8 mM/l).

Performance of horses treated has been slightly but not significantly better.


?For some times it has been know that ozone, an allotropic oxygen form, performs numerous activities when administered with autoemotherapy technique (Razumoskii and Zaikov, 1984; Viebahn, 1985):

    • Increasing of pyruvate oxidation, thus increasing Acetil-Coa production;
    • Activation of the cellular enzymal protection system against peroxides, oxygen and ozone free radicals;
    • Activation of the erythrocyte metabolism;
    • Increasing the 2,3-diphosphogliceric acid cycle;
    • Increasing of the redox function of the mitochondrial respiratory chain.
  • Ozone effects are not restricted to the amount of blood taken for autohemotherapy (~ 4 ml/kg BW): reactions with blood that come in touch with ozone continue in the body for a period varying from a few hours to a few days (Vogelsberger and Herget, 1983).

    ?Treatment with ozone autohemotherapy caused a decrease in blood lactate concentration after a maximal effort. This effect can be interpreted in 2 different ways: 

    1. Increase of aerobic ability during a maximal effort, therefore anaerobic metabolism delay with a consequent less lactate production;
  • Increase of pyruvate regeneration speed, starting from lactate produced by anaerobic glicolysis.
  • We believe of the significant decrease of blood lactate concentration is caused by both factors. In fact, a larger quantity of oxygen available for tissues, the shift towards the right of dissociation hemoglobin curve together with a more efficient red cell, improves aerobic metabolism; while the effect on mitochondrial respiration cause to a larger availability of oxidised nicotinamide adenine dinucleotide (NAD+), essential coenzyme to change lactic acid in pyruvic acid.

    We know that measurement of blood lactate concentration after a maximal effort is interpreted as performance index in race horse (Kubo et al., 1984; Evans et al., 1993).

    In accordance with Rushall (1991) and Roneus (1994), we are convinced that this kind of relief doesn’t depict the index of anaerobic work done by the animal. Other subjective factors, such as type of muscular fibres, diet, mental state, race perception, environmental conditions and pain resistance, play an important role on energetic metabolism.

    With this experience, we have demonstrated that after treatment with ozone autoemotherapy, blood lactate concentration after a short maximal effort, regardless of its source, significantly decrease. Likewise, the return of lactate 60 minutes after the race towards the pre-race average concentration is fasten in subjects treated with ozone.


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  • ChartObject FIG. 1: Blood lactate concentration 15 mins before 800 meters race.

    ChartObject FIG. 2: Blood lactate concentration 2 mins after 800 meters race. *Significant difference (P=0,011) compared to the control group.

    ChartObject FIG. 3: Blood lactate concentration 5 mins after 800 meters race. *Significant difference (P=0,045) compared to the control group.

    ChartObject FIG. 4: Blood lactate concentration 10 mins after 800 meters race. *Significant difference (P=0,003) compared to the control group.

    ChartObject FIG. 5: Blood lactate concentration 30 mins after 800 meters race. *Significant difference (P=0,005) compared to the control group.

    ChartObject FIG. 6: Blood lactate concentration 2 mins after 800 meters race. *Significant difference (P=0,000) compared to the control group.

    ChartObject FIG. 7: Mean blood lactate concentration in the control group and in the treated group. *Significant difference (P=0,011) compared to the control group.