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NUTRITIONAL ASPECTS OF FERTILITY IN STALLIONS

Author : CATHERINE DUNNETT

Categories : Vets

Date : September 1, 2008

CATHERINE DUNNETT registered nutritionist explains how advances in nutrition-based research

into stallion sperm may provide the impetus needed to improve fertility rates

THE past century has seen great improvements in the management and care of breeding
horses, and increased practical use of advances in reproductive technology and nutrition.

Despite this, inherent fertility rates, while good, are not exceptionally high. In well-managed
thoroughbred flat and National Hunt mares and stallions, where fertility is considered to be
relatively high, the fertility rate was reported to be 63 per cent in flat horses to 65 per cent in
National Hunt horses (Allen et al, 2007). In this retrospective study, pregnancy loss accounted for
13.5 per cent (flat mares) to 16.2 per cent (National Hunt mares) of pregnancies. The authors cited
advancing maternal age as the single most significant factor to consider with the rate of fertility in
this study.

In other species, including cattle, pigs, sheep and humans, the effects of diet on reproductive
success have been studied fairly extensively. However, in horses, this remains a relatively
unexplored area. This article will discuss research that has been published with respect to fertility in
stallions.

The area of nutrition-related research that has received most attention in stallions is the effect of
diet on sperm characteristics, either within a natural environment or when using artificial
insemination (AI). Lipids, including phospholipids, saturated fatty acids, polyunsaturated fatty acids
and sterols contained within the sperm body and sperm membrane are central to sperm quality, as
these components can affect the flexibility and functional characteristics of the sperm cell.

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However, the high lipid and polyunsaturated fatty acid (PUFA) content of sperm cell membranes
makes them a prime target for lipid peroxidation.

Sperm quality
Due to the high level of PUFA docosahexaenoic acid (DHA) in sperm cell membranes, and its
association with membrane fluidity and its effect on sperm quality in other species, there has been
interest in increasing the DHA content of the equine diet.

In sub-fertile men with reduced sperm motility and/or a reduced sperm count, the level of DHA and
the ratio of total omega-3 and omega-6 fatty acids found in sperm cells is reduced compared to
normal controls (Aksoy et al, 2006). The DHA content of a stallion’s diet may be low unless there
is significant access to fresh grazing. Traditional concentrate rations are normally rich in omega-6
fatty acids and typically low in omega-3 fatty acids, including DHA. The inclusion of ingredients
such as linseed may increase the intake of alpha linolenic acid (omega- 3). This is the main
precursor for long-chain omega-3 fatty acids, although the rate of conversion to DHA is thought to
be relatively low. Horse studies have typically focused on the effect of supplement ing the stallion’s
diet with long-chain omega-3 fatty acids, such as DHA and eicosapentaenoic acid (EPA). These
are usually derived from fish, such as tuna or salmon, and studies have investigated their effect on
both fresh and cooled semen.

Positive results were reported by Brinsko et al (2005), who investigated the effect of
supplementation with a nutraceutical product rich in DHA. Supplementation with DHA for 14 weeks
resulted in a three-fold increase in semen DHA levels and a 50 per cent increase in the ratio of
DHA to an omega-6 marker fatty acid docosapentaenoic acid (DPA). While there was no significant
effect of supplementation on fresh semen, the results suggest that the motion characteristics of
cooled semen were improved, especially in stallions with marginal fertility under AI conditions. A
similar study published in the same year reported a 46 per cent significant increase in daily sperm
output over a 90-day period of supplementation with 29g of longchain omega-3 PUFA, compared to
control stallions. However, while the long-chain omega-3 content of sperm membranes increased,
there was no difference in motility characteristics with either fresh or cold-stored semen (Harris et
al, 2005).

Thus , while supplementation of the diet with omega - 3 fatty acids may seem promising, the
optimum level of their intake requires further definition, as does the importance of the ratio of
omega-3 to omega-6 fatty acids in the diet.

A study of mares has attempted to begin to address this issue in horses. This study involved
supplementation with DHA and EPA (10g/day to 40g/day for 28 days) using a stepwise study
design (King et al, 2008).

Unsurprisingly, the trial revealed that plasma levels of these two omega-3 fatty acids increased in

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line with the level of supplementation, reaching peak concentrations of 10 × DHA and 13 × DPA,
compared to baseline values following only seven days of supplementation. The authors noted that
the circulating level of DHA and EPA appeared to be affected by the availability of fresh forage and
that the elevation in these two long-chain omega-3 fatty acids was lost in a relatively short time,
with values returning to baseline figures within 42 days following cessation of supplementation.

Oxidative stress
The high lipid content of sperm leaves it vulnerable to oxidative stress and associative damage to
the constituent lipids, proteins and DNA, which has been proposed as a contributory factor in sub-
fertility in humans.

Oxidative status appears to improve during the active breeding season in sperm samples and
seminal plasma, in both normal and sub-fertile stallions, compared to the non-breeding season
(Morte et al, 2008).

This was associated with increased sperm production during the active breeding season, but was
also coupled to a reduction in semen quality when assessed by classical parameters. Additionally,
in the non-breeding season, markers for lipid and protein peroxidation in both sperm and seminal
plasma provided a good indicator for sperm damage in sub-fertile stallions.

These results obviously emphasise the importance of adequate dietary antioxidant intake and that
of their co-factors, including vitamin E, vitamin C, beta carotene, selenium, copper, zinc and
manganese. This has inevitably led to the introduction of products that combine omega-3 fatty
acids with antioxidant micronutrients. One combination product was investigated in pony stallions
by Deichsel et al (2008). The supplement in question combined L-carnitine 4g/day and folic acid
12mg/day with an anti-oxidant cocktail including tocopherol 300mg/day and ascorbic acid
300mg/day. It was fed to pony stallions for 16 weeks and resulted in a slight, but significant,
reduction in morphologically abnormal sperm.

In contrast, no change in sperm motility, progressive motility, membrane integrity or semen

longevity for 24 hours was detected as a result of the supplementation. However, the level of
supplementation, certainly with respect to vitamin E and vitamin C, was relatively low in
comparison to published requirements (NRC, 2007) and other higher recommendations. In
addition, the form of vitamin C provided has been shown to have low availability in horses
compared to other sources (Snow et al, 1990; Deaton et al, 2003).

Carnitine
Carnitine, a betaine derivative of beta-hydroxybutyrate, and salt acetyl carnitine, are compounds
that have been investigated for their potential benefit in sub-fertile men, due to their reputed effect
in supporting mitochondrial function and antioxidant action within spermatozoa. A meta-analysis of

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nutrition trials in men, with either L-carnitine or L-acetyl carnitine, revealed an improvement in
pregnancy rate and sperm mobility, but no change in sperm concentration or semen volume (Zhou
et al, 2007).

Similarly, supplementation of boars with 500mg of L-carnitine per day for five weeks had a positive
effect on sperm quality (Jacyno et al, 2007). The total ejaculate volume and sperm-rich fraction
volume increased by 11 per cent and 10 per cent respectively, while the total ejaculate sperm
count increased significantly by 11.5 per cent. In addition, the number of spermatozoa with major
and minor morphological changes decreased.

The carnitine content of the horse’s diet as a herbivore is quite low, and biosynthetic capacity from
lysine and methionine may be limiting. Therefore, carnitine is often regarded as a conditionally
essential nutrient. Early work by Foster et al (1989) reported that L-carnitine is absorbed in horses,
albeit with low efficiency, with plasma levels responding to an increasing intake of dietary carnitine.
More recently, L-carnitine and its derivatives have been suggested as useful markers of semen
quality in horses. Stradaioli et al (2000) reported a good positive correlation between L-carnitine,
acetylcarnitine and spermatozoa concentration, and also between acetyl carnitine and total motile
morphologically normal spermatozoa (TMMNS). These authors also suggest that L-carnitine may
be involved in the maintenance of functionality in cooled semen samples. Further work by these
authors concluded that supplementation of stallions with questionable fertility using 20g of L-
carnitine daily offered potential benefit in terms of sperm motility (Stradaioli et al, 2004).

Supplementation of stallions with a number of nutraceutical- type ingredients, such as long-chain

omega-3 PUFA, antioxidants and L-carnitine, has thus far yielded some interesting, but
inconclusive, results.

It is likely that the outcome from such supplementation will depend on many factors, including the
content of the basal diet PUFA intake, omega-3 to omega-6 ratios, as well as the dietary
antioxidant intake.

However, it is also likely from the studies to date that the relative success of such supplementation
may be limited to those stallions that are sub-fertile.

References
Aksoy Y, Aksoy H, Altinkaynak K, Aydin H R and Ozkan A (2006). Sperm fatty acid
composition in sub-fertile men, Prostaglandins Leukotrienes and Essential Fatty Acids
75(2): 75-79.
Allen W R, Brown L, Wright M and Wilsher S (2007). Reproductive efficiency of flat race
and National Hunt thoroughbred mares and stallions in England, Equine Veterinary Journal
39(5): 438-445.
Brinsko S P, Varner D D, Love C C, Blanchard T L, Day B C and Wilson M E (2005). Effect

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of feeding a DHA-enriched nutriceutical on the quality of fresh, cooled and frozen stallion
semen, Theriogenology 63(5): 1,519-1,527.
Deaton C M, Marlin D J, Smith N C, Roberts C A, Harris P A, Kelly F J and Schroter R C
(2003). Pulmonary bioavailability of ascorbic acid in an ascorbate-synthesising species, the
horse, Free Radical Research 37(4): 461-461.
Deichsel K, Palm F, Koblischke P, Budik S and Aurich C (2008). Effect of a dietary
antioxidant supplementation on semen quality in pony stallions, Theriogenology 69(8):
940-945.
Foster C V and Harris R C (1989). Plasma carnitine concentrations in the horse following
oral supplementation using a triple dose regime, Equine Veterinary Journal 21(5): 376-377.
Harris M A, Anderson C W, Webel S K, Godbee R, Sanders S R, Schurg W A, Baumgard L
H and Arns M J (2005). Effects of feeding an omega- 3-rich supplement on the fatty acid
composition and motion characteristics of stallion spermatozoa, Proceedings of the 19th
Equine Science Society Symposia, Tuscon, Arizona, Equine Science Society.
Jacyno E et al (2007). Effect of L-carnitine supplementation on boar semen quality, Acta
Veterinaria Brno 76(4): 595-600.
King S S, Abugha z a leh A A, Webel S K and Jones K L (2008). Circulating fatty acid
profiles in response to three levels of dietary omega-3 fatty acid supplementation in horses,
Journal of Animal Science 86(5): 1,114-1,123.
Morte M I, Rodrigues A M, Soares, D, Rodrigues A S, Gamboa S and Ramalho-Santos J
(2008). The quantification of lipid and protein oxidation in stallion spermatozoa and seminal
plasma: seasonal distinctions and correlations with DNA strand breaks, classical seminal
parameters and stallion fertility, Animal Reproduction Science 106(1-2): 36-47.
NRC (2007). Nutrient Requirements of Horses (sixth edn). The National Academies Press,
Washington DC. Snow D H and Frigg M (1990). Bioavailability of ascorbic acid in horses,
Journal of Veterinary Pharmacology and Therapeutics 13(4): 393-403.
Stradaioli G, Sylla L, Zelli R, Chiodi P and Monaci M (2004). Effect of L-carnitine
administration on the seminal characteristics of oligoasthenospermic stallions,
Theriogenology 62(3-4): 761-777.
Stradaioli G, Sylla L, Zelli R, Verini Supplizi A, Chiodi P, Arduini A and Monaci M (2000).
Seminal carnitine and acetyl carnitine content and carnitine acetyl transferase activity in
young Maremmano stallions, Animal Reproduction Science 64(3 and 4): 233-245.
Zhou X, Liu F and Zhai S (2007). Effect of L-carnitine and/or L-acetyl-carnitine in nutrition
treatment for male infertility: a systematic review, Asia Pacific Journal of Clinical Nutrition
16 (supplement one): 383-390.

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Photo: ISTOCKPHOTO/MURAT KOC.

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 The carnitine content of the horse’s diet as a herbivore is quite low, and biosynthetic
capacity from lysine and methionine may be limiting. Therefore, carnitine is often regarded
as being a conditionally essential nutrient.

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