WO2014131422A2

WO2014131422A2 - Fermented rapeseed feed ingredient

Info

Publication number: WO2014131422A2
Application number: PCT/DK2014/050043
Authority: WO
Inventors: Jens Høffner Legarth
Original assignee: Fermentationexperts AS
Current assignee: Fermentationexperts AS
Priority date: 2013-02-28
Filing date: 2014-02-28
Publication date: 2014-09-04
Anticipated expiration: 2015-08-28
Also published as: WO2014131422A3

Abstract

The present invention relates to a fermented rapeseed product. The rapeseed product may be used as a medicament e.g. in the treatment or prevention of salmonella infections in poultry. In addition, the invention relates to an efficient production process including a cooling step, which avoids agglomeration of the fermented product.

Description

Fermented rapeseed feed ingredient

Technical field of the invention

The present invention relates to a fermented rapeseed feed product. In particular the present invention relates to a fermented rapeseed feed product for use a medicament, e.g. in the treatment of salmonella and other infections in poultry or other species.

Background of the invention

There is a need in animal production units to minimize the use of antibiotics but at the same time to maximize the yield from production units. Furthermore, a stable infected with e.g. a salmonella infection may result in that all animals must be sacrificed without any profit. WO 2008/006382 describes a process for producing homofermented products and also discloses homofermented products. Furthermore, it is described that feeding the animals with such products may reduce the risk of infection with

Campylobacter and Salmonella spp. Glucosinolates are secondary plant metabolites that occur naturally in

Brassicaceae, a plant family that has given rise to important crops such as oilseeds (Brassica napus) and cabbages (Brassica oleracea). They are known to be harmful to humans and animals at high concentration and therefore over the years rape crops have been developed with low levels of glucosinolates to avoid high intake of these compounds.

Hence, an improved method for avoiding antibiotics would be advantageous, and in particular a more efficient and/or reliable feed assisting in prevention or removal of infections would be advantageous. Summary of the invention

The present inventors have surprisingly found that a fermented rapeseed product may function as a medicament in the treatment of bacterial and yeast infections e.g. in poultry. This was not the case for controlled liquid fermented products. WO 5 2008/006382 as mentioned above is silent in respect of the beneficial medical effect of fermented rapeseed.

Thus, an object of the present invention relates to providing a feed product, which may function as a medicament in the treatment or prevention of bacterial and/or0 yeast infections.

Glucosinolates are known to be present in rape at different concentration depending on the specific sort. Without being bound by theory, it is believed that that the reduction in glucosinolates by fermentation, generates one or more5 degradation products which are beneficial to humans and animals. Thus, contrary to the general opinion of using "double low" rapeseeds to avoid the glucosinolates, the present invention proposes to use rape (or other plants) with a relatively high content of glucosinolates (e.g. "single low" rape) in the production of fermented food/feed. The content of unhealthy glucosinolates is then reduced by lactic acid0 fermentation producing new product beneficial for the health (see example

section).

In particular, it is an object of the present invention to provide a product that solves the above mentioned problems with overuse of antibiotics.

5

Thus, one aspect of the invention relates to a fermented dry food/feed ingredient comprising

- fermented rapeseed in an amount of 5-50% by weight; and

- one or more probiotic bacteria strains in a total amount in the range 10⁵-0 10¹² CFU per gram.

A further aspect relates to a process for producing a fermented food/feed ingredient, said process comprising

a) providing a plant product to be fermented having a total glucosinolate5 content of at least 5 μη-iol/g (dry weight, DW); b) providing one or more lactic acid bacteria strains or one or more inoculums comprising lactic acid bacteria;

c) combining the products from step a) and b), and fermenting the plant product of step a) with the one or more lactic acid bacteria of step b); providing a fermented food/feed ingredient, having a total glucosinolate content of at less than 1 μη-iol/g (DW);

Yet an aspect relates to a food/feed ingredient obtained/obtainable by a process according to the invention. Yet another aspect relates to a fermented food/feed ingredient

• comprising a fermented plant product comprising one or more of

o plants from the genus Brassica such as oilseeds, rape, such as

rapeseed, rapeseed cake, rapeseed meal, cabbages, plants of the order Brassicales, such as families Brassicaceae/Cruciferae,

Capparidaceae, and Caricaceae, and plants from the genus

Drypetes, such as from the family Euphorbiaceae;

• having a total glucosinolate content below 1 mol/g (DW); and

• one or more lactic acid bacteria strains in a total amount in the range 10⁵- 10¹² CFU per gram.

An aspect also relates to a food/feed product comprising a fermented food ingredient according to the invention.

Another aspect of the present invention relates to the food/feed product according to the invention for use as a medicament.

Yet another aspect of the present invention is to provide a food product according to the invention for use as a medicament in the treatment and/or prevention of bacterial and yeast infections in an animal.

Still another aspect of the present invention is to provide a process for producing a dry food ingredient comprising

a) providing a food/feed product to be fermented;

b) providing one or more lactic acid bacteria strains or one or more

inoculum comprising predominantly lactic acid bacteria; c) fermenting the food/feed product with the one or more lactic acid bacteria strains, providing a fermented food/feed ingredient; d) introducing said fermented food/feed ingredient into a dryer chamber; e) contacting said fermented food/feed ingredient with a stream of drying gas in said dryer chamber;

f) cooling the fermented food/feed ingredient exiting the drying chamber by instantly exposing the dried fermented food/feed ingredient to a cooling gas with a temperature in the range 5-30°C, such as 5-25°C, such as 10-25°C or such as 15-25°C; and

g) providing a dry fermented food/feed ingredient.

In yet an aspect the invention relates to a dry fermented food/feed ingredient obtainable by a process according to claim the invention. Brief description of the figures

Figure 1

Figure 1 shows the three treatment groups: Left) Control, Middle) Fermented feed, Right) Fermented Rapeseed, each with three replicate pens.

Figure 2

Figure 2 shows the diet setup for the three treatment groups. Left) Control, Middle) Fermented feed, Right) Fermented Rapeseed,

Figures 3-5

Figures 3-5 show a schematic representation of the study design for the three treatment groups Figure 3 (control), Figure 4 (FC), and Figure 5 (FR).

Figure 6

Figure 6 shows a graphic representation of the percentage of positive Salmonella swaps during the study (see also table 2).

Figure 7

Figure 7 shows a schematic overview of a spin flash The present invention will now be described in more detail in the following.

Detailed description of the invention

The present inventors have surprisingly found that a fermented rapeseed product may function as a medicament in the treatment of bacterial and yeast infections e.g. in poultry. This was not the case for other controlled fermented products. See example section. This effect is even further pronounced when fermenting plant material comprising a high amount of glucosinolates, such as rapeseed cake compared to plant materials comprising low amounts of glucosinolates, such as rapeseed meal.

Process for producing a fermented food/feed ingredient

Thus, an aspect of the invention relates to a process for producing a fermented food/feed ingredient, said process comprising

a) providing a plant product to be fermented having a total glucosinolate

content of at least 3 μη-iol/g (dry weight, DW);

b) providing one or more lactic acid bacteria strains or one or more inoculums comprising lactic acid bacteria;

d) optionally,

o introducing said fermented food/feed ingredient into a drying

chamber;

o contacting said fermented food/feed ingredient with a stream of drying gas in said dryer chamber;

o cooling the dried fermented food/feed ingredient, exiting the drying chamber, by instantly exposing the dried fermented food/feed ingredient to a cooling gas with a temperature in the range 5-30°C, such as 5-25°C, such as 10-25°C or such as 15-25°C; and providing a dry fermented food/feed ingredient. The food/feed ingredient according to the invention is to be understood in its broadest sense. The food/feed ingredient may comprise one or more of mature and/or immature plants and parts thereof, such as cereals, e.g. wheat, barley, rye, rice, maize (cob maize silage (CCM) or ripe), triticale, oat; vegetables (e.g. potatoes, beans, peas, maize, soy; whey, curd, skim milk and the like.

Glucosinolates

Glucosinolates are secondary plant metabolites that occur naturally in

Brassicaceae, a plant family that has given rise to important crops such as oilseeds (Brassica napus) and cabbages (Brassica oleracea). Based on the amino acid origin of the side chain, glucosinolates are divided into aliphatic, aromatic, and indole glucosinolates, which are derived from methionine, phenylalanine, and tryptophan, respectively. Thus, glucosinolates are natural components of many pungent plants such as mustard, cabbage and horseradish. The pungency of those plants are due to mustard oils produced from glucosinolates when the plant material is chewed, cut or otherwise damaged. It is generally believed that the use of high content glucosinolate-containing crops as primary food source for animals and humans has negative effects. Glucosinolates are well known for their toxic effects (mainly as goitrogens) in both humans and animals at high doses.

Probiotics

Different strains of probiotic bacteria may be used according to the present invention. In general, two types of fermentation processes can take place, namely "heterofermentative fermentation" by which the fermentation results in the formation of lactic acid, acetic acid, butyric acid, C02, water and ethanol, and "homofermentative fermentation" by which the fermentation results in the formation of practically only lactic acid. In a preferred embodiment the one or more lactic acid bacteria is homofermentative lactic acid bacteria.

According to the invention, fermentations comprise lactic acid-producing bacteria. Thus, in the broadest aspect, the present invention relates to lactic acid-producing bacteria. The lactic acid bacteria comprise a clade of Gram positive, low-GC, acid tolerant, non-sporulating, non-respiring rod or cocci that are associated by their common metabolic and physiological characteristics. These bacteria, usually found in decomposing plants and lactic products produce lactic acid as the major metabolic end product of carbohydrate fermentation. This trait has historically linked lactic acid bacteria with food fermentations as acidification inhibits the growth of spoilage agents. Proteinaceous bacteriocins are produced by several lactic acid bacteria strains and provide an additional hurdle for spoilage and pathogenic microorganisms. Furthermore, lactic acid and other metabolic products contribute to the organoleptic and textural profile of a food item. The industrial importance of the lactic acid bacteria is further evidenced by their generally regarded as safe (GRAS) status, due to their ubiquitous appearance in food and their contribution to the healthy microflora of human mucosal surfaces. The genera that comprise the lactic acid bacteria are Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, and Streptococcus, Aerococcus, Carnobacterium, Enterococcus, Oenococcus, Teragenococcus, Vagococcus, and Weisella; these genera belong to the order Lactobacillales. In the present invention, the lactic acid-producing bacteria used for fermentation are mainly and non-exclusively lactic acid bacteria of the genus Enterococcus, Lactobacillus, Pediococcus or Lactococcus, or combinations thereof. A starter culture or inoculum according to the invention can also comprise lactic acid bacteria selected from the group consisting of one or more Enterococcus spp., Lactobacillus spp., Lactococcus spp., and Pediococcus spp.. In another

embodiment of the invention, the lactic acid bacteria are selected from the group consisting of one or more one Enterococcus faecium, Lactobacillus rhamnosus, Lactobacillus plantarum, Pediococcus acidililactili, and Pediococcus pentosaceus. In further embodiment, the lactic acid producing bacteria are of the order

Lactobacillales. The lactic acid-producing bacteria can also be selected from

Lactobacillus spp., Pediococcus spp., Enterococcus spp., and Lactococcus spp.. or a combination thereof. In yet another embodiment, the lactic acid- producing bacteria comprise Pediococcus pentosaceus, Pendiococcus acidilactici and

Lactobacillus plantarum, Lactobacillus rhamnosus, and Enterococcus faecium, or a combination thereof. In still another embodiment, the lactic acid bacteria comprise Enterococcus faecium and/or Lactobacillus rhamnosus. In a further embodiment, the lactic acid bacteria comprise one or more of Enterococcus faecium MCIMB 30122, Lactobacillus rhamnosus NCIMB 30121 , Pediococcus pentosaceus HTS (LMG P-22549), Pendiococcus acidilactici NCIMB 30086 and/or Lactobacillus plantarum LSI (NCIMB 30083). In another embodiment the lactic acid bacteria is a Lactobacillus or Lactococcus. In yet a further embodiment the lactic acid bacteria is selected from the group consisting of Lactobacillus plantarum, Lactococcus Lactis, Lactococcus Cremoris, 5 Lactococcus Diacetylactis and Leuconostoc Cremoris. The amount of probiotic bacteria may influence the efficiency of the feed. Thus, in an embodiment the fermented dry food/feed ingredient according to the invention comprises probiotic bacteria in a total amount in the range 10⁶-10¹² CFU per gram, such as 10⁷ - 10¹² CFU/gram, such as 10⁸ - 10¹² CFU/gram, such as 5¹⁰ - 9¹⁰ CFU/gram, or such as 10 7¹⁰ - 9¹⁰ CFU/gram. In the example section feed with CFU's of approximately 10⁷ were used, however this may be adapted to different types of feed.

The terms "fermented product" or "fermented feed" indicate any product or feed that has been fermented or is in the process of being fermented.

15

The term "animal(s)" as used herein is intended to include mammals such as humans, pigs, piglets, cattle, and horses, poultry such as chickens, turkeys, hens, geese and ducks, and fish such as salmon and trout. Monogastric animals, such as humans, pigs, horses, dogs, and cats, have a simple single chambered stomach. 20 In contrast, ruminant animals or ruminants have a 10 multi-chambered complex stomach. Ruminants digests their food in two steps, first by eating the raw material and regurgitating a semi-digested form known as cud, then eating (chewing) the cud, a process called ruminating. Ruminants include for example cattle, goats, sheep and deer.

25

Homofermentation

The fermentation process provided by the starter culture according to the invention is preferably essentially a homofermentative process. "Essentially homofermentative" means, that the predominant bacterial flora driving the

30 fermentation is homofermentative. In one embodiment, 99% or more of the

bacteria are homofermentative. In another embodiment of the invention, 95% or more of the bacteria are homofermentative. In yet another embodiment, 90% or more of the bacteria are homofermentative. "Essentially homofermentative" indicates also that the major fermentation product is lactic acid, and the levels of

35 acetic acid and ethanol are either below taste threshold, around taste threshold or slightly above taste threshold. Alternatively, "essentially homofermentative" indicates a ratio of lactic acid to acetic acid or lactic acid to ethanol (mM/mM) of 10: 1 or more, 20: 1 or more, 50: 1 or more, or 100: 1 or more. As shown in the example section positive effects have been observed when using fermented rapeseed, such as rapeseed meal or rapeseed cake. Thus, in an embodiment the plant product in step a) comprises a rapeseed product, such as rapeseed meal or rapeseed cake, preferably rapeseed cake.

Rape

Rape (or Rapa, oilseed rape, rapa, rappi, rapeseed, canola)

Rape (such as Brassica campestris and B. napus) is a member of the Brassica genus, which include cabbage, radish, kale, mustard and cauliflower. Oilseed rapes were grown in India over 3,000 yr ago, and at least 2,000 yr ago in China and Japan. Rape is believed to have originated in the Mediterranean area. The rapeseed contains about 40% semi-drying oil, which may be used as salad and cooking oil, in the manufacture of margarine and in the production if bio-fuel. Rapeseed meal/cake and canola meal/cake are the by-product of rapeseed and canola oil production. Canola is the name given to rape cultivars with a low glucosinolate content.

Rape may be "double low" / "double zero" / "rapeseed 00". These names all relate to rape cultivars with low content of erucic acid and low content of glucosinolates (an example is canola). Erucic acid may be harmful at high concentrations therefore sorts with low content is normally appreciated ("double low" sorts). "Double low" rapeseed, as defined in here has less than 25 pmol/g (dry weight) of total glucosinolates, such as 1-25 μη-iol/g, such as 3-25 μη-iol/g, such as 10-25 μη-iol/g, or such as 15-25 μη-iol/g.

Rape may also be "single low". The term "single low" rapeseed refers to high glucosinolate rapeseed with low erucic oil. "Single low" rapeseed, as defined in here has at least 25 μη-iol/g (dry weight) of total glucosinolates, such as at least 50 μη-iol/g, such as at least 75 μη-iol/g, such as at least 100 μη-iol/g, such as at least 150 μη-iol/g, such as in the range 35-250 μη-iol/g, such as in the range 50- 150 μη-iol/g, such as in the range 75-150 μη-iol/g, or such as in the range 80-120 μη-iol/g. Rapeseed cake

The term "rapeseed cake" refers to the product obtained after removing most of the oil from the rapeseed by physical pressing, resulting in a rapeseed "cake" having an oil content around 10-12% by weight. Rapeseed "cake" is mainly an intermediate product, since there is an interest to extract the remaining oil, resulting in the by-product rapeseed meal.

Rapeseed meal

Rapeseed meal is obtained by further removal of oils from the rapeseed cake preferably by hexane extraction of the remaining oil in the press-cake. Rapeseed meal has an oil content of around 1% by weight. The hexane treatment and heating or drying also result in the destruction/degradation of glucosinolates. Thus, it is generally acknowledged that "cake" has a higher content of

glucosinolates than "meal".

In relation to the present invention, rape is preferably provided as rape cake. Even more preferably, the rape meal cake is from a "single low" rape which has at least 25 μη-iol/g (dry weight) of total glucosinolates, such as at least 50 μη-iol/g, such as at least 75 μη-iol/g, such as at least 100 μη-iol/g, such as at least 150 μη-iol/g, such as in the range 35-150 μη-iol/g, such as in the range 50-150 μη-iol/g, such as in the range 75-150 μΓηοΙ/g, or such as in the range 80-120 μΓηοΙ/g.

Thus, in an embodiment the rapeseed product is derived from one or more "single low" rapeseeds, such as Bienvenu or Midas. The skilled person will be able to find others "single low" rapeseeds and is well aware of the term. In yet an

embodiment the plant product in step a) comprises rapeseed or rapeseed cake.

As just mentioned "single low" rape is preferred due to their higher content of glucosinolates. Therefore, in an embodiment the plant product in step a) does not comprise rape from "double low" sorts of rape(seed), such as rapeseed meal from double low rapeseeds. As also mentioned above, it is preferred to use rapeseed cake. Thus, in an embodiment the plant product in step a) does not comprise rapeseed meal. In yet an embodiment the ratio between fermented rape (in total) to fermented rapeseed meal (by weight) in the fermented food/feed ingredient is at least 2: 1 such as at least 5: 1 such as at least 10: 1, such as in the range 2: 1 to 10000: 1, such as in the range 5: 1 to 10000: 1, such as in the range 10: 1 to 10000: 1, such as in the range 2: 1 to 1000: 1, such as in the range 2: 1 to 500: 1, such as in the range 2: 1 to 100: 1.

Rapeseed cake has a higher content of oil. Thus, in an embodiment the plant product provided in step a), has an oil content (w/w) in the range 5-20%, such as 8-20%, such as 8-15%. In another embodiment, the oil content in the plant product in step a) is at least 5% (w/w), such as in the range 5-60% such as 5- 40%, such as 5-20% such as 8-20% or such as 8-15%. Rapeseed meal does not comprise such amount of oils. In a preferred embodiment the amounts of oil is rape oil.

Glucosinolates are also present in other plants than rape. Thus, in an embodiment the plant product in step a) comprises one or more plants selected from the group consisting of oilseeds, cabbages, plants of the order Brassicales, such as families Brassicaceae/Cruciferae, Capparidaceae, and Caricaceae, and plants from the genus Drypetes, such as from the family Euphorbiaceae.

From the experiments performed (see example 6), it is shown that fermentation is able to completely remove the glucosinolates from rapeseed. Thus, in yet an embodiment the plant product of step a) has a total glucosinolate content of at least 5 μη-iol/g (dry weight, DW), such as at least 8 μη-iol/g, such as at least 10 μη-iol/g, such as at least 15 μη-iol/g, such as at least 20 μη-iol/g (dry weight, DW), such as at least 25 μη-iol/g such as at least 50 μη-iol/g, such as at least 100 μη-iol/g, such as in the range 20-200 μη-iol/g, such as in the range 20-150 μη-iol/g, such as in the range 20-100 μΓηοΙ/g such as in the range 20-75 μΓηοΙ/g, or such as in the range 20-50 μΓηοΙ/g. In another embodiment the fermented food/feed ingredient provided in step c), has a total glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g.

In some preferred embodiments, - the plant product of step a) has a total glucosinolate content of at least 10 μη-iol/g (dry weight, DW), such as at least 15 μηιοΙ/g, such as at least 20 μη-iol/g (dry weight, DW), such as at least 25 μηιοΙ/g and the fermented food/feed ingredient provided in step c) has a total glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g or such as less than

- the plant product of step a) has a total glucosinolate content of at least 10-200 μη-iol/g (dry weight, DW), such as 25-200 μη-iol/g, such as 50-200 μη-iol/g (dry weight, DW), and the fermented food/feed ingredient provided in step c) has a total glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g or such as less than 0.001 μη-iol/g;

Moisture content

The dry food/feed ingredient may have different moisture contents. Thus, in an embodiment the moisture content of the ingredient is in the range 3-19% such as in the range 3-15%, such as in the range 3-10%, such as in the range 6-10%, such as in the range 8-19%, such as in the range 10-19%, such as in the range 10-15%. Advantages of the dry product are increased shelf life and better preservation of the probiotic bacteria. Examples of dry food/feed products are meal, silage, crumbles, pellets and other agglomerated meals. Such products can be supplied in packages like (big) bags or in bulk.

As described in example 8, the moisture content during fermentation has an influence on how efficient the end-product is as an ingredient. Thus, in a related embodiment the fermentation process is performed at a moisture content in the range 25-60%, such as 30-50%, such as 30-50%, such as 30-40%, such as 40- 60% or such as 50-60%.

The fermentation of course has to run for a certain period to be able to remove all glucosinolates from the plant material. In yet an embodiment the fermentation step c) is continued for a period of at least 1 day, such as at least 2 days, such as at least 4 days, such as 1-20 days, such as 1- 10 days, such as 2-10 days, or such as 2-5 days. To protect the food/feed ingredient for contamination, it may be advantageously that the pH is lowered in the food/feed ingredient. Thus, in a further embodiment the provided fermented food/feed ingredient in step c) has a pH in the range 3.5- 4-2, such as 3.5-4.0, such as 3.5-3.8, such as 3.7-4.2, such as 3.7-4.0, or such as 3.8-4.2. In another embodiment said lactic acid bacteria are

homofermentative. In yet another embodiment, the fermented food/feed ingredient provided in step c) has a lactic acid concentration of at least 50 mM, such as at least 100 mM, such as 100-1000 mM, such as 100-500 mM, such as 100-300 mM, such as 100-200 mM, such as 150-500 mM, such as 200-500 mM or such as 300-500, mM lactic acid.

Food/feed ingredient obtained/obtainable by process

As previously mentioned it is believed that the degradation glucosinolates during fermentation results in degradation products having a desired effect.

Thus, in a further aspect the invention relates to a food/feed ingredient

obtained/obtainable by a process according to the invention as outlined above.

Fermented food/feed ingredient

In yet an aspect the invention relates to a fermented dry food/feed ingredient comprising

· fermented rapeseed in an amount of 5-80% by weight; and

• one or more probiotic bacteria strains in a total amount in the range 10⁵- 10¹² CFU per gram.

In yet another aspect the invention relates to a fermented food/feed ingredient · comprising a fermented plant product comprising one or more of

o fermented plants from the genus Brassica such as oilseeds, rape, such as rapeseed, rapeseed cake, rapeseed meal, cabbages, plants of the order Brassicales, such as families Brassicaceae/Cruciferae, Capparidaceae, and Caricaceae, and plants from the genus

Drypetes, such as from the family Euphorbiaceae;

in an amount of 5-99% by weight;

• having a total glucosinolate content below 1 mol/g (DW); and

• one or more lactic acid bacteria strains in a total amount in the range 10⁵- 10¹² CFU per gram. As mentioned above fermented rape (or parts thereof) are preferred plant material for the ingredients according to the invention. Thus, in an embodiment the fermented plant product is from the genus Brassica such as oilseeds, rape, such as rapeseed, rape seed cake, or rapeseed meal is derived from "single low" rape. In yet an embodiment the fermented plant product comprises rapeseed or rapeseed cake.

As also mentioned above sorts of "double low" rape are less appropriate due to the low glucosinolates content. Therefore, in an embodiment the fermented plant product does not comprise rapeseed from "double low" rapeseeds, such as rapeseed meal from double low rapeseeds.

In yet an embodiment the ratio between fermented rape (in total) to fermented rapeseed meal (by weight) in the fermented food/feed ingredient is at least 2: 1 such as at least 5: 1 such as at least 10: 1, such as in the range 2: 1 to 10000: 1, such as in the range 5: 1 to 10000: 1, such as in the range 10: 1 to 10000: 1, such as in the range 2: 1 to 1000: 1, such as in the range 2: 1 to 500: 1, such as in the range 2: 1 to 100: 1.

Since rapeseed meal also has a low content of glucosinolates (e.g. due to hexane treatment and heating during processing), meal is also less preferred. Thus, in an embodiment the fermented plant product does not comprise rapeseed meal. In an embodiment, the one or more plant materials are present in an amount of 5-80% by weight, such as 20-80%, such as 40-80%, such as 10-60%, such as 20-40%, or such as 20-30%.

In yet an embodiment at least one lactic acid strain is Lactobacillus such as Lactobacillus plantarum. In a further embodiment, the concentration of lactic acid in the fermented food/feed ingredient is at least 100 mM measured on a dry matter basis, such as 100-1000 mM, such as 100-500 mM, such as 100-300 mM, such as 100-200 mM, such as 150-500 mM, such as 200-500 mM or such as 300- 500 mM. In yet a further embodiment the pH of said ingredient is in the range 3.5-4-2, such as 3.5-4.0, such as 3.5-3.8, such as 3.7-4.2, such as 3.7-4.0, or such as 3.8-4.2. In another embodiment, the food/feed ingredient has a protein content above 15% by weight, such as 15-50%.

Other protein sources may be added to the food/feed ingredient. Therefore, in an embodiment the sources of protein is constituted of at least two, such as three at least four different protein sources selected from plant sources, algae sources, meat sources, fish sources, slaughter house waste material sources, feather sources and/or seaweed sources, such as at least five, such as at least six or such as at least seven different protein sources. In a further embodiment the sources of protein is constituted of at least four different plant protein sources selected from the group consisting of mature and/or immature plants and parts thereof, such as cereals, e.g. wheat, barley, rye, rice, maize such as cob maize silage (CCM) or ripe, triticale, oat; vegetables such as potatoes, beans, peas, maize, soy; whey, curd, skim milk and the like.

If desired the food/feed ingredient may be dried. Thus, in an embodiment the ingredient is a dry ingredient. In a related embodiment the dry food/feed ingredient has a moisture content in the range 3-19% (w/w) such as in the range 3-15%, such as in the range 3-10%, such as in the range 6-10%, such as in the range 8-19%, such as in the range 10-19%, such as in the range 10-15%.

As described above, it is a goal of the present invention to ferment a plant product having a relatively high content of glucosinolates to a product with a low content. Thus, in an embodiment the food/feed ingredient has a total glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g, such as less than 0.005.

A food/feed product

In another aspect the invention relates to a food/feed product comprising a food ingredient according to the invention.

The term "product" according to the invention is to be understood in its broadest sense. Commonly, the products are food or feed related. "Product(s)" and "feed product(s)" may suitably be obtained from the dairy industry, the agricultural industry, the wine industry, the spirit industry, or beer industry, or combinations thereof. Examples of suitable "products" and "feed products" comprise one or more of mature and/or immature plants and parts thereof, such cereals, e.g. wheat, barley, rye, rice, maize (cob maize silage (CCM) or ripe), triticale, oat; vegetables (e.g. potatoes, beans, peas, maize, soy; whey, curd, skim milk and the like.

The amount of the ingredient in the food product may vary. In an embodiment the food/feed product comprises 3-15% (w/w) of the of the food/feed ingredient according to the invention, such as 3-10%, such as 4-10%, such as 4-8%, such as 4-6%. In a related embodiment, the fermented food/feed product comprises in the range 50-100% of the food/feed ingredient. In an embodiment the fermented food/feed product comprises in the range 20-100% of the food/feed ingredient, such as in the range 30-100%, such as in the range 40-100%, such as in the range 50-100%, such as in the range 70-100%, such as in the range 80-100%, such as in the range 30-80%, such as in the range 30-60%, such as in the range 30-50%. In a particular embodiment, the fermented food/feed product comprises 100% of the food/feed ingredient. This means that the ingredient is the final product. In yet an embodiment the food/feed product comprises 3-15% by weight of the of the food/feed ingredient according to the invention, such as 5-15%, such as 7.5-12%.

In yet an embodiment the fermented dry food/feed ingredient according to the invention comprises in the range 2-6% by weight lactic acid, such as in the range 2-5.5%, such as 2-4%, or such as 2-3%. In another embodiment the food/feed ingredient according to the invention has a concentration of lactic acid in the fermented dry food/feed ingredient of at least 10 mM, such as at least 50 mM, such as at least 100 mM, such as 100-1000 mM, such as 100-500 mM, such as 100-300 mM, such as 100-200 mM, such as 150-500 mM, such as 200-500 mM or such as 300-500 mM. To produce a food product with rich sources of proteins, the food/feed product preferably comprises at least three sources of protein, such as at least four or such as at least five or such as 5-7 sources of protein. In example 3 recipes with four and five sources of protein are provided. In yet an embodiment the food/feed product has a total glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g, or such as less than 0.005 μη-iol/g. Medicament

As described in the example section the produced feed has a positive effect on pigs. Thus, an aspect of the invention relates to the food/feed product according to the invention, for use as a medicament. In another aspect, the invention relates to the food/feed product according to the invention, for use as a medicament in the treatment and/or prevention of yeast and/or bacterial infections in an animal and/or in humans. In an embodiment, the food/feed product is for use as a medicament in the treatment and/or prevention of Porcin Proliferativ Enteropati and/or Oedema disease. Porcin Proliferativ Enteropati may be caused by a lawsonia infection whereas Oedema disease may be caused by E. coli.

In a related embodiment, the bacterial infection is selected from the group consisting of salmonella infection, E. coli infection and lawsonia infection.

In yet an embodiment the animal is poultry such as chicken. Other animals may also be cow, pig, sheep, cat, dog, turkey. In an embodiment, the animal is a mammal. In a different embodiment, the subject may also be a human. The term "animal(s)" as used herein is intended to include mammals such as pigs, piglets, cattle, and horses, poultry such as chickens, turkeys, hens, geese and ducks, and fish such as salmon and trout. Monogastric animals, such as humans, pigs, horses, dogs, and cats, have a simple single chambered stomach. In contrast, ruminant animals or ruminants have a multi-chambered complex stomach.

Ruminants digests their food in two steps, first by eating the raw material and regurgitating a semi-digested form known as cud, then eating (chewing) the cud, a process called ruminating. Ruminants include for example cattle, goats, sheep and deer.

Process for producing a dry food ingredient

The product may be obtained by a particular process, which provides a high content of probiotic bacteria. See also example 2. Thus, aspect of the invention relates to a process for producing a dry food ingredient comprising a) providing a food/feed product to be fermented;

b) providing one or more lactic acid bacteria strains or one or more

inoculum comprising predominantly lactic acid bacteria;

c) fermenting the food/feed product with the one or more lactic acid

bacteria strains, providing a fermented food/feed ingredient; d) introducing said fermented food/feed ingredient into a dryer chamber; e) contacting said fermented food/feed ingredient with a stream of drying gas in said dryer chamber;

g) providing a dry fermented food/feed ingredient. In an embodiment, the stream of drying gas in said dryer chamber is performed by a spin flash dryer. The effect on CFU obtained by spin flash drying is shown in example 2.

In yet further embodiments the drying chamber is in a configuration comprising a heater, which is capable of heating the said gas, and/or a collector having a discharge valve and optionally being equipped with an exhaust fan.

A conventional configuration comprising a drying chamber as used in the present invention is shown in Figure 7. The configuration comprises drying gas inlet (A), drying chamber (B), and optionally rotor (C), heater (D), collector (E), discharge valve (F), exhaust fan (G), slit-shaped opening in the drying chamber (H), tangentially arranged outlet (J), and snail transporter for transporting the fermented feed into the drying chamber (K). In the method according to the invention, the moisture content of the product obtained in step (d) can be controlled by adjusting the amount of fermented feed product feed to said dryer chamber in step (b) and/or adjusting the drying capacity of said drying gas. In order to obtain sufficient dehydration of the fermented feed product provided in step (a) it may be preferred that the moisture content of the excess drying gas is 0.4 kg moisture per kilogram dry gas or higher, such as 0.5 kg moisture per kilogram dry gas or higher, e.g. 0.6 kg moisture per kilogram dry gas or higher.

Thus, typically the moisture is evaporated at a rate in the range of 600 to 900 kg moisture per hour, such as 650 to 850 kg moisture per hour, e.g. 700 to 800 kg moisture per hour, preferably 725 to 775 kg moisture per hour. Most preferably the moisture is evaporated at a rate at 735 kg/h or around 735 kg/h.

In an embodiment the flow rate of which the drying gas flows through the drying chamber is in the range of 30000 to 40000 m³ drying gas per hour, such as in the range of 31000 to 39000 m³ drying gas per hour, e.g. in the range of 32000 to 38000 m3 drying gas per hour, such as in the range of 33000 to 37000 m3 drying gas per hour, e.g. in the range of 34000 to 36000 m³ drying gas per hour, such as in the range of 35000 to 35500 m³ drying gas per hour, preferably in the range of 33500 to 34500 m³ drying gas per hour. In a preferred embodiment the flow rate of which the drying gas flows through the drying chamber is 34000 m³ drying gas per hour or around 34000 m³ drying gas.

The term drying gas is to be understood in its broadest context, thus the term covers any gas including air applicable for drying. From a financial point of view it may be preferred that the drying gas is air. In a preferred embodiment the drying gas it hot air. Air is mainly composed of nitrogen, oxygen, and argon, which together constitute the major gases of the atmosphere.

Thus, preferably the inlet temperature of the drying gas is in the range of 120 to 250°C, such as 160 to 220°C or 120 to 160°C, such as in the range from 125 to 155°C, such as in the range from 130 to 150°C, e.g. in the range from 135 to 145°C, such as in the range from 140 to 160°C. In a preferred embodiment, the inlet temperature of the drying gas is in the range of 135 to 145°C. In another embodiment, the inlet temperature of the drying gas is in the range 240-270°C such as 251-270°C or such as 251-260°C. Clearly, it is important to use an inlet temperature of the drying gas which simultaneously (i) provides a sufficient dehydration of the fermented feed product provided in step (a) and (ii) retain viable lactic acid bacteria in the dry fermented feed product obtained in step (f).

As the inlet temperature of the drying gas affect the temperature of the dry fermented feed product obtained in step (f) it may be preferred that the

temperature of the product obtained in step (f) is in the range of 30 to 55°C, such as 40 to 55°C or 50 to 55°C. In a preferred embodiment the temperature of the product obtained in step (f) is measured at the outlet of the drying chamber (before cooling).

In yet an embodiment the spin flash drying is performed for a period of 1-15 seconds, such as 3-12 seconds, such as 3-10 seconds, such as 5-10 seconds, such as 7-10, or such as around 10 seconds. In yet an embodiment the

temperature of the input stream of drying gas in step e) has a temperature in the range 120 to 200°C. In the present context, the period defining the spin flash drying refers to the period the fermented product is present in the drying chamber (B). The effect of keeping the temperature in step f) at maximum 55°C is that the viability of the lactic acid bacteria is severely lowered when the temperature exceeds 55°C and in particular at temperatures above 60°C or 65°C. If fast drying is important the temperature at step f) may be e.g. 55-65°C such as 60-65°C. The obtained food/feed ingredient may be mixed or assembled with other ingredients to provide a food/feed product. Thus, in an embodiment the process further comprises mixing the provided dry fermented food/feed ingredient with one or more food ingredients, thereby providing a food product.

In a preferred embodiment the input stream of drying gas in step e) has a temperature in the range 120 to 160°C, the stream of drying gas is in contact with the fermented food/feed for a period of maximum 10 seconds and the provided dry fermented food/feed ingredient in step f) has a maximum

temperature of 55°C.

In another preferred embodiment the input stream of drying gas in step e) has a temperature in the range 200 to 260°C, the stream of drying gas is in contact with the fermented food/feed for a period of maximum 10 seconds and the provided dry fermented food/feed ingredient in step f) has a maximum

temperature of 65°C. In yet an aspect the invention relates to a dry fermented food/feed ingredient obtainable by a process according to the invention.

Dryer chamber and auxiliary elements

Reference is made to fig. 7, which shows schematically and in a 2-dimensional cross sectional view a dryer according to the present invention. The dryer is embodied as a spin flash dryer and comprising a dryer chamber B, formed by cylindrically shaped wall I with a rounded top, inside which drying chamber B the fermented product is dried. The dryer comprising a drying gas inlet A through which gas at elevated temperature is fed into the dryer chamber B. As indicated in fig. 1, the drying gas inlet comprises a tube encircling a distal end of the cylindrically shape wall of the dryer B having a diminishing cross section similar in tangential direction to provide an even inflow of drying gas through the slit-shaped opening H provided at the bottom of the cylindrically shaped wall. The inlet A is furthermore arranged so that the drying gas enters into the drying chamber B with a tangential velocity component so as to generate a swirling flow pattern inside the dryer chamber B.

The dryer further comprises a tangentially arranged outlet J through which the dry fermented product leaves the dryer. Fermented product to be dried is transported by the snail transporter K into the drying chamber B above the inlet A. Fermented product introduced into the drying chamber gets in contact with the drying gas swirling inside the drying chamber. The fermented product is typically particulate material and lumps of fermented product above a certain size and weight will, depending on the swirling velocities, be transported upwardly and to the outlet J, whereas heavier lumps of fermented products will fall towards to the bottom of the drying chamber B.

In the bottom of the drying chamber B, a rotor may be provided. The rotor is formed with vanes supporting the swirling motion of the material and gas inside the drying chamber B and cutting heavier lumps of fermented material into smaller lumps, which then will be transported upwardly due to the swirling motion, which also includes an upwardly going velocity component. The swirling motion will force heavier lumps of material, which are too light to fall to the bottom, upwardly and towards the cylindrically shaped wall of the dryer. To avoid such particles from escaping the drying chamber (as they often are not fully dried due to their relative large sizes), a discriminator L in the form of a downwardly inclined disc is provided below the outlet. The discriminator will force such heavier lumps of material out to the periphery of the drying chamber B where the entrainment velocity is so small that the action of the gravity will transport the lumps of material towards the bottom of the drying chamber and into contact with the rotor, which in turn will cut the lumps into smaller ones. Dried material leaves the drying chamber A together with gas. A collector E optionally in the form of a cyclone is provide at the outlet J and being operated so that dried fermented material is extracted from the process at the bottom of the collector through a discharge valve F. The flow through the collector may be assisted by an exhaust fan G.

The temperature of the drying gas is elevated by use of the heater D which typically is embodied as a heat exchanger.

The dimensioning of the dryer and the auxiliary elements disclosed in figure 7 are made in accordance with the desired capacity of the dryer. Control of the drying process is typically performed by measuring the temperature of the dried fermented product at the outlet J and adjusting the amount of fermented product being fed into the drying chamber to obtain a pre-selected temperature such as maximum 55°C of the dried material leaving the drier; other process parameters, such as air flow, rotational speed of the rotor C is kept constant while the amount of feed of fermented product is adjusted to match the preselected temperature at the outlet. Further, the moisture content in the dried fermented product is advantageously around 10-15 wt % to make it possible for the bacteria to survive in the dried feed. In further embodiments of the invention the dried, fermented product is cooled instantly after exiting the spin flash dryer and therefore prior to storage. To accomplish this, a cooling section (not shown) is provided downstream of the collector E and may utilise a stream of cold air directed towards and into the dried fermented product. Such downstream cooling element may be in the form of a cyclone. However other types of cooling elements may also be used. Preferably the temperature of the air is in the range 5-30°C, such as 10-30°C, such as 15- 30°C, such as 5-25°C, such as 5-20°C, such as 10-25°C, such as 15-25°C.

Preferably a cooling step is performed very quickly such as within 5 seconds to 5 minutes, such as 10 seconds to 3 minutes, such as 30 seconds to 3 minutes.

Thus, in an embodiment the fermented food/feed ingredient exiting the drying chamber is instantly exposed to a cooling gas with a temperature in the range 5- 30°C, such as 5-25°C, such as 10-25°C or such as 15-25°C. The effect of the cooling step is that the temperature of the product exiting the spin flash dryer is lowered to a temperature in the range 5-30°C, such as 15-25°C shortly after exiting the spin flash dryer.

The inventors have discovered that this cooling step is important for at least two reasons. Firstly the CFU of the probiotics present in the product is maintained since the viability of the bacteria is heavily impacted if stored at longer periods above 30°C. Another very important effect of the cooling step is the impact on the stored products e.g. when stored in "big bags". If not instantly cooled, the product will aggregate into a large pile and/or form big clumps. Overall, if not instantly cooled the product will be difficult to handle and it may even be difficult to get it out of the storage material wherein it is stored. Without being bound by theory this aggregation may be due to the protein content of the food/feed.

Furthermore, as explained in example 8, if the cooling step after drying is not included and the fermented product comprises fermented glucosinolates, the cooling step is essential for maintaining the effect of the fermented glucosinolates. In an additional embodiment, the moisture content of the feed product obtained in step (d) is in the range of 10 to 15%. In yet an embodiment the moisture content of the feed provided in step (a) is in the range of 25 to 95%. In another embodiment, the fermented feed product provided in step (a) is provided in the form of a liquid feed or a paste. In yet a further embodiment said fermented feed product provided in step c) is obtained essentially by homo-fermentative or hetero-fermentative fermentation. In another embodiment said fermented feed product provided in step c) and/or g) has a pH of 4.2 or lower, such as in the range 3.5-4-2, such as 3.5-4.0, such as 3.5-3.8, such as 3.7-4.2, such as 3.7-4.0, or such as 3.8-4.2, such as between 4.2 and 3.5, or around 3.8. In yet another embodiment said fermented feed product provided in step (c) and or g) has a lactic acid concentration in the range of 50-100 mM, 100-150 mM, 150-200 mM, 200-250 mM, 250-300 mM, or 300 mM or more. In an embodiment said

fermented feed product provided in step (a) has been fermented at a temperature between 10-50°C, 15-40°C, 18-30°C, 20-25°C, or 22-24°C or around 23°C.

In an further embodiment, the provided fermented food/feed ingredient in step c) and/or step g) has a total amount of lactic acid bacteria in the range 10⁵-10¹² CFU per gram, such as 10⁷-10¹², such as 10⁸-10¹², such as 10⁹-10¹², such as 10¹⁰-10¹² CFU per gram. In a further embodiment, the provided fermented food/feed ingredient in step c) and/or step g) has a protein content in above 15% (w/w), such as 15-50%. In a further embodiment the provided fermented food/feed ingredient in step c) and/or step g) has sources of protein from at least four different protein sources selected from plant sources, algae sources, meat sources, fish sources, slaughter house waste material sources, feather sources and/or seaweed sources, such as at least five, such as at least six or such as at least seven different protein sources.

In an additional embodiment, the provided food/feed ingredient in step c) and/or step g) comprises fermented rapeseed meal and/or fermented rape seed cake. In an embodiment, said fermentation step c) results in a product having within 24 hours:

- a pH of 4.2 or less, such as less that 4 or less than 3.8; and a lactic acid content of at least 100 mM, such as in the range 100-500 mM.

In an embodiment, the inlet temperature of the drying gas is in the range of 120 to 250°C. In a related embodiment the temperature of the product exiting the dryer, but before cooling, is in the range of 30 to 55°C, such as 40 to 55°C or 50 to 55°C.

In another embodiment, the spin flash drying in step e) is performed for a period of 1-15 seconds, such as 3-12 seconds, such as 3-10 seconds, such as 5-10 seconds, such as 7-10 seconds, or such as around 10 seconds. In a related embodiment

In an embodiment the provided food/feed ingredient in step c) and/or step g) has a total glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g and wherein the plant product of step a) has a total glucosinolate content of at least 20 μη-iol/g (dry weight, DW), such as at least 25 μηιοΙ/ such as at least 50 μη-iol/g, such as at least 100 μη-iol/g, such as in the range 20-200 μη-iol/g.

In the example section, the inventors show that a fermented rapeseed meal cake and whole rape feed surprisingly assist in the clearance of a salmonella infection in poultry. This is in contrast to another fermented feed where rapeseed is substituted with soy.

Medicament

As shown in the example section the fermented rapeseed product has beneficial effect compared to control fermented products when it comes to e.g. clearance of salmonella infections. Thus, an aspect of the present invention relates to the food product according to the invention for use as a medicament.

It should be noted that embodiments and features described in the context of one of the aspects of the present invention also apply to the other aspects of the invention. All patent and non-patent references cited in the present application, are hereby incorporated by reference in their entirety. The invention will now be described in further details in the following non-limiting examples.

Examples

Example 1 Effect of fermented rapeseed on salmonella infections in chickens Trial design

A trial was conducted to investigate the effect of a fermented rapeseed diet (FR) and a fermented conventional diet (FC) on salmonella shedding in chickens compared with a control non-fermented pelleted diet (CP).

Fifty one day old White leghorn chicks were randomly allocated to FR, FC or CP diets. Chicks in the FR group received the FC diet for during the first 10 days and thereafter the FR diet. At 7 days of age chicks were infected with 106 CFU Salmonella Typhimurium Sal 1344 nalr by oral gavage. Cloacal swabs were taken from all chicks at 9, 13, 16, 20, 23 and 27 days of age and tested for the presence of S. Typhimurium Sal 1344.

Diets

Conventional diets comprising either rapeseed or soy were fermented. An un- fermented control diet with soy was also included in the trial. Fermentation of the two diets (not the control diet) was done by incubating the diets with

homofermentative lactic acid bacteria {Lactobacillus plantarum) and exposed to spin flash drying. For further details see example 2.

Fermented rapeseed diet (FR)

Content Amount (by weight

Rapeseed 25%

Estimated CFU LAB 100.000 - 10.000.000 / gram Fermented conventional diet (FC)

Content Amount

HP soya 25%

Estimated CFU LAB < 100.000 - 10.000.000 / gram

Study Plan Certified disease free White leghorn eggs were purchased from Valo Germany and eggs incubated at 37.7 °C for 21 days. On hatching 50 chicks were randomly allocated to one of three dietary treatments with three pens per treatment (Figure 1). Each treatment comprised three pens of either 5 or 6 chicks giving a total of 17 chicks for the control and experimental diets FC and 16 chicks for experimental diet FR. The detail of each treatment is given in Figure 2.

• The control diet was in pelleted form

• Treatment FC consisted of a conventional diet of the same formulation as the control diet, fermented, dried and provided in meal form.

• Treatment FR consisted of the same diet as FC for the first 10 days, then in the grower phase chicks were fed on a fermented rapeseed meal diet.

This was also supplied in meal form.

Chicks were fed starter diets from day 1 - day 10, then grower diets from day 10 until the end of the trial.

• The trial was conducted according to a randomized block design with chicks and feed treatments randomly allocated to blocks of 3 pens.

• On day 2 of the trial one chick in treatment T2 died, this was not treatment related. There were no further mortalities.

• Fresh water and feed were supplied daily and chicks were weighed weekly.

• On day 6 cloaca I swabs were taken and tested for the presence of Salmonella.

• On day 7 chicks were infected with 10⁶ colony form ing units (cfu) Salmonella Typhimurium Sal 1344 nalr by oral gavage. On days 9, 13, 16, 20, 23 and 27 cloacal swabs were taken and tested for the presence of S. Typhimurium Sal 1344 by plating onto Brilliant green agar containing 15Mg/ml nalidixic acid, followed by enrichment in Rappaport Vassiliadis broth.

• At the end of the trial chicks were killed by cervical dislocation.

Microbiology

Incidence of Salmonella

Prior to infection with Salmonella Typhimurium Sal 1344 nalr all cloacal swabs were negative for salmonella. Table 1 shows the number of Salmonella negative swabs out of the number of chicks per pen, with all other swabs being positive for Salmonella. Over the course of the trial the number of negative swabs tended to increase. However, there was a good deal of fluctuation and no individual chick was Salmonella negative throughout the trial. This suggests that for some chicks the infection cleared and they became re-infected from pen mates.

Table 1 Salmonella negative swabs (negative/total chicks per pen)

Day post pen Control FC FR

2 1 0/6 0/5 0/5

2 0/6 1/6 0/4

3 0/5 1/6 0/6

6 1 0/6 0/5 1/5

2 0/6 0/6 1/4

3 0/5 1/6 0/6

9 1 0/6 0/5 2/5

2 1/6 0/6 0/4

3 0/5 1/6 0/4

13 1 0/6 0/5 2/5

2 0/6 1/6 0/4

3 0/5 1/6 3/6

16 1 0/6 0/5 2/5

2 1/6 2/6 1/4

3 1/5 2/6 1/6

20 1 0/6 1/5 2/5

2 0/6 0/6 0/4

3 0/6 2/6 4/6 As the number of chicks per pen varied the number of positive swabs was expressed as a % of the total. On any one swabbing event there were no significant differences between feed treatments in the number of salmonella positive swabs. However, when the overall number of % positive swabs was considered chicks fed experimental diet FR had significantly fewer positive swabs than chicks fed the control diet (Table 2, Figure 6).

Table 2 Percentage of Salmonella positive swabs

Conclusion

The chicks fed with the fermented rape feed showed the highest reduction in the number of Salmonella positive swabs when compared to a non-fermented control diet and a fermented control diet.

Furthermore, in the contents of the ileum there were significantly higher numbers of lactic acid bacteria (LAB) in chickens fed the fermented diets compared to the control diet. However, there was no significant difference in the number of LAB between the two fermented diets. Without being bound by theory, this indicates that the drop in salmonella infections is not only due to the number of LAB's but is due to the combination of LAB and rapeseed. It is noted that such drop is not seen when unfermented rapeseed is used. Again, without being bound by theory the effect may be due to products generated by the fermentation of the rapeseed, such as degradation or modification of glycosylates.

Example 2

CFU of dry fermented rape feed following spin flash drying

Methods Fermented feed comprising large amounts of lactic acid producing bacteria was obtained by providing an inoculum comprising lactic acid producing bacteria and mixing the inoculum with a feed material (including rape) and fermenting the mixture to obtain a fermented feed (liquid feed). Samples of the fermented feed were subjected spin flash drying and conventional drying in the form of toasting, respectively.

Results

The counts of viable bacteria in the feed were measured. The data are presented in table 1 below:

Conclusion

The above data demonstrate that lactic acid producing bacteria in fermented feed are sensitive to the drying method applied to produce dry fermented feed from (semi)liquid feed. The data surprisingly demonstrate that spin flash drying may successfully be applied in order to obtain a dry fermented feed with a high count of viable lactic acids producing bacteria. Example 3

Positive effects of shifting to a fermented rapeseed feed.

The farmer observed the following changes after shifting to a fermented rapeseed feed :

- The mortality among pigs was reduced by approximately 50%;

- The expenses to medicine dropped by 70-80%;

- A drop in the number of pigs with diarrhoea. Corresponding observations have been reported from farmers who were selected to test the product.

Example 4

Recipes of feeds comprising fermented rapeseed meal and or rapeseed cake.

Optimized for pigs (10-20 kg)

Ingredient Amount by weight

Spring Barley 10 %

Wheat 55.8%

soybean meal 8.5%

Fermented rapeseed meal or cake 12%

Potato protein 2.5%

Fat 3.3%

Landmix (mineral concentrate) 7.9%

Total 100%

Optimized for pigs (6.5-10 kg)

Ingredient Amount by weight

Wheat 59%

Fermented rapeseed meal or cake 8%

Fishmeal 5%

Fat 5%

Landmix (mineral concentrate) 23%

Total 100% Example 5

Comparison of products with or without a cooling step included. Methods

Fermented products according to the invention were produced and dried in a spin flash drier as described previously.

Example 4 provides examples of feed compositions comprising fermented rapeseed (meal or cake).

Results

Product without a final cooling step:

These products had a low CFU relative to the desired levels. In addition, after storage in a big bag the products were agglomerated making it very difficult to handle.

The effects described in examples 3 and 7 could not be repeated if the cooling step was omitted. Products with final cooling step included :

These products had an acceptable CFU relative to the desired levels (around 10⁹ CFU per gram). In addition, after storage in a big bag the products were still in a powder form making it easy to handle and measure. The effects described in examples 3 and 7 required the cooling step.

Conclusion

Products having the desired number of CFU and the effect described in example 3 and 7 can only be obtained if the cooling step is included.

In addition, the agglomeration results in long term heat which may degrade the beneficial products present in the fermented product.

Example 6

Amount of glucosinolates in rape before and after fermentation Methods

Rapeseed was fermented for a period of around 4-5 using an inoculum comprising lactic acid bacteria.

Before and after fermentation the amount glucosinolates where determined Results

The results provided in the table below show that by fermentation of the rapeseed it is possible to reduce the amount of glucosinolates from around 9 Mmol/gram to practically 0.

LOD= Below limit of detection Conclusion

Fermentation is surprisingly able to remove practically all glucosinolates from rape.

Example 7

The effect of fermented rapeseed cake and fermented rapeseed meal has been tested in a feed for pigs.

Results

Farm experiments - relative low microbial pressure Comparative studies showed that the same effect could be obtained using a feed comprising only 5% (w/w) fermented rapeseed cake as compared to a feed comprising 7% (w/w) fermented rapeseed meal. Farm experiments - relative high microbial pressure

Comparative studies showed that the same effect could be obtained using a feed comprising only 8-10% (w/w) fermented rapeseed cake as compared to a feed comprising 15%(w/w) fermented rapeseed meal. The above results are based on

- The mortality among pigs;

- The expenses to medicine; and

Drop in the number of pigs with diarrhoea. Conclusion

Without being bound by theory, it is believed that the higher amounts of glucosinolates in rapeseed cake (before fermentation) is responsible for the above mentioned effects can be obtained (compared to "meal") with a lower amount of added rapeseed cake in the complete feed. As previously mentioned, it is believed that the fermentation process results in the production of "healthy" degradation products from the glucosinolates, while the unhealthy glucosinolates are removed. Since the rapeseed cake has a higher amount of glucosinolates than the rapeseed meal, it is preferred to use the "cake". Thus, a reduction of almost 30% of the fermented rapeseed in a complete feed is obtainable by using fermented "cake" instead of meal.

It is to be understood that different pig stable have different microbial pressure in relation to bacterial and yeast infection in pigs. Thus, in some stables a higher amount of fermented rapeseed was required to withstand bacterial and yeast infections in pigs

It is also noted that this effect cannot be obtained if the cooling step during drying is not included, further supporting the importance of this step. However, the skilled person may be able to find alternative drying and cooling steps when knowing the content of this application. Example 8

Fermented rapeseed cake was produced with fermentation under different moisture conditions, 25%, 30%, 50% and 70%.

Results

The effect of the fermented rapeseed (as described in example 6) was only obtainable when the fermented rapeseed was produced at a moisture content of 30% and 50%, whereas the effect could not be obtained with 25% and 70%.

Conclusion

The moisture content during fermentation is important when trying to get the desired effect.

Claims

1. A process for producing a fermented food/feed ingredient, said process comprising

a) providing a plant product to be fermented having a total glucosinolate content of at least 3 μη-iol/g (dry weight, DW);

d) optionally,

o introducing said fermented food/feed ingredient into a drying

chamber;

o cooling the dried fermented food/feed ingredient, exiting the drying chamber, by instantly exposing the dried fermented food/feed ingredient to a cooling gas with a temperature in the range 5-30°C, such as 5-25°C, such as 10-25°C or such as 15-25°C; and providing a dry fermented food/feed ingredient.

2. The process according to claim 1, wherein the plant product in step a) comprises a rapeseed product, such as rapeseed meal or rapeseed cake, preferably rapeseed cake.

3. The process according to claim 2, wherein the rapeseed product is derived from one or more "single low" rapeseeds, such as Bienvenu or Midas.

4. The process according to claim 1, wherein the plant product in step a) comprises one or more plants selected from the group consisting of oilseeds, cabbages, plants of the order Brassicales, such as families

Brassicaceae/Cruciferae, Capparidaceae, and Caricaceae, and plants from the genus Drypetes, such as from the family Euphorbiaceae.

5. The process according to any of the preceding claims, wherein the plant product in step a) comprises rapeseed or rapeseed cake.

6. The process according to any of the preceding claims, wherein the plant

5 product in step a) does not comprise rapeseed from "double low" rapeseed, such as rapeseed meal from double low rapeseeds.

7. The process according to any of the preceding claims, wherein the plant product in step a) does not comprise rapeseed meal.

10

8. The process according to any of the preceding claims, wherein plant product provided in step a) and/or c), has an oil content (w/w) in the range 5-20%, such as 8-20%, such as 8-15%, preferably derived from rape.

15 9. The process according to any of the preceding claims, wherein the fermentation process is performed at a moisture content in the range 25-60%, such as 30- 50%, such as 30-50%, such as 30-40%, such as 40-60% or such as 50-60%.

10. The process according to any of the preceding claims, wherein the plant 20 product of step a) has a total glucosinolate content of at least 5 μη-iol/g (dry

weight, DW), such as at least 8 μη-iol/g, such as at least 10 μη-iol/g, such as at least 15 μη-iol/g, such as at least 20 μη-iol/g (dry weight, DW), such as at least 25 μηιοΙ/ such as at least 50 μη-iol/g, such as at least 100 μη-iol/g, such as in the range 20-200 μη-iol/g, such as in the range 20-150 μη-iol/g, such as in the range 25 20-100 μΓΤΐοΙ/g such as in the range 20-75 μΓηοΙ/g, or such as in the range 20-50

11. The process according to any of the preceding claims, wherein the fermented food/feed ingredient provided in step c), has a total glucosinolate content of less

30 than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g, such as less than 0.005 μη-iol/g, or such as less than 0.001 μη-iol/g.

12. The process according to any of the preceding claims, wherein the

fermentation step c) is continued for a period of at least 1 day, such as at least 2 days, such as at least 4 days, such as 1-20 days, such as 1-10 days, such as 2-10 days, or such as 2-5 days.

13. The process according to any of the preceding claims, wherein the provided fermented food/feed ingredient in step c) has a pH in the range 3.5-4-2, such as

3.5-4.0, such as 3.5-3.8, such as 3.7-4.2, such as 3.7-4.0, or such as 3.8-4.2.

14. The process according to any of the preceding claims wherein said lactic acid bacteria are homofermentative.

15. The process according to any of the preceding claims, wherein the ratio between fermented rape (in total) to fermented rapeseed meal (by weight) in the fermented food/feed ingredient is at least 2: 1 such as at least 5: 1 such as at least 10: 1, such as in the range 2: 1 to 10000: 1, such as in the range 5: 1 to 10000: 1, such as in the range 10: 1 to 10000: 1, such as in the range 2: 1 to 1000: 1, such as in the range 2: 1 to 500: 1, or such as in the range 2: 1 to 100: 1.

16. The process according to any of the preceding claims, wherein the oil content in the plant product in step a) is at least 5% (w/w), such as in the range 5-60% such as 5-40%, such as 5-20% such as 8-20% or such as 8-15%.

17. A food/feed ingredient obtained/obtainable by a process according to any of claims 1-16.

18. A fermented food/feed ingredient

• comprising a fermented plant product comprising one or more of

o plants from the genus Brassica such as oilseeds, rape, such as

Capparidaceae, and Caricaceae, and plants from the genus

Drypetes, such as from the family Euphorbiaceae;

• having a total glucosinolate content below 1 mol/g (DW); and

19. The fermented food/feed ingredient according to claim 18, wherein the fermented plant product is from the genus Brassica such as oilseeds, rape, such as rapeseed, rape seed cake, or rapeseed meal is derived from "single low" rape.

5 20. The fermented food/feed ingredient according to claim 18 or 19, wherein the fermented plant product comprises rapeseed cake.

21. The fermented food/feed ingredient according to any of claims 18-20, wherein the fermented plant product does not comprise rapeseed from double low

10 rapeseeds, such as rapeseed meal from double low rapeseeds.

22. The fermented food/feed ingredient according to any of claims 18-21, wherein the fermented plant product does not comprise rapeseed meal.

15 23. The fermented food/feed ingredient according to any of claims 18-22, wherein the one or more fermented plant materials are present in an amount of 5-80% by weight, such as 20-80%, such as 40-80%, such as 10-60%, such as 20-40%, or such as 20-30%.

20 24. The fermented food/feed ingredient according to any of claims 18-23, wherein at least one lactic acid strain is Lactobacillus such as Lactobacillus plantarum.

25. The fermented food/feed ingredient according any of claims 18-24, wherein the concentration of lactic acid in the fermented food/feed ingredient is at least

25 100 mM measured on a dry matter basis, such as 100-1000 mM, such as 100-500 mM, such as 100-300 mM, such as 100-200 mM, such as 150-500 mM, such as 200-500 mM or such as 300-500 mM.

26. The fermented food/feed ingredient according to any of claims 18-25, wherein 30 the pH of said ingredient is in the range 3.5-4-2, such as 3.5-4.0, such as 3.5-3.8, such as 3.7-4.2, such as 3.7-4.0, or such as 3.8-4.2.

27. The fermented food/feed ingredient according to any of claims 18-26, having a protein content above 15% by weight, such as 15-50%.

35

28. The fermented food/feed ingredient according to any of claims 18-27, wherein the sources of protein is constituted of at least two, such as three at least four different protein sources selected from plant sources, algae sources, meat sources, fish sources, slaughter house waste material sources, feather sources and/or seaweed sources, such as at least five, such as at least six or such as at least seven different protein sources.

29. The fermented food/feed ingredient according to any of claims 18-28, wherein the sources of protein is constituted of at least four different plant protein sources selected from the group consisting of mature and/or immature plants and parts thereof, such as cereals, e.g. wheat, barley, rye, rice, maize such as cob maize silage (CCM) or ripe, triticale, oat; vegetables such as potatoes, beans, peas, maize, soy; whey, curd, skim milk and the like.

30. The fermented food/feed ingredient according to any of claims 18-29, wherein the ingredient is a dry ingredient.

31. The fermented food/feed ingredient according to any of the preceding claims, wherein the ratio between fermented rape (in total) to fermented rapeseed meal (by weight) in the fermented food/feed ingredient is at least 2: 1 such as at least 5: 1 such as at least 10: 1, such as in the range 2: 1 to 10000: 1, such as in the range 5: 1 to 10000: 1, such as in the range 10: 1 to 10000: 1, such as in the range 2: 1 to 1000: 1, such as in the range 2: 1 to 500: 1, or such as in the range 2: 1 to 100: 1.

32. The fermented food/feed ingredient according to any of claims 18-31, having a total glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g, or such as less than 0.005 μη-iol/g.

33. A food/feed product comprising a fermented food ingredient according to any of claims 18-31.

34. The food/feed product according to claim 33, comprising 3-15% (w/w) of the of the food/feed ingredient according to any of claims 12-24, such as 3-10%, such as 4-10%, such as 4-8%, such as 4-6%.

35. The food/feed product according to claim 33 or 34, having a total

glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g.

36. The food/feed product according to any of claims 33-35 for use as a medicament.

37. The food/feed product according to any of claims 33-36 for use as a medicament in the treatment and/or prevention of yeast and/or bacterial infections in an animal and/or in humans.

38. The food/feed product according to any of claims 33-37 for use as a medicament in the treatment and/or prevention of Porcin Proliferativ Enteropati and/or Oedema disease.

39. The food/feed product according to claim 38, wherein the bacterial infection is selected from the group consisting of salmonella infection, E. coli infection and lawsonia infection.

40. A process for producing a dry food ingredient comprising

a) providing a food/feed product to be fermented;

b) providing one or more lactic acid bacteria strains or one or more

inoculum comprising predominantly lactic acid bacteria;

c) fermenting the food/feed product with the one or more lactic acid

cooling the fermented food/feed ingredient exiting the drying chamber by instantly exposing the dried fermented food/feed ingredient to a cooling gas with a temperature in the range 5-30°C, such as 5-25°C, such as 10-25°C or such as 15-25°C; and

g) providing a dry fermented food/feed ingredient.

41. The process according to claim 40, wherein the provided fermented food/feed ingredient in step c) and/or step g) has a total amount of lactic acid bacteria in the range 10⁵-10¹² CFU per gram, such as 10⁷-10¹², such as 10⁸-10¹², such as 10⁹-10¹², such as 10¹⁰-10¹² CFU per gram.

42. The process according to claim 40 or 41, wherein the provided fermented food/feed ingredient in step c) and/or step g) has a lactic acid concentration of at least 100 mM, such as 100-1000 mM, such as 100-500 mM, such as 100-300 mM, such as 100-200 mM, such as 150-500 mM, such as 200-500 mM or such as 300- 500, mM.

43. The process according to any of claims 40-42, wherein the provided

fermented food/feed ingredient in step c) and/or step g) has a pH in the range 3.5-4-2, such as 3.5-4.0, such as 3.5-3.8, such as 3.7-4.2, such as 3.7-4.0, or such as 3.8-4.2.

44. The process according to any of claims 40-43, wherein the provided

fermented food/feed ingredient in step c) and/or step g) has a protein content in above 15% (w/w), such as 15-50%.

45. The process according to any of claims 40-44, wherein the provided

fermented food/feed ingredient in step c) and/or step g) has sources of protein from at least four different protein sources selected from plant sources, algae sources, meat sources, fish sources, slaughter house waste material sources, feather sources and/or seaweed sources, such as at least five, such as at least six or such as at least seven different protein sources.

46. The process according to any of claims 40-45, wherein the provided food/feed ingredient in step c) and/or step g) comprises fermented rapeseed meal and/or fermented rape seed cake.

47. The process according to any of claims 40-46, wherein said fermentation step c) results in a product having within 24 hours:

48. The process according to any of claims 40-47, wherein the inlet temperature of the drying gas is in the range of 120 to 250°C.

49. The process according to any of claims 40-48, wherein the temperature of the product exiting the dryer, but before cooling, is in the range of 30 to 55°C, such as 40 to 55°C or 50 to 55°C.

50. The process according to any of claims 40-49, wherein the spin flash drying in step e) is performed for a period of 1-15 seconds, such as 3-12 seconds, such as

3-10 seconds, such as 5-10 seconds, such as 7-10 seconds, or such as around 10 seconds.

51. The process according to any of claims 40-50, wherein the dryer is a spin flash dryer.

52. The process according to any of claims 40-51, wherein the provided food/feed ingredient in step c) and/or step g) has a total glucosinolate content of less than 0.5 μη-iol/g (DW), such as less than 0.01 μη-iol/g, such as 0.005 μη-iol/g and wherein the plant product of step a) has a total glucosinolate content of at least 20 μη-iol/g (dry weight, DW), such as at least 25 μηιοΙ/ such as at least 50 μη-iol/g, such as at least 100 μη-iol/g, such as in the range 20-200 μη-iol/g.

53. A product obtained/obtainable by a process according to any of claims 40-52.