Author: Judith Schmidt, Product Manager On Farm Solutions

Alarm in the gut! Horses have a susceptible digestive system that can quickly become unbalanced. Intestinal disorders in horses are usually associated with colic. Many factors can be responsible for intestinal issues. Have you ever thought about mycotoxins? What can horse owners do to support their horse´s gut health?
The equine stomach is not robust at all. Depending on their age and use, more than half of all horses suffer from stomach pain. Their digestive system is very sensitive and very different from that of other mammals: Horses cannot vomit and often suffer from severe abdominal pain, diarrhea, or cramps if they overeat or ingest spoiled feed.

The horse´s digestive system is complex and sensitive

The horse´s stomach has a relatively small capacity of around twelve to fifteen liters. Depending on the feed’s consistency and composition, it remains in the stomach for around one to five hours before it is pressed through the stomach outlet (pylorus) into the small intestine. The horse´s entire intestine is about ten times its body length.

Figure 1: The horse’s digestive tract

The horse´s gastrointestinal tract is a complex network, reacting extremely sensitively to changes and, therefore, highly susceptible to disorders. It essentially consists of the head intestine (lips, oral cavity, teeth, and esophagus), stomach (blind pouch, fundus, and stomach outlet), small intestine (duodenum, jejunum, and ileum), and large intestine (caecum, colon and rectum). Each section plays a crucial role in the digestive process; any disruption can lead to health issues. Understanding this structure is key to maintaining a horse’s digestive health.

Digestive disorders can have various reasons

Intestinal problems in horses can stem from diverse causes, often a complex interplay of multiple factors. By understanding these causes more deeply, horse owners can be better equipped to prevent and manage these issues. In the following, we delve into several of these causes.

1.   Too long time between the feedings

Usually, a feeding break should be at most four to six hours, as, in nature, a horse is busy eating for at least 18 hours a day. In contrast to humans, who produce stomach acid only after food intake, the horse’s stomach produces gastric acid around the clock. The continuous intake of roughage, intensive chewing, and high saliva production (a horse produces 5 to 10 L of saliva per day) is, therefore, essential to protect the stomach mucosa by neutralizing excess gastric acid.

A too-long time between feedings and, therefore, no saliva production leads to an accumulation of gastric acid in the stomach. Four hours without roughage can already cause inflammation of the mucosa and probably ulcers.

2.   Excessive amounts of concentrated feed

Excessive amounts of concentrates such as wheat or rye, conditioned by less chewing activity, increase gastric acid and histamine production, and the stomach lining can be attacked. Also, in this case, the development of stomach ulcers is possible.

Furthermore, the possibly resulting hyperacidity of the organism can lead to malfunctions of the organs, the skin, and the hooves.

3.   Stress

Stress can also lead to a higher production of gastric acid and, therefore, to gastric ulcers. The horse is a flight animal.  When it is under stress, it prepares for the impending escape, and the muscles are preferably supplied with blood, resulting in a lower blood flow to the mucous membranes. Furthermore, the rising cortisone level reduces the hydrochloric acid-suppressing prostaglandin E. As a result, more stomach acid is produced, irritating the gastric mucosa.

Stress can be triggered, e.g., by transportation, competitions, training, a change of house, a new rider, unsuitable equipment, or poor posture.

4.   Dental diseases

The teeth are essential for digestion. When feed is chewed, it is broken down and mixed with saliva. Chipped teeth cannot chew well, and the feed is not sufficiently salivated or crushed, which has a detrimental effect on digestion.

For this reason, an expert vet should check the horse´s teeth at least once a year.

5.   Administration of painkillers/medication

As with humans, long-term medication administration can promote the formation of stomach ulcers. For this reason, it is essential to ensure that horses are fed a gentle diet on the stomach, especially when using oral pain therapy, and to add stomach protection if necessary.

6. Endotoxins

If pathogens such as E. coli or clostridia proliferate extremely or are killed by an antibiotic, endotoxins can be released. These toxins can cause transformation or inflammation of the gut mucosa. In drastic cases, whole areas of the mucosa can die off. 

7. Mycotoxins – the hidden danger in horse feed

Mycotoxins in plants and horse feed are a common but often unnoticed danger to horses’ health. Mycotoxins are natural, secondary metabolites of molds that have a toxic effect on humans and animals and can trigger mycotoxicosis. Contaminated feed can severely affect the horse’s health and, in the worst case, lead to death.

Over 90 % of the world´s feed production is estimated to be contaminated with at least one mycotoxin (see also Global Mycotoxin Report 2023, EW Nutrition. The intake of mycotoxins via hay, grain, silage, or compound feed can hardly be avoided. Mycotoxins are an increasing problem for all horse owners. Scientific studies show that the mycotoxins DON and ZEA are most frequently found in horse feed and, therefore, are also frequently detected in sports horses’ urine and blood samples.

Due to the highly toxic metabolic products, feed contaminated with molds can lead to severe liver and kidney diseases in horses, affect fertility, trigger colic, and promote digestive issues (diarrhea and watery stools).

Figure 2: Mycotoxins and their impact on horses

How to protect the horse from mycotoxins?

The first measure against the ingestion of mycotoxins is prevention. Correct pasture management and adequate barn and feed hygiene can contribute to preventing the ingestion of toxins.

However, despite the best prophylactic measures, it is impossible to prevent mycotoxin contamination of feed completely. As mycotoxins are not visible, analyzing the feed regarding mycotoxin contamination is recommended.

To protect your horse from mycotoxins, EW Nutrition developed MasterRisk, a tool for evaluating the risk of mycotoxins. Additionally, EW Nutrition has developed a complementary feed specifically for your horse´s needs in the form of granules. The sophisticated formulation of “Toxi-Pearls” is designed to bind mycotoxins and mitigate the adverse effects of mycotoxin contamination.

The pearls contain a mixture of mycotoxin binder, brewer’s yeast, and herbs:

• The contained mycotoxin binder effectively controls the most important feed myco- and endotoxins. It additionally supports the liver and immune system and strengthens the intestinal barrier.
• Brewer´s yeast supports the natural strength of the gastrointestinal tract. Due to its high natural content of beta-glucans and mannan-oligosaccharides (MOS), unique surface structure, and the associated high adsorption power, brewer´s yeast has a prebiotic effect on the intestinal microbiome.
• The additional unique herbal mixture consists of the typical gastrointestinal herbs oregano, rosemary, aniseed, fennel, and cinnamon. The processed beetroot is a true all-rounder. Literature shows that it has an antioxidant effect and strengthens the immune system. It also promotes bile secretion and, therefore, supports fat digestion.

Conclusion

The horse’s digestive tract is highly sensitive and must be supported by all means. Besides failures in management, such as too long breaks between feedings or too high amounts of feed concentrate, mycotoxins present a high risk in horse nutrition. To prevent horses from intestinal issues, feed and stress management, dental care, and medication in the case of disease must be optimized. Particular attention should be paid to possible mycotoxin contamination. Effective toxin risk management, which consists of analysis, risk evaluation, and adequate toxin risk-managing products, should be implemented.

Part 2: Beak/mouth lesions

by Marisabel Caballero and Inge Heinzl, EW Nutrition

The second part of this series will focus on oral lesions as signs of mycotoxin exposure. In this segment, we will delve into the appearance and development of oral lesions, their specific locations based on the type of mycotoxin, and how toxin levels and duration of exposure impact these lesions.

A bit of history: oral lesions in poultry and their association with mycotoxin exposure

Exposure to trichothecenes, a specific group of mycotoxins that includes T-2 toxin and scirpenols- such as monoacetoxyscirpenol (MAS), diacetoxyscirpenol (DAS), and triacetoxyscirpenol, has been associated with oral lesions since the early studies related with mycotoxins:

• After reports of toxicosis in farm animals, Bamburg’s group (1968) aimed to isolate the toxins produced by Fusarium tricintum, then considered the most toxic fungus found in moldy corn in Wisconsin (USA). Their experiments led to the discovery of the T-2 toxin, named after the strain of F. tricintum from which it was isolated. Today, we know that this fungus was wrongly identified; it was F. sporotrichioides (Marasas et al., 1984). However, the toxin remained known as T-2.
• Wyatt’s group (1972) already described yellowish-white lesions in the oral cavity of commercial broilers in a case report from 1972. The birds also presented lesions on the feet, shanks, and heads, which raised the possibility of contact with the toxin from the litter.
• In some of the earliest experimental works regarding T-2 toxin in poultry, Christensen (1972) noted the development of oral necrosis in turkey poults consuming increasing levels of feed invaded by tricintum; also Wyatt (1972) found a linear increase in lesion size and severity with increasing toxin concentrations of T-2 in broilers, starting with 1 ppm. He noted that oral lesions occurred without exception in all birds receiving T-2 toxin.
• Later, Chi and co-workers (1977) tested what later were considered sub-acute levels of T-2 in broiler chickens, finding oral lesions from 0.4 ppm after 5 to 6 weeks of exposure. At higher levels, the lesions appeared after two weeks. In the same year, Speers’ group (1977) concluded that adult laying hens are more tolerant to T-2 than young chicks and also found that another mycotoxin can produce oral lesions in poultry: monoacetoxyscirpenol (MAS).
• Fast forward, scientific research continued and the effects of T-2 and scirpenols, either alone or in combinations, on performance and oral lesions in poultry are today well known, as studied by Kubena et al. (1989), Ademoyero & Hamilton (1991), Kubena et al. (1994), Diaz et al. (1994), Brake et al. (2000), Schuhmacher-Wolz et al. (2010), Verma & Swamy (2015), Vaccari (2017), and reviewed by Sokolovic et al. (2008), Minafra et al. (2018), Puvača & Ljubojević Pelić (2023), and Vörösházi et al. (2024).

What are oral lesions and how do they develop?

Oral lesions caused by feed contaminated by T-2 toxin or scirpenols first occur as yellow plaques that develop into raised yellowish-gray crusts with covered ulcers (Hoerr et al., 1982). They also have been described as white in color and sometimes caseous in nature, as well as round and small, pin-point-sized, or large sheets covering a wider part of the mouth (Wyatt et al., 1972; Ademoyero and Hamilton, 1991).

Under the microscope, the lesions show a fibrinous surface layer and intermediate layers with invaginations full of rods and cocci, suggesting that the surrounding microbiota quickly colonizes the lesion. Inflammation immediately ensues as Wyatt’s team (1972) found the underlying tissues filled with granular leukocytes.

Why do T-2 toxins and other trichothecenes cause such lesions?

T-2 toxin and other trichothecenes are known for their caustic nature (evidenced by studies of Chi and Mirocha, 1978; Marasas et al., 1969), and for incidents involving accidental exposure by laboratory personnel (Bamburg et al., 1968, cited in Wyatt et al., 1972).

Induction of necrosis has been proposed as the main toxicity effect based on in vitro experiments on human skin fibroblast models. The findings were a reduction of ATP production in the cell line together with disruption of mitochondrial DNA (mtDNA) but without an increase in reactive oxygen species (ROS) or activity of caspase-3 and caspase-7, which would be the case for apoptosis (Janik-Karpinsa et al., 2022). A further study (Janik-Karpinsa et al., 2023) found that T-2, on the same cell line, reduced the number of mtDNA copies, damaging several genes and hindering its function; consequently, ATP production is inhibited, and cell necrosis ensues.

Meanwhile, an inflammatory response is triggered, and the lesions are colonized by the surrounding microbial flora (Wyatt et al., 1972). Supporting this notion, Hoerr et al. (1981) observed no mouth lesions after directly administering toxins via crop gavage. Enterohepatic recirculation, facilitating the return of toxins to the oral cavity through saliva, can amplify their toxic effects (Leeson et al., 1995).

Oral lesions depend on…

…the toxin

Oral lesions vary depending on the type of toxin involved. The location of lesions is influenced by the specific mycotoxin in the feed. For instance, research by Wyatt et al. (1972) revealed that with T-2 toxin, lesions initially manifest on the hard palate and along the tongue’s margins. Over two weeks, these lesions progress to affect the lingual papillae at the tongue’s root, the underside of the tongue, and the inner side of the lower beak near the midline.

In contrast, Ademoyero and Hamilton (1991) found that scirpenols present a different pattern. A study including 4 mycotoxins at 5 different levels found, after three weeks of exposure, that the lesions caused by triacetoxyscirpenol (TAS) predominantly occurred in the angles of the mouth (53% of the birds in the study), sparing the tongue. On the other hand, diacetoxyscirpenol (DAS) primarily induces lesions inside the upper beak (shown 47% of the broilers), followed by the inside of the lower beak (in 32% of the birds). The lesion distribution for scirpentriol mirrors that of TAS, while monoacetoxyscirpenol (MAS) resembles DAS in its impact.

Chi and Mirocha (1978) conducted a comparative analysis of lesions caused by T-2 toxin and DAS (both 5 ppm). They observed that the severity of DAS-induced lesions was higher, leading to difficulties in mouth closure for some chicks due to encrustations in the mouth angles.

…the contamination level

Different findings regarding the dose dependency of the lesions are available. Wyatt et al. (1972) (Figure 1) showed a relationship between the lesion size and the toxin level. A clear relationship between the severity and incidence of lesions and the amount of T-2 toxin was also demonstrated by Chi et al. (1977) and Speers et al. (1976). This linear relationship in the case of T-2 toxin could be confirmed for the scirpenols TAS, STO, MAS, and DAS by Ademoyero and Hamilton (1991). They demonstrated a distinct dose-response relationship in a trial with the scirpenols STO, TAS (at 5 levels between 0-8 µg/g), MAS, and DAS (at 5 levels between 0-4 µg/g).

Sklan et al. (2001) tested T-2 toxin at more likely levels (0, 110, 530, and 1,050 ppb) in male chickens and found lesions in 90% of the chickens fed 500 ppb T-2 and in 100% of the ones fed 1,000 ppb of T-2 after 10 to 15 days; the higher dosage provoked the lesions of higher severity. When feeding 100 ppb of T-2, mild lesions appeared in 40% of the chickens after 25 and 35 days. Another group led by Sklan (2003) studied four groups of 12 one-day-old male turkey poults fed mash diets with 0 (control), 241, 485, or 982 ppb T-2 toxin for 32/33 days. Feed intake and feed efficiency were not affected, but oral lesions were apparent on day 7. The severity of the lesions plateaued after 7–15 days, and the lesion score was dose-related (see Figure 2). In the same trial, they also tested DAS (0, 223, 429, or 860 ppb) and found a similar dose relationship.

Figure 2: Lesion scores in poults fed T-2 toxin at different inclusion rates and lengths of exposure (Sklan et al., 2003)

A different result is found in the trial conducted by Hoerr et al. (1982), who observed lesions 2-4 days after initiating toxin exposure (T-2 toxin and DAS; 4 and 16 ppm for 21 days) and comparable lesions when feeding 50, 100, or 300 ppm of the same toxins for 7 days. They asserted that the toxin concentration did not influence the time to onset of lesions nor their severity. Most research, however, shows a clear dose-response relation.

…the duration of exposure

On one hand, chronic exposure to low levels of toxins often requires a specific duration before noticeable effects emerge. And on the other hand, symptoms may also diminish due to hormesis, an adaptive response of the organism to moderate, intermittent stress.

With high toxin levels, lesions appear very soon after exposure. For example, Diaz et al. (1994) exposed hens to a diet containing 2 mg DAS/kg feed, finding lesions in 40% of the birds after only 48 h of exposure. Chi and Mirocha (1978) noted lesions after five days with a T-2 level of 5 ppm. At a comparable level (4 ppm), Chi et al. (1977) reported lesions emerging in the second week of exposure, with nearly 75% of chicks experiencing oral lesions by the third week. Sklan et al. (2003) saw lesions already on day 7 when feeding T-2 toxin or DAS at 1 ppm.

When testing lower levels (200 ppb), Sklan et al. (2001) found lesions after 10 days. They became more severe after 15 to 20 days and then, their severity decreased. Hoerr et al. (1982) also confirmed this by reporting that the number and size of the lesions increased until day 14 but decreased thereafter. Both studies confirm the phenomenon of hormesis.

… animal factors

In general, lesions appear with lower levels of toxins in broilers compared with layers and in layers compared with breeders. Turkeys are also less sensitive than broilers (Puvača & Ljubojević Pelić (2023).

Age also has an influence: young birds usually still have a maturing immune system, and the detoxification processes might not be entirely in place. However, their feed intake is lower and for this reason, in studies like Wang and Hogan (2019), higher impact of mycotoxins is found in older chicks.

Furthermore, additional stress factors influence the impact of mycotoxins in animals. Stress factors are cumulative and, when different factors concur, the severity of mycotoxin effects can increase.

Are oral lesions key indicators for implementing effective toxin risk management?

Oral lesions are painful for the animals, distract them from eating, and deteriorate growth performance. Often they are related with mycotoxins; however, when they appear, an investigation of different factors should take place, including mycotoxin analysis, as oral lesions may have other causes. Some of the known causes of oral lesions in poultry are also very fine feed particle size, deficiency of Vitamins A, E, B6 and Biotin, excessive levels of copper sulphate, and some parasite infections.

This article aimed to help with the differential diagnosis by providing a summary of the knowledge we have about the type and shape of the lesions related to mycotoxin contamination, which can help on a differential diagnosis. Checking the feed for mycotoxins and implementing effective toxin management helps prevent their negative effects, keeps the animals healthy, and contributes to animal welfare and, consequently, performance.

References

Ademoyero, Adedamola A., and Pat B. Hamilton. “Mouth Lesions in Broiler Chickens Caused by Scirpenol Mycotoxins.” Poultry Science 70, no. 10 (October 1991): 2082–89. https://doi.org/10.3382/ps.0702082.

Bamburg, J.R., N.V. Riggs, and F.M. Strong. “The Structures of Toxins from Two Strains of Fusarium Tricinctum.” Tetrahedron 24, no. 8 (January 1968): 3329–36. https://doi.org/10.1016/s0040-4020(01)92631-6.

Bamburg, J.R., N.V. Riggs, and F.M. Strong. “The Structures of Toxins from Two Strains of Fusarium Tricinctum.” Tetrahedron 24, no. 8 (January 1968): 3329–36. https://doi.org/10.1016/s0040-4020(01)92631-6.

Brake, J., P.B. Hamilton, and R.S. Kittrell. “Effects of the Trichothecene Mycotoxin Diacetoxyscirpenol on Feed Consumption, Body Weight, and Oral Lesions of Broiler Breeders.” Poultry Science 79, no. 6 (June 2000): 856–63. https://doi.org/10.1093/ps/79.6.856.

Chi, M.S., and C.J. Mirocha. “Necrotic Oral Lesions in Chickens Fed Diacetoxyscirpenol, T—2 Toxin, and Crotocin.” Poultry Science 57, no. 3 (May 1978): 807–8. https://doi.org/10.3382/ps.0570807.

Chi, M.S., C.J. Mirocha, H.J. Kurtz, G. Weaver, F. Bates, and W. Shimoda. “Subacute Toxicity of T-2 Toxin in Broiler Chicks ,.” Poultry Science 56, no. 1 (January 1977): 306–13. https://doi.org/10.3382/ps.0560306.

Christensen, C. M., R. A. Meronuck, G. H. Nelson, and J. C. Behrens. “Effects on Turkey Poults of Rations Containing Corn Invaded by            Fusarium Tricinctum            (CDA.) Sny. & Hans.” Applied Microbiology 23, no. 1 (January 1972): 177–79. https://doi.org/10.1128/am.23.1.177-179.1972.

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Hoerr, F, W Carlton, B Yagen, and A Joffe. “Mycotoxicosis Caused by Either T-2 Toxin or Diacetoxyscirpenol in the Diet of Broiler Chickens.” Fundamental and Applied Toxicology 2, no. 3 (May 1982): 121–24. https://doi.org/10.1016/s0272-0590(82)80092-4.

Hoerr, F. J., W. W. Carlton, and B. Yagen. “Mycotoxicosis Caused by a Single Dose of T-2 Toxin or Diacetoxyscirpenol in Broiler Chickens.” Veterinary Pathology 18, no. 5 (September 1981): 652–64. https://doi.org/10.1177/030098588101800510.

Janik-Karpinska, Edyta, Michal Ceremuga, Magdalena Wieckowska, Monika Szyposzynska, Marcin Niemcewicz, Ewelina Synowiec, Tomasz Sliwinski, and Michal Bijak. “Direct T-2 Toxicity on Human Skin—Fibroblast HS68 Cell Line—in Vitro Study.” International Journal of Molecular Sciences 23, no. 9 (April 29, 2022): 4929. https://doi.org/10.3390/ijms23094929.

Janik-Karpinska, Edyta, Michal Ceremuga, Marcin Niemcewicz, Ewelina Synowiec, Tomasz Sliwiński, and Michal Bijak. “Mitochondrial Damage Induced by T-2 Mycotoxin on Human Skin—Fibroblast HS68 Cell Line.” Molecules 28, no. 5 (March 6, 2023): 2408. https://doi.org/10.3390/molecules28052408.

Kubena, L.F., R.B. Harvey, T.S. Edrington, and G.E. Rottinghaus. “Influence of Ochratoxin A and Diacetoxyscirpenol Singly and in Combination on Broiler Chickens.” Poultry Science 73, no. 3 (March 1994): 408–15. https://doi.org/10.3382/ps.0730408.

Kubena, L.F., R.B. Harvey, W.E. Huff, D.E. Corrier, T.D. Phillips, and G.E. Rottinghaus. “Influence of Ochratoxin A and T-2 Toxin Singly and in Combination on Broiler Chickens.” Poultry Science 68, no. 7 (July 1989): 867–72. https://doi.org/10.3382/ps.0680867.

Leeson, Steven, Gonzalo J. Diaz, and John D. Summers. Poultry metabolic disorders and Mycotoxins. University Books, 1995.

Marasas, W.F.O., J.R. Bamburg, E.B. Smalley, F.M. Strong, W.L. Ragland, and P.E. Degurse. “Toxic Effects on Trout, Rats, and Mice of T-2 Toxin Produced by the Fungus Fusarium Tricinctum (Cd.) Snyd. Et Hans.” Toxicology and Applied Pharmacology 15, no. 2 (September 1969): 471–82. https://doi.org/10.1016/0041-008x(69)90045-3.

Minafra, Cibele, Denise Russi Rodrigues, Isabel Cristina Mores Vaccari, Vinícius Duarte, Fabiana Ramos dos Santos, Weslane Justina da Silva, Alison Batista Vieira Silva Gouveia, Lorrayne Moraes de Paulo, Janaina Borges dos Santos, and Júlia Marixara Souza Silva. “Oral Lesions in Broilers Caused by Corn Mycotoxins: Review – Original: Lesões Orais Em Frangos de Corte Provocadas Por Micotoxinas Do Milho: Revisão.” Pubvet 12, no. 07 (July 17, 2018). https://doi.org/10.31533/pubvet.v12n7a134.1-11.

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Puvača, Nikola, and Dragana Ljubojević Pelić. “Problems and Mitigation Strategies of Trichothecenes Mycotoxins in Laying Hens Production.” Journal of Agronomy, Technology and Engineering Management (JATEM) 7, no. 2 (April 1, 2024): 1074–87. https://doi.org/10.55817/isad5453.

Riahi, Insaf, Virginie Marquis, Anna Maria Pérez-Vendrell, Joaquim Brufau, Enric Esteve-Garcia, and Antonio J. Ramos. “Effects of Deoxynivalenol-Contaminated Diets on Metabolic and Immunological Parameters in Broiler Chickens.” Animals 11, no. 1 (January 11, 2021): 147. https://doi.org/10.3390/ani11010147.

Schuhmacher-Wolz, Ulrike, Karin Heine, and Klaus Schneider. “Toxicity of HT-2 and T-2 Toxins.” European Food Safety Authority, 2010. https://www.efsa.europa.eu/en/supporting/pub/en-65.

Sklan, D., E. Klipper, A. Friedman, M. Shelly, and B. Makovsky. “The Effect of Chronic Feeding of Diacetoxyscirpenol, T-2 Toxin, and Aflatoxin on Performance, Health, and Antibody Production in Chicks.” Journal of Applied Poultry Research 10, no. 1 (March 2001): 79–85. https://doi.org/10.1093/japr/10.1.79.

Sklan, D., M. Shelly, B. Makovsky, A. Geyra, E. Klipper, and A. Friedman. “The Effect of Chronic Feeding of Diacetoxyscirpenol and T-2 Toxin on Performance, Health, Small Intestinal Physiology and Antibody Production in Turkey Poults.” British Poultry Science 44, no. 1 (March 2003): 46–52. https://doi.org/10.1080/0007166031000085373.

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Wyatt, R. D., J. R. Harris, P. B. Hamilton, and H. R. Burmeister. “Possible Outbreaks of Fusariotoxicosis in Avians.” Avian Diseases 16, no. 5 (October 1972): 1123. https://doi.org/10.2307/1588839.

Over the past decade, the food industry witnessed a surge in the popularity of alternative proteins, driven by growing consumer awareness of environmental issues, animal welfare concerns, and health considerations. However, recent trends indicate a decline in both consumer interest and investment in alternative proteins. This article explores the challenges in producing viable replacements for traditional meat, the status of sales investments, and the global outlook for protein consumption.

Figure 1. Uncompetitive prices of artificial meat are a critical factor in the market downturn

Challenges in artificial meat production

Producing artificial meat, also known as cultured or lab-grown meat, has been widely hyped and substantially funded over the last decade. However, many challenges remain on several levels.

Cell Culturing and Growth

Cell Source: Obtaining high-quality animal cells is crucial. Researchers typically use muscle cells (myocytes) from animals like cows, pigs, or chickens.

Cell Proliferation: Culturing cells in the lab requires precise conditions, including the right nutrients, temperature, and oxygen levels. Ensuring rapid and efficient cell growth is essential.

Scaffold Development

3D Structure: Creating a meat-like texture involves growing cells on a scaffold that mimics the natural 3D structure of muscle tissue. Developing suitable scaffolds is challenging.

Biocompatibility: The scaffold material must be biocompatible and support cell attachment, proliferation, and differentiation.

Nutrient Supply

Medium Formulation: The nutrient-rich medium used to feed the cells must provide essential amino acids, vitamins, and minerals. Designing an optimal medium is complex.

Cost Efficiency: Developing cost-effective and sustainable nutrient solutions is critical for large-scale production.

Scaling Up Production

Bioreactors: Moving from small-scale lab experiments to large-scale bioreactors is a significant challenge. Bioreactors must maintain consistent conditions for cell growth.

Energy Consumption: Scaling up production while minimizing energy consumption and environmental impact is essential.

Flavor and Texture

Taste and Aroma: Artificial meat would be expected to taste and smell like traditional meat. Achieving the right flavor profile is an ongoing challenge.

Texture: Mimicking the texture of different meat cuts (e.g., steak, ground beef) requires precise engineering.

Safety and Regulation

Food Safety: Ensuring that cultured meat is safe for consumption is critical. Contamination risks, such as bacterial growth, must be minimized.

Regulatory Approval: Cultured meat faces regulatory hurdles related to labeling, safety assessments, and consumer acceptance.

Cost Reduction

High Initial Costs: Currently, producing artificial meat is expensive due to research, development, and infrastructure costs. Reducing these costs is essential for commercial viability.

Acceptance and Perception

Consumer Perception: Convincing consumers that cultured meat is a viable and ethical alternative to traditional meat remains a challenge.

Cultural and Social Factors: Cultural preferences and traditions play a role in consumer acceptance.

Challenges in alternative protein production

As opposed to artificial meat, which still involves animal cells, alternative proteins usually designate plant-based meat imitations. However, producing alternative proteins comes with its own set of challenges.

Diverse protein sources are one challenge that is not easy to overcome. It turns out, it is quite hard to replicate the availability, as well as the diversity of health and nutritional benefits of traditional meat. While plant-based proteins have made significant progress, there’s still room for improvement in terms of variety and availability.

Procuring the technology needed to extract protein efficiently and sustainably is another hurdle. Innovations in extraction methods are essential for scaling up alternative protein production.

Lower nutritional benefits of alternative proteins represent a major hurdle. Not only is it hard to mimic the entirety of meat’s benefits, but plant nutritional values are notoriously fickle depending on region, soil, production type, season, and so on.

Flavor and texture remain extremely elusive. Contenders are closer to a meat-feel than before, yet this remains a major factor skewing negative in consumer perception.

Figure 2: Alternative protein producers have been unable to replicate the taste and texture of traditional meat

Scaling and Supply Chain Challenges are getting more, not less complicated. Achieving affordability at scale is essential for alternative meats to compete with traditional meat products. Additionally, ensuring a robust and efficient supply chain for alternative proteins is a concern that has not found a sustainable solution.

The Status of Alternative Protein Sales and Investment

Sales Trends

According to the Plant Based Foods Association (PBFA), overall plant-based meat units have declined by 8.2% in 2022, while dollar sales decreased by 1.2% following a significant growth phase in previous years. Similarly, Euromonitor International reported that global sales of plant-based meat substitutes grew by only 1% in 2022, a stark contrast to the double-digit growth rates seen earlier in the decade.

Beyond Meat, one of the market leaders, reported a decline in net revenues of 13.9% in the third quarter of 2022 compared to the same period in 2021. This decline reflects broader market trends where consumer enthusiasm appears to be waning.

Figure 3. Rabobank indicates a downward trend in both sales and investments

Investment Trends

Investment in alternative protein startups also shows signs of slowing, with funding of sustainability food and agriculture startups dramatically declining (see Figure 2 below). According to the Good Food Institute (GFI), global investment in alternative proteins dropped 42% year-over-year to $1.2 billion in 2022, a significant decrease from the $3.1 billion invested in 2021.

The financial challenges faced by some high-profile companies have led to increased caution among investors. For instance, Beyond Meat and Oatly have both experienced substantial stock price declines, leading to a reassessment of the market’s growth potential.

Figure 4: 2023 funding variation for climate and sustainability technologies

Factors Contributing to the Decline

Market Saturation and Competition

The initial surge in demand led to rapid market saturation. Numerous companies entered the market, resulting in intense competition and a proliferation of products. This saturation has made it difficult for individual brands to maintain market share and grow sales.

In the US, for example, plant-based milk remains the largest category, while plant-based meat and seafood sales declined substantially in 2023.

Consumer Preferences and Expectations

While early adopters of alternative proteins were driven by ethical and environmental considerations, mainstream consumers remain price-sensitive and often prefer traditional meat products (to the extent they may choose smaller meat portions over alternative proteins). Additionally, taste and texture remain critical factors. Despite advancements, many consumers still find plant-based alternatives lacking in these areas.

Figure 5: Seems fake? Consumers find it hard to believe the claims of identical taste and texture in non-meat products

Economic Factors

The global economic downturn and inflation have impacted consumer spending power. As a result, many consumers are prioritizing affordability over sustainability, leading to reduced purchases of typically more expensive plant-based products.

Regulatory and Supply Chain Challenges

Regulatory hurdles and supply chain disruptions have also played a role. The COVID-19 pandemic exacerbated supply chain issues, affecting the availability and cost of raw materials needed for alternative protein production.

Conclusion: Global Outlook for Protein and Alternative Proteins

Traditional meat consumption continues to grow, particularly in emerging markets. According to the Food and Agriculture Organization (FAO), global meat consumption is projected to increase by 14% by 2030, driven by population growth and rising incomes in developing countries.

Advances in food technology, such as precision fermentation and cell-cultured meat, offer the potential to create products that more closely mimic traditional meat. However, the recent decline in interest in alternative proteins reflects a complex interplay of market saturation, economic factors, and consumer preferences.

High prices, lack of scalability, sustainability concerns, and an inability to recreate the nutritional content, texture, and taste of meat are hurdles that cannot be easily overcome. Instead, perhaps a more accessible long-term solution might be improved sustainability in the livestock sector, accompanied by continued innovation and improvements in the production of both traditional protein and alternative proteins.

The European Environment Agency (EEA) has recently published a briefing detailing the impact of veterinary antimicrobials on Europe’s environment. Positive developments are to be applauded, however they do not tell the whole story.

The use of antimicrobials for farmed animals and in aquaculture decreased by around 28% between 2018 and 2022. Nonetheless, the rate of antimicrobial resistance continues to rise around the world, including as an important cause of death in the European Economic Area (the EEA includes all EU countries, as well as Norway, Lichtenstein, and Iceland). At present, antimicrobial-resistant infections are estimated to caused 35,000 deaths per year in the European Union. For reference, in the EU, traffic accidents cause around 20,000 deaths per year.

A large number of EU guidelines, policies, and regulations attempt to control and monitor the use of antimicrobials in food-producing animals. This makes the regulatory landscape somewhat confusing, especially that many implementation details are still left to the states.

Figure 1. Overview of the EU regulatory framework applicable to antimicrobials used in food-producing animals

One of the results of unequal implementation is that there is no standardized way of tracking the actual use of antimicrobials in food-producing animals. To collect the numbers, the EEA has used proxy numbers, especially antimicrobial sales data and self-reported data. With such broad strokes, it is to be expected that the actual figures might be higher.

Lower numbers…

According to the European Surveillance of Veterinary Antimicrobial Consumption (ESVAC) database, in the European Economic Area countries, plus Switzerland and the UK, total antimicrobial consumption for farmed animals and aquaculture was estimated at 73.9 mg/PCU* in 2022. This signifies a 30% reduction over 5 years.

In numbers: 4,458 tons of active substances were sold in one years for farmed animals & aquaculture.

*PCU represents a population correction unit (PCU). The PCU takes into account the population and relative weight of animals and “is used the normalize antimicrobial sales data for the size of the animal population that could potentially be treated with these substances. Using this methodology, 1 PCU corresponds to 1 kilogram of animal biomass”.

…But higher risks

In 2021, total antimicrobial consumption in humans – measured in 28 European countries – was estimated at 125.0 mg/kg. This number has unfortunately not gone down. What is worse: a much larger volume of antimicrobials is sold for food-producing animals than for human medicine. Which means that, relative to the total population, the impact of veterinary-use antimicrobials remains disproportionately large.

Moreover, two outliers (Poland and Lithuania) exhibited a worrying increasing trend, showing that no good development is irreversible. The EEA also highlights this danger, in the context of growing global consumption of animal protein. Increased demand “may put pressure on farmers to adopt intensive production practices that require increased use of antimicrobials”. The use of antimicrobials elsewhere in the world may lead to impacts in Europe, “not just by theoretically exposing consumers to antimicrobial residues but also by contributing to rising global rates of drug-resistant pathogens and infections”.

Figure 2. Sales by EU Member States in 2018 vs. 2022

Because of declining livestock populations in the last few years, while demand remained constant, EU-27 imports of animal products more than doubled between 2002 and 2022. There are no reliable global data on the veterinary use of antimicrobials, however it is generally believed that over 70% of antimicrobials sold globally may be used in for animal protein production.

Data collected by the World Organization for Animal Health (WOAH) from its member countries suggest that, between 2017-2019, “the use of antimicrobials in animals decreased by 25% in the Asia, Far East and Oceania regions, while it increased in Africa (+45%) and the Americas (+5%). Despite these partial improvements, a recent study forecasted that global use of antimicrobials in food-producing animals could rise by 8% in 2030, compared to 2020 levels (Mulchandani et al., 2023)”.

Quick summary

Reduction in Antimicrobial Use: There has been a significant reduction in the use of antimicrobials in farming and aquaculture across the EU. From 2018 to 2022, there was a decrease of approximately 28%, which aligns with the EU’s Farm to Fork strategy targeting a 50% reduction by 2030.

Antimicrobial Resistance (AMR): Despite the decrease in use, antimicrobial resistance remains a severe public health threat, causing an estimated 35,000 deaths annually in the European Economic Area. The resistance is attributed to the use of antimicrobials, which – as has been widely documented and discussed – can promote the evolution of resistant microorganisms.

Environmental Impact: The briefing underscores significant knowledge gaps in monitoring antimicrobial residues, resistant bacteria, and resistance genes in the environment. Improved surveillance could help identify pollution hotspots and assess the impact of reduction measures.

Regulatory and Policy Framework: The EU has implemented several policies to regulate the use of antimicrobials, including banning their use as growth promoters and setting stricter conditions for prescriptions. These measures are crucial for managing the risk of AMR.

Further efforts are needed to decrease the reliance on antimicrobials in food production. These include enhanced monitoring, promoting alternative practices in animal farming, and better animal welfare and biosecurity measures.

While improvements are clear and commendable in the EU-27 states, increased antimicrobial usage in some EU countries and in various areas around the world represent a significant concern.

For further details on the use of veterinary antimicrobials in Europe’s environment, you can refer to the EEA’s full report.

Author: Ajay Bhoyar, Global Technical Manager, EW Nutrition

Exogenous feed enzymes are increasingly utilized in poultry diets to manage feed costs, mitigate the adverse effects of anti-nutritional factors, and enhance nutrient digestion and bird performance. These enzymes are primarily employed to bolster the availability of nutrients within feed ingredients. Among the various enzymes utilized, those capable of breaking down crude fiber, starch, proteins, and phytates are commonly integrated into animal production systems.

In monogastric animals such as poultry and swine, a notable deficiency exists in the endogenous synthesis of enzymes necessary for the hydrolysis of non-starch polysaccharides (NSPs) like xylan (McLoughlin et al., 2017). This deficiency often manifests in poultry production as a decline in growth performance, attributed to increased digesta viscosity arising from the prevalence of NSPs in commonly utilized poultry feed ingredients. Without sufficient endogenous enzymes to degrade xylan, NSPs can increase digesta viscosity, encase essential nutrients, and create a barrier to their effective digestion. In response to this issue, monogastric animal producers have implemented exogenous enzymes such as xylanases into the feeds for swine and poultry to degrade xylan to short-chain sugars, thus reducing intestinal viscosity and improving the digestive utilization of nutrients (Sakata et al., 1995; Aragon et al., 2018)

Understanding Xylanase Enzymes

Xylanase enzymes belong to the class of carbohydrases that specifically target complex polysaccharides, such as xylan, a backbone nonstarch polysaccharide (NSP) prevalent in plant cell walls. These enzymes catalyze the hydrolysis of xylan into smaller, more digestible fragments, such as arabino–xylo-oligosaccharides (AXOs) and xylo-oligosaccharides (XOs), thereby facilitating the breakdown of dietary fiber in poultry diets.

Mechanism of action

It is generally agreed that the beneficial effects of feed xylanase are primarily due to the reduction in viscosity. Studies have shown that supplementing xylanases to animal feeds reduces digesta viscosity and releases encapsulated nutrients, thus improving the overall feed digestibility and nutrient availability (Matthiesen et al., 2021). The reduction in digesta viscosity by adding xylanase is achieved by the partial hydrolysis of NSPs in the upper digestive tract, leading to a decrease in digesta viscosity in the small intestine (Choct & Annison, 1992).

GH10 vs. GH11 Xylanases

Well-characterized xylanases are mostly grouped into glycoside hydrolase families 10 (GH10) and 11 (GH11) based on their structural characteristics (amino acid composition), mode of xylan degradation, the similarity of catalytic domains, substrate specificities, optimal conditions, thermostability, and practical applications.

Why are GH10 xylanases more efficient in animal production?

While both GH10 and GH11 xylanases act on the xylan main chain, these two enzyme types have different folds, substrate specificities, and mechanisms of action (Biely et al., 2016). The GH10 xylanases are more beneficial in animal feed production due to their efficient mechanism of action, broader substrate specificity, and better thermostability, as discussed below.

GH10 xylanase exhibits broader substrate specificity

Generally, the GH10 xylanases exhibit broader substrate specificity and can hydrolyze various forms of xylan, including soluble and insoluble substrates. On the other hand, GH11 xylanases have a narrower substrate specificity and are primarily active on soluble xylan substrates. GH10 xylanases exhibit higher catalytic versatility and can catalyze the cleavage of the xylan backbone at the nonreducing side of substituted xylose residues, whereas GH11 enzymes require unsubstituted regions of the xylan backbone (Collins et al., 2005; Chakdar et al., 2016).

As a result, GH10 xylanases generally produce shorter xylo-oligosaccharides than members of the GH11 family (Collins et al., 2005). Moreover, as shown in Fig.1, the GH10 xylanase can rapidly and effectively break down xylan molecules.

Fig.1.: Activity of a bacterial GH10 xylanase against soluble and insoluble arabinoxylans

Higher thermostability

Enzymes are proteins, and the protein’s primary structure determines their thermostability. The enzyme protein tends to denature at higher than tolerable temperatures, rendering it inactive. An enzyme’s high-temperature tolerance ensures its efficacy throughout the pelleted feed manufacturing. This results in consistent enzyme activity in the finished feed, subsequent gut health, and predictable performance benefits.

Xylanases with higher thermostability are more suitable for applications requiring high-temperature processes. An intrinsically heat-stable bacterial xylanase maintains its activity even under high-temperature feed processing conditions, such as pelleting.

A study conducted at the University of Novi Sad, Serbia (Fig. 2), with three pelleting temperatures (85 °C, 90 °C, and 95 °C) and conditioning times of 4 and 6 mins, showed that Axxess XY, an intrinsically thermostable GH10 xylanase, demonstrated more than 85% recovery even at 4 to 6 mins conditioning time and 95 °C temperature.

Fig.2: Optimum recovery of Axxess XY at elevated conditioning time and temperatures

Maintaining consistently optimum enzyme activity is crucial for realizing the benefits of enzyme inclusion in feed under challenging feed processing conditions.

Conclusion

In conclusion, exogenous feed enzymes, including xylanase, have gained widespread recognition for their pivotal role in poultry nutrition. The increasing use of xylanase is attributed to its ability to effectively manage feed costs while incorporating high-fiber ingredients without compromising poultry performance. However, the efficacy of xylanase is based on several factors, including its mode of action, substrate specificity, catalytic efficacy, and thermostability. Selecting the appropriate xylanase enzyme tailored for specific needs is crucial to harnessing its full benefits.

A GH10 xylanase, such as Axxess XY, designed explicitly as a feed enzyme, offers distinct advantages in poultry production. Its efficient mechanism of action, broader substrate specificity, and superior thermostability make it a preferred choice for optimizing animal performance. Notably, Axxess XY exhibits exceptional activity against soluble and insoluble arabinoxylans, thereby enhancing nutrient utilization, promoting gut health, and ultimately elevating overall performance levels in poultry.

Incorporating specialized GH10 Xylanase enzymes like Axxess XY represents a strategic approach to unlocking the nutrients in feedstuffs, ensuring optimal performance, and maximizing profitability in the poultry business.

References

Aragon, Caio C., Ana I. Ruiz-Matute, Nieves Corzo, Rubens Monti, Jose M. Guisán, and Cesar Mateo. “Production of Xylo-Oligosaccharides (XOS) by Controlled Hydrolysis of Xylan Using Immobilized Xylanase from Aspergillus Niger with Improved Properties.” Integrative Food, Nutrition and Metabolism 5, no. 4 (2018). https://doi.org/10.15761/ifnm.1000225.

Bedford, Michael R., and Henry L. Classen. “Reduction of Intestinal Viscosity through Manipulation of Dietary Rye and Pentosanase Concentration Is Effected through Changes in the Carbohydrate Composition of the Intestinal Aqueous Phase and Results in Improved Growth Rate and Food Conversion Efficiency of Broiler Chicks.” The Journal of Nutrition 122, no. 3 (March 1992): 560–69. https://doi.org/10.1093/jn/122.3.560.

Biely, Peter, Suren Singh, and Vladimír Puchart. “Towards Enzymatic Breakdown of Complex Plant Xylan Structures: State of the Art.” Biotechnology Advances 34, no. 7 (November 2016): 1260–74. https://doi.org/10.1016/j.biotechadv.2016.09.001.

Chakdar, Hillol, Murugan Kumar, Kuppusamy Pandiyan, Arjun Singh, Karthikeyan Nanjappan, Prem Lal Kashyap, and Alok Kumar Srivastava. “Bacterial Xylanases: Biology to Biotechnology.” 3 Biotech 6, no. 2 (June 30, 2016). https://doi.org/10.1007/s13205-016-0457-z.

Choct, M., and G. Annison. “Anti‐nutritive Effect of Wheat Pentosans in Broiler Chickens: Roles of Viscosity and Gut Microflora.” British Poultry Science 33, no. 4 (September 1992): 821–34. https://doi.org/10.1080/00071669208417524.

Collins, Tony, Charles Gerday, and Georges Feller. “Xylanases, Xylanase Families and Extremophilic Xylanases.” FEMS Microbiology Reviews 29, no. 1 (January 2005): 3–23. https://doi.org/10.1016/j.femsre.2004.06.005.

Matthiesen, Connie F., Dan Pettersson, Adam Smith, Ninfa R. Pedersen, and Adam. C. Storm. “Exogenous Xylanase Improves Broiler Production Efficiency by Increasing Proximal Small Intestine Digestion of Crude Protein and Starch in Wheat-Based Diets of Various Viscosities.” Animal Feed Science and Technology 272 (February 2021): 114739. https://doi.org/10.1016/j.anifeedsci.2020.114739.

McLoughlin, Rebecca F, Bronwyn S Berthon, Megan E Jensen, Katherine J Baines, and Lisa G Wood. “Short-Chain Fatty Acids, Prebiotics, Synbiotics, and Systemic Inflammation: A Systematic Review and Meta-Analysis.” The American Journal of Clinical Nutrition 106, no. 3 (March 2017): 930–45. https://doi.org/10.3945/ajcn.117.156265.

Sakata, T., M. Adachi, M. Hashida, N. Sato, and T. Kojima. “Effect of N-Butyric Acid on Epithelial Cell Proliferation of Pig Colonic Mucosa in Short-Term Culture.” DTW – Deutsche Tierärztliche Wochenschau 102, no. 4 (1995): 163–64.

By Dr. Ajay Bhoyar, Global Technical Manager, EW Nutrition

In modern, intensive poultry production, the imminent threat of resistant Eimeria looms large, posing a significant challenge to the sustainability of broiler operations. Eimeria spp., capable of developing resistance to our traditional interventions, has emerged as a pressing global issue for poultry operators. The resistance of Eimeria to conventional drugs, coupled with concerns over drug residue, has necessitated a shift towards natural, safe, and effective alternatives.

Several phytogenic compounds, including saponins, tannins, essential oils, flavonoids, alkaloids, and lectins, have been the subject of rigorous study for their anticoccidial properties. Among these, saponins and tannins in specific plants have emerged as powerful tools in the fight against these resilient protozoa. In the following, we delve into innovative strategies that leverage the potential of these compounds, particularly saponins and tannins, to prevent losses by mitigating the risk of resistant Eimeria in poultry production.

Understanding resistant Eimeria in broiler production

The World Health Organization Scientific Group (World Health Organization, 1965) developed the definition of resistance in broad terms as ‘the ability of a parasite strain to survive and/or to multiply despite the administration and absorption of a drug given in doses equal to or higher than those usually recommended but within the limits of tolerance of the subject’.

The high reproduction rate of Eimeria spp. allows them to evolve quickly and develop resistance to drugs used for their control. Moreover, the resistant strains of Eimeria can persist in the environment due to their ability to form resistant oocysts, leading to the re-infection of animals and further spread of resistant strains.

Resistant Eimeria strains present many challenges in modern poultry farming, significantly impacting overall productivity and economic sustainability. However, one of the primary challenges is the reduced efficacy of traditional anti-coccidial drugs.

Eimeria resistance occurs in different types

There are different possibilities as to why Eimeria are resistant to specific drugs.

Acquired resistance results from heritable decreases in the sensitivity of specific strains and species of Eimeria to drugs over time. There are two types of acquired resistance: partial and complete. These types depend upon the extent of sensitivity lost. There is a direct relationship between the concentration of the drug and the degree of resistance. A strain controlled by one drug dose may show resistance when a lower concentration of the same drug is administered.

Cross-resistance is the sharing of resistance among different compounds with similar modes of action (Abbas et al., 2011). This, however, may not always occur (Chapman, 1997).

Multiple resistance is resistance to more than one drug, even though they have different modes of action (Chapman, 1993).

Natural substances can bring back the efficacy of anticoccidial measures

It was found that if a drug to which the parasite has developed resistance is withdrawn from use for some time or combined with another effective drug, the sensitivity to that drug may return (Chapman, 1997).

Botanicals and natural identical compounds are well renowned for their antimicrobial and antiparasitic activity, so they can represent a valuable tool against Eimeria (Cobaxin-Cardenas, 2018). The mechanisms of action of these molecules include degradation of the cell wall, cytoplasm damage, ion loss with reduction of proton motive force, and induction of oxidative stress, which leads to inhibition of invasion and impairment of Eimeria spp. development (Abbas et al., 2012; Nazzaro et al., 2013). Natural anticoccidial products may provide a novel approach to controlling coccidiosis while meeting the urgent need for control due to the increasing emergence of drug-resistant parasite strains in commercial poultry production (Allen and Fetterer, 2002).

Saponins and Tannins: Nature’s Defense against Eimeria Challenge

Phytogenic solutions, specifically those based on saponins and tannins, have recently surfaced as promising alternatives to mitigate the Eimeria challenge in poultry production. By harnessing the power of these natural compounds, poultry producers can boost the resilience of their flocks against the Eimeria challenge, promoting both the birds’ welfare and the industry’s sustainability.

Saponins are glycosides found in many plants with distinctive soapy characteristics due to their ability to foam in water. In the context of Eimeria, saponins can disrupt the integrity of the parasites’ cell membranes. When consumed, saponins can interfere with the protective outer layer of Eimeria, weakening the parasite and rendering it vulnerable to the host’s immune responses. This disruption impedes the ability of Eimeria to attach to the intestinal lining and reproduce, effectively curtailing the infection.

Tannins are polyphenolic compounds with astringent properties, occurring in various plant parts, such as leaves, bark, and fruits. Choosing the proper tannin at the right level and time is crucial to realize the benefits of tannin-based feed additives.

In the context of Eimeria, tannins exhibit several mechanisms of action. Firstly, they bind to proteins within the parasites, disrupting their enzymatic activities and metabolic processes. This interference weakens Eimeria, hindering its ability to cause extensive damage to the intestinal lining. Secondly, tannins are anti-inflammatory, reducing the inflammation caused by Eimeria infections. Additionally, tannins act as antioxidants, protecting the intestinal cells from oxidative stress induced by the parasite.

When incorporated into broilers’ diets, saponins and tannins create an unfavorable environment for Eimeria, inhibiting their growth and propagation within the host. Moreover, these compounds fortify the broiler’s natural defenses, enhancing its ability to resist Eimeria infections. By leveraging the innate properties of saponins and tannins, the impact of resistant Eimeria strains can effectively be managed and mitigated, fostering healthier flocks and sustainable poultry production.

What is Pretect D?

Pretect D is a unique proprietary blend of phytomolecules, including saponins and tannins, that supports the control of coccidiosis challenges in poultry production. It can be used alone or in combination with coccidiosis vaccines, ionophores, and chemicals as part of a shuttle or rotation program.

Fig.1. Key active ingredients of Pretect D

Modes of action of Pretect D

Pretect D exhibits multiple modes of action to optimize gut health during challenging times. Due to its anti-protozoal, anti-inflammatory, immunomodulatory, and antioxidant properties, it

1. effectively decreases oocyst excretion and disease spread
2. promotes restoring the mucosal barrier function and improves intestinal morphology
3. protects the intestinal epithelium from inflammatory and oxidative damage.

The beneficial effects of Pretect D

The beneficial effects of Pretect D’s inclusion in the coccidiosis control program include improving overall gut health and broiler production performance.

In a challenge study with Cobb 500 broiler chicks under a mixed Eimeria inoculum challenge, it was evident that the group receiving Pretect D (@500g/ton) in the feed throughout the 35-day rearing period had less coccidia-caused lesions (D27) than the broilers challenged and fed control diets.

Fig. 2: Pretect D reduced coccidia-caused lesions in broilers

In another field study, a traditional anticoccidial program (Starter and Grower I feeds: Narasin + Nicarbazin, Grower II feed: Salinomycin, Finisher/ withdrawal feeds: No anticoccidial) was compared with a program combining anticoccidials with Pretect D (Starter and Grower I feeds: Narasin + Nicarbazin, Grower II and Finisher feeds: Pretect D). The addition of Pretect D significantly reduced OPG count and lowered the coccidiosis lesion score compared to the control (Fig. 3).

Fig.3. Pretect D reduced broilers’ coccidiosis lesion score and OPG count

Consequently, broilers receiving Pretect D showed better overall production performance.

Fig. 4. Overall improved production performance by Pretect D

Pretect D: Application Strategies

The introduction of an effective phytogenic combination in the coccidiosis control program can help mitigate the drug resistance issue. Such a natural anticoccidial solution can be used as a standalone, preferably in less challenging months, as well as in combination with chemicals (shuttle/ rotation) or a coccidiosis vaccine (bio-shuttle), reducing the need for frequent drug use.

Shuttle programs are commonly employed for managing coccidiosis, and they yield a satisfactory level of success. Within these programs, multiple drugs from distinct classes of anticoccidials are administered throughout a single flock. For instance, one class of drug is utilized in the starter feed, another in the grower stage, reverting to the initial class for the finisher diet and concluding with a withdrawal period.

In rotation programs, anticoccidial drugs are alternated between batches rather than within a single batch.

Conclusions

Coccidiosis is considered one of the most economically significant diseases of poultry and the development of anticoccidial resistance has threatened the profitability of the broiler industry. Therefore, regularly monitoring Eimeria species to develop resistance against different anticoccidial groups is crucial to managing resistance and choosing an anticoccidial. It would be rewarding to use an effective phytogenic solution in the coccidiosis control program as a strategic and tactical measure and to focus on such integrated programs for drug resistance management in the future.

References:

Abbas, R.Z., D.D. Colwell, and J. Gilleard. “Botanicals: An Alternative Approach for the Control of Avian Coccidiosis.” World’s Poultry Science Journal 68, no. 2 (June 1, 2012): 203–15. https://doi.org/10.1017/s0043933912000268.

Abbas, R.Z., Z. Iqbal, D. Blake, M.N. Khan, and M.K. Saleemi. “Anticoccidial Drug Resistance in Fowl Coccidia: The State of Play Revisited.” World’s Poultry Science Journal 67, no. 2 (June 1, 2011): 337–50. https://doi.org/10.1017/s004393391100033x.

Allen, P. C., and R. H. Fetterer. “Recent Advances in Biology and Immunobiology ofEimeriaSpecies and in Diagnosis and Control of Infection with These Coccidian Parasites of Poultry.” Clinical Microbiology Reviews 15, no. 1 (January 2002): 58–65. https://doi.org/10.1128/cmr.15.1.58-65.2002.

Chapman, H. D. “Biochemical, Genetic and Applied Aspects of Drug Resistance inEimeriaParasites of the Fowl.” Avian Pathology 26, no. 2 (June 1997): 221–44. https://doi.org/10.1080/03079459708419208.

Chapman, H.D. “Resistance to Anticoccidial Drugs in Fowl.” Parasitology Today 9, no. 5 (May 1993): 159–62. https://doi.org/10.1016/0169-4758(93)90137-5.

Cobaxin-Cárdenas, Mayra E. “Natural Compounds as an Alternative to Control Farm Diseases: Avian Coccidiosis.” Farm Animals Diseases, Recent Omic Trends and New Strategies of Treatment, March 21, 2018. https://doi.org/10.5772/intechopen.72638.

Nazzaro, Filomena, Florinda Fratianni, Laura De Martino, Raffaele Coppola, and Vincenzo De Feo. “Effect of Essential Oils on Pathogenic Bacteria.” Pharmaceuticals 6, no. 12 (November 25, 2013): 1451–74. https://doi.org/10.3390/ph6121451.

Pop, Loredana Maria, Erzsébet Varga, Mircea Coroian, Maria E. Nedișan, Viorica Mircean, Mirabela Oana Dumitrache, Lénárd Farczádi, et al. “Efficacy of a Commercial Herbal Formula in Chicken Experimental Coccidiosis.” Parasites & Vectors 12, no. 1 (July 12, 2019). https://doi.org/10.1186/s13071-019-3595-4.

World Health Organization Technical Report Series No. 296, (1965) pp:. 29.

by Marisabel Caballero, Global Technical Manager, and Madalina Diaconu, Global Manager Gut Health, EW Nutrition

In practical poultry production, multiple stress factors occur simultaneously: nutrition, management, environment, etc.. The effects of these factors are additive, leading to chronic stress, a condition in which animals cannot regain homeostasis and continuously deviate the use of resources to inflammation and restoring the gut barrier-function (Das et al., 2011). As a result, the gut microbiome is altered and oxidative stress ensues (Mishra et al., 2019). In this situation, health and productivity are compromised.

The feed supplied to production animals is designed to help them express their genetic potential. However, some feed components are also continuous inflammatory triggers. Anti-nutritional factors, oxidized lipids, and mycotoxins induce a low-grade inflammatory response (Cardoso Del Pont et al., 2020). Other factors that trigger gut health issues include the environment, management, and pathogens.

Feed interventions have shown to increase productivity and improve gut-related biomarkers, demonstrating a mitigation effect over the challenge factors (Deminicis et al., 2020; Latek et al., 2022).

Meta-analysis of broiler studies shows consistent results

As broilers are continuously challenged during the production period, the effects of an in-feed phytogenic (Ventar D – EW Nutrition GmbH) were extensively researched in broiler meat production. 21 trials in different locations (7 in Europe, 6 in the USA, 4 in Japan, 3 in Middle East, and 2 in India), with different production levels (grouped by EPEF) and challenges were analyzed to establish Ventar D’s benefits for the broiler production industry in terms of performance and sustainability. In all trials, the treatment group consisted of a supplementation of the basal feed with Ventar D at a dose 100 g/ton. The control groups were not supplemented with any gut health improvement feed additive.

Of these 21 trials, 14 had corn/soybean meal-based diets and 7 had high fiber diets (based on wheat and rye, which constituted a challenge as no NSP-enzymes were included). Reused litter (by 12 to 14 flocks, previous to the trial) also was used as a challenge. 18 trials were performed in research facilities and 3 in commercial farms.

Consistency in the results from Ventar D could be demonstrated as 19 out of 21 trials showed an improvement in FCR, lowering 3.4 points on average; 18 /21 trials showed higher body weight, with an average of 64 grams more; and 17 trials showed lower mortality than the control group, averaging 1.19 percentual points of reduction. The phenolic compounds included in Ventar D, such as thymol, possess antioxidant, anti-inflammatory, and antibacterial activities, which account for improving gut health and thus increasing performance in production animals.

The European Poultry Efficiency Factor (EPEF) was used to establish the performance level of each flock. This index is based on the average daily weight gain, mortality, and feed conversion, and takes in consideration the age of the flock at collection, allowing to make comparisons on performance within and between farms.

Of the 21 trials, 10 control groups had an EPEF lower than 375, and were considered of low performance level, in 8 the EPEF was between 375 and 425 and considered of medium performance, and for 3 the performance was considered high having an EPEF of 425 or more.

Ventar D increased performance at all levels (Figure 1). However, the effects were challenge-dependent:
Low performing flocks averaged an 8% increase in EPEF, and high performing flocks increased 4%, indicating that Ventar D can help broilers to overcome challenges commonly found in poultry production, and boost performance even with excellent farm and management conditions. These results concur with a meta-analysis by Valle Polycarpo and collaborators (2022), finding that a microbial challenge can influence the performance of phytogenic feed additives.

Overall, this analysis demonstrates that effective nutritional interventions can give consistent results and constitute effective tools to help production animals overcome stress and enhance productivity.

Part 1: Impact on Feathering

By Dr. Inge Heinzl, Editor, and Marisabel Caballero, Global Technical Manager, EW Nutrition

Mycotoxins are known to decrease health and performance in poultry production. Their modes of action, such as reducing protein synthesis and promoting oxidative stress and apoptosis, lead to cell destruction and lower cell replacement, affecting several organs and tissues.

When different stress factors collude, such as high temperatures and humidity, poor ventilation, high stocking density, and management events, the effects of in-feed mycotoxins can reach a higher level, which may include external signs.

The most common and recognized external sign of mycotoxicosis is mouth lesions caused by trichothecenes, which are highly associated with the presence of T-2 in the feed. However, other signs may appear, such as paleness of combs, shanks, and feet, as well as leg problems, ruffled feathers and poor feather coverage, feed passage, and abnormal feces.

In a series of articles, we want to report on external signs facilitating a differential diagnosis of mycotoxin contamination. This is necessarily followed by feed or raw material mycotoxin analysis and strategies to avoid or mitigate the effects of mycotoxin contamination in poultry production. In the first article, we will cover feathers.

A healthy plumage is crucial for growth and reproduction

Feathering is a crucial aspect of poultry health and productivity. Feathers are essential for thermoregulation, locomotion, adequate skin protection, and reproductive success, protecting hens from injury during mating. Inadequate feathering can lead to lower feed efficiency (Leeson and Walsh, 2004) as well as loss in fertility and chick production (Fisher, 2016). Mycotoxins in poultry feed can compromise feather quality in poultry production animals. This first article delves into the relationship between mycotoxins and poor feathering, exploring different mycotoxins and their mechanisms of action.

In which way do mycotoxins compromise feathering?

On the one hand, chronic mycotoxin exposure impairs the digestive process, hindering the absorption and utilization of vital nutrients essential for feather growth. This disruption can lead to malnutrition, directly impacting the quality and health of feathers. On the other hand, mycotoxins also interfere with metabolic processes critical for feather development, such as keratin synthesis (Wyatt et al., 1975;  Nguansangiam, 2004). Enzymatic pathways involved in synthesizing keratin, the protein building block of feathers, are particularly vulnerable to mycotoxin-induced disruptions. The presence of mycotoxins in feed has been associated with the manifestation of sparse feathering and the sticking out of feathers at an unnatural angle (Emous and Krimpen, 2019). In the case of multiple mycotoxins occurring in the feed, even at singularly unimportant concentrations, a negative impact on feathering is possible. Different mycotoxins have different target organs and consequences for the animal, so their ways of compromising feathering also vary. As feathering needs protein availability, all mycotoxins affecting the protein metabolism or the absorption of nutrients also impact the feathering process. Let us look at the most prominent mycotoxins.

1.   T-2 toxin

Due to climate change, T-2 toxins are on the rise. In the US, more than 50% of the tested samples contained T-2 toxin; in Europe, we found it in 31%, and in China, in 82% of the samples (EW Nutrition, 2024). The highest level was found in Europe, with 850 ppb.

Adverse effects of T-2 toxin in goslings were shown by Gu et al. (2023), who exposed the animals to 6 different levels of T-2 toxin, from 0.2 to 2.0 mg T-2 toxin/kg of feed. The goslings showed a sparse covering with short, dry, rough, curly, and gloss-free feathers on their back with dosages ≥0.8 mg/kg. When zooming on, T-2 can cause necroses of the layer of regenerative cells in the feather base, implying malformation or absence of new feathers, as well as structural damage to existing feathers on the base of the ramus and barb ridges (Hoerr et al. (1982), Leeson et al. (1995)).

The effects in feather regenerative cells are dose-dependent, as confirmed by Hoerr et al. (1982), who applied different doses of T-2 toxin (1.5, 2, 2.5, and 3 mg/kg body weight/day) to 7-day-old broilers for 14 days. Delayed feather development, especially at high dosages, was noticed, as well as malformations and opaque bands in the feathers, the latter probably caused by a segmental reduction in diameter.

Manafi et al. (2015) noticed feather malformations when broiler chickens were challenged with 0.5 ppm T-2 toxin in the feed in combination with an inoculation of 2.4×10⁸ cfu Mycoplasma gallisepticum. When the chickens were challenged only with T-2 toxin, the feathers were ruffled, showing that a coincidence of stress factors even aggravates the symptoms.

2.   Aflatoxins

Aflatoxins, produced by certain Aspergillus species, are among the most notorious mycotoxins. Looking at test results of the last year, Aflatoxin shows incidences between 25 (USA) over 40-65% (Europe, LATAM, MEA, and SEAP) up to 84-88% (China and South Asia) with average levels up to 42 ppb in South Asia (EW Nutrition, 2023). However, more information about the concrete impact of aflatoxins on feathering is needed. They may indirectly affect feathering because they impact digestion and the utilization of nutrients or trace minerals such as zinc, which is essential for the feather construction process. Damage to the liver impacts protein metabolism, and keratin is also necessary for feather production.

In other studies, Muhammad et al. (2017) fed 5 mg AFB1/kg to Arbor Acres broilers, and the birds showed ruffled feathers. A significantly lower feather shine was noticed by Saleemi et al. (2020) when they gave the animals 300 μg AFB1/kg of feed, and the birds of Zafar et al. (2017) showed ruffled, broken, dull, and dirty feathers after six weeks of feeding an aflatoxin-contaminated diet.

3.   Ochratoxin

Ochratoxins, commonly produced by Aspergillus and Penicillium fungi, also pose a significant threat to poultry. When looking at the mycotoxin report, this mycotoxin was found in 16% (Europe) to 70% (SEAP) of the samples (EW Nutrition, 2023). Ochratoxins primarily affect feathering by compromising the structural integrity of feathers and causing delayed feathering in broilers (Leeson, 2021).

Several trials have shown the negative impact of ochratoxin on feather quality. Hassan et al. (2010) fed OTA to laying hens and saw a dose-dependent (dosages from 0 to 10 mg/kg feed) occurrence of ruffled and broken feathers in the OTA group, whereas the plumage of the control group was shiny and well-formed. Hameed et al. (2012) also realized dull feathers when feeding 0.4 and 0.8 mg OTA per kg of feed. A further dose-dependent decrease in feather quality was described by Khan et al. (2023) in broiler chicks. He injected them with dosages from 0.1 to 1.7 mg/kg body weight on day 5 of age and saw a deterioration of feather appearance (rippled feathers) in the groups with the higher dosages of 1.3 and 1.7 mg/kg. Abidin et al. (2016) observed a similar dose-dependent deterioration of the feather quality in white Leghorn cockerels when feeding 1 or 2mg OTA/kg feed.

Combinations of aflatoxins and ochratoxins were also tested. Khan et al. (2017) fed moldy feed naturally containing 56 µg OTA and 136 µg AFB1 per kg to layer hens and saw a deterioration of feather quality with increasing feeding time. Qubih (2017) noticed ruffled feathers when feeding a diet naturally contaminated with 800 ppb of OTA and 100 ppb of AFB1.

4.   Scirpenol mycotoxins

Parkhurst et al. (1992) examined the effects of different scirpenol mycotoxins. After feeding graded levels of fusarium mycotoxins to broiler chicks until three weeks of age, they discovered that the impact of scirpenols stretched across the entire feathered body parts and that the degree of feather alteration is dose-dependent. The main alteration was a frayed or even missing web on the medial side of the outer end of the feather due to poor development of the barbs, barbules, and barbicels, and the tip of the feathers became square instead of rounded—the thinner and weaker shafts of the feathers inclined to show an accentuated medial curve.

Parkhurst et al. (1992)

Figure 1: Feathering affected by scirpenol mycotoxins

In their trial, Parkhurst and Hamilton realized that 15-monoacetoxyscirpenol (15-MAS) caused the most severe alterations of feathers, and they determined a minimum effective dose (MED) of 0.5 µg/g diet. The MEDs for 4,15-diacetoxyscirpenol (4,15-DAS) and 3,4,15-triacetoxyscirpenol (TAS) were higher, 2 µg/g and > 8 µg/g, respectively.

How can we enable adequate feathering in poultry?

Adequate feathering of poultry is necessary for the animal’s health and welfare and to ensure fertility and productivity. The occurrence of mycotoxins in the feed – and the probability is high! – can cause poor feathering or the development of malformed feathers.

To best equip broilers, layers, and breeders, their feed must contain all nutrients essential for healthy growth and appropriate feathering. As the risk of contamination of the feed materials is very high (see EW Nutrition’s mycotoxin report 2023), it is of crucial importance to have an efficient mycotoxin risk management in place, which includes sampling, analysis of samples, and the use of mycotoxin binders. EW Nutrition offers MasterRisk, an online tool where farmers and feed millers can feed the results of their feed analysis concerning mycotoxins and get a risk management recommendation.

In the next part of the series, we will report on beak lesions and skin paleness, two other external signs of mycotoxin contamination.

References:

Abidin, Zain ul, Muhammad Zargham Khan, Aisha Khatoon, Muhammad Kashif Saleemi, and Ahrar Khan. “Protective Effects Ofl-Carnitine upon Toxicopathological Alterations Induced by Ochratoxin A in White Leghorn Cockerels.” Toxin Reviews 35, no. 3–4 (August 22, 2016): 157–64. https://doi.org/10.1080/15569543.2016.1219374.

Emous, R. A., and M. M. Krimpen. “Effects of Nutritional Interventions on Feathering of Poultry – a Review.” Poultry Feathers and Skin: The Poultry Integument in Health and Welfare, 2019, 133–50. https://doi.org/10.1079/9781786395115.0133.

Fisher, Colin. “Feathering in Broiler Breeder Females – Aviagen.” https://aviagen.com/, 2016. http://en.aviagen.com/assets/Tech_Center/Broiler_Breeder_Tech_Articles/English/Feathering-in-Broiler-Breeeder-Females-EN-2016.pdf.

Gu, Wang, Qiang Bao, Kaiqi Weng, Jinlu Liu, Shuwen Luo, Jianzhou Chen, Zheng Li, et al. “Effects of T-2 Toxin on Growth Performance, Feather Quality, Tibia Development and Blood Parameters in Yangzhou Goslings.” Poultry Science 102, no. 2 (February 2023): 102382. https://doi.org/10.1016/j.psj.2022.102382.

Hameed, Muhammad  Raza, Muhammad Khan, Ahrar Khan, and Ijaz Javed. “Ochratoxin Induced Pathological Alterations in Broiler Chicks: Effect of Dose and Duration.” Pakistan Veterinary Journal Pakistan Veterinary Journal 8318, no. 2 (December 2012): 2074–7764.

Hassan, Zahoor-Ul, M. Zargham Khan, Ahrar Khan, and Ijaz Javed. “Pathological Responses of White Leghorn Breeder Hens Kept on Ochratoxin A Contaminated Feed.” Pakistan Veterinary Journal 30, no. 2 (2010): 118–23.

Hoerr, F. J., W. W. Carlton, and B. Yagen. “Mycotoxicosis Caused by a Single Dose of T-2 Toxin or Diacetoxyscirpenol in Broiler Chickens.” Veterinary Pathology 18, no. 5 (September 1981): 652–64. https://doi.org/10.1177/030098588101800510.

Hoerr, F.J., W.W. Carlton, B. Yagen, and A.Z. Joffe. “Mycotoxicosis Produced in Broiler Chickens by Multiple Doses of Either T‐2 Toxin or Diacetoxyscirpenol.” Avian Pathology 11, no. 3 (January 1982): 369–83. https://doi.org/10.1080/03079458208436112.

Khan, Ahrar, Muhammad Mustjab Aalim, M. Zargham Khan, M. Kashif Saleemi, Cheng He, M. Noman Naseem, and Aisha Khatoon. “Does Distillery Yeast Sludge Ameliorate Moldy Feed Toxic Effects in White Leghorn Hens?” Toxin Reviews, January 25, 2017, 1–8. https://doi.org/10.1080/15569543.2017.1278707.

Khan, Shahzad Akbar, Eiko N. Itano, Anum Urooj, and Kashif Awan. “Ochratoxin-a Induced Pathological Changes in Broiler Chicks.” Pure and Applied Biology 12, no. 4 (December 10, 2023): 1608–16. https://doi.org/10.19045/bspab.2023.120162.

Leeson, S., and T. Walsh. “Feathering in Commercial Poultry II. Factors Influencing Feather Growth and Feather Loss.” World’s Poultry Science Journal 60, no. 1 (March 1, 2004): 52–63. https://doi.org/10.1079/wps20045.

Leeson, Steve. “Effects of Nutrition on Feathering.” Poultry World, May 22, 2021. https://www.poultryworld.net/specials/effects-of-nutrition-on-feathering/.

Leeson, Steven, Gonzalo J. Diaz Gonzalez, and John D. Summers. Poultry metabolic disorders and Mycotoxins. Guelph, Ontario, Canada: University Books, 1995.

Manafi, M., N. Pirany, M. Noor Ali, M. Hedayati, S. Khalaji, and M. Yari. “Experimental Pathology of T-2 Toxicosis and Mycoplasma Infection on Performance and Hepatic Functions of Broiler Chickens.” Poultry Science 94, no. 7 (July 2015): 1483–92. https://doi.org/10.3382/ps/pev115.

Muhammad, Ishfaq, Xiaoqi Sun, He Wang, Wei Li, Xinghe Wang, Ping Cheng, Sihong Li, Xiuying Zhang, and Sattar Hamid. “Curcumin Successfully Inhibited the Computationally Identified CYP2A6 Enzyme-Mediated Bioactivation of Aflatoxin B1 in Arbor Acres Broiler.” Frontiers in Pharmacology 8 (March 21, 2017). https://doi.org/10.3389/fphar.2017.00143.

Nguansangiam, Sudarat, Subhkij Angsubhakorn, Sutatip Bhamarapravati, and Apichart Suksamrarn. The Southeast Asian J of Tropical Medicine 34, no. 4 (2004): 899–905.

Parkhurst, Carmen R., Pat B. HamiltonON, and Adedamola A. AdemoyeroERO. “Abnormal Feathering of Chicks Caused by Scirpenol Mycotoxins Differing in Degree of Acetylation.” Poultry Science 71, no. 5 (May 1992): 833–37. https://doi.org/10.3382/ps.0710833.

Qubih, T. S. “Relationship between Mycotoxicosis and Calcium during Preproduction Period in Layers.” Iraqi Journal of Veterinary Sciences 26, no. 1 (June 28, 2012): 11–14. https://doi.org/10.33899/ijvs.2012.46888.

Saleemi, M. Kashif, Kamran Ashraf, S. Tehseen Gul, M. Noman Naseem, M. Sohail Sajid, Mashkoor Mohsin, Cheng He, Muhammad Zubair, and Ahrar Khan. “Toxicopathological Effects of Feeding Aflatoxins B1 in Broilers and Its Amelioration with Indigenous Mycotoxin Binder.” Ecotoxicology and Environmental Safety 187 (January 2020): 109712. https://doi.org/10.1016/j.ecoenv.2019.109712.

Wyatt, R.D., P.B. Hamilton, and H.R. Burmeister. “Altered Feathering of Chicks Caused by T-2 Toxin.” Poultry Science 54, no. 4 (July 1975): 1042–45. https://doi.org/10.3382/ps.0541042.

Zafar, Roheena, Farhat Ali Khan, and Muhammad Zahoor. “In Vivo Amelioration of Aflatoxin B1 in Broiler Chicks by Magnetic Carbon Nanocomposite.” Pesquisa Veterinária Brasileira 37, no. 11 (November 2017): 1213–19. https://doi.org/10.1590/s0100-736×2017001100005.

By Dr. Ajay Awati, Director of Enzymes, EW Nutrition

The use of by-products and high-fiber ingredients in feed formulations has increased in swine operations.  Driven by both economic and sustainability goals, this shift has emphasized the importance of understanding the role of dietary fibers and carbohydrases in swine nutrition and health (Petry & Patience, 2020). These feeds rich in fiber are generally considered to have low nutritional value due to the lower digestive energy or amino acid levels when compared to concentrated feeds with high starch or proteins (Woyengo et al., 2014).

Dietary fiber is vital in pig nutrition, necessitating a baseline inclusion to support regular digestive tract functions (Wenk, 2001). Incorporating fiber into the diets of monogastric animals raises concerns due to its correlation with reduced nutrient utilization and diminished net energy levels (Noblet; Le Goof, 2001). High-fiber diets can present challenges for inclusion in monogastric animals’ feeds, especially young animals, due to their bulky nature and restricted ability to ferment fiber, impacting nutrient uptake based on fiber type, the age of the pig, and diet composition (Bach Knudsen et al., 2012).

Moreover, the apparent ileal digestibility (AID) of nutrients is adversely affected by dietary fiber, attributed to the small intestine’s deficiency in endogenous enzymes necessary for breaking down these bonds (Bach Knudsen et al., 2012).

This article aims to demonstrate how enzymatic degradation of arabinoxylans, particularly through xylanase enzymes, can mitigate anti-nutritional effects and enhance the nutritional value of high-fiber swine diets, thereby improving animal health and performance.

Into the World of Arabinoxylans

In plants classified as monocotyledonous, such as cereals, the main non-starch polysaccharides (NSP’s) components of the cell wall are arabinoxylans, cellulose and β-glucan (Bach Knudsen, 1997). Arabinoxylans represent a complex group of dietary fibers with significant implications for swine nutrition and health. Their structural heterogeneity can influence physicochemical properties, biological activities, and affect pigs’ gut microbiota and immune system. Present in both soluble and insoluble forms, it consists of a backbone of xylose residues with arabinose side chains, playing a crucial role in the nutritional dynamics within swine diets (Mudgil & Barak, 2013).

The fermentability of corn-based dietary fiber is limited by its insoluble fraction and complex branched structure; impacting the digesta transit rate and reducing the digestibility of nutrients (Gutierrez et al., 2013). Supplementing exogenous carbohydrases offers a viable approach to enhance the utilization of fiber that is otherwise difficult to ferment, potentially amplifying its positive effects.

Xylanase’s Impact on Fiber and Gut Health

Non-digestible carbohydrates may be fermented by microbial populations along the gastrointestinal tract to synthesize short-chain fatty acids that may be absorbed and metabolized by the pig. Such indigestible carbohydrates consist of specific disaccharides, oligosaccharides, resistant starches, and non-starch polysaccharides. The presence and composition of these indigestible carbohydrates in pig diets vary based on the types of feed ingredients incorporated into their meals (Navarro et al., 2019). Xylanase works on the hydrolysis of the arabinoxylan fraction of NSPs. The NSPs present in the walls of plant cells encapsulate nutrients, making them unavailable for the action of the animal’s own digestive enzymes. Moreover, NSPs exhibit a high affinity for water within the gastrointestinal lumen, leading to elevated digesta viscosity. This increased viscosity reduces gastrointestinal motility, facilitating an environment conducive to the proliferation of pathogenic microflora (Choct, 1998).  The advantageous outcomes of enzyme supplementation arise from the enzymatic disruption of intact cellular membranes, leading to the release of sequestered nutrients, or are a consequence of modifying the physicochemical properties of non-starch polysaccharides, due to changes in viscosity and water-holding capacity and/or changes in the composition and content of bacteria in the intestine (Bedford, M. R., & Classen, 1992).

Arabinoxylans in Cereal Grains and Their By-products

Factors such as genetics, climate, maturity stage, fertilizer use, and post-harvest storage time influence the proportion of total cell wall polysaccharides in cereal grains. These factors vary across production systems and countries, depending on the availability of feed resources (Paloheimo et al., 2010).

Cereal grains and their by-products, including wheat bran, corn distillers dried grains with solubles (DDGS), and rice husks, serve as significant sources of arabinoxylans. Their incorporation into swine diets is growing due to economic advantages.

The ethanol industry’s growth has increased the availability of distillery by-products. Brazil alone generates an estimated 366 million tons of DDGS annually (USDA, 2017). Among these by-products, distiller-dried grains are prevalent, especially in the U.S. pork industry as feed ingredients.

Corn, wheat, and barley, as staple ingredients in swine feed, exhibit significant variations in their NSP and arabinoxylans content. In grain form, corn contains 4.7% AX with a soluble component of 0.5%, while wheat has a higher arabinoxylans content at 7.3% with 1.8% being soluble. Barley stands out with the highest arabinoxylans content at 8.4%, of which 1.2% is soluble, reflecting its rich fiber composition. The processing into flour results in a reduction of arabinoxylans content across all three cereals, highlighting the impact of processing on dietary fiber availability (Knudsen, 2014).

Rice distillers’ by-product is recognized as a valuable protein source, boasting high crude protein levels. Nonetheless, its high fiber content can restrict usage (Huang et al., 2017). Wheat bran is particularly rich in arabinoxylans, enhancing its dietary fiber content. DDGS also contain significant amounts of both soluble and insoluble arabinoxylans, resulting from the corn kernel’s residual non-starch polysaccharides (Agyekum & Nyachoti, 2017).

It is essential to understand the specific levels of arabinoxylans in these components to create balanced diets that optimize nutritional benefits while minimizing potential anti-nutritional effects.

Addressing Arabinoxylan Degradation

Xylanases target specific substrates, necessitating the presence of arabinoxylans for their effective action. The complex structure of arabinoxylans makes them resistant to degradation by the swine’s endogenous enzymes, presenting a dual challenge: how to harness the beneficial effects of soluble arabinoxylans while mitigating the negative impacts of their insoluble counterparts.

These enzymes specifically cleave the 1,4-β-D-xylosidic bonds in arabinoxylans, randomly targeting xylose linkages within the xylan structure. Each enzyme type has a unique pattern of degradation (Collins et al., 2005) and GH 10 xylanases specialize in breaking down arabinoxylans with high arabinose substitution into smaller oligosaccharides. These oligosaccharides are valuable for fermentation, serving as energy sources or prebiotics.

Also, this group of enzymes action not only reduces gut viscosity but can lead to enhanced feed efficiency, growth performance, and overall health of swine by improving the digestibility of fibrous components in feed (Lærke et al., 2015). GH 10 xylanases often have optimal activity at pH levels found in the animal gut, and their thermal stability ensures they retain activity under feed processing temperatures.  Lei et al. (2016) highlighted the efficacy of xylanase in improving nutrient digestibility and overall feed efficiency. By targeting the arabinoxylans present in swine diets, xylanase enzymes facilitate a more efficient conversion of feed into energy, contributing to improved growth rates and performance metrics.

As detailed by Tiwari, Singh, & Jha (2019), arabinoxylans undergo fermentation in the gut, leading to the production of short-chain fatty acids (SCFAs) that beneficially alter the gut microbial ecology. The application of GH 10 xylanases has been highlighted for its potential to significantly enhance the degradation of arabinoxylans, thereby improving the fermentation process and increasing the yield of SCFAs. This enzymatic breakdown facilitates more efficient nutrient absorption and overall better gastrointestinal health, directly influencing swine growth and performance positively.

A study reveals that xylanase supplementation significantly reduces mortality rates in pigs in a dose-dependent manner. With mortality rates dropping from 4.16% in the control group to as low as 2.25% with the highest xylanase dosage, the results highlight xylanase’s potential to improve gut health and increase survival rates. This suggests a promising approach for boosting pig well-being and reducing the reliance on enteric antibiotics, marking a significant stride in sustainable animal nutrition practices.(Zier-Rush et al., 2016).

The research conducted by Petry et al. (2020) demonstrated that xylanase increased the digestibility of non-starch polysaccharides, particularly arabinoxylan, in diets high in insoluble corn fiber. This improvement in nutrient absorption highlights xylanase’s role in optimizing the use of high-fiber ingredients in swine diets, thereby enhancing animal health and performance. Due to its cost-effectiveness and nutrient profile, xylanase supplementation enhances the nutritional value of DDG in swine diets.

The strategic implementation of xylanase in swine diets represents a promising approach to the challenges posed by high-fiber feed ingredients. By improving the digestibility of arabinoxylans and other complex carbohydrates, xylanase supplementation can mitigate the anti-nutritional effects of insoluble fibers, enhance feed efficiency, and support optimal growth and health outcomes in swine.

Enhancing Swine Productivity with Enzyme Solutions

With the growing incorporation of co-products in swine feed, there arises a crucial need to transform the high fiber content into a beneficial asset for the animals. The strategic incorporation of enzyme solutions, particularly xylanase enzymes, into swine feed formulations emerges as a scientifically supported approach to significantly enhance the digestibility of high-fiber diets. This method effectively addresses the nutritional intricacies posed by arabinoxylans, facilitating improved feed utilization. Moreover, the action of xylanase enzymes extends beyond enhancement of nutrient absorption; it plays a pivotal role in promoting the health and performance of swine. Such targeted nutritional strategies are vital in the context of swine production systems, highlighting the necessity of integrating these enzymatic solutions to achieve optimal animal health, growth, and productivity.

References:

1. Agyekum, K. A., & Nyachoti, C. M. (2017). Nutritional and metabolic consequences of feeding high-fiber diets to swine: A review. Engineering, 3(5), 716-725. https://doi.org/10.1016/J.ENG.2017.03.010
2. Bach Knudsen, K. E. (1997). Carbohydrate and lignin contents of plant materials used in animal feeding. Animal Feed Science and Technology, 67, 319-338.
3. Bach Knudsen, K. E., Hedemann, M. S., & Laerke, H. N. (2012). The role of carbohydrates in intestinal health of pigs. Animal Feed Science and Technology, 173, 41–53.
4. Bedford, M. R., & Classen, H. L. (1992). Reduction of intestinal viscosity through manipulation of dietary rye and pentosanase concentration is effected through changes in the carbohydrate composition of the intestinal aqueous phase and results in improved growth rate and food conversion efficiency of broiler chicks. The Journal of Nutrition, 122, 560-569.
5. Choct, M. (1998). The effect of different xylanases on carbohydrate digestion and viscosity along the intestinal tract in broilers. Australian Poultry Science Symposium, 10.
6. Collins, T., Gerday, C., & Feller, G. (2005). Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiology Reviews, 29(1), 3–23. https://doi.org/10.1016/j.femsre.2004.06.005
7. Gutierrez, N. A., Kerr, B. J., & Patience, J. F. (2013). Effect of insoluble-low fermentable fiber from corn-ethanol distillation origin on energy, fiber, and amino acid digestibility, hindgut degradability of fiber, and growth performance of pigs. Journal of Animal Science, 91, 5314–5325. https://doi.org/10.2527/jas.2013-6328
8. Huang, Y. F., Gao, X. L., Nan, Z. B., & Zhang, Z. X. (2017). Potential value of the common vetch (Vicia sativa L.) as an animal feedstuff: A review. Journal of Animal Physiology and Animal Nutrition, 101, 807-823. https://doi.org/10.1111/jpn.12617
9. Lærke, H. N., Arent, S., Dalsgaard, S., & Bach Knudsen, K. E. (2015). Effect of xylanases on ileal viscosity, intestinal fiber modification, and apparent ileal fiber and nutrient digestibility of rye and wheat in growing pigs. Journal of Animal Science, 93(9), 4323-4335.
10. Lei, Z., Shao, Y., Yin, X., Yin, D., Guo, Y., & Yuan, J. (2016). Combination of xylanase and debranching enzymes specific to wheat arabinoxylan improve the growth performance and gut health of broilers. Journal of Agricultural and Food Chemistry, 64(24), 4932-4942.
11. Mudgil, D., & Barak, S. (2013). Composition, properties and health benefits of indigestible carbohydrate polymers as dietary fiber: a review. International Journal of Biological Macromolecules, 61, 1-6. https://doi.org/10.1016/j.ijbiomac.2013.06.044
12. Navarro, D. M. D. L., Abelilla, J. J., & Stein, H. H. (2019). Structures and characteristics of carbohydrates in diets fed to pigs: a review. Journal of Animal Science and Biotechnology, 10, 39. https://doi.org/10.1186/s40104-019-0345-6
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15. Petry, A., Huntley, N., Bedford, M., Zijlstra, R. T., & Patience, J. (2020). Supplementing xylanase increased the digestibility of non-starch polysaccharides, particularly arabinoxylan, in diets high in insoluble corn fiber fed to swine with a 36-d dietary adaptation period. Journal of Animal Science, 98(52-52).
16. Petry, A. L., & Patience, J. F. (2020). Xylanase supplementation in corn-based swine diets: a review with emphasis on potential mechanisms of action. Journal of Animal Science, 98(11), skaa318. https://doi.org/10.1093/jas/skaa318
17. Tiwari, U., Singh, A., & Jha, R. (2019). Fermentation characteristics of resistant starch, arabinoxylan, and β-glucan and their effects on the gut microbial ecology of pigs: A review. Animal Nutrition, 5, 217-226.
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21. Zier-Rush, C. E., Groom, C., Tillman, M., Remus, J., & Boyd, R. D. (2016). The feed enzyme xylanase improves finish pig viability and carcass feed efficiency. Journal of Animal Science, 94(suppl_2), 115. https://doi.org/10.2527/msasas2016-244

By Dr. Ajay Awati, Global Director Enzymes, EW Nutrition

In recent years, the scientific understanding of xylanase inhibitors (XIs) and their impact on animal nutrition has grown significantly. Xylanase, a crucial enzyme used to enhance nutrient availability in feed, can face challenges from XIs present in cereal grains. This article explores the evolution of plant protection mechanisms, the economic impact of XIs, and the development of a novel xylanase, Axxess XY, resistant to these inhibitors.

Xylanase inhibitors – an evolutionary protection mechanism of plants

Xylanase inhibitors (XI) are a classic example of the evolutionary development of protection mechanisms by cereal plants against pathogens. Microorganisms, such as fungal pathogens, involve the degradation of xylan as one of the mechanisms in pathogenesis (Choquer et al., 2007). There are also other mechanisms by which microorganism-produced xylanases affect plants.

To protect themselves, plants evolved xylanase inhibitors to prevent the activities of xylanases. XIs are plant cell wall proteins broadly distributed in monocots. There are three classes of XIs with different structures and inhibition specificities (Tundo et al., 2022):
1. Triticum aestivum xylanase inhibitors (TAXI)
2. Xylanase inhibitor proteins (XIP), and
3. Thaumatin-like xylanase inhibitors (TLXI).

Xylanase inhibitors have an economic impact

In animal nutrition, xylanases are widely used in diets containing cereal grains and other plant materials to achieve a higher availability of nutrients. The inhibitory activity of XIs prevents this positive effect of the enzymes and, therefore, makes them economically relevant. Studies have reported that higher levels of XIs negatively impact broiler performance. For example, in one of the studies, broilers fed with grains of a cultivar with high inhibitory activity showed a 7% lower weight on day 14 than broilers fed with grains of a cultivar with less inhibitory activity (Madesen et al., 2018). Another study by Ponte et al. (2004) also concluded that durum wheat xylanase inhibitors reduced the activity of exogenous xylanase added to the broiler diets.

Xylanase inhibitors can withstand high temperatures

Even though XIs can impact the performance of exogenous xylanase in different ways, only minor attention was paid to the reduction of xylanase’s susceptibility to xylanase inhibitors during the xylanase development in the last decades. Firstly, the issue was ignored mainly through the assumption that XIs are denatured or destroyed during pelleting processes. However, Smeets et al. (2014) showed that XIs could sustain significant temperature challenges. They demonstrated that after exposing wheat to pelleting temperatures of 80°C, 85°C, 92°C, and 95°C, the recovery of inhibitory activity was still 99%, 100%, 75%, and 54%, respectively. Furthermore, other studies also confirmed that conditioning feed at 70-90°C for 30 sec followed by pelleting had little effect on the XI activity in the tested feed, showing that xylanase inhibitors are very likely present in most xylanase-supplemented feeds fed to animals.

Do we only have the problem of xylanase inhibitors in wheat?

No. After first reports of the presence of xylanase inhibitors in wheat by Debyser et al. (1997, 1999), XIs were also found in other cereal grains (corn, rice, and sorghum, etc.), and their involvement in xylanase inhibition and plant defense has been established by several reports (Tundo et al., 2022).

In most of the countries outside Europe, exogenous xylanase is used not only in wheat but also in corn-based diets. Besides broiler feeds, also other animal feeds, such as layer or swine feed being part of more mixed-grain diets, are susceptible to the inhibitory activity of XIs. Nowadays, the situation is getting worse with all the raw material prices increasing and nutritionists tending to use other feed ingredients and locally produced cereals. They need a xylanase which is resistant to xylanase inhibitors.

Xylanases’ resistance to XIs is crucial – Axxess XY shows it

To prevent xylanases from losing their effect due to the presence of xylanase inhibitors, the resistance of new-generation xylanases to these substances is paramount in the development process, including enzyme discovery and engineering.

In the past 25 years, scientists have learned much about XI-encoding genes and discovered how xylanase inhibitors can block microbial xylanases. Additionally, there has been a significant increase in understanding the structural aspects of the interaction between xylanases and XIs, mainly how xylanase inhibitors interact with specific xylanases from fungi or bacteria and those in the GH10 or GH11 family. With such understanding, a new generation xylanase, Axxess XY, was developed. Besides showing the essential characteristics of intrinsic thermostability and versatile activity on both soluble and insoluble arabinoxylan, it is resistant to xylanase inhibitors.

Axxess XY takes xylanase application in animal feeds to the next level.

Axxess XY outperforms other xylanases on the market

Recent scientific developments (Fierens, 2007; Flatman et al., 2002; Debyser, 1999; Tundo et al., 2022; Chmelova, 2019) and internal research can be summarized as follows:

Figure 1: Schematic summary of the susceptibility of different xylanase to xylanase inhibitors from three main groups.

The high resistance to xylanase inhibitors is one of the reasons that a novel xylanase with bacterial origin and from the GH-10 family was chosen to be Axxess XY. EWN innovation, together with research partners, made an interesting benchmark comparison between xylanases that are commercially sold by different global suppliers and Axxess XY. For these trials, all xylanase inhibitors from wheat were extracted. The inhibitors, together with the respective xylanase, were incubated at 40⁰C (to mimic birds’ body temperature) for 30 mins. Then, the loss of xylanase activity was calculated by analyzing remaining activity after incubation. Results are shown below in Figure 2. There were varying levels of activity loss observed in the different commercially sold xylanases. In some xylanases, the losses were alarmingly high. However, Axxess XY was not inhibited at all.

Fig. 2: Extracted total xylanase inhibitors from wheat incubated with the respective xylanase at 40°C for 30 mins. – Loss of activity after incubation with xylanase inhibitors

Conclusion:

Xylanase inhibitors are present in all cereal grains and, unfortunately, heat tolerant (up to 90⁰C, still 75% of inhibition activity was retained). Regardless of the diets used, there is a possibility that the xylanase used may come across xylanase inhibitors, resulting in a loss of activity. More importantly, this can lead to inconsistent performance.

For effective, consistent, and higher performance of NSP enzyme application, it is a must to use xylanase that is resistant to xylanase inhibitors.

Literature:

Chmelová, Daniela, Dominika Škulcová, and Miroslav Ondrejovic. “Microbial Xylanases and Their Inhibition by Specific Proteins in Cereals.” KVASNY PRUMYSL 65, no. 4 (2019). https://doi.org/10.18832/kp2019.65.127. LINK

Choquer, Mathias, Elisabeth Fournier, Caroline Kunz, Caroline Levis, Jean-Marc Pradier, Adeline Simon, and Muriel Viaud. “Botrytis CinereaVirulence Factors: New Insights into a Necrotrophic and Polyphageous Pathogen.” FEMS Microbiology Letters 277, no. 1 (2007): 1–10. https://doi.org/10.1111/j.1574-6968.2007.00930.x. LINK

Debyser, W, WJ Peumans, EJM Van Damme, and JA Delcour. “Triticum Aestivum Xylanase Inhibitor (Taxi), a New Class of Enzyme Inhibitor Affecting Breadmaking Performance.” Journal of Cereal Science 30, no. 1 (1999): 39–43. https://doi.org/10.1006/jcrs.1999.0272. LINK