Here we take a look at iron and its impact in the horse. An earlier article attempted to explain the bioavailability of iron and why this was an important issue in the nutrition of horses. Returned comments was that the article was confusing and it is apparent that for many people iron is just one chemical and the complexities of iron metabolism were not fully explained. In fact they were only touched on as the route of iron from soil to mammalian metabolism is so incredibly complex it has not been fully explained.
Iron has an essential role in the metabolism of all life. It has the ability to form complex chelates (groupings of organic molecules like the heme molecule in blood) across all life forms and in many cases these complexes are ways of locking away its activity. I am fully aware that potential toxicity from iron is a worry to people as iron, changing its form from ferric to ferrous within the body can release free radicals that have a devastating effect on tissue.
So I would like to start the story from the soil. Iron exists in two forms; ferric and ferrous ions. In the soil they are found mainly as inorganic, ionic salts such as ferric oxide and ferrous sulphate, although complex salts and organic chelates (usually two organic chains surrounding the iron molecule) can be found. However the main criteria for absorption by the roots of plants is the solubility of these compounds. Iron oxide, for example, is insoluble and therefore unavailable whilst one of the many hydrated forms of iron sulphate is reasonably soluble and is potentially bioavailable.
There are two strategies for the absorption of iron (Hell & Stephan 2003). One, for the higher plants, is a creation of an iron chelate that changes ferric iron to ferrous and also increases the solubility of ferrous iron (by increasing level of hydration). The second – exclusive to the grasses – is the formation and absorption of a ferric chelate. Both strategies end up with both ferric and ferrous iron being incorporated in a chelate with Nicotianamine, which is the major compound for transporting iron to the shoots, leaves and storage area s via the phloem and xylem. Phloem and xylem forms may be different but the target tissues will receive iron as nicotianamine (Fe-NA), and then become incorporated in protein, heme, phytoferritin, iron sulphide clusters or precipitate. Phytoferritin and precipitate are the only two fates if iron is absorbed in excess, and the precipitate by its very nature – insoluble – will be unavailable.
Phytoferritin is a complex of excess iron and tends to accumulate in the chloroplasts and plastid stomata of leaves. Basically large quantities of iron are bound in protein sinks and are released in times of iron deficiency to avoid reduction of photosynthesis and chlorosis.
So, basically, because iron is relatively unavailable to the plant it has developed a sophisticated absorption, transport and storage system to:
• Ensure there are reserves for the plant to continue to photosynthesise during iron deficiency
• Keep the iron in a stable form so it does not destroy cell membranes.
Because an approximate 3,000,000,000 people are defined as anaemic (WHO 1998) a tremendous amount of work has been conducted to try to increase iron concentrations in plants and this is leading to genetic modification of mainly cereal crops, with variable success (Fossard et al. 2000, Zehng et al. 2010). Under general circumstances, however, plants tend to absorb sufficient iron for their immediate and potential needs irrespective of the iron content of the soil.
Studies on South African soils, which have iron contents in excess of those theoretically needed for wheat growth (Materachera 1999), showed weak correlation between iron levels and iron uptake. In house data at British Horse Feeds show that South African Teff, Lucerne and Oat straw all have iron levels that are lower than NRC (and therefore presumably U.S.) quoted values despite high iron levels in the soil. It is not a simple matter to equate iron levels in the soil with those in plants and illustrates one reason why iron deficiency is a greater problem than iron excess.
The ability of any plant to absorb and utilise iron is dependent on the form and solubility of that form in the soil.
So why do some plants such as lucerne, copra and sugar beet have higher levels of iron than other plant species? In the case of beet it is because of its ability to produce sugar. Iron sulphide clusters are integral in photosynthesis and the production of sugar that is then stored in the roots. As sugar beet has very high levels of sugar generation it needs a strong mechanism for iron absorption. The root tips contain high levels of NADH (an energy generator involved in Strategy 1 iron uptake) and it has been shown that adding iron to the soil increases sugar yield from beets (Yarnia et. al. 2008). It is logical to assume that beet will store excess iron – as with all other plants – as an insoluble precipitate in the roots to maintain a reserve to transport to the leaves when needed, to maintain the high level of sugar production.
The high levels of iron in beet root systems are stored as an insoluble precipitate and are maintained to transport to leaves and shoots.
Plants – with different strategies for iron accumulation between grasses/cereals and higher plants – will have iron present across all tissues but the form and availability of this will vary between plants and areas. Leafy material contains the more “active” types (heme, iron clusters) and its storage mechanism involves phytoferritin. Seed heads tend to have more iron protein chelates and roots more insoluble iron.
So what does this mean? And how does it translate into absorption of iron into the horse? And how does the horse cope with deficiency/excess of iron. The first step in this story is see how animals deal with all the various forms of iron it ingests.
Bioavailability is a term used to describe the amount of a mineral or trace element that can be incorporated into an organic system. Bioavailability of iron for plants will be different to bioavailability of iron for animals and this will vary between species and individuals.
However the mechanism that determines bioavailability is the same across animal species and this is down to the capability of the gut to absorb minerals.
Much is known about the sites and processes of absorption. Mineral ions (the metal parts of salts such as sodium, zinc and iron) can be absorbed both actively (requiring energy expenditure and biochemical/physiological changes from the gut wall) and passively (straight transfer across a concentration gradient) along the length of the gut. These absorption sites are areas of conflict as some minerals compete with each other and high levels of one will reduce the absorption of the other. For example high levels of copper will reduce the absorption of iron, although the reverse does not occur. Zinc, manganese, phosphorus and copper all affect iron absorption, while iron affects phosphorus and possibly manganese. In addition organic minerals – such as chelates – can be absorbed actively along the small intestine (the molecules tend to be too large to cross the hindgut membrane) and by pass the sites of competition. Chelated minerals, increasingly found in commercial mineral premixes, capitalise on this effect to allow more precise administration of essential elements.
Work on human subjects, for example, have shown that soya ferritin is absorbed with almost the same availability as ferrous sulphate – the most soluble of the iron salts (Davila-Hicks et al. 2004), although both crossed the gut by different pathways.
Two other factors affect the bioavailability of the minerals and these are the physical conditions of the gut – its acidity along its length – and the presence of micro-organisms in the gut. Plant chelated iron is at its most stable in neutral conditions whilst soluble iron salts will likely dissociate in the stomach acid. Insoluble iron salts, however, will only slightly dissociate – if at all – in the stomach, which is why iron oxide does not become a more available iron chloride.
The second factor for bioavailability is the presence of the gut microflora. Generally speaking, as iron deficiency is more common than iron excess in all environments, there is competition for iron. Animal iron sources have been shown to be bound by gut microflora with varying degrees on strength (Styriak 2004) and it has been shown that fungi have a number of strategies to capture the iron from plant chelates (Nevitt 2011). Microbes, including fungi can absorb and utilise free iron, as in iron salts, whilst utilising chelates is a specialist function where the microbes break down the chelate and use the iron. However some chelates have antimicrobial properties and so can affect microbial populations (Holbein et al. 2010). This occurs in all environments including the soil, plants the gut and within the animal itself (Jung et al. 2008).
The animal gut can absorb iron in the form of soluble salts and some chelates, although the latter only in the small intestine. The stomach may improve the solubility of some iron salts but there will be competition along the gut from the microflora. Chelated iron has a more complex interaction; some will be broken down and utilised by microbes, some will be absorbed. However insoluble iron salts will not be absorbed and will pass through the gut unchanged.
Although the mechanisms described above hold true for all species of animals there are variations between species. Carnivores for example do not usually suffer from deficiencies as they can readily absorb the heme and animal ferritin chelates, whilst herbivores such as the black rhino (selective herbivore) has developed strategies to utilise the reasonable levels of iron in sub-tropical leaves. In comparison the horse appears to have a higher efficiency of absorption, but grazes on forage which has lower iron content (Clauss et al. 2007).Tapirs have a lower efficiency (Clauss et al 2009).
Foals, as with all mammals, receive iron from their mother’s milk as lactoferrin, which is assumed to be easily digested. As it is based around the ferritin chelate (similar to both phytoferritin and animal ferritin) it probably has similar bioavailability to iron sulphate.
When it comes to the weaned horse the source of iron is more varied. Phytoferritin, iron clusters etc. are all ingested and so the type of feed will impact on the proportion of these forms. In addition the horse will also ingest iron salts from the soil. Iron deficiency has been observed in stabled Dutch Warmblood foals fed fresh cut grass, but not in their grazing counterparts. The soluble iron salts in the soil would be the only difference (Brommet et al. 2001).
Another source of iron that is perhaps overlooked is that found in mineral premixes. Traditionally this will be an iron salt – usually ferrous sulphate – although increasingly manufacturers are using chelated iron (in this case a specific chelate based on amino acids that can be actively and efficiently absorbed).
So when it comes down to it, what can the horse utilise, and why is iron deficiency not a major problem? Firstly it is down to the efficiency of iron metabolism within the horse which is very good at keeping iron in the body. Unlike humans, the exercising horse does not lose increasing amounts of iron in sweat and urine, and the exercise itself improves iron absorption (Inoue et al. 2005). The horses in that trial were fed a ration of Lucerne, oats, timothy hay and a vitamin/mineral premix. Measurement of iron availability was 17% which was higher than previous reports of 12% (Schriver et al. 1986).
Trials on horses fed similar diets of alfalfa, fescue and bluegrass, but no supplements, gave iron digestibility of a negative value (Crozier et al. 1997). Presumably this is down to experimental error but the point to make is that without supplementation absorption of iron decreased.
The NRC quote bioavailability of feeds at around 15% which is similar to those values quoted in technical research for horses.
Why is this so? In a previous article I wrote… “Iron is bound, to various degrees, with various fibre sources. It has a strong binding with cellulose and hemicellulose, and a relatively weak bind with pectins. However these ligands (binding arrangements) are acid dependent and iron is less susceptible to acid than other minerals such as calcium, zinc, copper and manganese. It means that the stomach acids will not release any appreciable iron from the pectins. As iron absorption mainly takes place in the foregut, very little is available if bound as fibre ligands. Once the pectins are fermented in the hindgut, the iron will be released and passed in the faeces.
Iron in sugar beet is mainly bound with the beet pectin, although appreciable levels are incorporated into cellulose and hemicellulose and technical research has give the in vitro availability of as little as 1.5%. This means that the available iron content of Speedi-Beet is 11mg/kg.”
What I maybe should have added is that root iron is mainly insoluble – there will be small amounts of the nicotianamine form but the majority of the phytoferritin, protein and clusters will be in the leafy material that is not present in beet pulp. As pectin and fibre is fermented in the hindgut insoluble iron hydroxide is released, and is incapable of being absorbed.
The horse ingests feeds through its diet, from the soil and via any supplementation. Deriving information from various sources it appears there is a scale of absorbability across species. Between species there is variation but for the herbivore absorbability and therefore bioavailability is, in descending order:
• Protein bound chelates, such as supplements, heme, lactoferrin.
• Soluble iron salts (Ferrous or ferric ions)
• Insoluble Precipitates such as Iron Hydroxide.
As this passes through the tract of the horse soluble iron salts may become more soluble by conversion to iron chloride in the stomach. Whilst chelates, phytoferrin and insoluble precipitates are mainly unaffected.
In the small intestine there is passive absorption of the soluble salts, depending on the level of competition from e.g. copper and manganese and gut microflora, whilst chelates are actively absorbed. As the iron bound in fibre is released by gut fermentation there will be limited absorption – depending on the form. However in the case of the insoluble precipitates there will be little, if any absorption.
Although the high levels of iron in beet pulp may look high, the figure is a chemically derived number and does not take into account any of the information offered above. As a root crop the bioavailability of the iron is very low.
Having said that iron excess can be a concern. Just as absorption efficiencies vary between species, so do they within species. There will also be a spectrum of efficiencies for the “safe keeping “ of iron within the body (ferritin etc.). A horse may well show signs of iron excess but it is far more likely to be a response to soluble iron salts, or even leafy material, than the inert iron in beet pulp.
Rinsing the iron out of beet pulp is not really a solution. As this iron is precipitated and mainly bound within the pectin, all that happens is pectin and soluble fibre being washed out of the beet. As these are the components that help towards the prebiotic effect of beet pulp it is no real help.
If the horse does suffer from iron overload there are more realistic culprits, although the actual iron content may seem lower. Iron present in stems of plants tend to be chelated with nicotianamine, whilst leaves have iron – citrate chelates, which are more readily absorbed across the gut wall. Leafy material therefore has iron of a greater bioavailability; seed heads also have more readily available iron (protein chelates) and so it is sensible to see what else the horse is eating.
Is the diet supplemented? What level & form is the iron in it? Is the supplement, on the basis of “a little is good, a bit more is better”, being given at too high levels? Are other trace elements conflicting with each other and iron, simply because of the total content, becomes an easy suspect.
And what about the soil itself? Just because it is high in iron, does this make it the culprit? It depends on the form. It needs to be a soluble salt.
So it may not be possible to prove the culprit in iron excess situations, but root crops – including beet – are amongst the most unlikely candidates.
By Dr Tom Shurlock, Consultant Nutritionist.
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