Strategies to Reduce Use of Zinc Oxide by

Modulating Gut Microbiota in Pigs

01 March 2015
Zinc (Zn) plays an important role in the metabolism of swine and as such is an essential trace
element for growing pigs, according to Nutriad. Applying a sound SANACORE EN strategy can
make a real difference for producers that want to limit the use of zinc oxide in pig feeds.
Zinc deficiency may result in reduced feed intake and growth, parakeratosis, impaired wound
healing, alopecia, thymus atrophy and impaired immune function.
Zinc recommendations for growing pigs vary among national institutes but decrease with
increasing body weight from approximately 100 to 50mg per kg. In practice, Zn is often
administered in excess to assure sufficient supply.
During the late 1980s, it was discovered that pharmacological concentrations (1,500 to
3,000ppm) of Zn oxide (ZnO) resulted in reduced diarrhoea and increased growth in weanling
pigs (Poulsen, 1989; Sales, 2013). Unfortunately, as Zn is poorly absorbed, it becomes highly
concentrated in manure. To minimise the risk of environmental pollution, European
regulations have reduced the maximal Zn concentration authorised in pig diets. In the EU, the
legal norm for Zn is 150mg per kg (ppm) of Zn in complete animal feed (EU Regulation
1334/2003). It should be noted that pig feed usually already contains levels of around 30 to
40ppm of Zn, as it is a natural component of grains in feed. This means that, on average,
110ppm of Zn can be added as an additive.
Since 2005, ZnO at pharmacological levels has been reallowed in some European countries as a
means to reduce the use of antibacterial compounds in the period shortly after weaning. As in
those countries, supplementation of ZnO at dosages higher than 150ppm Zn in complete feed
falls under the Veterinary Medicines Regulations, the use of ZnO is only possible on veterinary
prescription.
Mode of Action of ZnO
The mechanisms behind the beneficial effects of ZnO to prevent diarrhoea and promote
growth in weaned piglets are not well understood.
Recent advances suggest that the effects of ZnO on piglet growth are achieved through
multiple regulatory pathways (Li et al., 2010).

Growth-promoting effect
As Zn absorption after weaning is about 30 per cent lower than before weaning and
bio-availability may be less than 20 per cent, administering higher levels of ZnO in feed
will increase plasma Zn levels which is needed to meet the requirements of the piglets.
Moreover, it has been shown that ZnO regulates secretion of brain-gut peptides that
stimulate feed intake (Li et al., 2010).

Improved intestinal barrier function

ZnO improves intestinal structure and function, alleviating the intestinal damage
caused by weaning, by favouring cell regeneration and stimulating epithelial growth.
Barrier function is strengthened by increasing the expression of intestinal insulin-like
growth factor-1 (IGF 1, regulates cell growth and development) and its receptors, in

this way speeding up the recovery of the damage induced by the changed diet at
weaning (Li et al., 2010). ZnO also strengthens the intestinal barrier function through
reducing paracellular permeability. This effect is the consequence of the upregulation
of tight junction proteins in the intestine (Zhang & Ghuo, 2009).

Immunomodulating effect
ZnO reduces the release of pro-inflammatory histamine, by inhibiting proliferation and
activation of intestinal mast cells (Kim et al., 2012).

Reduced ion secretion

ZnO reduces secretion of ions to the intestinal lumen, thereby enhancing water
resorption and preventing diarrhoea.

Effects on bacteria
Supplementation of weaning diets of piglets with ZnO during a short period of time at
relatively high doses (2,500ppm) stabilizes intestinal microbiota and prevents
attachment of pathogenic bacteria to the intestinal villi, which may prevent many
problems associated with weaning diarrhoea.
o

Research on the in vitro sensitivity to ZnO of a range of reference strains of

intestinal origin has shown that Zn sensitivity is very variable. Liedtke & Vahjen
(2012) have concluded that we cannot assume a generalised antibacterial
effect of ZnO as MIC values differ greatly among but also within different
bacterial species.

Roselli et al (2003) demonstrated that ZnO reduces adhesion and invasion of

ETEC in an enterocyte cell culture and that it recovers an optimal antiinflammatory cytokine balance after ETEC infection. Also in in vivo studies with
weaned piglets it was demonstrated that a diet with pharmacological
concentrations of ZnO diminishes the paracellular permeability (Huang et al.,
1999; Zhang & Guo, 2009) and prevents the translocation of (pathogenic)
bacteria such as E. coli and Enterococcus spp. in the mesenteric lymph nodes
of the small intestine (Huang et al., 1999).

High levels of Zn enhance stability and diversity of the microbiota after the
change of diet in weaned piglets (Starke et al., 2012).

It is clear that the use of high levels of ZnO (1,500 to 3,000ppm) has many positive effects on
piglet health and performance. However, such use of pharmacological ZnO dosages has
potentially some negative consequences as well.
Downsides of Zn Usage
1. Zinc toxicity
Although ZnO promotes health and performance in piglets, Zn remains a heavy metal and is as
such toxic to most living organisms, including pigs. Pharmacological usage of ZnO may benefit
piglets post-weaning, but according to the US National Research Council (NRC), it may affect
piglet health and performance (marked depression in feed intake) if used for longer periods. In
countries where pharmaceutical concentrations of ZnO in weaning diets are allowed, so far no
severe negative consequences for the health of the animal have been reported, provided the
supplementation is done for short periods (maximum 14 days). After three to four 4 weeks of
overdosing, health problems have been reported.

2. Nutritional interactions
High levels of Zn result in overproduction of metallothionein, an intestinal transporter protein
that binds several heavy metals such as copper, iron, zinc and selenium. When higher levels of
these transport proteins are present there is a risk for sub-deficiencies of some of these
minerals. For example, reduced absorption of iron and copper (which has a role in iron
transport) might lead to anaemia (Sandstrm, 2001).
Studies about interactions between Zn and phytase suggest that pharmacological dosages of
Zn have a negative effect on phytase activity and consequently on phytate-phosphorus (P)
liberation: by complex formation of Zn with P-phytate, the phosphorus cannot be released by
the phytase. This implies a reduced efficacy in phytase functions, and possible phosphorus
deficiency for affected animals (Lizardo, 2004).
3. Contamination by heavy metals
Impurities in commercial ZnO can be a considerable problem when Zn quality is not strictly
controlled. For example, tissues contaminated during the post-weaning period with cadmium
(known for of its organ toxicity), can result in elevated levels of cadmium in tissues at slaughter
time due to its long half-life. A study from the French institute IFIP indicated that cadmium
concentration in kidneys exceeds the regulatory limit (1mg per kg) for human consumption
when pigs are fed contaminated diets (0.5mg cadmium per kg diet) between 42 and 160 days.
In Thailand, where pharmacological Zn dosages are used, a recent study on 214 pork kidneys
showed that more than 25 per cent of pork kidneys contained cadmium concentrations
exceeding the regulatory limits. Contaminated ZnO was most probably the major source of
dietary cadmium.
4. Zinc and microbial resistance
Although bacterial sensitivity to Zn is not always defined, acquired Zn resistance does seem to
occur. Little is known about this resistance.
Much more alarming, however, intensive usage of Zn in animal diets may favour the
development of bacterial resistance against other antimicrobials. This can be caused by
different mechanisms.

Bacteria regulate intracellular Zn concentration with a system of efflux pumps. These

pumps can be specific to Zn or can evacuate other molecules such as antibiotics. High
levels of Zn tend to increase their synthesis, and the use of ZnO at pharmacological
dosages may reduce the sensitivity of bacteria to antibiotics.

A genetic coupling can also be observed, as genes of heavy metal resistance and those
of antibiotic resistance are sometimes associated. Consequently, the selection of
bacteria resistant to Zn leads to the co-selection of bacteria resistant to some
antibiotics.

5. Environmental concerns
Zn as a heavy metal tends to accumulate in soil after Zn-rich manure from piggeries is applied
to the fields. High levels of Zn in soil, and in water reservoirs due to run-off, are considered to
be an environmental pollutant and health hazard.
Search for Alternatives: SANACOREEN

Because of these negative aspects of ZnO usage, producers are looking to find a suitable
replacement.
When developing an antimicrobial support product, Nutriad has focused on products with a
similar mode of action as ZnO. This research has resulted in the development of
SANACORE EN, a multifunctional product with a broad spectrum antibacterial approach. The
aim of using the product is to reduce the use of ZnO and/or antimicrobials while supporting
health and welfare of the animals, as well as to improve production results.
SANACORE EN is a product based on a balanced, well-researched formula of different active
components. Each component has a specific mode of action and thanks to the specially
developed production process of SANACORE EN, the various active components are delivered
to the correct places in the intestine. A specifically designed coating, for example, ensures that
butyrate, one of the important components in SANACORE EN, is delivered to the more distal
regions of the intestine. This all results in the unique antimicrobial and gut empowering effect
of SANACORE EN.
The different components in SANACORE EN have comparable effects as described above for
ZnO:

Growth promoting effect

Butyrate is a strong stimulator of several intestinal growth factors in humans and
animals, for example glucagon-like peptide-2 (GLP-2). There is a great number of GLP2-induced effects in the intestine that promote growth and performance: decrease in
gastric motility, enhancement of intestinal nutrient transport, stimulation of intestinal
blood flow and increase in intestinal cell proliferation.

Improved intestinal barrier function

Besides providing epithelial cells with energy, butyrate markedly enhances
proliferation, differentiation and maturation of enterocytes in the small intestine
(Wang et al., 2005; Manzanilla et al., 2006; Sengupta et al., 2006). Through its
influence on gene expression and protein synthesis, butyrate speeds up intestinal
mucosa maturation during the development or repair after injury (Kotunia et al., 2004;
Guilloteau et al., 2010). Butyrate also up-regulates tight junction proteins in the
intestine and in this way decreases intestinal permeability (Guilloteau et al., 2010).

Immunomodulating effect
butyrate has been shown to reduce inflammation through its effects on several types
of immune cells. It is for these anti-inflammatory properties that butyrate is used in
human medicine in patients with Crohns disease, which is characterized by excessive
intestinal inflammation.

Reduced ion secretion

Butyrate modulates ion absorption and may alleviate the severity of the diarrhoea
(Vidyasagar & Ramakrishna, 2002; Binder, 2010).

Effect on bacteria
o

For the development of SANACORE EN several components with a distinct

antibacterial spectrum were selected ensuring that their combination resulted
in a broad antibacterial spectrum (Gram-negative and Gram-positive).

Butyrate also stimulates antimicrobial host defense peptides secretion in the

avian GIT (Sunkara et al. 2011).

Studies done by Galfi and coworkers (Galfi et al., 1991) have shown that
butyrate increases the number of intestinal lactic acid and lactobacilli in
butyrate-fed pigs, while decreasing the number of coliforms and E. coli, hereby
ensuring a healthier and stable intestinal microbiota.

A trial done at the University of Bologna (Bosi et al., 2009) looked into the
possible impact of butyrate (coated and uncoated) on E. coli K88 (ETEC)
infection in piglets. The piglets receiving the uncoated sodium butyrate
showed a lower mortality rate (5.0 per cent) and a higher growth than the
challenged control group. In the group receiving the coated sodium butyrate,
none of the piglets died and the growth rate was even slightly higher in
comparison with the piglets that were not challenged.

In studies of Boyen and co-workers it was shown that butyrate, when present
in the intestinal tract, is able to alter virulence properties of Salmonella
Typhimurium and decrease intestinal colonisation in pigs (Boyen et al., 2008).
The mode of action of this activity of butyrate is hypothesized to be at least
partially mediated through modulation of bacterial gene expression. Butyrate
specifically down-regulates Salmonella Pathogenicity Island 1 (SPI-1) gene
expression, hereby preventing invasion of intestinal epithelial cells, one of the
important steps of Salmonella pathogenesis in the bird (Gantois et al., 2006).
Van Immerseel and colleagues (2005) also demonstrated the importance of an
effective coating in order to get significant reduction in Salmonella
colonisation in the ceca and internal organs in vivo.

Recently, several field trials have indeed shown that the use of SANACORE EN allows for
reduction in the use of ZnO in piglets in the post-weaning phase.
Filed Trials with SANACORE EN
In a first trial, 68 crossbred piglets (LD-LW Pietrain), weaned at 28 days of age, were allocated
to one of two dietary treatments based on live weight (see below).
The following dietary treatment groups were evaluated:

It is important to allow some time before relying completely on the effects of SANACORE EN:
strengthening intestinal barrier function and shifting a microbiota in a stable, healthy direction
takes some time. For this reason, ZnO was only left out in the starter phase.

Zootechnical parameters were measured at the end of pre-starter phase and starter phase,
when piglets were 42 and 76 days old, respectively.
Performance parameters during the trial:

SANACORE EN inclusion in pre-starter feed (on top of ZnO), resulted in a significant increase in
feed intake of 48 g per day and an increase in ADG of 10g per day. These results indicate that
using the product on top of normal ZnO usage can proof to be beneficial.
SANACORE EN inclusion in starter feed (no ZnO) lead to higher performance in comparison
with the control group (standard ZnO inclusion): a significant increase in feed intake with 56g
per day, a significant increase in ADG of 94g per day and a reduced feed conversion ratio (0.29
lower).
When looking at the performance results throughout the complete trial period (pre-starter +
starter period), SANACORE EN application in pre-starter (on top of ZnO) and starter (no ZnO)
feeds, resulted in a significant increase in feed intake of 56g per day, a significant higher ADG
of 71g per day, and lower FCR (-0.2) in comparison with a standard ZnO programme (3kg per
tonnes throughout pre-starter and starter). At the end of the trial period at 76 days of age,
piglets on the SANACORE EN weighed on average 3.6kg heavier than the piglets from the
straight ZnO programme.
Faecal consistency was monitored throughout the trial period and no severe diarrhoea was
observed in the piglets of both trial groups.
This study demonstrated the potential of SANACORE EN in weaning piglet diets with reduced
levels of ZnO and instigated further field trials using the same strategy. As the goal of these
field trials was to include larger numbers of animals, the set-up was done differently to the
first trial in the sense that comparison was done over time, not in one trial (historical
comparison).
Trial with 1,000 piglets per group, weaned at 28 days

In this field trial, feed intake and ADG was increased and FCR and mortality was decreased
when using the SANACORE EN strategy.
Trial results from 3.750 sows, 19.000 piglets

Results of this trial show and increase in weight at 76 days (end of starter period) and lower
mortality with SANACORE EN programme.
Trial results from 16,000 piglets per group

In the above field trial, a financial comparison was made taking into account all performance
benefits but also the cost of SANACORE EN and of medication per piglet.
Next to the benefits of being able to reduce ZnO usage, applying the SANACORE EN
programme resulted in a financial profit for the pig producer.
Summary
In conclusion, several field trials have proven the potential of SANACORE EN to be used in
programmes with reduced ZnO levels. Not only does this programme diminish the dependency
of ZnO, it also results in a financial profit, making it an economical solution.
The positive results seen can easily be explained when comparing the mode of action of
SANACORE EN with that of ZnO.
Applying a sound SANACORE EN strategy can make a real difference for producers that want to
limit the use of ZnO.
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March 2015
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