WO2021260198A1

WO2021260198A1 - Agaricus blazei fermented grains against lawsonia intracellularis infection

Info

Publication number: WO2021260198A1
Application number: PCT/EP2021/067553
Authority: WO
Inventors: Petra Johanna ROUBOS VAN DEN HIL
Original assignee: Nutreco Ip Assets B.V.
Priority date: 2020-06-26
Filing date: 2021-06-25
Publication date: 2021-12-30

Abstract

The present disclosure is in the field of animal nutrition, in particular for use in animal husbandry. The disclosure relates to an animal feed, or an ingredient, premix or supplement therefor, comprising Agaricus blazei or extract thereof, or Agaricus blazei fermented grains or extract thereof. Animals fed with the feed, ingredient, premix or supplement are less susceptible to Lawsonia intracellularis infection and/or experience less symptoms thereof, allowing for an increased average daily weight gain of said animals.

Description

Title: Agaricus blazei fermented grains against Lawsonia intracellularis infection

TECHNICAL FIELD

The present disclosure is in the field of animal nutrition, in particular for use in animal husbandry. The disclosure particularly relates to Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains, said composition being, for example, an animal feed, ingredient, premix or supplement, for use in reducing the risk of contracting Lawsonia intracellularis infection and/or alleviating Lawsonia intracellularis infection and/or alleviating symptoms of Lawsonia intracellularis infection, in an animal or animal population.

In another aspect, the disclosure pertains to use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains, said composition being, for example, an animal feed, ingredient, premix or supplement, for prophylaxis and/or treatment of proliferative enteropathy (also known as ileitis), such as porcine proliferative enteropathy or equine proliferative enteropathy, (porcine) intestinal adenopathy, necrotic enteritis, (regional) ileitis and proliferative (haemorrhagic) enteropathy.

BACKGROUND ART

Around the globe, animal performance is impeded by occurrence of diseases, such as the occurrence of Lawsonia intracellularis infections, resulting in economic losses.

Lawsonia intracellularis is a bacterial pathogen causing intestinal diseases including (porcine) intestinal adenopathy, necrotic enteritis, (regional) ileitis and proliferative (haemorrhagic) enteropathy. The pathogen can infect a wide range of animals including pigs and horses, and can lead to symptoms such as (hemorrhagic) diarrhea, fever, lack of appetite, weight loss, decreased feed conversion ratio (FCR), growth retardation, and even death. Characteristic of the pathogen are intestinal lesions caused by proliferation of immature enterocytes in the ileal intestinal crypts; these infected cells usually contain Lawsonia intracellularis within their apical cytoplasm. Antibiotics may be used to control the clinical impact of Lawsonia intracellularis infections. However, concerns regarding multi-drug resistant bacterial infections have led to restrictions on antibiotics available for treatment in animal husbandry.

It is an objective of the present invention to reduce the risk of contracting Lawsonia intracellularis infection and/or alleviating Lawsonia intracellularis infection and/or symptoms due to Lawsonia intracellularis infections, as well as associated disorders, in an animal, preferably a farming animal, more particularly a monogastric farming animal, preferably a pig.

SUMMARY OF THE INVENTION

In certain aspects, the present disclosure relates to use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains, for reducing the risk of contracting Lawsonia intracellularis infection and/or alleviating Lawsonia intracellularis infection and/or symptoms of Lawsonia intracellularis infection, preferably in an animal such as a farming or companion animal, preferably a farming animal, more preferably a monogastric farming animal.

In another aspect, the present disclosure pertains to use of Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains for prophylaxis and/or treatment of proliferative enteropathy (also known as ileitis), such as porcine proliferative enteropathy or equine proliferative enteropathy, (porcine) intestinal adenopathy, necrotic enteritis, (regional) ileitis and proliferative (haemorrhagic) enteropathy.

In another aspect, the present disclosure provides for use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains, for preventing or alleviating L intracellularis-assocated lesions, e.g., L intracellularis-assoc\atedi lesions, and/or damage, e.g., L. intracellularis- induced damage, to the intestinal wall in an animal, e.g., a non-human animal, e.g., a pig or a horse.

In a preferred embodiment, the animal is a pig, preferably nursery pig, grower pig, or finisher pig. In certain embodiments, the animal is at least 25, 50, 75, 100, 125, 150 days of age and/or the Agaricus blazei or an extract thereof, or the composition comprising Agaricus blazei or an extract thereof, e.g., the Agaricus blazei fermented grains, the composition comprising Agaricus blazei fermented grains, the extract of Agaricus blazei fermented grains or the composition comprising an extract of Agaricus blazei fermented grains, are administered during gestation period, lactation period, grower period, finisher period, and/or slaughter age periods.

In a preferred embodiment, the Agaricus blazei is provided in the form of fermented Agaricus blazei, e.g. solid-state fermented Agaricus blazei. Preferably, the Agaricus blazei is fermented on grains. In an embodiment, the Agaricus blazei is provided in the form of Agaricus blazei fermented grains. Also, an extract of Agaricus blazei, fermented Agaricus blazei, solid-state fermented Agaricus blazei, or Agaricus blazei fermented grains may be used in the present disclosure.

In an embodiment, the Agaricus blazei or the extract thereof or the Agaricus blazei fermented grains or extract thereof are comprised in a composition, e.g., an animal feed, in an amount of 0.2 - 10 kg, preferably 0.5 -5 kg per ton, preferably of animal feed. The grain may be chosen from any type of grain, e.g., corn, wheat bran, barley, whole wheat, oat, sorghum, rye, soy beans, maize, preferably oat or rye.

In a further aspect of the disclosure, the Agaricus blazei or an extract thereof, e.g., in the form of Agaricus blazei fermented grains, or an extract thereof are provided in a composition comprising the same, e.g., as an ingredient, premix or supplement for an animal feed. The Agaricus blazei or an extract thereof, or the Agaricus blazei fermented grains or an extract thereof may, for example, be combined with vitamins, trace elements, minerals and/or organic acids.

In yet another aspect, the present disclosure provides for a method for increasing average daily body weight gain in an animal population having Lawsonia intracellularis infection or at risk of contracting a Lawsonia intracellularis infection, said method comprising administering said Agaricus blazei or an extract thereof, or said Agaricus blazei fermented grains or extract thereof, or composition comprising said Agaricus blazei or an extract thereof, said Agaricus blazei fermented grains, said extract of said Agaricus blazei fermented grains, or said composition comprising said extract of said Agaricus blazei fermented grains to said animal population. In a preferred embodiment, the population comprises at least 10, 25, 50, 100 animals. The animals may be farming animals or companion animals, more preferably monogastric animals. In a preferred embodiment, the animals are pigs, preferably nursery pigs, grower pigs, or finisher pigs.

GENERAL DEFINITIONS

In the following description and examples, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, including the scope to be given to such terms, the following definitions are provided. Unless otherwise defined herein, all technical and scientific terms used have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

The terms “Agaricus blazei” and “Agaricus blazei fermented grains” are known by the skilled person. Agaricus blazei, also referred to as Agaricus subrufescens, Agaricus brasiliensis or Agaricus rufotegulis, can be seen as a species of mushroom. Agaricus blazei fermented grains can for example be obtained as described in WO2013171194. In particular, Agaricus blazei fermented grains can be seen as material comprising grain and Agaricus blazei, preferably Agaricus blazei mycelium, wherein the grain preferably comprises the Agaricus blazei mycelium in an amount of between 10 and 50% by weight on dry matter, and/or the material has a moisture content of less than 10% (relative to dry matter). In an embodiment, the Agaricus blazei fermented grains do not comprise Agaricus blazei fruiting bodies. Several commonly known grain types can be used, such as for example corn, wheat bran, barley, whole wheat, oat, sorghum, rye, soy beans, maize and the like, or mixtures of any of these. In a preferred embodiment, the grain is rye or oat. A suitable method to obtain Agaricus blazei fermented grains comprises: a) fermenting Agaricus blazei on grain, preferably until 10-50% mycelium content is achieved, wherein the grain preferably has a moisture content between 10 and 90%; b) drying the fermented grain, preferably to a moisture content of between 2 and 8 wt% (with respect to the dry material); c) milling the so obtained fermented and dried material, e.g., to a particle size with a D50 between 0.01 and 20 mm. In an embodiment, the Agaricus blazei fermented grains are obtainable using such method.

The Agaricus blazei may be allowed to ferment on said grain during a time period of at 4 weeks, such as at least 5, 6, 7, 8, 9, 10, 11, 12, or more weeks, in order to allow a mycelium content to be formed of at least 10%, preferably at least 15%, even more preferably at least 20%.

The drying may take place at any suitable temperature using any suitable means known to the skilled person. Similarly, the milling may take place using any suitable means known to the skilled person.

The term “extract” as used herein refers to a substance or composition obtained from e.g. Agaricus blazei or Agaricus blazei fermented grains, or from a composition comprising Agaricus blazei or Agaricus blazei fermented grains. The extract may for example be obtained by chemical or mechanical action, e.g. by treating with solvent, or by applying pressure, distillation, sublimation or evaporation. In an embodiment, the extract may for example comprise ergosterol, extra-cellular enzymes like cellulases and amylases, and/or 1,3- and I,q-b-glucans. Lawsonia intracellularis (LI) infection is an infection which can be detected in an animal or animal population by a veterinarian. An LI infection can be detected for example by PCR on faecal or tissue samples as for example described in Veterinary Microbiology 102(3- 4): 189-201. Alternatively, LI infection can be detected in (such) a sample by using a commercial L intracellularis detection kit, for example the VetMAX™ L. intracellularis Kit (Thermo Fisher Scientific, Qiagen).

“Proliferative enteropathy (ileitis)” is an infectious enteric disease characterized by thickening of the mucosa of the intestine due to hyperplasia of the crypt enterocytes. The disease in pigs includes several acute and chronic clinical manifestations, including proliferative hemorrhagic enteropathy and acute hemorrhagic diarrhea with sudden death of pigs close to market age, and porcine intestinal adenomatosis, a chronic mild diarrhea with reduced performance of growing pigs. Other terms used to describe this disease include necrotic enteritis, regional or terminal ileitis, garden-hose gut, and subclinical ileitis. All forms of the disease have in common the presence of proliferating crypt epithelial cells caused by an obligately intracellular bacterium, Lawsonia intracellularis. Proliferative enteropathy has been reported in various other animal species, including hamsters, horses, ferrets, foxes, rabbits, deer, some avian species, and non-human primates.

In horse, L intracellularis infections may cause diarrhea, depression, fever, inappetance (anorexia), weight loss, edema (fluid swelling) on the abdomen or lower limbs, a poor hair coat, and intermittent colic. With pigs suffering from Lawsonia intracellularis infections, the pig may appear clinically normal and initially eats well but there is a chronic diarrhoea, gradual wasting and loss of condition, followed in some cases by a potbellied appearance. Acute disease may be manifest by bloody gut or proliferative haemorrhagic enteropathy (PHE) and the pig may die suddenly or appear very pale and anaemic and pass black bloody faeces. Secondary bacterial infections often increase the severity of the disease. PHE occurs frequently in young gilts, particularly within 4 to 6 weeks of arrival on the farm, at the point of service and up to the middle stage of pregnancy. Pigs with the chronic form of porcine enteropathy may recover over a period of four to six weeks, however there can be considerable losses in feed efficiency and daily gain of up to 0.3 and 80g/day respectively. As a consequence there can be marked variations in sizes of pigs. Often the signs are subclinical, with only a variation in pig performance noted. Mortality is low and most pigs recover, though there will be a reduction in average weight gain. The acute form (proliferative hemorrhagic enteropathy) occurs more commonly in young adults 4 to 12 months of age, including late finishing pigs and breeding gilts. In this form, an acute diarrhea with black, tarry, bloody feces occurs. Often, sudden death is the first sign. Mortality is high, with around half of clinically affected animals dying. The term “animal” as used herein includes all non-human animals. The animals may, for example, be selected from farming animals and companion animals. Non-limiting examples of farming animals, also known as livestock animals, include poultry (e.g., chickens), swine (pig), beef cattle, milk cows, veal, pigs, goats, sheep, bison. Suitable companion animals include, but are not limited to, cats, dogs, horses, rabbits, rodents (e.g., mice, rats, hamsters, gerbils, and guinea pigs), hedgehogs, and ferrets. The animals may further be selected from monogastric animals and ruminant animals. A monogastric animal has a simple single-chambered stomach, compared with a ruminant organism, like a cow, goat, or sheep, which has a four-chambered complex stomach. Examples of monogastric farming animals include poultry, swine, and the like. The animals may further be selected from swine or pig, e.g. sow or piglet, preferably (post-)weaner pig (up to 25 kg), grower pig (25-70 kg), or finisher pig (more than 70 kg). The animal may be at least 25, 50, 75, 100,

125, 150 days of age.

As used herein, the term “test animal” refers to an animal that received Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains as taught herein as part of its diet. In contrast, a “control animal” refers to an animal of the same species that did not receive Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains as taught herein as part of its diet. Suitably, both the test animals and control animals were fed the same diet, albeit with the exception of the presence of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains as taught herein, and were kept under similar climatic conditions.

The term “diet” as used herein refers to the sum of feed and beverages or drinks or drinking water consumed by an animal.

As used herein, “feed conversion ratio (FCR)” is a ratio measuring the efficiency with which the bodies of livestock convert animal feed into the desired output, e.g., body weight. For grower-finisher pigs, for example, the output may be the body mass gained by the animal. FCR is the mass of the input divided by the output (thus kg feed / kg growth or body weight gain). Conversely, feed efficiency, which is the output divided by the input (i.e. the inverse of FCR), may be used to express the efficiency with which the bodies of livestock convert animal feed into growth.

An animal feed as taught herein is any substance or combination of substances used for feeding animals. It may comprise only bulk feed ingredients or it may be a complete feed. The term “complete feed” as used herein refers to a formulated feed which is balanced in all the essential nutrients needed for normal growth and production of animals. It may be in the form of mash, pellets, crumbles or any other form known to the skilled person.

An ingredient for an animal feed is meant to be any substance that is or may be used in producing an animal feed.

Premixes are compositions for addition to bulk feed ingredients, e.g., to make up complete feed, and may be composed of ingredients such as vitamins, minerals, chemical preservatives, organic acids, antibiotics, fermentation products, and other beneficial ingredients. Premixes are often used for blending into bulk feed products or complete feeds, but may sometimes may be added to animal feed in the form of a top dress, or may be added to the drinking water.

A supplement is an ingredient or combination of ingredients added to animal feed, e.g., to fulfil a specific need. Supplements are usually used in relatively small quantities and requires careful handling and mixing (AAFCO, 2000). Such a supplement may, optionally, provide further nutrients such as vitamins, minerals, chemical preservatives, antibiotics, fermentation products, and other essential ingredients in order to increase the quantity of their consumption. Supplements may be fed separately from other feed products.

An ingredient, premix or supplement according to the disclosure may be suitable for feeding mammals, in particular non-human mammals, particularly pigs or swine.

Dry matter represents everything contained in a feed sample except water; this includes protein, fiber, fat, minerals, etc. In practice, it is the total weight of feed minus the weight of water (moisture content) in the feed, expressed as a percentage. It is determined by drying the feed sample in an oven (e.g. at 40, 60, 80, 100 degrees Celsius) until the sample reaches a stable weight.

The terms “comprising” or “to comprise” and their conjugations, as used herein, refer to a situation wherein said terms are used in their non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. It also encompasses the more limiting verb “to consist essentially of” and “to consist of”.

The term "alleviating" can, apart from its generally accepted meaning, that is to say the alleviation of the cause and/or of the symptoms of an affliction, herein also be used in the sense of the generally accepted meanings of the term of its properties in terms of nutrition physiology and pro-"treatment". i.e. preventing obviating pushing back, alleviating, improving, slowing, stopping or reversing the progression, or severity of a pathological condition or of the afflictions therewith. In particular, apart from the alleviation of the cause and/or of the symptoms of a Lawsonia intracellularis infection and disorders associated therewith, the treatment and prevention of the respective disorder can in this case also be comprised. Vice versa, the terms “prevention” and “prophylaxis” encompass reduction of the risk of contracting an affliction, and the term “treatment” also encompasses alleviation of (symptoms) of an affliction.

Reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article "a" or "an" thus usually means "at least one".

DETAILED DESCRIPTION

The present inventors have surprisingly found that epithelial tissue damage observed in epithelial tissue challenged with Lawsonia intracellularis can be prevented by Agaricus blazei as set out in the examples herein. Thus, Agaricus blazei prevents and/or epithelial tissue damage by Lawsonia intracellularis. In particular, administering Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains to farming animals reduces their risk of contracting Lawsonia intracellularis infection and/or alleviates Lawsonia intracellularis infection and/or any symptoms of a Lawsonia intracellularis infection. Symptoms of Lawsonia intracellularis infection may include (hemorrhagic) diarrhea, fever, lack of appetite, weight loss, decreased feed conversion ratio (FCR), growth retardation, and even death. The present disclosure also teaches a method of feeding a farming animal, said method comprising the step of administering to said farming animal Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains, in order to reduce the risk of contracting Lawsonia intracellularis infection and/or to alleviate Lawsonia intracellularis infection and/or symptoms of Lawsonia intracellularis infection. Thus, conditions and/or diseases associated with Lawsonia intracellularis infection, such as proliferative enteropathy (also known as ileitis), such as porcine proliferative enteropathy or equine proliferative enteropathy, (porcine) intestinal adenopathy, necrotic enteritis, (regional) ileitis and proliferative (haemorrhagic) enteropathy, may be prevented and/or treated. An aspect of the present disclosure relates to Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains for use in the prevention and/or treatment of Lawsonia intracellularis infection and/or associated conditions and/or diseases, such as proliferative enteropathy (also known as ileitis), such as porcine proliferative enteropathy or equine proliferative enteropathy, (porcine) intestinal adenopathy, necrotic enteritis, (regional) ileitis and proliferative (haemorrhagic) enteropathy, or for use in reducing risk of contracting Lawsonia intracellularis infection and/or alleviating Lawsonia intracellularis infection and/or alleviating symptoms of Lawsonia intracellularis infection. The use according to the present disclosure may concern a single animal, or a population of animals, for example a population of at least 2, 5, 10, 25, 50, 75, 100, 150, 200 animals. Common observations associated with Lawsonia intracellularis infection in such a population include variation in animal weight, decreased growth rates, and apathy, which can be countered according to the present disclosure.

In another aspect, the present disclosure provides for use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains, for preventing or alleviating L intracellularis-assocated lesions, e.g., L intracellularis-assoc\atedi lesions, and/or damage, e.g., L intracellularis- induced damage, to the intestinal wall in an animal, e.g., a non-human animal, e.g., a pig or a horse.

In a further aspect, the present disclosure teaches the use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains for increasing (average) body weight or (average) body weight gain and/or average daily (weight) gain in an animal, in particular a farming animal, particularly a population of at least 2, 5, 10, 25, 50, 75, 100, 150, 200 animals. The body weight or body weight gain of a test animal (population) may be increased relative to the body weight or body weight gain of a control animal (population). The body weight gain of a test animal, for example a pig, may be determined using any standard methodology known to the skilled person. For example, the animals (both the control and the test animals) may be weighed daily or weekly to keep track of the body weight gain. The body weight gain may be increased when the body weight gain of the test animals throughout the test period is higher than the body weight gain of the control animals. In another aspect, the present disclosure teaches the use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains for reducing variation in body weight in an animal population, e.g. a population of at least 2, 5, 10, 25, 50, 75, 100, 150, 200 (farming) animals of similar age, e.g. differing at most 50, 25, or 10 days in age. Accordingly, the standard deviation of body weight values of a test animal population may be reduced relative to the standard deviation of body weight values of a control animal population. The variation of body weight of a test animal population, for example pigs, may be determined using any standard methodology known to the skilled person. For example, the animals (both the control and the test animals) may be weighed daily or weekly to keep track of the body weight variation. The body weight variation may be reduced when the body weight variation of the test animals throughout the test period is lower than the body weight variation of the control animals.

The test period may be the period from 25 days from birth or 50 days from birth up to the time of slaughter. For example, the test period may be from 0, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 (days of birth) up to about the age of 110, 120, 130, 140, 150, 160, 170, 180,

190, 200 or more days, depending upon the desired time of slaughter. For calculating the average daily gain, the body weight gain is divided by the number of days of the test period. It will be clear that average daily gain and body weight gain are closely related.

In another aspect, the present disclosure teaches the use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains, for decreasing (i.e. improving) feed conversion ratio (FCR), i.e., improving feed efficiency, in an animal, in particular a farming animal. The FCR may be decreased in a test animal relative to the feed conversion ratio observed in a control animal. The FCR may be determined using any standard methodology known to the skilled person. For example, feed intake (in, e.g., grams) may be recorded over a period, e.g., a week, as well as body weight gain (or mass gain) (in, e.g., grams) over the same period, and FCR can be calculated by dividing the total feed intake over that period by the body weight gain over that period.

In another aspect, the present disclosure teaches the use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains for increasing feed efficiency in an animal, in particular a farming animal. The skilled person is capable of converting FCR into feed efficiency.

In another aspect, the present disclosure teaches the use of Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains to increase daily feed intake in an animal, preferably a farming animal. The daily feed intake may be determined by measuring the amount of feed that an animal eats each day. The daily feed intake may be averaged over several days to calculate an average daily feed intake.

In embodiments of the disclosure, the animal (population) concerns a farming or companion animal, preferably a monogastric animal, such as a pig or a horse. Preferably the animal (population) concerns a pig, preferably a (post-)weaner pig, nursery pig, grower pig, or finisher pig. In the context of the present disclosure, a (post-)weaner pig preferably is a pig weighing 5-25 kg. A grower pig can be seen as a pig weighing 25-75 kg. A finisher pig can be seen as a pig weighing more than 70 kg. The animal or population thereof according to the present disclosure may concern animal(s) at least 25, 50, 75, 100, 125, 150 days of age and/or between 25-250 days, or 50-250, 50-200 days of age.

The Agaricus blazei, the extract thereof, or the composition comprising Agaricus blazei or the extract thereof, e.g., Agaricus blazei fermented grains, the composition comprising Agaricus blazei fermented grains, the extract of Agaricus blazei fermented grains, or the composition comprising the extract of Agaricus blazei fermented grains may be administered to said animals in any way known to the skilled person. The Agaricus blazei, the extract thereof, or the composition comprising Agaricus blazei or the extract thereof, e.g., Agaricus blazei fermented grains, the composition comprising Agaricus blazei fermented grains, the extract of Agaricus blazei fermented grains, or the composition comprising an extract of Agaricus blazei fermented grains is preferably administered orally. The Agaricus blazei, the extract thereof, or the composition comprising Agaricus blazei or the extract thereof, e.g., Agaricus blazei fermented grains, the composition comprising Agaricus blazei fermented grains, the extract of Agaricus blazei fermented grains, or the composition comprising an extract of Agaricus blazei fermented grains may be administered during post birth period, weaning period, juvenile period, fattening period, gestation period, lactation period, grower period, finisher period and/or slaughter age period. The Agaricus blazei, the extract thereof, or the composition comprising Agaricus blazei or the extract thereof, e.g., Agaricus blazei fermented grains, the composition comprising Agaricus blazei fermented grains, the extract of Agaricus blazei fermented grains, or the composition comprising an extract of Agaricus blazei fermented grains may be incorporated in starter, grower and/or finisher diets. The composition of such diets is adjusted depending on the need of an animal, depending on its age. Such diets may comprise about 23%, about 20% and about 18% protein, such as respectively between 21 and 25%, between 18 and 22%, and between 16 and 20% protein.

The present disclosure also teaches an animal feed comprising Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains e.g. in an amount of 0.2 - 10 kg, preferably 0.5 -5 kg per ton animal feed. The animal feed may comprise the Agaricus blazei, an extract thereof, or the composition comprising Agaricus blazei or the extract thereof, e.g., Agaricus blazei fermented grains, the composition comprising Agaricus blazei fermented grains, the extract of Agaricus blazei fermented grains, or the composition comprising an extract of Agaricus blazei fermented grains or the animal feed may be obtained by mixing a commercially obtainable animal feed with an ingredient, premix or supplement for an animal feed suitable for feeding animals, comprising the Agaricus blazei, the extract thereof, or the composition comprising Agaricus blazei or the extract thereof, e.g., Agaricus blazei fermented grains, the composition comprising Agaricus blazei fermented grains, the extract of Agaricus blazei fermented grains, or the composition comprising an extract of Agaricus blazei fermented grains e.g. to obtain animal feed with Agaricus blazei or an extract thereof or Agaricus blazei fermented grains or an extract thereof in an amount of 0.2 - 10 kg, preferably 0.5 -5 kg per ton animal feed. The animal feed may be in the form of mash, pellets, crumbles or any other form known to the skilled person.

In yet another aspect, the disclosure provides for an ingredient, premix or supplement, comprising Agaricus blazei, an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, e.g., Agaricus blazei fermented grains, a composition comprising Agaricus blazei fermented grains, an extract of Agaricus blazei fermented grains, or a composition comprising an extract of Agaricus blazei fermented grains as taught herein, wherein the dosage of said Agaricus blazei, an extract thereof or said Agaricus blazei fermented grains or extract thereof in said ingredient, premix or supplement is preferably 50, 100, 250 g/kg or higher, preferably 50 - 500 g/kg, based on the total weight of said ingredient, premix or supplement.

In an embodiment, the animal feed, the ingredient, premix or supplement according to the present disclosure further comprises vitamins, trace elements, minerals and/or organic acids.

In another aspect, the disclosure provides for a method for increasing (average) body weight gain, average daily gain, (average) daily intake, (average) feed efficiency, in an animal or animal population having or at risk of having Lawsonia intracellularis infection or any one of the diseases or symptoms mentioned herein, particularly such population of at least 2, 5, 10, 25, 50, 75, 100, 150, 200 animals, said method comprising feeding said animal or said animal population with the ingredient, premix or supplement, or with the animal feed taught herein.

In another aspect, the disclosure provides for a method for reducing variation in body weight in an animal population, e.g. a population of at least 2, 5, 10, 25, 50, 75, 100, 150,

200 (farming) animals, said method comprising feeding said animal or said animal population with the ingredient, premix or supplement, or with the animal feed taught herein. Preferably the animals in the population are of similar age, e.g. differing at most 50, 25, or 10 days in age.

In preferred embodiments, the animal (population) concerns a farming or companion animal, preferably a monogastric animal, such as a pig or a horse. Preferably the animal (population) concerns a pig, preferably a (post-) weaner pig, nursery pig, grower pig, or finisher pig. It may also be a sow.

Agaricus Blazei as taught herein (also called Agaricus Subrufescens, Agaricus Blazei Brasiliensis, or Agaricus subrufescens, or Agaricus rufoteguiis) preferably refers to Agaricus Blazei Murill. Herein, the abbreviation ABM for Agaricus Blazei murill is used interchangeably. Agaricus Blazei fermented grains are known as such, and are for example described in WO2013171194. However, the use as taught in the present disclosure is neither described, nor suggested therein.

The Agaricus species as taught herein, preferably Agaricus Blazei or ABM can be grown as mycelium on a grain source, and subsequently harvested as such, either in combination with the grain source or separate from the grain source. For example, fruiting bodies may be harvested as such, or both fruiting bodies, grain source and mycelium may be harvested, or the emergence of fruiting bodies may substantially be prevented and grain source and mycelium may be harvested. Thus, optionally, mycelium growth can be executed until till fruiting bodies emerge, but this is not necessary. It is also not necessary, although optional, to separate the growth medium, e.g., grain source or solid-state substrate, from the mycelium. Thereby, a non-laborious and economically effective way of providing useful feed is provided.

In the current disclosure, several commonly known grain types can be used, such as for example corn, wheat bran, barley, whole wheat, oat, sorghum, rye, soy beans, maize and the like, or mixtures of any of these. To increase growth of the mycelium, addition of a carbon source, ammonia source or calcium compounds like chalk is foreseen. Rye or oat are particularly preferred grain types because ABM grows particularly well thereon. For obtaining Agaricus Blazei or ABM fermented grain as taught herein, it is preferred that the grain source has a high moisture content, for example between about 10-80% moisture (measured as weight of the moist product minus a dried product, divided by the moist product weight, when drying is preferably performed in an oven (e.g. at 40, 60, 80, 100 degrees Celsius, e.g. at atmospheric pressure until a stable weight is reached). A preferred moisture content is between about 30 and 70%, or between about 40 and 60%, like for example about 50 %.

The grain source is preferably sterilized before inoculation with the Agaricus blazei or ABM mycelium. The inoculation, and the preparation of the inoculate can be performed according to standard techniques, such as described in US4204364. The fermentation typically takes place in containers of 20 to 50 litre size, such as in bags or trays, for example 25 or 30 litre bags. Fermentation preferably is conducted in a conditioned environment. Generally, the time for fermentation can range between 15 and 75 days, or between 15 and 55 days, preferably between 20 and 45 days, in order to obtain optimum results with respect to mycelium content. The temperature during fermentation preferably ranges between 20 and 35 °C, and most preferably between 28 and 30 °C. The humidity preferably ranges between 40% and 90% RH, and may be, for example, 50 or 60% RH.

The amount of mycelium may range preferably between about 10 and 50 % (on dry weight of the mixture of grain and mycelium), preferably between 20 and 30 %. It is possible to measure the amount of mycelium indirectly, for example based on the ergosterol content, as is well known to the skilled person and as has been, for example, described in WO2013171194, which is incorporated herein by reference. Any alternative method for determining the mycelium content may be used, for example, the mycelium content can be measured based on the amount of chitin using methods well known to the skilled person.

After the fermentation, the Agaricus blazei fermented grain is preferably dried. The moisture content of the dried fermented grain preferably is less than about 10%, preferably less than 8% and more preferably less than about 6 (measured as weight of the dried product minus a further dried product, divided by the dried product weight, when further drying is preferably performed in an oven at e.g. 40, 60, 80, or 100 degrees Celsius at e.g. atmospheric pressure until a stable weight is reached). A lower moisture content was found to be beneficial for good storage stability. Generally, the moisture content may be about 1- 5%, or 2-4% preferably about 2% or more, or about 3% or more. A feed with very low moisture content has no disadvantages from technical perspective, but may be more costly to produce.

Any drying method may be suitable for the drying purposes as taught herein. A suitable drying device includes a belt dryer, bulb dryer, tumble dryer or fluid bed dryer. Preferably, the device is such that it is able to perform the drying at a reduced pressure, e.g. less than 40, 50, 60, 70, 80, 90, 100% atmospheric pressure. In general, it is preferred that the temperature during drying remains below 100 °C. For example, the fermented grain may be dried by air drying at between 5 and 100 °C, preferably 15 between 25 and 90 °C, more preferably below 63°C, and even more preferably below 50 °C. Drying at a temperature below about 50 °C has the advantage that extra-cellular enzymes, like cellulases keep their activity. However, in case the enzymes that are of interest have been modified to be resistant to higher temperatures, a higher temperature for drying is preferred because the drying speed is increased. If a temperature is chosen below 63 °C, 45 °C, or in particular at about 35 °C or lower, it is preferred to apply reduced pressure, e.g. less than 40, 50, 60, 70, 80, 90, 100% atmospheric pressure. The low temperature applied during the drying step has the further advantage that heat sensitive compounds produced during fermentation remain in an active form in the feed.

The dried fermented grain as taught herein can be storage stable for several months up to at least one year (e.g. for two years), with little reduction of the nutritive value and beneficial effects of the animal feed. The fermented dried grain (animal feed) can be used as such, as or in pig feed.

For improving mixing characteristics with common feedstock, and to allow reproducible administration to the animals, it is preferred to crush or mill the fermented dried grain to a particle size (D50) between 0.01 and 20 mm, preferably lower than 15 mm, and even more preferably between 0.1 and 10 mm. The particle size range preferably is between 0.1 and 10 mm for about 90% or more of the animal feed, preferably of about 95% or more of the animal feed, and more preferably of about 98% or more of the animal feed. Preferably, grain size (absolute) is smaller than 25 mm.

Animal feed as taught herein may for example be produced by a method comprising a) fermenting Agaricus Blazei on grain, preferably until 10-50% mycelium content is achieved, wherein the grain preferably has a moisture content between 10 and 90%, and wherein the grain is preferably rye or oat; b) optionally, drying the fermented grain, preferably to a moisture content of between 2 and 8 wt% of the dried material; c) optionally, milling the so obtained fermented and dried material, e.g., to a particle size with a D50 between 0.01 and 20 mm.

The fermented, optionally dried and optionally milled fermented grain can be used as such as feed, as an additional feed source, or can be comprised in a premix, or it can be mixed with other common feedstock, for example as ingredient or supplement for an animal or swine feed. Preferably, the animal feed is mixed with the general feedstock, as that allows better standardization and/or reproducibility.

The animal feed as taught herein may contain between 10 and 50 wt% of mycelium (based on dry weight), preferably between 20 and 30 wt%. The amount of mycelium can be measured based on the content of ergosterol (and/or content of chitin), as e.g. described in the Example section of WO2013171194.

The fermentation will produce a number of compounds that can be useful for the animal. The Agaricus Blazei species produces for example, ergosterol, extra-cellular enzymes like cellulases and amylases, and 1,3- and I,d-b-glucans. The amount of ergosterol in the obtained animal feed typically will be between 0.05 and 0.5 g ergosterol/kg animal feed. Ergosterol can be measured with standard techniques, like HPLC or GC. Preferably, quantitative extraction is performed on pulverized dry fermented grain, which can be done with hot 80% ethanol.

It is preferred to use such an amount of animal feed as taught herein, that the mixed feed comprises between 50 and 1000 microgram ergosterol per kg animal feed, preferably in an amount of 100-600 microgram ergosterol per kg animal feed.

The amount of mycelium (as pure dry matter) given to the animals per day, can be be between 0.1 and 1.0 kg per ton feed, preferably between 0.2 and 0.8 kg per ton feed. The amount of animal feed as taught herein can be about 0.2 kg per ton or more, preferably about 0.5 kg or more, and more preferably about 1 kg per ton feed or more. Generally, the amount will be about 10 kg per ton feed or less, preferably about 5 kg per ton feed or less. Suitable examples include 2, 3 or 5 kg per ton feed.

The animal feed as taught herein is preferably mixed with other feed components. Common feedstock are grains, soybean meal, sesame meal, fish meal, cottonseed meal and the like. It is not necessary to use the animal feed as taught herein every day. Intermittent administration, like for example every other day, once every three days, or once a week may be suitable as well. It is thought that every day, or every other day is most effective.

The present disclosure is further illustrated, but not limited, by the following examples. From the above description and these examples, one skilled in the art can ascertain the essential characteristics of the present invention, and without departing from the teaching and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions. Thus, various modifications of the invention in addition to those shown and described herein will be apparent to those skilled in the art from the foregoing description.

BRIEF DESCRIPTION OF FIGURE

Figure 1 : Percentage of healthy epithelium covering In vitro organ culture (explants) challenged with Lawsonia intracellularis at early (A) and late (B) time points. Box plots for each challenge group depict 25-75 percentiles, with whiskers representing the minimum and maximum points and the horizontal line denoting the median. EXAMPLE 1 - production of Agaricus blazei fermented grains

A bag of 15 L with 3 kg wet rye (50% moisture) was inoculated with 80 g ABM mycelium and cultivated for 42 days at 28 °C at 50% humidity. The resulting product was dried (to a moisture content of less than 5%) with a vacuum rotary vapor drum at 35 °C at 20mmHg pressure. The dried material was coarsely milled at d50 1000 micron (1 mm).

EXAMPLE 2 - in vitro screening of Agaricus blazei fermented grains to mitigate intestinal lesions caused by Lawsonia intracellularis in swine

The present inventors considered that in-feed delivery of Agaricus blazei fermented grains may support beneficial bacterial communities in the large intestine while helping maintain epithelial lining integrity. This, in turn, may help mitigate intestinal lesions associated with bacterial infections.

In vitro organ culture is a technique used to preserve fully developed organs in a controlled environment (explant). This example aimed to evaluate the in vitro efficacy of Agaricus blazei fermented grains to mitigate lesions associated with colon exposure to Lawsonia intracellularis.

Material and methods

Spiral colon collection and explant culture

A total of 15 commercial crossbred male pigs, between 5-6 weeks of age, from a high health commercial farm were used as tissue donors. Animals were euthanized by exsanguination after cranial captive bolt concussion. Distal spiral colon collection and culture followed a previously described protocol (Costa et al Methods Mol Biol. 2018; 1817:185-195). For each pig, following gastrointestinal post-mortem examination, a 10 cm segment of the spiral colon was aseptically collected and transported in a container with precooled (6°C - 10°C) Hank’s balanced salt solution (HBSS, Gibco, Canada) and transported within 10 minutes to a biosafety cabinet. Colon segments were washed with approximately 200 mL of the transport solution to remove luminal contents. This was followed by separation of the colonic serosa from the mucosa on a refrigerated surface. Next, the mucosa was divided into 2 cm x 2 cm segments (explants), which were individually placed on a 70 pm cell strainer (Fisher Scientific, Hanover Park, IL, US) in a six well plate (Millipore Sigma®, St. Louis, MO, US) containing 3 ml of culture media (KBM-Gold calcium and phenol red free Bullet Kit, Lonza, Walkersville, MD). Plates were placed in a modular incubator chamber (Billups Rothenberg INC, MIC101 , San Diego, CA, US) and gassed for 2 minutes with 99% oxygen (O2), 1% carbon dioxide (CO2) gas mix, and cultured at 37 °C. Inocula and compounds preparation

Explants were randomly allocated into groups and challenged with Lawsonia intracellularis. For L intracellularis, a live vaccine (Enterisol® Ileitis, Boehringer Ingelheim, Mississauga, ON) was used as inoculum. Immediately before challenging the explants, inocula were retrieved from the vaccine vials and an aliquot was collected for quantification by quantitative PCR (qPCR, sample kept frozen at -80°C until processing). Inoculum averaged 1x10⁴ cells/m L for L intracellularis. Phase contrast microscopy was used to verify that bacterial motility (when suitable), as an indicator of viability at inoculation.

Compound dilution was calculated based on in vivo guidelines: 2 kg of compound/tone of feed. The average daily intake of a 15 kg pig is 0.800 kg, thus each animal would have ingested 1.6 g of compound/day/live weight. Intestinal explants have 4 cm2 of surface area and weight in average 1g. Based on this, 160 pg of compound was delivered in vitro to each explant. All compounds were diluted in phosphate buffered saline (PBS). Explants in the compound + pathogen group could only receive a maximum of 100 pi within the inoculation ring. Thus, concentrated (2x) aliquots of the compounds and bacterial inocula were prepared by centrifuging 1 ml at 10.000 g for 5 minutes. The supernatant was discarded and the pellet was resuspended in 50 mI of 0.1 M, pH 7.0 sterile PBS.

Inoculation trials

Five pigs were used (n=15) and a total of 18 explants/compound/pig were randomly exposed to one of the following inoculum combinations: 1) Lawsonia intracellularis only (positive control, inoculated with Lawsonia intracellularis only, n=6 explants; “Pathogen only”), 2) Agaricus blazei fermented grain only (negative control, inoculated with ABM fermented grains only, n=6 explants; “Compound only”), 3) treatment group (co-exposure to Lawsonia intracellularis and Agaricus blazei fermented grain combination, n=6 explants; “Compound + pathogen”). To confine the inoculum within the luminal aspect of the explants, a polystyrene ring (1 cm diameter ^c 1 cm height) was attached to the mucosal side of each explant using a cyanoacrylate adhesive (3M Vetbond Tissue Adhesive, Saint Paul, MN). Explants were cultured for the following periods: L intracellularis for 2 (early, n=3) and 8 hours (late, n=3).

At each time-point, 1 explant was fixed in 10% buffered formalin until processing for histopathology and the remaining 2 were immersed in RNA-later® (Giagen, Germantown,

MD, US) at 4°C for 24 hours and then stored at -80°C until qPCR analysis. In addition, two technical control explants were immediately sampled for histopathology after processing to confirm that lesions did not result from manipulation or other ante-mortem factors.

Histopathology analysis Explants fixed in formalin were sectioned and stained using hematoxylin and eosin (H&E). Healthy epithelium was defined as the superficial layer of cells covering the luminal aspect of the explants displayed a simple columnar fashion, not having signs of metaplasia (abnormal cell shape), edema (increased intercellular or intracellular space), or apoptosis and necrosis (pyknotic nuclei).

RT- qPCR assays

Explant mRNA levels evaluated the glyceraldehyde-3-phosphate dehydrogenase (GAPDH, housekeeping reference gene), interleukin-1 a (I L-1 a), interferon-y (IFN-g), tumor necrosis-a (TNF-a) genes. Total RNA was extracted from explants preserved in RNAIater using a commercial kit (RNeasy animal cell and tissue kit, Qiagen, Mississauga, ON) and quantified by NanoDrop (ThermoFisher Scientific, Madison, Wl).

RT-qPCR was performed using the PowerUp SYBR Master Mix® (Thermo Fisher Scientific, Hanover Park, IL). Next, genomic DNA contaminants were removed and cDNA was transcribed (QuantiTect Reverse Transcription® Kit, Qiagen, Germantown, MD, US). Samples were diluted to 500 ng/pL with nuclease-free water, resulting in 10 ng/2 pl_ of template cDNA. Quantitative PCR reactions were performed using an ABI 7500 Fast Real- Time PCR System (Applied Biosystems, ThermoFisher Scientific, Madison, Wl). Each 20 pi- reaction contained 10 pL of PowerUp SYBR Master Mix, 1 mI_ each forward and reverse primers (10 mM), 6 mI of nuclease free water and 2 mI_ of cDNA template. Reactions were incubated at 50°C for 2 min and 95°C for 2 min, followed by 40 cycles of 15 s at 95°C, 30 s at 60°C and 1 min at 72°C and a melt curve step of 95°C for 15 s, 60°C for 1 min and 95°C for 15 s. Every reaction was performed in duplicates, alongside negative extractions and no template controls in each run. Samples where duplicates differed by more than 1 Ct were re analyzed.

Statistical analysis

ANOVA with donor pig as a random effect was used. Percentage of healthy epithelium were compared between all-time points (early and late) and between groups by generalized estimating equations (GEE) using an unstructured correlated working matrix while clustering by pig. The data followed a normal distribution. The mRNA levels were compared between groups using the MCMC qPCR package (one-way design) with naive statistical model was used. Analyses were performed using R version 3.5.3 and IBM SPSS v24 (San Diego, CA, USA).

Results

Infection model evaluation Histopathology data were used to compare the compound only (negative control) group to the pathogen only group (positive control). Explant exposure to a given pathogen (positive control) led to histopathological lesions and a detectable response, when compared to the compound-only exposed group (negative control).

Early time-point

Explants challenged with L intracellularis treated with Agaricus blazei fermented grains had numerically higher epithelial coverage than untreated, challenged explants (Figure 1A).

Late time-point

Explants challenged with L intracellularis and treated with Agaricus blazei fermented grains had higher epithelial coverage than explants challenged but not treated (P =0.003, Figure 1B). It is noteworthy that, despite the use of a vaccine strain, epithelial lesions and differences between groups could still be detected.

Conclusion

Simultaneous exposure of swine intestinal explants to L intracellularis and Agaricus blazei fermented grains demonstrated the usefulness of Agaricus blazei fermented grains to prevent and/or counteract the damaging effect L intracellularis had on the epithelial tissue. Based on the present example, Agaricus blazei, in particular Agaricus blazei fermented grains, seem to be particularly effective in preventing or alleviating L intracellularis- associated lesions; i.e. , damage to the intestinal wall.

Claims

1. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use in reducing risk of contracting Lawsonia intracellularis infection and/or alleviating Lawsonia intracellularis infection and/or alleviating symptoms of Lawsonia intracellularis infection.

2. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof, for use in prophylaxis and/or treatment of proliferative enteropathy, preferably porcine proliferative enteropathy or equine proliferative enteropathy, (porcine) intestinal adenopathy, necrotic enteritis, (regional) ileitis or proliferative (haemorrhagic) enteropathy.

3. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to any one of claims 1-2, wherein said Agaricus blazei or said extract thereof is in the form of Agaricus blazei fermented grains or an extract of Agaricus blazei fermented grains.

4. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to any one of claims 1-3, wherein the use is in an animal, preferably a farming animal, more preferably a monogastric animal.

5. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to claim 4, wherein the animal is a pig, preferably a nursery pig, grower pig, or finisher pig.

6. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to any one of claims 4-5, wherein the animal is at least 25, 50, 75, 100, 125, 150 days of age.

7. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to any one of claims 4-6, wherein the Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof are administered during gestation period, lactation period, grower period, finisher period, and/or slaughter age periods.

8. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to any one of claims 3-7, wherein the grain is chosen from corn, wheat bran, barley, whole wheat, oat, sorghum, rye, soy beans, maize, preferably oat or rye.

9. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to any one of the previous claims, wherein said Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof are comprised in an animal feed, preferably in an amount of 0.2 - 10 kg, preferably 0.5 -5 kg per ton animal feed.

10. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to any one of the previous claims, wherein said Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof are provided in an ingredient, premix or supplement for an animal feed.

11. Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof for use according to any one of the previous claims, wherein the Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof are combined with vitamins, trace elements, minerals and/or organic acids.

12. Method for increasing average daily body weight gain in an animal population having Lawsonia intracellularis infection, said method comprising administering to said animal population Agaricus blazei or an extract thereof, or a composition comprising Agaricus blazei or an extract thereof.

13. Method according to claim 12, wherein said Agaricus blazei or said extract thereof is in the form of Agaricus blazei fermented grains or an extract of Agaricus blazei fermented grains.

14. Method according to any of claims 12-13, wherein the population comprises at least 10, 25, 50, 100 animals.

15. Method according to any one of claims 12-14, wherein the animals are farming animals, more preferably monogastric animals.

16. Method according to any one of claims 12-15, wherein the animals are pigs, preferably nursery pigs, grower pigs, or finisher pigs.