Unique blends of plant-derived components providing support for piglets during post-weaning feeding period

Abstract

A plant-derived composition having an essential oil core and a herbal powder is used to feed weaned piglets. PW piglets provided with a basal feed supplemented with these plant-derived compositions have improved growth performance. The essential oil core comprises three essential oils.

Description

[0001] Cross-references to related applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 518,692, filed August 10, 2023, which is incorporated herein by reference in its entirety. Technical Field

[0003] This disclosure relates to feeding piglets, and more specifically to plant-derived compositions provided to weaned piglets. Background Technology

[0004] Post-weaning diarrhea (PWD) is a problem faced by pig farmers worldwide, characterized by diarrhea, dehydration, death, and stunted growth in surviving pigs. The impact ranges from a slight decrease in growth performance to severe weight loss and death. The pathogen commonly associated with PWD is the enterotoxin-producing pathogenic strain of *Escherichia coli*.

[0005] Many factors, including livestock farming, environment, nutrition, and genetic susceptibility, play a key role in the severity of this syndrome. Therefore, control of PWD typically begins at different levels: livestock practices with appropriate hygiene and vaccination programs on sow farms and nurseries; and reducing post-weaning stress by providing newly weaned piglets with fresh and readily available feed and water, as well as a suitable environment (temperature and humidity) and space.

[0006] Nutritional program management is also a crucial aspect of controlling PWD. Conventional swine farming systems rely on feeds containing antibiotics, pharmacological levels of zinc, fiber, highly digestible protein, spray-dried plasma, dietary acidification, and functional feed additives to help weaned pigs overcome post-weaning challenges. Due to the development of bacterial resistance to clinically important antibiotics and increasing concerns about antibiotic accumulation in manure and the environment, antibiotics and other antimicrobial growth promoters in feed, such as zinc oxide, are being phased out globally. Therefore, the combination of regulatory requirements and increasing consumer pressure for more “natural” solutions makes finding alternatives to traditional antimicrobial growth promoters (AGPs) essential for maintaining efficient swine production in the future. Plant-derived bioactive compounds (hereinafter referred to as plant-derived) are particularly promising in this regard. Summary of the Invention

[0007] PWD is a syndrome that affects piglets and has a significant impact on the welfare, growth performance, and cost-effectiveness of pig farming. The present specification relates to a unique blend of plant-derived components that provide support to post-weaning (PW) piglets. The plant-derived components specifically interact with the gut and its microbiome, targeting microorganisms to reduce pathogenicity and increase intestinal barrier integrity. The ability of bacteria such as pathogenic Escherichia coli (E. coli), a most common pathogen associated with PWD, to express virulence factors or invade the intestinal epithelium is affected. Successful prevention and / or mitigation of pathogenicity supports antibiotic-free management of growth performance, animal welfare, and livestock production, whether in terms of use as a growth promoter or in terms of reducing the need for medication to treat disease.

[0008] The present specification relates to a modern livestock production method that provides support to piglets during the critical PW period. Thus, the methods and compositions described herein are part of a modern health and nutrition management strategy aimed at providing support to piglets during the critical weaning period.

[0009] In one aspect, the present disclosure provides a method of feeding a livestock animal. The method includes providing a post-weaned piglet with a basal feed supplemented with a plant-derived composition, wherein the plant-derived composition comprises an essential oil core, an herbal powder, and preferably a filler. The essential oil core comprises an essential oil, wherein the essential oil comprises carvacrol, eugenol, and star anise oil, wherein the PW piglet has improved growth performance relative to a PW piglet provided with a basal feed that does not contain the plant-derived composition.

[0010] The plant-derived composition preferably comprises carvacrol at 20% to 60% by weight of the essential oil mixture. The plant-derived composition more preferably comprises carvacrol at 40% by weight of the essential oil. The plant-derived composition preferably comprises eugenol at 10% to 50% by weight of the essential oil. The plant-derived composition more preferably comprises eugenol at about 40% of the essential oil. The plant-derived composition preferably comprises star anise oil at 10% to 30% by weight of the essential oil. The plant-derived composition more preferably comprises star anise oil at about 20% of the essential oil. One or more of the essential oils are microencapsulated, wherein the microencapsulated oil comprises the essential oil and an encapsulation matrix. The filler in the plant-derived composition can be limestone.

[0011] The basal feed is preferably supplemented with carvacrol at a dose of 10 ppm to 50 ppm, eugenol at a dose of 10 ppm to 50 ppm, and star anise oil at a dose of 5 ppm to 50 ppm. The basal feed is more preferably supplemented with carvacrol at a dose of about 20 ppm, eugenol at a dose of about 20 ppm, and star anise oil at a dose of about 9 ppm in the basal feed.

[0012] The herbal powder in the plant-derived composition preferably comprises fenugreek seed powder, more preferably fenugreek seed powder, licorice powder, and / or turmeric powder. The dosage of fenugreek seed powder in the basal feed is preferably from 200 ppm to 1000 ppm, more preferably about 660 ppm. The dosage of turmeric powder in the basal feed is 5 ppm to 50 ppm, more preferably about 25 ppm. The dosage of licorice powder in the basal feed is preferably from 1 ppm to 50 ppm, more preferably about 14 ppm.

[0013] The basal feed in this method is preferably supplemented with 0.25 kg / t to 2.5 kg / t of plant-derived composition, more preferably with about 1 kg / t of plant-derived composition.

[0014] The method preferably provides the plant-derived composition to PW piglets from day 18 to day 75, more preferably from day 20 to day 75, and even more preferably from day 21 to day 70 and any range therebetween. The method provides the plant-derived composition to healthy PW piglets and / or PW piglets infected with pathogenic microorganisms. The method preferably improves growth performance, wherein growth performance preferably includes improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG), and / or improved fecal score.

[0015] In another aspect, this disclosure also provides PW piglet feed. The PW piglet feed comprises a basal feed and a plant-derived composition, wherein the plant-derived composition comprises an essential oil core and herbal powder, wherein the essential oil core comprises essential oils, wherein the essential oils include carvacrol oil, eugenol, and star anise oil, wherein the PW piglet feed contains the plant-derived composition at a content rate of 0.25 kg / t to 2.5 kg / t, and wherein the PW piglet feed contains carvacrol at a dosage of 10 ppm to 50 ppm, eugenol at a dosage of 10 ppm to 50 ppm, and star anise oil at a dosage of 5 ppm to 50 ppm, wherein the PW piglet feed improves growth performance compared to PW piglet feed without the plant-derived composition. Preferably, the PW piglet feed contains the plant-derived composition at a content rate of 1 kg / t, and wherein the PW piglet feed contains 20 ppm carvacrol, 20 ppm eugenol, and 10 ppm star anise oil. PW piglet feed preferably further comprises fenugreek seed powder, more preferably fenugreek seed powder, turmeric powder, and licorice powder. PW piglet feed preferably further comprises 200 ppm to 1000 ppm of fenugreek seed powder, 5 ppm to 50 ppm of turmeric powder, and 1 ppm to 50 ppm of licorice powder. PW piglet feed more preferably comprises about 660 ppm of fenugreek seed powder, 25 ppm of turmeric powder, and 14 ppm of licorice powder. PW piglet feed preferably comprises a basal feed supplemented with about 1 kg / t of the plant-derived composition. Preferably, PW piglet feed is provided to PW piglets from day 18 to day 70 and anywhere in between. This method provides the plant-derived composition to healthy PW piglets and / or PW piglets infected with pathogenic microorganisms. PW piglet feed preferably provides improved growth performance, including improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG), and / or improved fecal score. One or more of the essential oils are microencapsulated, wherein the microencapsulated oil comprises the essential oil and an encapsulating matrix. The plant-derived composition includes a filler, preferably limestone.

[0016] In another aspect, this disclosure also provides a plant-derived composition comprising an essential oil core and an herbal powder, wherein the essential oil core comprises an essential oil, wherein the essential oil comprises 30% to 50% by weight of carvacrol, 30% to 50% by weight of eugenol, and 10% to 30% by weight of star anise oil, and wherein the herbal powder comprises fenugreek seed powder, turmeric powder, and / or licorice powder. Preferably, the essential oil comprises 40% by weight of carvacrol, more preferably, the essential oil comprises 40% by weight of carvacrol and 40% by weight of eugenol, and even more preferably, the essential oil comprises 40% by weight of carvacrol, 40% by weight of eugenol, and 20% by weight of star anise oil. The plant-derived composition preferably comprises about 66% by weight of fenugreek seed powder, about 2.4% to about 2.5% by weight of turmeric powder, and about 1.4% to about 1.5% by weight of licorice powder. The plant-derived composition is preferably used in a basal feed at a rate of 0.25 kg / t to 2.5 kg / t, more preferably at a rate of 1 kg / t. The plant-derived composition is preferably added to PW piglet feed at a dosage of 10 ppm to 50 ppm of carvacrol, 10 ppm to 50 ppm of eugenol, and 5 ppm to 50 ppm of star anise oil, more preferably at a dosage of about 20 ppm of carvacrol, about 20 ppm of eugenol, and about 10 ppm of star anise oil. The plant-derived composition is preferably added to PW piglet feed at a dosage of 200 ppm to 1000 ppm of fenugreek seed powder, more preferably at a dosage of 660 ppm. The plant-derived composition is preferably added to PW piglet feed at a dosage of 5 ppm to 50 ppm turmeric powder, more preferably at a dosage of 25 ppm. The plant-derived composition is preferably added to PW piglet feed at a dosage of 1 ppm to 50 ppm licorice powder, more preferably at a dosage of 14 ppm licorice powder. The plant-derived composition is preferably added to PW piglet feed at a dosage of 660 ppm fenugreek seed powder, 25 ppm turmeric powder, 14 ppm licorice powder, 20 ppm carvacrol, 20 ppm eugenol, and 10 ppm star anise oil. One or more of the essential oils are microencapsulated oils, wherein the microencapsulated oils comprise essential oils and an encapsulating matrix. The plant-derived composition also comprises a filler, preferably limestone. The plant-derived composition preferably comprises about 4% to 5% by weight of the composition, more preferably about 4.8% to 5% by weight of the composition of essential oils. The plant-derived composition preferably contains about 65% to 70% by weight, more preferably about 67% by weight, of herbal powder.

[0017] In another aspect, this disclosure provides a method for alleviating symptoms in piglets with pyeloneps (PW) caused by PWD-associated microorganisms. The method includes providing the PW piglets with a basal diet supplemented with a plant-derived composition, wherein the plant-derived composition comprises an essential oil core, herbal powder, and preferred fillers, wherein the essential oil core comprises essential oils containing carvacrol, eugenol, and star anise oil. The PW piglets exhibit a lower incidence of diarrhea and / or reduced fecal excretion compared to PW piglets fed a basal diet without the plant-derived composition. The PWD-associated microorganisms can be pathogenic Escherichia coli. The Escherichia coli can be enterotoxigenic Escherichia coli (ETEC). Attached Figure Description

[0018] This patent or application contains at least one color-drawn drawing. A copy of this patent or patent application publication with a color drawing will be provided by the Patent Office upon request and payment of the necessary fees.

[0019] The accompanying figures illustrate, in a manner that is not restrictive, various aspects described herein.

[0020] Figure 1A This is a graph showing the effect of carvacrol on biofilm formation in Escherichia coli strains O143:H4 and O88:H8 and on violacein production in C. violaceum. (A) Results after 18 h incubation are expressed as mean ± standard deviation of absorbance values, normalized relative to the positive control (growth without the test substance).

[0021] Figure 1B This is a graph showing the effect of PFA core 1 on biofilm formation of Escherichia coli strains O143:H4 and O88:H8 and on violacein production of Chromobacterium violaceum. (A) Results after 18 h incubation are expressed as mean ± standard deviation of absorbance values, normalized relative to the positive control (growth without test substance).

[0022] Figure 1C This is a graph showing the effect of PFA core 2 on biofilm formation of Escherichia coli strains O143:H4 and O88:H8 and on violacein production of Chromobacterium violaceum. (A) Results after 18 h incubation are expressed as mean ± standard deviation of absorbance values, normalized relative to the positive control (growth without test substance).

[0023] Figure 1D This is a graph showing the effect of tea tree oil on biofilm formation of Escherichia coli strains O143:H4 and O88:H8 and on violacein production of Chlorobacterium violaceum. (A) Results after 18 h incubation are expressed as mean ± standard deviation of absorbance values, normalized relative to the positive control (growth without test substance).

[0024] Figure 1E This is a graph showing the effect of ZnO on biofilm formation of Escherichia coli strains O143:H4 and O88:H8 and on violacein production of Chromobacterium violaceum. (A) Results after 18 h incubation are expressed as mean ± standard deviation of absorbance values, normalized relative to the positive control (growth without test substance).

[0025] Figure 2 This is a graph of mucus-bound F4+ Escherichia coli carrying bacilli, expressed as a percentage of the introduced labeled bacteria.

[0026] Figure 3 This is a schematic diagram of an exemplary formulation of PFA1.

[0027] Figure 4 This is a schematic diagram of an exemplary formulation of PFA2.

[0028] Figure 5 This is a schematic diagram of another exemplary formulation of PFA2. Detailed Implementation

[0029] Reference will now be made specifically to certain aspects of the subject matter disclosed herein, examples of which are partially illustrated in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it should be understood that the illustrated subject matter is not intended to limit the claims to the disclosed subject matter.

[0030] In this document, unless the context clearly requires otherwise, the terms "an," "a," or "the" are used to include one or more. Unless otherwise indicated, the term "or" is used to mean a non-exclusive "or." All publications, patents, and patent documents cited in this document are incorporated herein by reference in their entirety as if individually cited. In the event of any inconsistency between the usage in this document and those documents so incorporated by reference, the usage in the incorporated references shall be considered supplementary to the usage in this document; in the case of irreconcilable inconsistencies, the usage in this document shall prevail.

[0031] Values ​​expressed in range format should be interpreted flexibly to include not only the values ​​explicitly listed as limits of the range, but also all individual values ​​or subranges covered within the range, as if each value and subrange were explicitly listed. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" should be interpreted to include not only about 0.1% to about 5%, but also individual values ​​(e.g., 1%, 2%, 3%, and 4%) and subranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the specified range. Unless otherwise stated, the statement "about X to Y" has the same meaning as "about X to about Y". Similarly, unless otherwise stated, the statement "about X, Y, or about Z" has the same meaning as "about X, about Y, or about Z".

[0032] Unless otherwise specified, ppm (parts per million), percentages, and ratios are based on weight. Percentages based on weight are also referred to below as weight% or (weight)%.

[0033] This disclosure relates to a method of feeding piglets during the post-weaning (PW) period by providing a basal diet supplemented with a plant-derived composition. The plant-derived composition is provided to PW piglets during the post-weaning phase to reduce post-weaning diarrhea (PWD) and improve growth performance. The plant-derived composition is provided to healthy PW piglets and / or PW piglets infected with pathogenic microorganisms. The basal diet is combined with the plant-derived composition at the stated inclusion rate and dosage to improve the growth performance of the PW piglets.

[0034] The plant-derived composition comprises an essential oil core, herbal powders, and preferred fillers. The essential oil core comprises a mixture of essential oils. Preferably, the essential oils comprise at least three essential oils in the disclosed percentages and / or ratios. The herbal powders comprise one or more herbal powders. In one aspect, the mixture of essential oils comprises carvacrol, eugenol, and star anise oil. In another aspect, the mixture of essential oils comprises tea tree oil, garlic oil, and lemon oil. The herbal powders included in the plant-derived composition preferably comprise fenugreek seed powder, licorice powder, turmeric powder, and combinations thereof. One or all of the essential oils may be microencapsulated. Microencapsulation may be performed before or after the essential oils are combined.

[0035] As used herein, the term "post-weaned piglet" or "PW piglet" refers to the period from weaning until the start of the fattening stage. The PW period, as referred to herein, is from approximately day 25 to approximately day 75 of life. It should be understood that the exact date of weaning can vary, and post-weaning refers to any day of life at which the piglet begins to be weaned. The terms "PW period," "PW stage," and "PW phase" are used interchangeably.

[0036] As used herein, the term "plant-derived" refers to plant-derived herbs, spices, or extracts, and / or bioactive compounds found in plants. Plant-derived includes natural products directly derived from plants. Plant-derived also includes synthetic and / or naturally identical compounds / compositions, which may be artificially prepared but whose chemical composition is identical to that of molecules found in plants. Plant-derived also includes synthetic and / or naturally identical compositions, which are formed by combining or formulating individual components (natural and / or synthetic components) to form naturally occurring compositions. This specification will refer to plant-derived compounds, but it should be understood that synthetic / naturally identical compounds or compositions may also be used.

[0037] As used herein, the term "plant-derived composition" refers to compositions comprising blends of essential oils and herbal powders. The plant-derived compositions described herein are standardized, specific, and science-based combinations of bioactive compounds found in plants that have proven efficacy and sustainable impacts on animals, humans, and / or the environment. Plant-derived compositions are combined or included in basal piglet feed to produce PW piglet feed.

[0038] As used herein, the terms “plant” and “plant derivative” refer to any part of a growing plant, including roots, stems, culms, leaves, branches, seeds, flowers, fruits, etc.

[0039] As used herein, the terms "essential oil blend" or "essential oil" refer to a combination of essential oils contained in the essential oil core of a plant-derived composition, preferably at least three essential oils. Essential oils and essential oil blends will be used interchangeably. As used herein, "essential oil" refers to an aromatic volatile liquid extracted from plant material. Essential oils are typically concentrated hydrophobic liquids containing volatile aroma compounds. The chemical components of essential oils can fall into a broad range of chemical categories, such as terpenes, terpenoids, phenols, aldehydes, ketones, and phenylpropanoids. Essential oils can be natural (i.e., derived from plants), synthetic, or composite to achieve the same essential oil as naturally occurring ones.

[0040] As used herein, the term "essential oil core" refers to a mixture of essential oils that also includes a carrier and / or an encapsulating matrix. One or more of the essential oils may be microencapsulated with an encapsulating matrix and / or combined with a carrier to form an essential oil core.

[0041] As used herein, the term "carvacrol" refers to 2-methyl-5-prop-2-ylphenol, which can be derived from plants, synthesized, or combined to achieve the same natural carvacrol. Carvacrol can be derived from, for example, oregano (Origanum vulgare) and thyme (Thymus vulgaris). Carvacrol derived from other plants may also be used, and is also within the scope of this disclosure.

[0042] As used herein, the term "eugenol" refers to 2-methoxy-4-prop-2-enylphenol, which can be derived from plants, synthesized, or combined to achieve the same natural eugenol. Eugenol can be derived from, for example, clove (Syzygium aromaticum), nutmeg (Myristica fragrans), cinnamon (Cinnamomum verum), basil (Ocimum basilicum), and / or bay leaf (Laurus nobilis). Eugenols derived from other plants may also be used, and are also within the scope of this disclosure.

[0043] As used herein, the term "star anise oil" refers to an essential oil consisting primarily of (>70%) 1-methoxy-4-[(E)-prop-1-enyl]benzene (trans-anesanthocyanin), which may be derived from plants such as star anise (Illicium verum). "Star anise oil" may also consist of synthetically produced trans-anesanthocyanin or a combination of natural and synthetic trans-anesanthocyanin. "Star anise oil" also includes combinations of the same natural composition with one or more natural or synthetic components found in star anise oil to achieve the effects of star anise oil. Essential oils derived from other plants and consisting primarily of trans-anesanthocyanin may also be used, and are also within the scope of this disclosure. As used herein, "star anise oil" also includes pure trans-anesanthocyanin.

[0044] As used herein, the term "tea tree oil" refers to an essential oil consisting primarily of terpinen-4-ol ([1S]-4-methyl-1-prop-2-ylcyclohex-3-en-1-ol) and γ-terpinene (1-methyl-4-prop-2-ylcyclohex-1,4-diene), which may be derived from plants such as Melaleuca alternifolia. "Tea tree oil" may also consist of synthetically produced terpinen-4-ol and γ-terpinene, or a combination of natural and synthetic terpinen-4-ol and γ-terpinene. "Tea tree oil" also includes combinations of the same natural composition with one or more natural or synthetic components found in tea tree oil to achieve the effects of tea tree oil. Essential oils derived from other plants and consisting primarily of terpinen-4-ol and γ-terpinene may also be used, and are also within the scope of this disclosure. As used herein, tea tree oil contains approximately 50% to 80% terpinen-4-ol ([1S]-4-methyl-1-prop-2-ylcyclohex-3-en-1-ol) and γ-terpinen (1-methyl-4-prop-2-ylcyclohex-1,4-diene).

[0045] As used herein, the term "garlic oil" refers to an essential oil primarily composed of allyl polysulfides such as diallyl disulfide (3-(prop-2-enyl disulfide)prop-1-ene) and diallyl trisulfide (3-(prop-2-enyl trisulfide)prop-1-ene), which may be derived from plants, such as garlic (Allium sativum). "Garlic oil" may also consist of synthetically produced allyl polysulfides or a combination of synthetic and naturally derived allyl polysulfides. "Garlic oil" also includes naturally occurring compositions in which one or more natural or synthetic components found in garlic oil are combined to achieve the effects of garlic oil. Essential oils derived from other plants and primarily composed of diallyl disulfide and diallyl trisulfide may also be used, and are also within the scope of this disclosure.

[0046] As used herein, the term "lemon oil" refers to an essential oil consisting primarily of D-limonene ([4R]-1-methyl-4-prop-1-en-2-ylcyclohexene), which can be derived from plants such as lemon (Citrus × limon) and other members of the Rutaceae family. "Lemon oil" can consist of synthetically produced D-limonene or a combination of natural and synthetic D-limonene. "Lemon oil" also includes naturally occurring compositions in which one or more natural or synthetic components found in lemon oil are combined to achieve the effects of lemon oil. Essential oils derived from other plants and consisting primarily of D-limonene may also be used, and are also within the scope of this disclosure.

[0047] As used herein, the term "PFA Core 2" or "PFA Core 2 essential oil blend" refers to an essential oil core. An essential oil core may comprise an encapsulating matrix, a carrier, or a combination thereof. The essential oil contains carvacrol, eugenol, and star anise oil. These terms will be used interchangeably.

[0048] As used herein, the terms "PFA Core 1" or "PFA Core 1 essential oil blend" refer to an essential oil core. An essential oil core may comprise an encapsulating matrix, a carrier, or a combination thereof. Essential oils include tea tree oil, garlic oil, and lemon oil. These two terms will be used interchangeably.

[0049] As used herein, the term “PFA 2 composition” or “PFA 2” refers to a plant-derived composition comprising: PFA core 2, herbal powders derived from fenugreek seeds, licorice and turmeric, and fillers such as limestone.

[0050] As used herein, the term “PFA 1 composition” or “PFA 1” refers to a plant-derived composition comprising: PFA core 1, herbal powders derived from fenugreek seeds, licorice and turmeric, and fillers such as limestone and wheat bran.

[0051] As used herein, the term "basal feed" refers to the feed given to PW piglets during the post-weaning period. It contains the ingredients and nutrients required by PW piglets. Basal feed does not contain plant-based compositions.

[0052] As used herein, the term “PW piglet feed” or “piglet feed” refers to a base feed supplemented with plant-derived compositions.

[0053] In one aspect, this specification includes a method for feeding livestock animals a combination of a basal feed and a plant-based composition. The animals are preferably piglets, more preferably PW piglets. At the start of the feeding method described herein, the PW piglets are preferably at least 21 days old, or more preferably at least 23 days old, or even more preferably 21 to 25 days old. The feeding method may also be started when the PW piglets are older than 25 days old.

[0054] The feeding method described herein may be initiated and / or introduced to PW piglets at any time during the PW period. The feeding method described herein is introduced to PW piglets from approximately day 21 to approximately day 75 of life. The feeding method is preferably applied directly to a grain-based diet after weaning (transition from breastfeeding to a grain-based diet), and preferably at least until day 14 of PW, but preferably at least until day 42 of PW. It should be understood that weaning time varies depending on the region and pig farming system, but the method described herein may be initiated immediately after weaning.

[0055] The feeding method described herein is provided to PW piglets from approximately day 21 to approximately day 75, preferably from day 21 to day 70, more preferably from day 21 to day 66, more preferably from day 21 to day 60, more preferably from day 23 to day 75, more preferably from day 23 to day 70, more preferably from day 23 to day 66, more preferably from day 23 to day 60, more preferably from day 25 to day 75, more preferably from day 25 to day 70, more preferably from day 25 to day 66, more preferably from day 25 to day 60, more preferably from day 27 to day 70, more preferably from day 27 to day 66, more preferably from day 27 to day 60.

[0056] In one aspect, a basal diet supplemented with a plant-derived composition is provided to healthy (e.g., without signs of disease) PW piglets. Preferably, the healthy piglets are not infected with pathogenic microorganisms. Alternatively, a basal diet supplemented with a plant-derived composition can be provided to PW piglets attacked by pathogen infection. The pathogen can be, for example, a pathogenic strain of Escherichia coli, such as enterotoxigenic Escherichia coli (ETEC). ETEC can be F4-carrying enterotoxigenic Escherichia coli (ETEC). A basal diet supplemented with a plant-derived composition can be provided to PW piglets with pathogen infection at any time during the PW period. Providing a basal diet supplemented with a plant-derived composition can alleviate the symptoms of pathogen infection.

[0057] This method includes providing a basal feed supplemented with a plant-derived composition / formulation. The basal feed can vary. Various basal feeds are known in the art and suitable for PW piglets. Examples of basal feeds for PW piglets formulated by Lindenberger Mühle (Germany) are disclosed in the following examples (Table 1). Basal feeds also include those manufactured by Agropodnik Mašovice, as (Mašovice, Czech Republic) or Purina (USA). Other basal feeds suitable for PW piglets are within the scope of this disclosure. The basal feed can be supplemented with the plant-derived composition described herein at the disclosed inclusion rates and dosages.

[0058] The plant-derived composition comprises an essential oil core, which preferably contains at least two essential oils, more preferably a combination of at least three essential oils. The essential oil core may contain more than three or more four essential oils. One or more of the essential oils contained in the essential oil core may be microencapsulated. Multiple combinations of essential oils may be used in the essential oil core. The essential oil core may contain, for example, carvacrol, eugenol, trans-anetinoside, terpinene-4-ol, γ-terpinene, diallyl disulfide, diallyl trisulfide, D-limonene, and combinations thereof. Essential oils included in the essential oil core include, for example, carvacrol, eugenol, and star anise oil. Essential oils included in another exemplary essential oil core include, for example, tea tree oil, garlic oil, and lemon oil.

[0059] In addition to the essential oil core, the plant-derived compositions described herein also contain herbal powders. In one aspect, the plant-derived compositions contain fenugreek seed powder. The plant-derived compositions may also contain turmeric powder, turmeric oil resin, licorice extract, and / or licorice root powder.

[0060] Plant-derived compositions may contain an essential oil core, wherein one or more of the essential oils are microencapsulated and / or premixed. The essential oils may be premixed with a carrier before being assembled into the essential oil core. The carrier may include sipernate, bentonite, sepiolite, etc. Plant-derived compositions may also contain one or more essential oils microencapsulated in an encapsulating matrix. The encapsulating matrix may include silica, proteins (such as wheat and yeast proteins), citric acid, natural polymers (such as starch, maltodextrin, glucose-fructose syrup), etc.

[0061] The plant-derived composition may also contain fillers. Fillers include, for example, wheat bran, limestone, corn cob, wheat husk, phyllite, glucose, wheat starch, corn starch, pea starch, potato starch, etc. Preferably, the plant-derived composition contains wheat bran and / or limestone as fillers.

[0062] In one aspect, a plant-derived composition is combined with a basal feed at a ratio to improve the growth performance of PW piglets. The content of the plant-derived composition in the basal feed can be from about 0.25 kg / t to about 2.5 kg / t, preferably from about 0.25 kg / t to about 2.0 kg / t, more preferably from about 0.25 kg / t to about 1.5 kg / t, more preferably from about 0.5 kg / t to about 2.5 kg / t, more preferably from about 0.5 kg / t to about 2.0 kg / t, more preferably from about 0.5 kg / t to about 1.5 kg / t, more preferably from about 0.8 kg / t to about 1.2 kg / t, more preferably from about 0.9 kg / t to about 1.1 kg / t, and more preferably from about 1 kg / t.

[0063] In one aspect, the essential oil core comprises carvacrol and / or eugenol. The essential oil core preferably also comprises star anise oil. An essential oil core comprising carvacrol, eugenol, and star anise oil will be referred to herein as PFA core 2. In one aspect, PFA core 2 consists essentially of carvacrol, eugenol, and star anise oil. "consistently of" means that PFA core 2 may not contain any other essential oils besides carvacrol, eugenol, and star anise oil.

[0064] PFA core 2 may contain microencapsulated essential oils, in which carvacrol, eugenol, and star anise oil are combined to form an essential oil mixture, and then in the presence of... Figure 5 Microencapsulation is performed in the case of the encapsulation matrix shown. Alternatively, carvacrol and / or eugenol can be microencapsulated before being mixed with star anise oil to form a microencapsulation matrix as shown. Figure 4 The image shows a partially microencapsulated essential oil mixture.

[0065] PFA core 2 contains the following amounts of carvacrol: 20% to 80% by weight of all essential oils in PFA core 2 prior to any microencapsulation, preferably 20% to 60% by weight of all essential oils in the core, preferably 30% to 70% by weight, more preferably 30% to 60% by weight, more preferably 30% to 50% by weight, more preferably 35% to 45% by weight, more preferably 38% to 42% by weight, more preferably 39% to 41% by weight, and more preferably about 40% by weight.

[0066] PFA core 2 may also contain the following amounts of eugenol: 0% to 80% by weight of all essential oils in PFA core 2 before microencapsulation, preferably 10% to 50% by weight, more preferably 20% to 50% by weight, more preferably 30% to 50% by weight, preferably more preferably 35% to 45% by weight, more preferably 38% to 42% by weight, more preferably 39% to 41% by weight, and more preferably about 40% by weight.

[0067] PFA Core 2 contains a combination of carvacrol and eugenol in the following amounts: about 70% to about 90% by weight of all essential oils in PFA Core 2 before microencapsulation, preferably about 75% to about 85% by weight of all essential oils in PFA Core 2 before microencapsulation, more preferably about 80% by weight.

[0068] PFA Core 2 also contains the following amounts of star anise oil: 10% to 30% by weight of all essential oils in PFA Core 2 before microencapsulation, preferably 15% to 25% by weight, preferably 18% to 22% by weight, more preferably 19% to 21% by weight, and more preferably about 20% by weight.

[0069] In one aspect, PFA core 2 comprises a combination of carvacrol and eugenol in amounts of about 70% to about 90% by weight of all essential oils in PFA core 2, preferably about 75% to about 85% by weight, more preferably about 80% by weight. PFA core 2 also comprises star anise oil in amounts of 10% to 30% by weight of all essential oils in PFA core 2, preferably 15% to 25% by weight of all essential oils in PFA core 2, preferably 18% to 22% by weight, more preferably 19% to 21% by weight, more preferably about 20% by weight.

[0070] PFA core 2 may contain microencapsulated essential oils or mixtures of essential oils. The matrix used for encapsulation may contain modified starch and maltodextrin. The essential oils may be microencapsulated as described, for example, in EP patent 1419811A1, which is incorporated herein by reference. Other encapsulation methods may also be used, and these methods are within the scope of this specification.

[0071] In one aspect, this specification includes a PFA 2 composition comprising a PFA core 2, herbal powders, and fillers as described herein. A variety of herbal powders may be used, and all of these are within the scope of this specification. The herbal powders are preferably fenugreek seed powder, licorice root powder, and / or turmeric powder, more preferably fenugreek seed powder, licorice root powder, and turmeric powder, and even more preferably, the turmeric powder is organic turmeric powder.

[0072] The PFA 2 composition comprises PFA core 2 in amounts of about 5% to about 30% by weight, preferably about 10% to about 20% by weight, more preferably about 12% to about 18% by weight, and even more preferably about 14% to about 15% by weight.

[0073] If the essential oil is microencapsulated, the PFA core 2 in the PFA 2 composition may contain an encapsulating matrix. The PFA 2 composition may contain the following encapsulating matrix by weight: about 5% to about 20% of the PFA 2 composition, preferably about 5% to about 15% of the PFA 2 composition, more preferably about 8% to about 12% of the PFA 2 composition, and even more preferably about 9% to about 10% of the PFA 2 composition.

[0074] The PFA 2 composition may also contain fillers. Various fillers are known in the art, and all are within the scope of this specification. Fillers may be, for example, limestone, wheat bran, etc. Preferably, the filler is limestone. The PFA 2 composition may contain fillers in amounts of: about 5% to about 40% by weight of the PFA 2 composition, preferably about 10% to about 30% by weight, more preferably about 10% to about 20% by weight, more preferably about 14% to about 16% by weight, and even more preferably about 15% to about 16% by weight.

[0075] The PFA 2 composition also comprises one or more herbal powders. The PFA 2 composition may contain fenugreek powder, turmeric powder, and licorice powder. The PFA 2 composition further comprises fenugreek powder in amounts of about 50% to 80% by weight of the PFA 2 composition, preferably 60% to 75% by weight, more preferably 60% to 70% by weight, more preferably 62% to 70% by weight, and even more preferably about 66% by weight.

[0076] The PFA 2 composition also contains the following amounts of turmeric powder: 2% to 3% by weight of the PFA 2 composition, preferably 2.2% to 2.8% by weight, more preferably 2.4% to 2.6% by weight, more preferably about 2.5% to 2.6% by weight, and more preferably 2.5% by weight.

[0077] The PFA 2 composition also contains licorice powder in amounts of 0.1% to 5% by weight, preferably 0.5% to 3.5% by weight, more preferably 1% to 2.5% by weight, more preferably about 1.2% to 1.8% by weight, and more preferably 1.4% by weight.

[0078] PW piglet feed contains the PFA 2 composition at the following proportions: about 0.25 kg / t to about 2.5 kg / t, preferably 0.25 kg / t to 2.0 kg / t, more preferably about 0.25 kg / t to about 1.5 kg / t, more preferably about 0.5 kg / t to about 2.5 kg / t, more preferably 0.5 kg / t to 2.0 kg / t, more preferably about 0.5 kg / t to about 1.5 kg / t, more preferably about 0.8 kg / t to about 1.2 kg / t, more preferably 0.9 kg / t to 1.1 kg / t, and more preferably about 1 kg / t.

[0079] The amount of PFA component 2 (e.g., essential oils and herbal powders in an essential oil blend) is suitable for improving the growth performance of PW piglets. In one aspect, fenugreek seed powder is included in the PW piglet feed in an amount of about 200 ppm to about 1500 ppm, preferably about 500 ppm to about 1000 ppm, more preferably about 600 ppm to about 750 ppm, more preferably about 640 ppm to about 680 ppm, and even more preferably about 660 ppm.

[0080] Turmeric powder is included in PW piglet feed in an amount of about 5 ppm to about 50 ppm, preferably about 15 ppm to about 35 ppm, more preferably about 20 ppm to about 30 ppm, more preferably about 22 ppm to about 28 ppm, and more preferably about 25 ppm.

[0081] Licorice root powder is included in PW piglet feed in an amount of about 1 ppm to about 50 ppm, preferably about 5 ppm to about 35 ppm, more preferably about 10 ppm to about 25 ppm, more preferably about 12 ppm to about 18 ppm, and even more preferably about 14 ppm.

[0082] Carvacrol is included in PW piglet feed in an amount of about 10 ppm to about 50 ppm, preferably about 15 ppm to about 50 ppm, more preferably about 15 ppm to about 40 ppm, more preferably about 15 ppm to about 35 ppm, more preferably about 15 ppm to about 30 ppm, more preferably about 15 ppm to about 25 ppm, more preferably about 10 ppm to about 30 ppm, more preferably about 10 ppm to about 25 ppm, and more preferably about 20 ppm.

[0083] Eugenol is included in PW piglet feed in an amount of about 10 ppm to about 50 ppm, preferably about 15 ppm to about 50 ppm, more preferably about 15 ppm to about 40 ppm, more preferably about 15 ppm to about 35 ppm, more preferably about 15 ppm to about 30 ppm, more preferably about 15 ppm to about 25 ppm, more preferably about 10 ppm to about 30 ppm, more preferably about 10 ppm to about 25 ppm, and more preferably about 20 ppm.

[0084] Star anise oil is included in PW piglet feed in an amount of about 5 ppm to about 50 ppm, preferably about 5 ppm to about 40 ppm, more preferably about 5 ppm to about 30 ppm, more preferably about 5 ppm to about 25 ppm, more preferably about 5 ppm to about 20 ppm, more preferably about 5 ppm to about 15 ppm, more preferably about 7 ppm to about 12 ppm, and more preferably about 10 ppm.

[0085] In one respect, the essential oil core contains tea tree oil, garlic oil, and lemon oil, and will be referred to herein as PFA Core 1. In another respect, PFA Core 1 is essentially composed of tea tree oil, garlic oil, and lemon oil. "Essentially composed of" means that PFA Core 1 may not contain any other essential oils besides tea tree oil, garlic oil, and lemon oil.

[0086] PFA core 1 contains tea tree oil in amounts ranging from about 50% to about 80% by weight of all essential oils in PFA core 1, preferably from about 55% to about 75% by weight, more preferably from about 62% to about 68% by weight, and even more preferably from about 65% by weight. PFA core 1 contains garlic oil in amounts ranging from about 20% to about 50% by weight of all essential oils in PFA core 1, preferably from about 32% to about 38% by weight, and even more preferably from about 34% to about 36% by weight. PFA core 1 contains lemon oil in amounts ranging from about 1% to about 2% by weight of all essential oils in PFA core 1, preferably from about 1.2% to about 1.8% by weight, and even more preferably from about 1.4% to about 1.6% by weight.

[0087] The PFA 1 composition comprises PFA core 1 as described above, herbal powder, and one or more fillers. The herbal powder is preferably fenugreek seed powder and turmeric powder, more preferably, the turmeric powder is organic turmeric powder. The filler is preferably wheat bran and limestone.

[0088] The PFA 1 composition comprises the following amounts of PFA core 1: about 5% to about 30% by weight of the PFA 1 composition, preferably about 10% to about 20% by weight, more preferably about 12% to about 18% by weight, more preferably about 15% to about 17% by weight, and more preferably about 16% to about 17% by weight.

[0089] The PFA 1 composition also contains fenugreek powder in amounts of about 40% to 60% by weight of the PFA 1 composition, preferably 45% to 55% by weight, more preferably 48% to 52% by weight, and even more preferably about 50% by weight.

[0090] The PFA 1 composition also contains the following amounts of turmeric powder: 2% to 8% by weight of the PFA 1 composition, preferably 3% to 7% by weight of the PFA 1 composition, more preferably 4% to 6% by weight, and more preferably about 5% by weight.

[0091] The PFA 1 composition further comprises a filler in an amount of about 15% to about 40% by weight of the PFA 1 composition, preferably about 20% to about 30% by weight, more preferably about 25% to about 30% by weight, and even more preferably about 28% to about 29% by weight. The PFA 1 composition may contain limestone as a filler in an amount of about 10% to about 20% by weight of the PFA 1 composition, preferably about 13% to about 18% by weight, and even more preferably about 15% to about 16% by weight. The PFA 1 composition may contain wheat bran as a filler in an amount of about 10% to about 20% by weight of the PFA 1 composition, preferably about 10% to about 15% by weight, and even more preferably about 13% by weight.

[0092] PW piglet feed contains PFA 1 composition at the following proportions: about 0.25 kg / t to about 2.5 kg / t, preferably 0.25 kg / t to 2.0 kg / t, more preferably about 0.25 kg / t to about 1.5 kg / t, more preferably about 0.5 kg / t to about 2.5 kg / t, more preferably 0.5 kg / t to 2.0 kg / t, more preferably about 0.5 kg / t to about 1.5 kg / t, more preferably about 0.8 kg / t to about 1.2 kg / t, more preferably 0.9 kg / t to 1.1 kg / t, and more preferably about 1 kg / t.

[0093] The amount of PFA component 1 (e.g., essential oils and herbal powders in an essential oil blend) is suitable for improving the growth performance of PW piglets. In one aspect, fenugreek seed powder is included in the PW piglet feed in an amount of about 400 ppm to about 600 ppm, preferably about 450 ppm to about 550 ppm, more preferably about 500 ppm. Turmeric powder is included in the PW piglet feed in an amount of about 30 ppm to about 70 ppm, preferably about 40 ppm to about 60 ppm, and more preferably about 50 ppm.

[0094] Tea tree oil is included in PW piglet feed at an amount of about 20 ppm to about 40 ppm, preferably about 25 ppm to about 35 ppm, and more preferably about 30 ppm. Garlic oil is included in PW piglet feed at an amount of about 5 ppm to about 25 ppm, preferably about 10 ppm to about 20 ppm, and more preferably about 15 ppm. Lemon oil is included in PW piglet feed at an amount of about 0.4 ppm to about 1 ppm, preferably about 0.5 ppm to about 0.8 ppm, and more preferably about 0.6 ppm.

[0095] In one aspect, this specification includes a method for feeding an animal (preferably a piglet) a basal feed supplemented with the plant-derived composition described herein. Preferably, the method includes supplementing the basal feed with a PFA 1 composition or a PFA 2 composition. The method may include providing the piglet with ad libitum access to the feed. The method may include providing the piglet with a limited amount of feed, for example, not ad libitum access. Preferably, the method includes providing the piglet with ad libitum access to a paste-like feed and / or pelleted feed. Preferably, the method includes providing the piglet with a paste-like feed. More preferably, the method includes providing the piglet with an ad libitum access to a paste-like feed. Water is always provided to the piglet.

[0096] The method of feeding PW piglets as described herein results in improved growth performance. This method includes, preferably, providing PW piglet feed, such as a basal diet supplemented with the plant-derived composition described herein, to PW piglets starting from day 21 to day 26. Piglet feed can be provided to PW piglets at any time during the PW period. Compared to PW piglets fed only a basal diet (e.g., without the plant-derived composition), the improved growth performance results in improved average daily gain (ADG), improved feed conversion ratio (FCR), increased average daily feed intake (ADFI), increased body weight (BW), and / or improved fecal score. Improved growth performance also includes a reduced incidence of PWD. Other improved growth performance indicators may also be achieved and are within the scope of this specification.

[0097] In the method described herein, a reduction in PWD-associated microorganisms, such as pathogenic Escherichia coli, is observed in the gut microbiome of PW piglets. This results in a reduction in the severity of PWD symptoms, namely, a decrease in diarrhea, dehydration, mortality, and growth retardation in surviving pigs. In PW piglets infected with PWD-associated microorganisms, this method reduces fecal excretion of PWD-associated microorganisms, decreases the incidence of diarrhea, and improves fecal scores in infected PW piglets. When the plant-derived composition described herein is administered to piglets during the post-weaning period, particularly under antibiotic-free conditions, improvements in growth performance, enhanced animal welfare, and reduced drug requirements are direct effects. Advantageously, the additional beneficial effect of using this unique blend of plant-derived compositions as an alternative to traditional antimicrobial growth promoters is the prevention of antibiotic resistance development through the use of antibiotics in livestock production.

[0098] Improvements in growth parameters will be described with reference to a basal diet supplemented with the PFA 2 composition; however, it should be understood that other plant-derived compositions described herein (such as the PFA 1 composition) also result in improved growth parameters in PW piglets. The PW phase typically lasts approximately 45 days, from approximately day 25 to approximately day 70 of piglet age. The PW period can vary by approximately + / - 7 days, for example, from day 18 to approximately day 77.

[0099] Unbound by any theoretical framework, plant-based compositions improve palatability and animal welfare. Adequate feed intake in early life contributes to the development of a normally functioning gut in piglets, thereby supporting their resilience to stress factors.

[0100] Compared to piglets not fed with the PFA 2 composition, piglets supplemented with the PFA 2 composition described herein may have improved ADG. The ADG of piglets fed with the PFA 2 composition is increased by at least 1%, preferably at least 2%, more preferably at least 4%, and even more preferably at least 6%, compared to piglets not fed with the PFA 2 composition.

[0101] Compared to piglets not fed with the PFA 2 composition, piglets supplemented with the PFA 2 composition described herein exhibit improved feed conversion ratios (FCRs). The FCR of piglets fed with the PFA 2 composition is increased by at least 2%, preferably at least 3%, and more preferably at least 4%, compared to piglets not fed with the PFA 2 composition.

[0102] Compared to piglets that are not provided with the PFA 2 composition, piglets supplemented with the PFA 2 composition described herein may have increased body size (BW). The BW of piglets supplemented with the PFA 2 composition increases by at least 2%, preferably at least 4%, and more preferably at least 5%, compared to piglets that are not provided with feed supplemented with the PFA 2 composition.

[0103] Compared to piglets not fed with the PFA 2 composition, piglets supplemented with the PFA 2 composition described herein exhibit improved ADFI. The ADFI of piglets fed with the PFA 2 composition is increased by at least 2%, preferably at least 4%, and more preferably at least 6%, compared to piglets not fed with the PFA 2 composition.

[0104] Piglets supplemented with the PFA 2 composition described herein maintain and / or improve normal fecal consistency. Compared to piglets not provided with the PFA 2 composition, piglets supplemented with the PFA 2 composition described herein exhibit improved fecal scores. The determination of fecal scores is known in the art and described, for example, in Jensen et al., 2006 or Fairbrother et al., 2017.

[0105] Piglets supplemented with the PFA 2 composition described herein may have a reduced tendency to develop PWD during the PW stage. Compared to PW piglets not supplemented with the PFA 2 composition, the incidence of PWD in PW piglets provided with the PFA 2 composition is reduced by at least 10%, preferably at least 25%, more preferably at least 50%, and even more preferably at least 75%.

[0106] Piglets supplemented with the PFA 2 composition described herein exhibit reduced mortality due to PWD during the PW stage. Compared to PW piglets not supplemented with the PFA 2 composition, the incidence of PWD-related mortality is reduced by at least 10%, preferably at least 25%, more preferably at least 50%, and even more preferably at least 75% in PW piglets provided with the PFA 2 composition.

[0107] PWD symptoms in piglets already infected with PWD-associated microorganisms and supplemented with the PFA 2 composition described herein may be reduced or eliminated. Possible affected PWD symptoms include a reduction in the amount and / or duration of fecal excretion of PWD-associated microorganisms. Possible affected PWD symptoms also include an increase in fecal consistency scores. The incidence of PWD in piglets already infected with PWD-associated microorganisms and supplemented with the PFA 2 composition is reduced by at least 10%, preferably at least 25%, more preferably at least 50%, and even more preferably at least 75%, compared to piglets not supplemented with the PFA 2 composition. The incidence of fecal excretion of infectious pathogens in piglets already infected with PWD-associated microorganisms and supplemented with the PFA 2 composition is reduced by at least 10%, preferably at least 25%, more preferably at least 30%, and even more preferably at least 40%, compared to piglets not supplemented with the PFA 2 composition.

[0108] In one aspect, this specification includes PW piglet feed. PW piglet feed comprises a basal feed as described above and a plant-based composition as described herein. Preferably, PW piglet feed comprises PFA core 2 and / or PFA core 1, more preferably, PW piglet feed comprises PFA core 2. Preferably, PW piglet feed is supplemented with a PFA 2 composition and / or a PFA 1 composition, more preferably, PW piglet feed is supplemented with a PFA 2 composition. PW piglet feed is supplemented with PFA 2 composition or PFA 1 composition at the following content rates: about 0.25 kg composition / ton of feed to about 2.5 kg / t, preferably 0.25 kg / t to 2.0 kg / t, more preferably about 0.25 kg / t to about 1.5 kg / t, more preferably about 0.5 kg / t to about 2.5 kg / t, more preferably 0.5 kg / t to 2.0 kg / t, more preferably about 0.5 kg / t to about 1.5 kg / t, more preferably about 0.8 kg / t to about 1.2 kg / t, more preferably 0.9 kg / t to 1.1 kg / t, and more preferably about 1 kg / t.

[0109] In one aspect, the PW piglet feed contains the following amounts of fenugreek seed powder: about 200 ppm to about 1000 ppm, preferably about 500 ppm to about 800 ppm, more preferably about 600 ppm to about 700 ppm, more preferably about 640 ppm to about 680 ppm, and even more preferably about 660 ppm in the PW piglet feed.

[0110] PW piglet feed contains the following amounts of turmeric powder: about 5 ppm to about 50 ppm, preferably about 15 ppm to about 35 ppm, more preferably about 20 ppm to about 30 ppm, more preferably about 22 ppm to about 28 ppm, and even more preferably about 25 ppm.

[0111] PW piglet feed contains the following amounts of licorice root powder: about 1 ppm to about 50 ppm, preferably about 5 ppm to about 35 ppm, more preferably about 10 ppm to about 25 ppm, more preferably about 12 ppm to about 18 ppm, and even more preferably about 14 ppm in the PW piglet feed.

[0112] PW piglet feed contains the following amounts of carvacrol: about 10 ppm to about 50 ppm, preferably about 15 ppm to about 50 ppm, more preferably about 15 ppm to about 40 ppm, more preferably about 15 ppm to about 35 ppm, more preferably about 15 ppm to about 30 ppm, more preferably about 15 ppm to about 25 ppm, more preferably about 10 ppm to about 30 ppm, more preferably about 10 ppm to about 25 ppm, and more preferably about 20 ppm.

[0113] PW piglet feed contains the following amounts of eugenol: about 10 ppm to about 50 ppm, preferably about 15 ppm to about 50 ppm, more preferably about 15 ppm to about 40 ppm, more preferably about 15 ppm to about 35 ppm, more preferably about 15 ppm to about 30 ppm, more preferably about 15 ppm to about 25 ppm, more preferably about 10 ppm to about 30 ppm, more preferably about 10 ppm to about 25 ppm, and more preferably about 20 ppm.

[0114] PW piglet feed contains the following amounts of star anise oil: about 5 ppm to about 50 ppm in PW piglet feed, preferably about 5 ppm to about 40 ppm, more preferably about 5 ppm to about 30 ppm, more preferably about 5 ppm to about 25 ppm, more preferably about 5 ppm to about 20 ppm, more preferably about 5 ppm to about 15 ppm, more preferably about 7 ppm to about 12 ppm, and more preferably about 10 ppm.

[0115] In one aspect, the PW piglet feed contains fenugreek seed powder in amounts of about 400 ppm to about 600 ppm, preferably about 450 ppm to about 550 ppm, more preferably about 500 ppm. The PW piglet feed contains turmeric powder in amounts of about 30 ppm to about 70 ppm, preferably about 40 ppm to about 60 ppm, and more preferably about 50 ppm. The PW piglet feed contains tea tree oil in amounts of about 20 ppm to about 40 ppm, preferably about 25 ppm to about 35 ppm, more preferably about 30 ppm. The PW piglet feed contains garlic oil in amounts of about 5 ppm to about 25 ppm, preferably about 10 ppm to about 20 ppm, more preferably about 15 ppm. Lemon oil is included in the PW piglet feed in amounts of about 0.4 ppm to about 1 ppm, preferably about 0.5 ppm to about 0.8 ppm, more preferably about 0.6 ppm.

[0116] Example

[0117] The invention is described in further detail with reference to the following experimental embodiments. Unless otherwise stated, these embodiments are provided for illustrative purposes only and are not intended to be limiting. Therefore, the invention should not in any way be construed as limited to the following embodiments, but should be construed as covering any and all variations that become apparent as a result of the teachings provided herein.

[0118] Example 1

[0119] Materials and methods

[0120] Bacterial strains and test compounds

[0121] Chlorobacterium violaceum DSM 30191 was purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ (Braunschweig, Germany)) and cultured in trypsin soybean broth (Biokar Diagnostics (Allone, France); TSB) at 30°C under aerobic conditions.

[0122] Field isolates of *Escherichia coli* O88:H8 and O143:H4 (both isolated from pigs) were provided by the Department of Veterinary Pathology, AGES GmbH Linz (Austria), and serotyped by the National Standard Laboratory for *Escherichia coli*, AGES GmbH Graz (Austria). Both strains were classified as pathogenic due to their high growth rates in pure cultures from various organs and their mucoid or hemolytic properties.

[0123] For the mucus adhesion assay, F4+ *Escherichia coli* strains carrying the bacteria were provided by Alimetrics Ltd (Espoo, Finland) as field isolates isolated in Finland. The *E. coli* field isolates were subjected to aerobic conditions at 37°C supplemented with 2 g / L glucose (Carl Roth GmbH + Co. KG (Karlsruhe, Germany); TSB). + The cells were cultured in Biokar Diagnostic casein soybean broth for biofilm assays.

[0124] Zinc oxide used for bacterial assays was purchased from Auhof Apotheke (Linz, Austria). Carvacrol and tea tree oil, as well as “PFA Core 1” and “PFA Core 2”, were supplied by Delacon Biotechnik GmbH (Engerwitzdorf, Austria).

[0125] For PFA Core 1, the essential oils were blended in a ratio of 32.571 (tea tree oil): 16.236 (garlic oil): 0.693 (lemon oil). In PFA Core 1, the essential oil blend comprised 65.8% by weight of tea tree oil, 32.8% by weight of garlic oil, and 1.4% by weight of lemon oil. The PFA Core 1 content in the feed was 49.5 ppm.

[0126] The PFA 1 composition is used in the basal feed at a dosage of 500 ppm fenugreek seed powder, 50 ppm turmeric powder, 33 ppm tea tree oil, 16 ppm garlic oil, and 0.7 ppm lemon oil.

[0127] For PFA Core 2, the essential oils are blended in a ratio of 20 (carvacrol): 20 (eugenol): 9 (star anise oil). The PFA 2 composition is used in the basal feed at a dosage of 660 ppm fenugreek seed powder, 25 ppm turmeric powder, 14 ppm licorice root powder, 20 ppm carvacrol, 20 ppm eugenol and 9 ppm star anise oil.

[0128] Determination of minimum inhibitory concentrations

[0129] The minimum inhibitory concentration (MIC) in each assay was determined optically as the lowest concentration of the test compound that completely inhibited visible growth (no obvious cell clumps or turbidity) after an incubation period (18 hours at 30°C for *Cyclophorus*, 18 hours at 37°C for *Escherichia coli* O88:H8 and O143:H4, and 24 hours at 37°C for F4+ *Escherichia coli* strains). The median of all replicates was defined as the MIC value for the corresponding test compound.

[0130] For MIC assessment of F4+ infected Escherichia coli strains, carvacrol and tea tree oil were serially diluted 2-fold in TSB containing 2% Tween 80.

[0131] Quorum sensing inhibition assay (P. aeruginosa)

[0132] The effects of plant-derived test compounds at concentrations ranging from 10,000 ppm to 5 ppm on *Chlorobacterium violaceum* were evaluated. The production of chlorophyll was quantified based on the protocol of Skogman et al. (2016). A series of dilutions ranging from 20,000 ppm to 10 ppm were prepared from a stock solution (200,000 ppm) of the plant-derived test compound in ≥99.8% ethanol (VWR International GmbH (Vienna, Austria)). Four technical replicates for each concentration were arranged in Nunc. ® In a polystyrene 96-well microtiter plate (Thermo Scientific (Waltham, USA)). Overnight culture of *Chlorella vulgaris* in 10 mL TSB was diluted in fresh TSB to approximately 1–2 × 10⁻⁶. 6 CFU mL -1 50 μL of aliquots were added to each test well and four wells containing only culture medium as positive growth controls (without the test compound). Four additional wells containing only TSB served as negative (sterile) controls. The initial bacterial concentration was verified using colony counting. The microtiter plates and colony counting plates were incubated at 30°C for 18 hours. After the incubation period, the MIC values ​​were determined before the cells and violet bacitracin pigment formed a precipitate by centrifugation at 3000 rpm for 10 minutes. The supernatant was removed from the wells by aspiration, and 200 μL of ≥99.8% ethanol (VWR International GmbH) was added to each well. The plates were sealed with a Microseal 'B' PCR plate sealing film (Bio-Rad (Hercules, USA)) and incubated overnight in the dark to dissolve the precipitated violet bacitracin pigment. The next day, the plates were centrifuged at 3000 rpm for 10 minutes to allow the destained cells to settle. Subsequently, 100 μL of the violet smeared supernatant was transferred to a new microtiter plate, and the absorbance at 570 nm was measured using a PHOmo microplate reader (Anthos Mikrosysteme GmbH (Friesoythe, Germany)).

[0133] Biofilm inhibition assay

[0134] The effects of plant-derived test compounds on biofilm formation in *Escherichia coli* O88:H8 and O143:H4 were evaluated in a microdilution assay in broth, followed by crystal violet staining with slight modifications according to Axmann et al. (2021). In short, the plant-derived test compounds were analyzed in TSB... + Serial dilutions were performed and arranged vertically in 96-well microtiter plates as described above in the "Quaternary Inhibition Assay (Chlorobacterium violaceum)". The outer columns of the microtiter plates were filled with sterile TSB. + To prevent evaporation from the center hole. (TSB) + Adjust the overnight culture of the corresponding E. coli to fresh TSB + Approximately 2×10 6 CFU mL -1 50 μL of the diluted solution was placed in each test well. Four test wells were prepared containing only bacterial cultures without plant-derived test compounds or only sterile TSB. + The wells of the culture medium were used as positive or negative controls. The plates were incubated at 37°C for 18 hours under aerobic conditions.

[0135] The antimicrobial activity of the plant-derived test compounds was evaluated as described in the "Determination of Minimum Inhibitory Concentration" section above. After removing the culture medium, the microtiter plate was washed twice with deionized H2O (diH2O) to remove unadhered cells and culture medium components, and air-dried for 30 minutes. The adhered biomass was stained with 130 μL of 0.1% (w / v) crystal violet in diH2O solution (Alfa Aesar, Thermo Fisher GmbH (Kandel, Germany)) at room temperature for 20 minutes. The microtiter plate was then washed three times with diH2O and allowed to air-dry completely. The adhered dye was dissolved in 130 μL of 30% (v / v) acetic acid (MerckKgaA (Darmstadt, Germany)) and transferred to a new microtiter plate. The absorbance at 570 nm was measured using a PHOmo microplate reader (AnthosMikrosysteme GmbH). The mean of four replicates was calculated, and then the negative control measurement was subtracted. The result was expressed as a percentage of biofilm relative to the positive control. Each test compound was measured three to four times. In this study, the entire adherent cell population stained with crystal violet solution was referred to as the biofilm, regardless of the stage of maturity.

[0136] In vitro mucus adhesion assay

[0137] In the preparation phase of the mucus adhesion assay, overnight cultures of F4+ *E. coli* carrying *Hymenopterus* were grown in TSB without the test compound. Starting from this seed culture, each treatment was cultured twice consecutively at a 10% inoculum. All treatments were cultured in triplet vials. Turbidity of the cultures was monitored by measuring absorbance at 600 nm to ensure cell density was close to that of the control culture without the test product. On day three, each culture was further diluted tenfold in growth medium containing the corresponding test product at a specific concentration (1 / 8 of the MIC, and for tea tree oil, a second concentration of 1 / 80 of the MIC to achieve a concentration level comparable to the tested carvacrol). Tritium-labeled thymidine was introduced into each vial to radiolabel the *E. coli* cells.

[0138] To prepare for the in vitro mucus adhesion assay, native intestinal mucus was obtained from euthanized piglets and used to coat the wells of microtiter plates. The mucus-coated microtiter plates were washed with HEPES-Hank's buffer, then labeled *E. coli* cells were introduced (nine replicates per treatment) and incubated at 37°C for 1 hour. Unbound cells were removed by washing the wells twice with HEPES-Hank's buffer. Scintillation solution was added before measuring the radioactivity of each well. Additionally, the total radioactivity of each labeled *E. coli* culture used for adhesion studies was measured to determine the proportion of attached cells.

[0139] Animals, diets and in vivo trial design

[0140] One thousand healthy weaned piglets (500 males; 500 females; DanBredx Duroc) at a weaning age of 25±2 days were divided into four treatment groups, totaling 100 pens (5 males & 5 females per pen, 25 pens per treatment group). The average pen temperature was maintained at approximately 30°C during the first week post-weaning and gradually decreased by 1.8°C weekly from day 53 until reaching 22°C. Throughout the experiment, the weaned piglets had free access to a paste-like diet and water supplied via water bowls.

[0141] The four dietary treatment groups included a negative control group (NC) receiving no supplemented basal diet, a positive control group whose diet was supplemented with pharmacological levels of zinc oxide (3 kg / t feed; ZnO), and treatment groups PFA 1 and PFA 2, which were fed a basal diet supplemented with either PFA 1 composition or PFA 2 composition at 1 kg / t feed, respectively.

[0142] Plant-derived prototype PFA1 ( Figure 3It contains 50% fenugreek seed powder, 5% turmeric root powder, 15.5% limestone and 13% wheat bran. Microencapsulated PFA core 1 is added to PFA 1 at 16.5% (4.6% essential oil).

[0143] Plant-derived prototype PFA2 composition ( Figure 4 The product contains 66% fenugreek seed powder, 2.5% turmeric root powder, 1.4% licorice root powder, and 15.4% limestone. PFA Core 2 is added to the PFA2 composition at 14.7% (4.9% of the total amount of essential oil and remaining encapsulating matrix and carrier). Carvacrol and eugenol are encapsulated using the encapsulating matrix as follows... Figure 4 Microencapsulation was performed at the concentration shown. Star anise oil was mixed with a carrier (sipernate, 40.5%) as follows... Figure 4 The premix is ​​shown. The final concentrations of carvacrol and eugenol in the PFA2 composition are each 1.99%, and the final concentration of star anise oil is 0.9%.

[0144] The 42-day observation period was divided into two feeding periods: a two-week initial period (from 25 to 38 days of age) and a four-week subsequent growth period (from 39 to 66 days of age). Except for zinc, the basal diet for each period was formulated according to the recommendations of the Nutritional Physiology Society (2006). The dietary composition and calculated nutrient content of the initial and growth diets are shown in Table 1 below. Plant-derived prototypes were supplied by Delacon Biotechnik GmbH (Austria). Zinc oxide (Spezialfutter Neuruppin GmbH & Co. KG (Neuruppin, Germany)) supplementation was achieved by reducing Tixosil (>97% silica), while the addition of PFA 1 and PFA 2 was achieved by reducing limestone.

[0145] Table 1

[0146]

[0147] The formulation of the basal feed / PFA 1 composition includes:

[0148] a. 500ppm fenugreek seed powder

[0149] b. 50ppm turmeric powder

[0150] c. 30ppm tea tree oil

[0151] d.15ppm garlic oil

[0152] e.0.625ppm lemon oil

[0153] The formulation of the basal feed / PFA 2 composition includes:

[0154] f. 660ppm fenugreek seed powder

[0155] g.25ppm turmeric powder (organic)

[0156] h.14ppm licorice root powder

[0157] i. 20ppm carvacrol

[0158] j.20ppm eugenol

[0159] 9ppm star anise oil

[0160] Table 2 provides the feed composition and calculation analysis for the starting diets (as is) from 25 to 38 days of age. Table 3 provides the feed composition and calculation analysis for the growing diets from 39 to 66 days of age. Treatment diets T1 consisted of non-nitrogenous (NC), T2 included zinc oxide, T3 included a PFA1 composition, and T4 included a PFA2 composition.

[0161] Table 2

[0162]

[0163] Table 3

[0164]

[0165] Determination of piglet growth performance and diarrhoea score

[0166] Throughout the 42-day experimental period, all piglets were observed twice daily for any abnormalities, unusual behaviors, and clinical signs of disease.

[0167] Weekly pen weight (BW) and the amount of feed supplied to each pen during the previous week were recorded. Piglet weight gain was calculated by dividing the average BW per pen at the end of each period by the average BW per pen at the beginning of each period and the number of piglets per pen. Feed intake per piglet was estimated as total feed supplied per pen and period, minus spilled / leftover feed and adjusted for the number of piglets per pen. Feed conversion ratio (FCR) was calculated as the weekly adjusted feed intake and the weight gain per piglet during that period. Diarrhea scores were based on pen measurements using a scale of 0 to 3 (0 = normal; 1 = soft stool; 2 = mild diarrhea; 3 = severe diarrhea).

[0168] Statistical analysis

[0169] The production of violacein by *Synthobacterium violaceum* and biofilm formation in *Escherichia coli* were analyzed using the SAS glimmix program (SAS 9.4, SAS Institute Inc. (Cary, NC, USA)). The concentration of the substance was treated as a fixed effect, and the measurement date (biological replicate) was treated as a random effect.

[0170] The in vivo study design was a completely randomized block design, where columns served as experimental units for statistical purposes of parameter recording. Growth performance was analyzed using the glimmix program in SAS (SAS 9.4, SAS Institute Inc.). Stool scores were analyzed using one-way ANOVA with SPSS software (IBM SPSS version 21). Treatments were used as fixed effects, and results are expressed as least squares mean and standard error of the mean. Body weight on day 1 was used as a cofactor in the analysis of body weight (BW) on day 15, BW on day 42, ADG, and ADFI.

[0171] Results

[0172] Effects of selected plant-derived substances on bacterial growth, pyocyanin production and biofilm formation

[0173] In this study, the effects of ZnO, two plant-derived prototype essential oil cores (PFA core 1 and PFA core 2), and their corresponding major compounds, tea tree oil and carvacrol, on bacterial susceptibility, biofilm formation in *Escherichia coli* strains O88:H8 and O143:H4, and violacein production in *Syntrophus violaceus* were initially investigated. The minimum inhibitory concentrations (ppm) of the test substances against the three *E. coli* strains and *Syntrophus violaceus* are shown in Table 4 below. Treatment with each substance had similar effects on the growth of all bacterial strains. In particular, carvacrol showed very strong bactericidal effects (MIC 150 ppm–600 ppm), while tea tree oil showed significantly lower antimicrobial activity against all three *E. coli* strains (MIC 2560 ppm–10000 ppm) and *Syntrophus violaceus* (MIC 2500 ppm). ZnO did not inhibit the growth of *E. coli* and *Syntrophus violaceus* strains at the highest test concentration up to 10000 ppm. The MIC values ​​of the combination of PFA core 1 and PFA core 2 substances are similar to those of the corresponding individual substances.

[0174] Table 4

[0175]

[0176] nt = Not detected

[0177] The values ​​of biofilm formation and violacein production observed at each treatment concentration were compared with the untreated positive control (PC) to identify potential sub-MIC effects. Both parameters responded to a dose-dependent decrease in all tested substances within the sub-MIC range. Specifically, carvacrol reduced biofilm formation in both E. coli strains to 80 ppm. (See [link to relevant documentation]) Figure 1A (See Table 3) Interestingly, although carvacrol showed the same effect on biofilm formation in both *E. coli* strains, the two strains responded significantly differently to carvacrol-based PFA Core 2. In fact, a minimum concentration of 150 ppm of PFA Core 2 was required to significantly reduce biofilm formation in *E. coli* O88:H8 (-69.2%), compared to 40 ppm for *E. coli* O143:H4 (-27.1%). (See Table 3) Figure 1B (and Table 4)

[0178] Although tea tree oil has weak antimicrobial potential, it significantly reduced biofilm formation in two strains of *E. coli* by -96.6% to -48.5% at three to four sub-MICs. (See also...) Figure 1C Similar to Table 5, PFA core 1 significantly reduced biofilm formation in *E. coli* O88:H8 and O143:H4 at two and three sub-MICs, respectively (-87.7% to -44%). (See Table 5) Figure 1D (See Table 6) Although ZnO does not inhibit the growth of any *E. coli* strain, it does show a significant effect on its biofilm-forming ability, with the lowest effective concentration range being 150 ppm (-81.4%) (for *E. coli* O88:H8) to 80 ppm (-41.7%) (for *E. coli* O143:H4). (See Table 6) Figure 1E (and Table 7).

[0179] Table 5

[0180]

[0181] Table 6

[0182]

[0183] Table 7

[0184]

[0185] Table 8

[0186]

[0187] Table 9

[0188]

[0189] Significant differences (P<0.05) between treatment concentrations within each bacterial model are indicated by different letters in Tables 3-7. Statistical analysis was performed using the SAS glimmix program.

[0190] Carvacrol and PFA core 2 at 20 ppm as the minimum effective concentration most effectively reduced violetin production in *Syntrophus violaceus*. Tea tree oil and PFA core 1 showed similar potential for inhibiting violetin production at the first three sub-MICs (-96.5% to -32.4% at 1250 ppm to 300 ppm). Interestingly, 10 ppm tea tree oil slightly increased violetin production in *Syntrophus violaceus*. Comparable to observations of biofilm formation, ZnO reduced the violetin signal to 40 ppm, although it had no effect on the growth of *Syntrophus violaceus*. (See [link to relevant documentation]) Figures 1A to 1E )

[0191] Effects of carvacrol and tea tree oil on in vitro adhesion of F4+ pilus-bearing E. coli to piglet small intestine mucus

[0192] The effects of carvacrol and tea tree oil on the mucus-binding properties of F4+ *Escherichia coli* strains were evaluated in an in vitro mucus adhesion assay. Approximately 20% of the radiolabeled F4+ *Escherichia coli* strains were introduced into the microtiter wells and adhered to the mucus-coated layer. Figure 2 This indicates that all receptor sites in the mucus were occupied by the introduced bacterial cells that grew in the absence of the test product. Treatment with tea tree oil and carvacrol showed opposite effects on the adhesion of F4+ *E. coli* cells: exposure to tea tree oil reduced the proportion of attached cells at both test concentrations (-52.6% at 320 ppm and -15.8% at 32 ppm), while carvacrol enhanced bacterial adhesion by 24.9%.

[0193] Effects of plant-derived prototypes on piglet growth performance and health parameters

[0194] In vivo studies were conducted without any adverse technical events. The effects of ZnO and plant-derived prototypes PFA 1 and PFA 2 on growth performance parameters of post-weaning piglets during the 42-day feeding trial are shown in Table 10. Piglets receiving a pharmacological dose of 3 kg ZnO / ton of feed showed the best growth performance, with an ADG increase of 12.56% during the 42-day trial period, resulting in a 9.19% difference in BW at the end of the trial compared to NC. The two plant-derived prototypes did not differ significantly in their effects on piglets. However, compared to NC, PFA 2 significantly improved ADG (7.05% from day 1 to day 42) and FCR (-4.4% from day 1 to day 42), resulting in a 5.25% increase in final BW at the end of the study. In addition, dietary supplementation with PFA 2 significantly increased ADFI during the first two weeks of the study compared to the negative control. On the other hand, PFA 1 supplementation only significantly improved FCR.

[0195] Table 10

[0196]

[0197] BW = Body weight [kg]; ADG = Average daily weight gain [g]; ADG = Average daily feed intake [g]; FCR = Feed conversion ratio [g feed / g body weight gain]

[0198] The mean values ​​with different superscript letters differed significantly (P < 0.05).

[0199] The mortality rates observed over the entire 42-day study period were: NC: 1.6%, ZnO: 0.4%, PFA 1: 0.8%, and PFA 2: 0.4%. The total dosing rates were: NC: 6.4%, ZnO: 5.6%, PFA 1: 7.2%, and PFA 2: 5.5%. Clinical signs of PWD were observed in 2.4% (6 animals) of piglets fed a non-supplemented basal diet (NC) and in 1.2% (3 animals) or 0.8% (2 animals) of piglets fed diets supplemented with PFA 1 or PFA 2, respectively. Piglets receiving ZnO were not affected by clinical signs of PWD. The remaining piglets exhibited normal activity and alertness, normal coat and eyes, and normal feces and urine.

[0200] Fecal scores were recorded on a scale of 0 to 3 (0 = normal; 1 = soft stool; 2 = mild diarrhea; 3 = severe diarrhea). The effects of ZnO and plant-derived prototype PFA 1 and PFA 2 on fecal scores in post-weaning piglets during the 42-day feeding trial are shown in Table 11. No signs of severe diarrhea (grade 3) were observed during the in vivo study. Compared with the unsupplemented treatment group (0.65 ± 0.35; NC), ZnO supplementation and lower levels of PFA 2 improved fecal consistency during the initial period (0.18 ± 0.16 and 0.32 ± 0.21, respectively). During the subsequent growth period from day 15 to day 42, ZnO reduced fecal scores compared with NC and PFA 1.

[0201] Table 11

[0202]

[0203] The mean values ​​with different superscript letters differed significantly (P < 0.05).

[0204] Discussion

[0205] Many challenges in livestock production are related to bacterial infections, especially during critical early stages of animal life. Piglets are among the most susceptible livestock species in this regard, particularly during the post-weaning period when young animals face significant environmental, dietary, and social changes. One of the most common strategies for managing post-weaning diarrhea is the administration of pharmacological concentrations of zinc oxide.

[0206] Microbroth dilution assays of individual substances showed that carvacrol exhibited significantly stronger antibacterial efficacy than tea tree oil across all tested strains. However, application of potentially lethal concentrations of the substance could be a strong inducer of resistance development. Therefore, we explored the potential of plant-derived substances to inhibit the expression of pathogenic traits without exerting selective pressure on resistance development or impairing the “healthy” functions of the microbiome. Several bacterial models associated with quorum sensing were employed in this study to investigate the potential of plant-derived compositions.

[0207] The first test model was a biosensor strain of *Syntrophus violaceus*, which produces the purple pigment violacein in response to QS activity (McClean 1997). At sub-MIC concentrations, treatment of *Syntrophus violaceus* with either carvacrol or tea tree oil inhibited or reduced the production of violacein pigment (see [link to relevant documentation]). Figures 1A-1E These results indicate that both test substances can interfere with quorum sensing in this model strain at different concentrations.

[0208] Quorum sensing has also been shown to participate in biofilm formation. The protective environment of biofilms plays an important role in bacterial infection by conferring resistance against antibiotics and host defense systems. In this study, carvacrol and tea tree oil showed significant effects on biofilm formation in two field isolates of *E. coli* at sub-MIC concentrations, but carvacrol exhibited these effects at much lower concentrations than tea tree oil (see [link to study]). Figures 1A-1E Although the plant-derived substance showed slightly stronger antimicrobial effects against *Syntrophus violaceus* than against the two pathogenic *Escherichia coli* field isolates, it reduced strychnine production by *Syntrophus violaceus* within a sub-MIC range similar to that observed in biofilm formation by the *E. coli* field isolates, and thus reduced quorum sensing. Therefore, it is hypothesized that the observed effects of carvacrol and tea tree oil on biofilm formation are related to their interference with quorum sensing.

[0209] The third model used in this study assessed the effects of tea tree oil and carvacrol on the ability of F4+ *Escherichia coli* strains to adhere to the intestinal mucus of piglets. This is particularly important in livestock farming because F4+ *Escherichia coli* is one of the major pathogens associated with post-weaning diarrhea, and the production of F4 pili is regulated through quorum sensing. Carvacrol treatment resulted in a greater number of adherent cells, which may indicate increased expression of adhesion factors. The chemical stress induced by carvacrol may lead to self-aggregation events. This type of bacterial behavior serves to protect the bacteria involved from external stress.

[0210] Based on the detection results of individual plant-derived substances (not shown), two prototypes were formulated: PFA Core 1 and PFA Core 2. PFA Core 1 is based on tea tree oil, while the second prototype, PFA Core 2, is based on carvacrol. Subsequent studies of the MIC values, QS, and biofilm inhibitory properties of these two prototypes showed that the effects of the two PFA cores are related to the effects of their respective individual major components (Table 2).

[0211] In vitro tests of ZnO, used as a positive control in in vivo feeding studies, showed no bactericidal activity within the applied concentration range. Literature evidence suggests that particle size has a significant impact on the antibacterial activity of ZnO, with zinc oxide nanoparticles exhibiting a particularly strong effect. The combination of high average particle size and low water solubility may explain the lack of bactericidal activity of the pharmaceutical-grade ZnO used in this study. Nevertheless, strong inhibition of E. coli biofilm formation and Chromobacterium violaceum ...

[0212] Side effects such as the onset of toxicity during long-term use and the accumulation of the heavy metal zinc in the environment prompted the European Union to ban the use of pharmaceutical-grade ZnO from June 2022. Therefore, this study evaluated the in vivo potential of a plant-derived feed additive prototype formulated during the nursery period as a ZnO substitute in piglets.

[0213] In a similar study, the beneficial effects of ZnO on growth parameters in piglets were reported by Molist et al. (2011). Compared to an unsupplemented control, piglets receiving 3000 ppm ZnO showed an 8.2% increase in body weight on day 12 post-weaning, which correlated with a 9.1% increase on day 15 post-weaning in this study. These results in this study indicate that pharmacological doses of ZnO show the expected improvement in animal growth performance, accompanied by a reduction in the incidence of post-weaning diarrhea. This effect is independent of high pathogen morbidity, as overall mortality and dosing rates, along with growth performance, indicate good health across the entire animal population observed in this study. The incidence of diarrhea, i.e., only 1.1% of all animals requiring treatment for PWD, was lower than the average reported in the literature. In piglets that did not receive ZnO or antibiotic supplementation, the reported prevalence of diarrhea (PWD) ranged from 34% to 51.1% in Danish independent herds (Eriksen 2021, Carstensen 2015), 24% in Australian commercial pig herds (Van Breda 2017), and 3.6% to 14.3% in standardized Indian farms (Vinodh Kumar 2019). Variations in diarrhea and PWD incidence may be a result of different pen conditions, as, for example, low hygiene standards may lead to increased susceptibility to stress and diarrhea in piglets (Bonetti 2021).

[0214] Due to the low overall morbidity rate in piglets, it was difficult to achieve significant differences using experimental settings, and these differences could only be discussed if they were medically relevant. Both ZnO and PFA continued to improve piglet condition in fecal scores from day 1 to day 14 of the experiment. This ranking of results was also observed in growth performance, where PFA 2 performed closer to the group receiving ZnO. PFA 1 supplementation significantly improved FCR. These results reflect in vitro observations (data not shown), where PFA core 2 showed effectiveness compared to PFA core 1 in E. coli biofilm inhibition assays and Chlorella violaceum quorum sensing inhibition assays at much lower concentrations.

[0215] Conclusions

[0216] The results of this study indicate that the inclusion of carvacrol-based plant-derived feed additives in the diet improved growth parameters and fecal scores in weaned piglets, similar to those of ZnO. In vitro assays used suggest that this effect may be related to, but is not limited to, the ability of carvacrol to influence bacterial behavior and may interfere with the expression of virulence factors. Therefore, carvacrol-based plant-derived feed additives provide an acceptable alternative to traditional antimicrobial growth promoters in livestock feed, particularly in the swine industry, to improve animal welfare and increase productivity.

[0217] Example 2 - Exploring dietary strategies for weaned piglets challenged with ETEC

[0218] Objective: To evaluate the effect of feed supplementation on weaned piglets against challenge with enterotoxigenic Escherichia coli carrying F4 fimbriae (F4-ETEC).

[0219] Materials and methods :

[0220] Thirty piglets, with a body weight of 9.4 ± 1.28 kg, were weaned at 30 days of age. They were mixed-sex piglets in a 1:1 ratio. Based on the SNP2 gene, the piglets were susceptible to F4. They were vaccinated with 3.0 × 10⁻⁶ styrosine phosphate on day 9 post-weaning. 10 CFU F4-ETEC. Three treatment groups were used, with 10 piglets per treatment group and 5 piglets per pen. Piglets were allowed free access to pelleted feed from weaning until 21 days post-weaning. Each pen contained two feed troughs and a drinking nipple.

[0221] Four dietary treatment groups included a negative control group (NC) receiving a basal diet without supplementation, and a positive control group receiving colistin via drinking water from days 8 to 14, in addition to days 0–3. Treatment group 3 was fed a basal diet supplemented with PFA 2 composition at a dose of 1 g / kg of feed.

[0222] Plant-derived prototype PFA2 composition ( Figure 4 The product contains 66% fenugreek seed powder, 2.5% turmeric root powder, 1.4% licorice root powder, and 15.4% limestone. PFA Core 2 is added to the PFA2 composition at 14.7% (4.9% of the total amount of essential oil and remaining encapsulating matrix and carrier). Carvacrol and eugenol are encapsulated using the encapsulating matrix as follows... Figure 4 The concentration shown is microencapsulated. Star anise oil and a carrier (sipernate) are then microencapsulated as follows. Figure 4 The premix is ​​shown. The final concentrations of carvacrol and eugenol in the PFA2 composition are each 1.99%, and the final concentration of star anise oil is 0.9%. The contents of the basal feed are as described in Tables 1-3 above.

[0223] The processing groups are summarized in Table 12.

[0224] Table 12

[0225]

[0226] From day 0 to day 3 post - weaning, all 30 piglets received colistin orally via drinking water. On day 9, the piglets were inoculated with F4 - ETEC. Fecal samples were collected on days 8, 10, 11, 12, 13, 14, 16, and 18. Fecal consistency was scored on days 0, 8, 10, 11, 12, 13, 14, 16, 18, and 21.

[0227] Data handling: Data lost due to two piglets reaching the humane endpoint were treated as missing values in the dataset. The performance of these animals was included until the day they were removed from the experiment.

[0228] Fecal sample qPCR: Each sample was analyzed in triplicate in the F4 - ETEC qPCR. The average cycle threshold (CT) was used to determine the F4 - ETEC concentration. For some samples, if one CT was not determined (≥40 cycles), the average of the other two CTs was used. If two CTs were not determined, the third CT (<40 cycles) was used to quantify F4 - ETEC. The limit of detection (LOD) was set at 200 CFU F4 - ETEC / gram of feces. Samples with an undetermined CT (≥40 cycles) were given a value of 0.5 times the LOD (100 CFU). Finally, the F4 - ETEC concentration was log10 - transformed before statistical analysis.

[0229] Statistical analysis: GenStat for Windows ® Version 23 (VSN International Ltd (Hemel Hempstead, UK)) was used to analyze the experimental data. Missing values were estimated. Analyses were performed with and without excluding outliers. An observation was labeled as an outlier if the residual (fitted observation) exceeded 2.5 times the standard error of the residuals of the dataset. One - way analysis of variance (ANOVA) was used to analyze the F4 - ETEC concentration (on each day separately), with replicates as random effects. The chi - square test was used to analyze the occurrence of F4 - ETEC diarrhea and excretion. There was a significant difference when P≤0.05, and a nearly significant trend when 0.05 < P≤0.10.

[0230] Proximate diet (crude protein and fat) analysis is shown in Table 13.

[0231] Table 13

[0232]

[0233] Results :

[0234] Table 14 shows the percentage of pigs that had diarrhea (fecal score ≤4) during the experiment (n=10 animals / treatment group).

[0235] Table 14

[0236]

[0237] Different superscripts (AB) within a row indicate the trend between processing groups (0.05 ≤ P < 0.10).

[0238] Table 15 shows the percentage of pigs that excreted F4-ETEC during the experiment (n=10 animals / treatment group).

[0239] Table 15

[0240]

[0241] Table 16 shows the logarithmic values ​​during the experiment. 10 CFU / g feces represents the result of F4-ETEC excretion in piglets. Vaccination was performed on day 9 post-weaning.

[0242] Table 16

[0243]

[0244] Following inoculation of pigs with the F4-ETEC strain on day 9 post-weaning, the overall percentage of piglets with diarrhea was less than 50% throughout the experiment (Table 14). No diarrhea was observed in the PC group. On day 13 post-weaning, pigs in the NC group showed a trend toward a higher proportion of diarrhea compared to the PC group. Piglets receiving PFA2 showed a numerically similar or lower proportion of diarrhea compared to the NC group, but a numerically higher proportion compared to the PC group. The percentage of pigs excreting F4-ETEC during the experiment was generally lowest in the PC group (Table 15). This is well consistent with the absence of signs of diarrhea observed in pigs in the PC group after inoculation with the F4 ETEC strain on day 9 post-weaning (Table 14). Animals excreting F4 ETEC in the PC group showed a lower average log [missing information - likely a missing word or phrase]. 10CFU / g fecal values ​​(Table 16). These results together demonstrate the significant effect of colistin treatment in protecting piglets from artificial F4 ETEC attack. Although the differences between the NC and PFA2 groups were not significant, the results still indicate the beneficial effects of the plant-derived additive. Although there was no statistical significance on a single day, on average, 24.3% of piglets in the NC group showed signs of diarrhea, compared to 13.5% in the PFA2 group (Table 14). This was observed from day 9 post-weaning after F4 ETEC administration until the end of the study on day 18. This is further supported by the data in Tables 15 and 16, where fewer animals in the PFA2 group shed F4 ETEC strains on days 13, 14, and 16, and the log of F4 ETEC strains throughout the study was lower. 10 The CFU / g fecal value was numerically lower, except on day 10 post-weaning. It is speculated that on day 10 post-weaning, more pigs in the PFA2 group excreted F4 ETEC, while also exhibiting log... 10 The trend in CFU / g fecal values ​​may be due to the product interfering with the ability of *E. coli* strains to establish larger colonies in the gut. This is based on information generated using different assays (biofilm formation, in vitro mucus adhesion, and *Synthobacterium violaceum* quorum sensing assay), demonstrating the potential of the active compound of PFA2 to influence bacterial behavior. It is hypothesized that F4 ETECs had more difficulty establishing colonies in PFA2-supplemented animals compared to the NC group, resulting in reduced adhesion to the intestinal surface and a larger proportion of inoculum excreted at day 10 post-weaning compared to the NC group. This would subsequently lead to a reduction in pathogen stress that would yield beneficial effects throughout the study period.

[0245] Representative features of the present invention are listed in the following clauses. These representative features may exist alone or may be combined in any combination with one or more features disclosed in the text of this specification.

[0246] This invention is as described in the following clauses:

[0247] Clause 1: A method for feeding livestock, said method comprising:

[0248] Providing post-weaning (PW) piglets with a basal diet supplemented with a plant-derived composition, wherein the plant-derived composition comprises an essential oil core, herbal powder, and preferred fillers, wherein the essential oil core comprises essential oils including carvacrol, eugenol, and star anise oil, wherein the PW piglets exhibit improved growth performance compared to PW piglets provided with the basal diet without the plant-derived composition.

[0249] Clause 2: The method according to Clause 1, wherein the carvacrol accounts for 20% to 60% by weight of the essential oil.

[0250] Clause 3: The method according to Clauses 1 to 2, wherein the carvacrol constitutes 40% by weight of the essential oil.

[0251] Clause 4: The method according to any one of the preceding clauses, wherein the eugenol accounts for 10% to 50% by weight of the essential oil.

[0252] Clause 5: The method according to any one of the preceding clauses, wherein the eugenol accounts for about 40% by weight of the essential oil.

[0253] Clause 6: The method according to any one of the preceding clauses, wherein the star anise oil accounts for 10% to 30% by weight of the essential oil.

[0254] Clause 7: The method according to any one of the preceding clauses, wherein the star anise oil accounts for about 20% by weight of the essential oil.

[0255] Clause 8: The method according to any one of the preceding clauses, wherein the dosage of carvacrol in the basal feed is 10 ppm to 50 ppm, the dosage of eugenol is 10 ppm to 50 ppm and the dosage of star anise oil is 5 ppm to 50 ppm.

[0256] Clause 9: The method according to any one of the preceding clauses, wherein in the basal feed, the dose of carvacrol is about 20 ppm, the dose of eugenol is about 20 ppm and the dose of star anise oil is about 9 ppm.

[0257] Clause 10: The method according to any one of the preceding clauses, wherein the base feed comprises fenugreek seed powder.

[0258] Clause 11: The method according to any one of the preceding clauses, wherein the basic feed comprises fenugreek seed powder, licorice powder and / or turmeric powder.

[0259] Clause 12: The method according to any one of the preceding clauses, wherein the base feed comprises fenugreek seed powder in a dose of 200 ppm to 1000 ppm.

[0260] Clause 13: The method according to any of the preceding clauses, wherein the basic feed comprises fenugreek seed powder in a dose of about 660 ppm.

[0261] Clause 14: The method according to any one of the preceding clauses, wherein the base feed contains turmeric powder in a dose of 5 ppm to 50 ppm.

[0262] Clause 15: The method according to any one of the preceding clauses, wherein the base feed contains turmeric powder in a dose of about 25 ppm.

[0263] Clause 16: The method according to any one of the preceding clauses, wherein the base feed contains licorice powder in a dose of 1 ppm to 50 ppm.

[0264] Clause 17: The method according to any one of the preceding clauses, wherein the basic feed comprises licorice powder at a dose of about 14 pm.

[0265] Clause 18: The method according to any one of the preceding clauses, wherein the basal feed is supplemented with the plant-derived composition at a concentration of 0.25 kg / t to 2.5 kg / t in the basal feed.

[0266] Clause 19: The method according to any one of the preceding clauses, wherein the basal feed is supplemented with about 1 kg / t of the plant-derived composition.

[0267] Clause 20: The method according to any one of the preceding clauses, wherein one or more of the essential oils are microencapsulated, wherein the microencapsulated oil comprises an essential oil and an encapsulating matrix, and wherein the filler is limestone.

[0268] Clause 21: The method according to any one of the preceding clauses, wherein the plant-derived composition is provided to the PW piglets during any period from day 18 to day 70 and thereafter.

[0269] Clause 22: The method according to any one of the preceding clauses, wherein the improved growth performance includes improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG), reduced fecal excretion of pathogenic microorganisms and / or improved fecal score.

[0270] Clause 23: A PW piglet feed, said PW piglet feed comprising:

[0271] Basic feed; and

[0272] A plant-derived composition comprising an essential oil core and herbal powder, wherein the essential oil core comprises essential oils, wherein the essential oils comprise carvacrol oil, eugenol, and star anise oil, wherein the PW piglet feed contains the plant-derived composition at a content of 0.25 kg / t to 2.5 kg / t, and wherein the PW piglet feed contains carvacrol at a dosage of 10 ppm to 50 ppm, eugenol at a dosage of 10 ppm to 50 ppm, and star anise oil at a dosage of 5 ppm to 50 ppm, wherein the PW piglet feed improves growth performance compared to PW piglet feed without the plant-derived composition.

[0273] Clause 24: The PW piglet feed as described in Clause 23, wherein the PW piglet feed contains 20 ppm carvacrol, 20 ppm eugenol and 10 ppm star anise oil.

[0274] Clause 25: PW piglet feed according to any one of Clauses 23 to 24, wherein the feed further comprises fenugreek seed powder.

[0275] Clause 26: PW piglet feed according to any one of Clauses 23 to 25, wherein the feed further comprises turmeric powder.

[0276] Clause 27: PW piglet feed according to any one of Clauses 23 to 26, wherein the feed further comprises licorice powder.

[0277] Clause 28: PW piglet feed according to any one of Clauses 23 to 27, wherein the feed further comprises 200 ppm to 1000 ppm of fenugreek seed powder, 5 ppm to 50 ppm of turmeric powder and 1 ppm to 50 ppm of licorice powder.

[0278] Clause 29: PW piglet feed according to any one of Clauses 23 to 28, wherein the feed further comprises about 660 ppm of fenugreek seed powder, 25 ppm of turmeric powder and 14 ppm of licorice powder.

[0279] Clause 30: PW piglet feed according to any one of Clauses 23 to 29, wherein the basal feed is supplemented with about 1 kg / t of the plant-derived composition.

[0280] Clause 31: PW piglet feed according to any one of Clauses 23 to 30, wherein the plant-derived composition is provided to the PW piglets during any period from day 18 to day 70 and thereafter.

[0281] Clause 32: PW piglet feed according to any one of Clauses 23 to 31, wherein the improved growth performance includes improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG), and / or improved fecal score.

[0282] Clause 33: PW piglet feed according to any one of Clauses 23 to 32, wherein one or more of the essential oils are microencapsulated, wherein the microencapsulated oil comprises an essential oil and an encapsulating matrix.

[0283] Clause 34: A plant-derived composition comprising an essential oil core and an herbal powder, wherein the essential oil core comprises an essential oil, wherein the essential oil comprises 30% to 50% by weight of carvacrol, 30% to 50% by weight of eugenol, and 10% to 30% by weight of star anise oil, and wherein the herbal powder comprises fenugreek seed powder, turmeric powder, and / or licorice powder.

[0284] Clause 35: The composition according to Clause 34, wherein the essential oil contains 40% by weight of carvacrol in the essential oil.

[0285] Clause 36: The composition according to any one of Clauses 34 to 35, wherein the essential oil comprises 40% by weight of eugenol in the essential oil.

[0286] Clause 37: A composition according to any one of Clauses 34 to 36, wherein the essential oil comprises 20% by weight of star anise oil.

[0287] Clause 38: The composition according to any one of Clauses 34 to 37, wherein the fenugreek seed powder accounts for about 66% by weight of the composition, the turmeric powder accounts for about 2.4% to about 2.5% by weight, and the licorice powder accounts for about 1.4% to about 1.5% by weight.

[0288] Clause 39: The composition according to any one of Clauses 34 to 38, wherein the composition is used in the basal feed at a rate of 0.25 kg / t to 2.5 kg / t.

[0289] Clause 40: The composition according to any one of Clauses 34 to 39, wherein the composition is used in a basal feed at a concentration of 1 kg / t.

[0290] Clause 41: The composition according to any one of Clauses 34 to 40, wherein the composition is used in the basal feed at a dosage of 10 ppm to 50 ppm carvacrol, 10 ppm to 50 ppm eugenol and 5 ppm to 50 ppm star anise oil.

[0291] Clause 42: The composition according to any one of Clauses 34 to 41, wherein the amount of carvacrol is about 20 ppm, the amount of eugenol is about 20 ppm, and the amount of star anise oil is about 10 ppm.

[0292] Clause 43: The composition according to any one of Clauses 34 to 42, wherein the composition is used in the feed at a dosage of 200 ppm to 1000 ppm of fenugreek seed powder.

[0293] Clause 44: The composition according to any one of Clauses 34 to 43, wherein the composition is used in the feed at a dose of 660 ppm fenugreek seed powder.

[0294] Clause 45: The composition according to any one of Clauses 34 to 44, wherein the composition is used in the feed at a dose of 5 ppm to 50 ppm of turmeric powder.

[0295] Clause 46: The composition according to any one of Clauses 34 to 45, wherein the composition is used in the feed at a dose of 25 ppm turmeric powder.

[0296] Clause 47: The composition according to any one of Clauses 34 to 46, wherein the composition is used in the feed at a dose of 1 ppm to 50 ppm of licorice powder.

[0297] Clause 48: The composition according to any one of Clauses 34 to 47, wherein the composition is used in the feed at a dose of 14 ppm of licorice powder.

[0298] Clause 49: The composition according to any one of Clauses 34 to 48, wherein the composition is used in the feed with 660 ppm fenugreek seed powder, 25 ppm turmeric powder, 14 ppm licorice powder, 20 ppm carvacrol, 20 ppm eugenol and 10 ppm star anise oil.

[0299] Clause 50: A composition according to any one of Clauses 34 to 49, wherein one or more of the essential oils are microencapsulated oils, wherein the microencapsulated oils comprise an essential oil and an encapsulating matrix.

[0300] Clause 51: The composition according to any one of claims 34 to 50, wherein the composition further comprises a filler.

[0301] Clause 52: The composition according to any one of claims 34 to 51, wherein the composition further comprises limestone.

[0302] Clause 53: A composition according to any one of Clauses 34 to 52, wherein the amount of the essential oil in the composition is about 4% to 5% by weight of the composition.

[0303] Clause 54: A composition according to any one of Clauses 34 to 53, wherein the amount of the essential oil in the composition is about 4.8% to 5% by weight of the composition.

[0304] Clause 55: A composition according to any one of Clauses 34 to 54, wherein the amount of the herbal powder in the composition is from about 55% to about 75% by weight.

[0305] Clause 56: A composition according to any one of Clauses 34 to 55, wherein the amount of the herbal powder in the composition is about 70% by weight.

[0306] Clause 57: The method according to any one of Clauses 1 to 22, wherein the PW piglets carry PWD-related microorganisms.

[0307] Clause 58: A method for alleviating symptoms caused by PWD-associated microorganisms in PW piglets, the method comprising providing the PW piglets with a basal diet supplemented with a plant-derived composition, wherein the plant-derived composition comprises an essential oil core, herbal powder, and preferred filler, wherein the essential oil core comprises essential oils including carvacrol, eugenol, and star anise oil, wherein the PW piglets have a lower incidence of diarrhea and / or reduced fecal excretion compared to PW piglets provided with the basal diet not containing the plant-derived composition.

[0308] Clause 59: The method described in Clause 58, wherein the microorganism is pathogenic Escherichia coli.

[0309] Clause 60, the method according to any one of Clauses 58 to 59, wherein the microorganism is enterotoxigenic Escherichia coli (ETEC).

Claims

1. A method for feeding livestock, the method comprising: Providing post-weaning (PW) piglets with a basal diet supplemented with a plant-derived composition, wherein the plant-derived composition comprises an essential oil core, herbal powder, and preferred fillers, wherein the essential oil core comprises essential oils including carvacrol, eugenol, and star anise oil, wherein the PW piglets exhibit improved growth performance compared to PW piglets provided with the basal diet without the plant-derived composition.

2. The method according to claim 1, wherein the carvacrol accounts for 20% to 60% by weight of the essential oil, preferably 40% by weight of the essential oil.

3. The method according to any one of the preceding claims, wherein the eugenol accounts for 10% to 50% by weight of the essential oil, preferably the eugenol accounts for about 40% by weight of the essential oil.

4. The method according to any one of the preceding claims, wherein the star anise oil accounts for 10% to 30% by weight of the essential oil, preferably the star anise oil accounts for about 20% of the essential oil.

5. The method according to any one of the preceding claims, wherein the dosage of carvacrol in the basal feed is 10 ppm to 50 ppm, the dosage of eugenol is 10 ppm to 50 ppm, and the dosage of star anise oil is 5 ppm to 50 ppm.

6. The method according to any one of the preceding claims, wherein the base feed comprises fenugreek seed powder, licorice powder and / or turmeric powder.

7. The method according to any one of the preceding claims, wherein the basic feed comprises fenugreek seed powder at a dose of 200 ppm to 1000 ppm, turmeric powder at a dose of 5 ppm to 50 ppm, and / or licorice powder at a dose of 1 ppm to 50 ppm.

8. The method according to any one of the preceding claims, wherein the basal feed is supplemented with the plant-derived composition at a concentration of 0.25 kg / t to 2.5 kg / t, preferably about 1 kg / t.

9. The method according to any one of the preceding claims, wherein one or more of the essential oils are microencapsulated, wherein the microencapsulated oil comprises an essential oil and an encapsulating matrix, and wherein the filler is limestone.

10. The method according to any one of the preceding claims, wherein the plant-derived composition is provided to the PW piglets during any period from day 18 to day 70 and therebetween.

11. The method according to any one of the preceding claims, wherein the improved growth performance includes improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG), and / or improved fecal score.

12. The method according to any one of the preceding claims, wherein the PW piglets carry PWD-related microorganisms.

13. A PW piglet feed, said PW piglet feed comprising: Basic feed; and A plant-derived composition comprising an essential oil core and herbal powder, wherein the essential oil core comprises essential oils, wherein the essential oils comprise carvacrol oil, eugenol, and star anise oil, wherein the PW piglet feed comprises the plant-derived composition at a content of 0.25 kg / t to 2.5 kg / t, preferably at a content of about 1 kg / t, and wherein the PW piglet feed comprises carvacrol at a dosage of 10 ppm to 50 ppm, eugenol at a dosage of 10 ppm to 50 ppm, and star anise oil at a dosage of 5 ppm to 50 ppm, wherein the PW piglet feed improves growth performance compared to PW piglet feed without the plant-derived composition.

14. The PW piglet feed according to any one of claims 13, wherein the feed further comprises fenugreek seed powder, turmeric powder and / or licorice powder.

15. The PW piglet feed according to any one of claims 13 to 14, wherein the feed further comprises 200 ppm to 1000 ppm of fenugreek seed powder, 5 ppm to 50 ppm of turmeric powder and 1 ppm to 50 ppm of licorice powder.

16. The PW piglet feed according to any one of claims 13 to 15, wherein one or more of the essential oils are microencapsulated, wherein the microencapsulated oil comprises an essential oil and an encapsulating matrix.

17. A plant-derived composition comprising an essential oil core and herbal powder, preferably, the herbal powder being present in an amount of about 55% to about 75% by weight, wherein the essential oil core comprises an essential oil, wherein the essential oil comprises 30% to 50% by weight of carvacrol, preferably 40% by weight of carvacrol; 30% to 50% by weight of eugenol, preferably 40% by weight of eugenol; and 10% to 30% by weight of star anise oil, preferably 20% by weight of star anise oil, and wherein the herbal powder comprises fenugreek seed powder, turmeric powder, and / or licorice powder.

18. The composition according to any one of claims 17, wherein the composition is used in a basal feed at a rate of 0.25 kg / t to 2.5 kg / t, preferably at a rate of 1 kg / t.

19. The composition according to any one of claims 17 to 18, wherein the composition is used in the basal feed at a dosage of 10 ppm to 50 ppm carvacrol, 10 ppm to 50 ppm eugenol and 5 ppm to 50 ppm star anise oil.

20. The composition according to any one of claims 17 to 19, wherein the composition is used in the feed at a dose of 200 ppm to 1000 ppm fenugreek seed powder, a dose of 5 ppm to 50 ppm turmeric powder, and / or a dose of 1 ppm to 50 ppm licorice powder.

21. The composition according to any one of claims 17 to 20, wherein the composition is used in the feed with 660 ppm fenugreek seed powder, 25 ppm turmeric powder, 14 ppm licorice powder, 20 ppm carvacrol, 20 ppm eugenol and 10 ppm star anise oil.

22. The composition according to any one of claims 17 to 21, wherein one or more of the essential oils are microencapsulated oils, wherein the microencapsulated oils comprise essential oils and an encapsulating matrix, preferably, the amount of the essential oils in the composition is from about 4.8% to 5% by weight of the composition.

23. A method for alleviating symptoms caused by PWD-associated microorganisms in PW piglets, the method comprising providing the PW piglets with a basal diet supplemented with a plant-derived composition, wherein the plant-derived composition comprises an essential oil core, herbal powder, and preferred filler, wherein the essential oil core comprises essential oils including carvacrol, eugenol, and star anise oil, wherein the PW piglets have a lower incidence of diarrhea and / or reduced fecal excretion compared to PW piglets provided with the basal diet without the plant-derived composition.

24. The method of claim 23, wherein the microorganism is pathogenic Escherichia coli.

25. The method according to any one of claims 23 to 24, wherein the microorganism is Escherichia coli F4-ETEC.

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