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Mycotoxins in silage

Penicillium Moulds in Silage. How they affect Rumen Health

Published: May 17, 2011
By: Dr. Anna Catharina Berge, Berge Veterinary Consulting
Introduction
Moulds and their toxins in feed are an increasing global challenge, and multiple environmental and pre-harvest and post-harvest agricultural practices may contribute to this increase. Mouldy feed and mycotoxins have been associated with lower feed intake, reduced digestibility and health disorders in ruminants. An array of mycotoxins can be present in mouldy silage, some of which are not normally found in concentrates. The mycotoxins of greatest concern are those produced by Penicillium, such as PR toxin, mycophenolic acid, roquefortine C and patulin. The clinical signs of mycotoxicosis are often subtle and the diagnosis is, therefore, often missed. The inclusion of a mycotoxin binder is commonly used in dairy production to safeguard against the potential production depressive effects of mycotoxins.
Moulds found in silage
Moulds are found everywhere and over 80 different fungal species have been identified in corn and grass silage. Silage is invaded by moulds that are adapted to the micro-aerobic, acid conditions of the silage (Storm et al., 2008). Air frequently invades silage and supports the growth of acid-tolerant yeast and moulds. Penicillium moulds are commonly found because they are acid-tolerant and have a low oxygen requirement. Moulds can produce mycotoxins depending on environmental conditions. Distribution and extent of spread of the moulds and toxin formation in silage is highly variable. There is an uneven growth of moulds in the silage and the growth rate, production and stability of various mycotoxins varies. Penicillium roqueforti is able to invade silage and considered an indicator of mould invasion. They can be seen as blue balls in corn silage and darker areas in grass silage. P. roqueforti destabilises silage and results in an increased silage temperature. The mould paves the way for other potentially pathogenic organisms. In the instable silage, the toxin-producing moulds and yeasts (Monascus ruber, Aspergillus fumigatus, Byssochlamys) may be found, together with potentially pathogenic bacteria, such as Listeria monocytogenes and Clostridia butyricum. P. roqueforti and A. fumigatus produce a wide range of secondary mycotoxins with antimicrobial or immunosuppressive effects. P. roqueforti comes later in the silage period, than some of the other mould toxins. This uneven temporal and spatial distribution of toxins creates challenges when samples are taken with the intent to detect a mycotoxin challenge.
The Mycotoxins
PR toxin has an antimicrobial effect and has been considered a marker for problem silages (Sumarah, et al., 2005). PR-toxin has been associated with reduced feed intake, rumen stasis, intestinal irritation, abortion, retained placenta and reduced fertility in cattle.
Penicillic acid has antimicrobial effects and can destabilise the rumen microbial flora
Roquefortine C is a neurotoxin. When P. roqueforti mould is used in cheese production the potential toxins are bound and inactivated by milk proteins and, therefore, cause no harm when ingested. However, in silage, these nerve toxins can have serious, adverse consequences.
Mycophenolic acid is a very strong immunosuppressant toxin, so strong that it has been used in therapy for kidney transplants as the strongest immunosuppressant drug available.
Patulin is a common mycotoxin in silage. It affects ruminal fermentation and is a nerve toxin (neurotoxin) and a cell toxin (cytotoxic). It has been implicated in deaths of cows but has received little attention in the literature.
The risk is that cattle may be exposed to the negative effects of a cocktail of mycotoxins that interact with each other. This risk is not taken into account by current legislation and recommendations, which are based on individual mycotoxins and not multi-contamination.

Rumen health effects
Cows are exposed to various moulds and mycotoxins both through inhalation and ingestion. The rumen has long been considered a very strong buffer against all these toxins because the rumen microflora were assumed to naturally detoxify mycotoxins (Schiefer, 1990). This ability makes ruminants relatively resistant to these toxins compared with non-ruminants. However, this is not the case for all mycotoxins. High-producing animals, such as lactating dairy cows have an increased rumen passage rate and this faster processing of contaminated feed may overwhelm the rumen microflora, so that they will not be able to denature all the toxins (Kiessling et al., 1984). Furthermore, young calves do not have fully developed rumens and are therefore, also more susceptible to mycotoxins. Mycotoxins create a cascade of events by destabilising the rumen environment, leading to endotoxin formation and ruminal wall leakage. Toxins with antibiotic effect can disturb the rumen microbiota to the point that they cannot properly handle other toxins. The silage-derived mycotoxins may, through their antibiotic impact on the rumen microflora, result in common pre-harvest mycotoxins, such as deoxynivalenol (DON), zearalenone and tremorgens becoming a health problem.
Systemic health effects
Mycotoxins can produce a variety of symptoms in dairy cattle that are vague and nonspecific. Mycotoxins absorbed into the systemic circulation will have various effects and result in an activation of the immune system (Sharma, 1993). Mycotoxins are considered one of the most important dietary factors that suppress immunity. Symptoms may include reduced feed intake, feed refusals, unthriftiness, rough hair-coat, poor body condition, lower fertility and increased susceptibility to disease. Conditions, such as altered rumen fermentation, digestive upsets, diarrhea, intestinal irritation, rumenitis, high somatic cell counts, mastitis, laminitis, skin erosions, , and reproductive problems are all associated with a mycotoxin challenge. Often, there are no clinical signs, but sub-clinical production losses, which have a serious financial impact on farm profitability. Mycotoxins in feed will often cause 10 to 15% loss in milk production and, in serious clinical cases, up to 50% loss in production can be seen.
Diagnostic methods
As mentioned, detection of moulds and toxins in feed is difficult and not very informative. A lot of research has been carried out to find diagnostic markers for a mycotoxin challenge in the animals (Zheng et al., 2006). Liver enzyme tests are only of value to detect an aflatoxin challenge and, although they have now been found in feed in southern Italy, they are not the major concern in Europe. Various markers of metabolic distress are not unique to mycotoxins but can assist in making a diagnosis. Research is on-going to find biomarkers, and researchers are currently developing a bovine gene signature that will be used to assist in the diagnosis of mycotoxicosis. Gene expression analysis may also be a good approach to present biomarkers of effect when animals are exposed to multiple mycotoxins.
When a mycotoxin challenge is suspected, a practical diagnostic method is to perform a diagnostic treatment with a broad-spectrum mycotoxin binder, such as Mycosorb®. If the clinical symptoms decrease or disappear following mycotoxin-binder administration, then it is very probable that there may have been a mycotoxin challenge through the feed. This approach avoids the issue with uneven sampling of materials and the difficulties to interpret the risks associated with mycotoxin levels when they are found in combination.
Silage management
Silage is a preserved feed where microbial processes have depleted the oxygen supply and lowered pH due to production of organic acids. Silage management recommendations are designed to achieve an anaerobic and acidic environment to prevent further microbial growth. Under these perfect conditions, undesirable spoilage microorganisms will not grow. Good silage management is very important to prevent moulds and mycotoxins. Plant stress needs to be minimized through optimised planting and harvesting times and proper moisture levels, packing and sealing are essential to ensure the exclusion of air. Inclusion of a silage inoculant, such as Sil-All (Alltech), optimises the fermentation and preservation of the forage. The feeding face should be cut cleanly to avoid deterioration and 15-30 cm should be removed daily to prevent deterioration of the feeding face and face cutters are recommended. Silage should be fed directly after removal from the silo and mouldy silage should be discarded.
Interventions to stabilise rumen function
Stable rumen fermentation is important to optimise mycotoxin degradation in the rumen. General recommendation to minimize the effect of mycotoxins on the rumen is to increase roughage inclusion in the feed. The use of live yeast product, such as Yea-Sacc® contributes to maintain a stable rumen pH and thereby optimises rumen fermentation. A good quality broad-spectrum mycotoxin binder, such as Mycosorb® (Alltech), should be used to bind mycotoxins. Mycosorb® is a yeast extract with ability to bind a broad range of mycotoxins in the feed. In a study determining mycotoxin-associated oxidative stress, 85% of the cows been supplemented with Mycosorb for two weeks showed reduced levels of oxidative stress, lower somatic cell counts and increased milk production (Santos & Fink-Gremmels, 2010)
Summary
Moulds in silage feeds and mycotoxin challenges to cows are common but are often neglected in terms of optimising cow health and production. Detection of moulds in feed is challenging due to the uneven spatial and temporal distribution of moulds and toxins in the feed. The antibiotic effects of some of these silage moulds can disturb the rumen microbial environment paving the way for other mould toxins, yeasts and bacterial hazards. Symptoms of mycotoxin exposure are non-specific and often sub-clinical but production losses can be significant. Moulds should be kept in mind dealing with any type of sub-optimal performance in ruminants and inclusion of an appropriate mycotoxin binder is recommended. For further information, the website www.knowmycotoxins.com is highly recommended and frequently visited by those involved in food animal production.
Written by Dr. Anna Catharina Berge, Berge Veterinary Consulting, Belgium, following Pr Johanna Fink-Gremmes's talk at Dairy Solutions Symposium 2010. Published in Dairy Solutions Newsletter, issue 7, 2011
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Authors:
Anna Catharina Berge
Berge Veterinary Consulting
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Ines Rodrigues
DSM-Firmenich
25 de mayo de 2011

Dear Dr. Anna Catharina Berge,
Thank you for sharing with us your very interesting article about Penicillium Moulds in Silage. 

I would like to share with you some information regarding the adsorption of mycotoxins and specifically on the adsorption of aflatoxins and the use of yeast or yeast-derived glucomannan products.
On a very interesting and comprehensive s scientific report submitted to EFSA (European feed Safety Authority) an overview is given on this type of substances. http://www.efsa.europa.eu/en/scdocs/doc/22e.pdf
In page 107 of this report it can be read:
In vitro studied reported that yeast-derived glucomannan bound 96.2%, but the subsequent in vivo study reported that this product at 0.5% of a diet containing 170 µg of AFB1/kg of feed was not effective in reducing milk AFM1 concentrations (-8%), aflatoxin excretion (-7%), or aflatoxin transfer (-4%) from feed to milk. Similarly, Kutz et al., Waltman et al., Battacone et al. found that the addition of different kinds of non-digestible yeast oligosaccharides were not effective in reducing the AFM1 concentrations in milk (Battacone et al., 2009; Kutz et al., 2009; Waltman et al., 2008). In the study of Waltman et al., experimental sequestering agents (10g/cow/daily) consisting of yeast-derived glucomannan did not affect AFM1 concentrations when cows were fed diets containing 80 to 100 µg of AFB1/kg of diet. Kutz et al. found that yeast-derived glucomannan (0.56%) was not effective in reducing milk AFM1 concentrations (-4%), AFM1 excretion (-5%), or aflatoxin transfer from feed to milk (-2.52%) in cows consuming a total mixed ration containing 112 µg of AFB1/kg of diet. Battacone et al. showed that a dried yeast culture product (which is marketed as a probiotic feed supplement for high-producing dairy ruminants) fed at 12 g/day per ewe did not affect absorption of the aflatoxin in the gastro-intestinal tract of dairy ewes fed diets naturally contaminated with 1-5 µg of AFB1/kg of feed.
If a comparison is to be done with clay binders, then here is what the same report (page 100) says about the use of this type of materials for the sequestration of aflatoxins:
Over the past two decades the use of smectite clays to suppress aflatoxins has been demonstrated for many farm animals. In a series of experiments on growing swine, Lindemann et al. demonstrated that the addition of sodium bentonite (0.5%) to diets contaminated with 800 ppb AFB1 improved average daily feed intake and increased average daily gain. Bentonite supplementation significantly improved concentrations of blood urea, total protein, albumin and activities of AST, ALP and GGT, which were significantly altered by AFB1 (Lindemann et al., 1993).
Furthermore and regarding the use of these products to counteract aflatoxins in milk, Pietri et al., 2009 reported a reduction on the AfM1 content in milk of more than 40 % with an inclusion rate of 50 g per cow and day. http://scialert.net/qredirect.php?doi=ijds.2009.34.42&linkid=pdf
For the other mycotoxins and due to the non-adsorbability of other mycotoxins, such as trichothecenes, zearalenone, ochratoxins and fumonisins, adsorption is known not to be effective, and therefore other strategies must be used as biological detoxification. This is regarded as the biotransformation or degradation of the toxin by microorganisms/enzymes to produce metabolites that are either non-toxic when ingested by animals or less toxic than the parent toxin molecule.
Kind Regards,
Inês Rodrigues

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Anna Catharina Berge
Berge Veterinary Consulting
25 de mayo de 2011
Dear Engormix readers. I have contacted expertise for Mycosorb concentrations. Mycosorb will bind Aflatoxin a1. At an inclusion rate of 0,0125% of mycosorb, it will bind 50% of Aflatoxins (200 ppb) in the feed. At a concentration of 0.05% they will bing 75% of aflatoxins (200 ppb in feed). Mycosorb in contrast to clay adsorbents furthermore binds a wide range of mycotoxins that are now found in feed. I will provide data on inclusion rates and reduction in milk (have requested data). The time before health is restored may vary depending in farm situation, the mycosorb is active within 25 minutes of ingestion, health improvements can be seen within 24 hours, but it may take longer depending on circumstances, I will request data on this. As regards mineral availability, I believe this is not affected, but have requested information on this, Kind regards, Anna Catharina
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Rk Walter
21 de julio de 2013
Dear Dr Swamy Could you throw light on those synthetic clay in preference to natural clay. Regards rk walter
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Muhammad Adeel
30 de abril de 2013
Dear Dr. Anna Catharina Berge, and Swamy Haladi Thank you for sharing with us your very interesting detail about Penicillium Moulds in Silage Regards dr adeel
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Swamy Haladi
Swamy Haladi
27 de mayo de 2011

Readers, this if Swamy from Alltech. The recovery from mycotoxins depends on the duration of exposure to mycotoxins. Silage mycotoxins affect rumen micro organisms, according to this study it may take 2 to 8 weeks to respond to any prevention procedures such as use of Mycosorb. Please note that to the best of my knowledge, this is the first study to look at mycotoxin binders to prevent the effects of silage mycotoxins. Regarding aflatoxin M1, Mycosorb dose is between 10 and 30 g per cow per day depending on the level of aflatoxin B1 in feed. If you need papers on this, please contact me. Mycosorb do not bind minerals as it does not work by charges. It binds by hydrogen and van der wall bonds. There are many products on the market which can take care of aflatoxins. Some of the good quality silicates and bentonites are also effective. However, silage mycotoxins and Fusarium mycotoxins are difficult to deal with. That is where Mycosorb has an upper hand. However, note that clays need to be included at higher levels and make sure you are buying from a repuated company who sell synthetic clays. Natural clays are poor binder of mycotoxins. The paper discussed here is an excellent example for why we see poor performance in cows even when aflatoxins and Fusarium mycotoxins are not detectable. We need to educate dairy farmers on this area. Thanks for the discussion.

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Dr. Karki Kedar
26 de mayo de 2011

Mycotoxin in ruminant cause an oxidative stress,apart of that it chans the elastin fiber to enzyme elastase,and collagen fiber to collagenes enzyme thus causing coagutative necrosis of mucosa in forstomach in ruminanants

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Dr.Thirumeignanam, D.,
25 de mayo de 2011

Thanks for your information About Penicillium Moulds in Silage. If you have data on reduction of AFLA M1 in milk with dose, kindly get back to us.
Regards

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Dr. Karki Kedar
25 de mayo de 2011

Thanks for very informatic information about Penicillium Moulds in Silage. I amalso working in this line but alone

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Drmuhammad Shafique
24 de mayo de 2011

Penicillium Moulds in Silage making is not an easy technique it require expertise and vast knowledge about the problem creating factors which deteriorate the whole effort of the silage making and cost of silage.in this article efforts are exploited to minimize the problems which pave the way for heath hazard effect on cattle and other small ruminants. Anna Catharina Berge efforts are appreciable and problem solvers.

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Dr.Thirumeignanam, D.,
24 de mayo de 2011

Very good article about Penicillium Moulds in Silage. I would like to know Mycotoxin binder (Mycosorb) can reduce the Aflatoxin M1 in milk. If so how much dose should be included per day per cow or per tone?.
What will the effect of dose on animal health and mineral availability?
Thanks

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