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Oxidative stress in sled dogs and effects of antioxidant supplementation

with polyphenols, Vitamin C and E

Oxidative stress is a major limiting factor for normal cell function in the organism.Its function is strictly connected to the intensity of the metabolism.To be able to control oxidative stress, allows for improved management and care of pets with a wide range of diseases where oxidative stress influences their development and progression.The evaluation of canine oxidative stress, is a recent issue in veterinary medicine. The aim of this clinical trial was to evaluate the effects of an antioxidant supplement with polyphenols, Vitamin C and E on oxidative stress.

Materials and MethodsThe dogs involved in this clinical trial were sled dogs involved in long distance training and racing. These dogs are therefore subjected to many hours of long-term strenuous exercise daily. The dogs have been bred and trained in the cold environment of the Swedish Lapland and are fed with a high-energy diet.

The oxidative stress of the dogs involved in this trial has been evaluated through blood samples, using d-ROMs test.

d-ROM (Reactive Oxygen Metabolites) test:The Reactive Oxygen Metabolites test assesses the total oxidant capacity in a sample of plasma or serum, using a reducing chromogenic reagent (N, N-diethyl-paraphenylen-diamine). The d-ROMs test uses the principle of Fenton’s reaction: by mixing a biological sample with an acidic buffer (Reagent R1). The newly created transition metal ion (iron or copper) catalyses the breakdown of an organic hydroperoxide, generating new radical species such as hydroperoxyl (ROO+) and alkoxyl (RO+). By adding a chromogenic (N, N-diethyl-paraphenylendiamine, Reagent R2) that has the ability to donate an electron and change colour when oxidized by free radicals, and using photometric reading available with the FRAS 4 analytical equipment, it becomes possible to quantify the level of hydroperoxides in the sample.

The d-ROMs test principle and reactions:

1 A) R-OOH + Fe2 + -> R-O* + Fe3+ + OH- 1 B) R-O* + A-N H2 -> R-O- + [A-NH2*]+A) R-OOH + Fe3+ -> R-OO* + Fe2+ + H+B) R-OO* + A-NH2 -> R-OO- + [A-NH2*]+

The test results are in conventional units, CARR UNITS, where 1 CARR= 0.08 mg/dl of H202.

MaterialsThe dogs included in this trial were sled dogs between the ages of 1 and 8 years, involved in strenuous activities and with normal biochemical and haematological blood tests. All the dogs live in their normal environment and are fed with the same food with high-energy content.

In order to evaluate the oxidative status of dogs in different periods of the season two stages were chosen:

Time 0, end of August: the first blood sample was taken, from all dogs, after a three month resting period.

Beginning of September: the dogs were split into two groups (one group of 7 dogs receiving antioxidants and one group of 5 dogs, defined as the “control group”). The antioxidant HF66* was administered daily, orally by tablet form, from the beginning of September to the study group.Dogs belonging to the two groups were subjected to the same conditions of environment, feeding and training. All dogs were from then on trained on the ground, pulling a four-wheeled loaded motorcycle. The routes were 5 km long initially, and 20 km long with temperatures around 0 Celsius towards the end of October.

Time 1, end of October: the second blood sample was taken from all dogs.

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R-OOH : hydroperoxide.R-O* : the alkoxyl radical of a hydroperoxide.R-OO* : the hydroperoxyl radical of a hydroperoxide. A-NH2 is N, N-diethyl-paraphenylendiamine, the chromogenic substrate of the d-ROMs test.[A-NH2*]+ is the colored radical cation of the chromogenic substrate.

ResultsTime 0: All dogs have values of d-ROMs in the range 70 to 128 CARR UNITS, Table 1.Time 1: The dogs in the study group having received the antioxidant HF66* all show reduced d-ROM values. Dogs in the control group show the same or slightly higher d-ROM values.

TABLE 1DOGS TIME 0 (d-ROM) TIME 1 (d-ROM)

TEST GROUPTEST GROUPTEST GROUPPATTY 79 64

LAKOTA 112 57SPIGA 96 55ARIEL 76 56SYRIA 72 55RAIN 105 65

RISSLA 128 83CONTROL GROUPCONTROL GROUPCONTROL GROUP

EMMA 80 93MIRO' 93 94NINA 78 76

FLOYD 85 85MISS X 86 79MEDIA 91,27 71,18MAX 128 94MIN 72 55

DEV STAND 17,57891 15,47784

ConclusionsThe markers of oxidative stress became significantly lower in the dogs treated with HF66*. The oxidative stress was 15% lower in the study group compared to the control group.

HF66* = Proage® 1200 (DRN), antioxidant in tablets containing polypenols, Vitamin E and Vitamin C.

This trial was sponsored by DRN.

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