Unique blend of plant-derived components to support piglets during nursery phase post-weaning

Abstract

Phytogenic compositions with an essential oil core and herbal powders are used for feeding post-weaning piglets. The PW piglets provided basal feed supplemented with the phytogenic compositions have improved growth performance. The essential oil core includes three essential oils.

Description

UNIQUE BLEND OF PLANT-DERIVED COMPONENTS TO SUPPORT PIGLETS DURING NURSERY PHASE POST-WEANINGCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 518,692, filed August 10, 2023, which is incorporated by reference herein in its entirety.FIELD

[0002] This disclosure relates to feeding piglets and more particularly relates to plant- derived compositions provided to post-weaning piglets.BACKGROUND

[0003] Post-weaning diarrhea (PWD) is a problem swine producers face worldwide, and it is characterized by diarrhea, dehydration, mortality, and growth retardation in surviving pigs. The impact ranges from mild reductions in the growth performance to severe weight loss and mortality. An etiological agent commonly associated with PWD are pathogenic strains of enterotoxin producing Escherichia coli.

[0004] Many factors, including husbandry, environment, nutrition, and genetic susceptibility, play a critical role in the severity of this syndrome. Therefore, the control of PWD usually has to be approached at different levels: husbandry practices at the sow farms and nursery sites with proper sanitation and vaccination programs; minimize the stress after weaning by providing a fresh and easily accessible feed and water and an adequate environment (temperature and humidity) and space to the newly weaned pigs.

[0005] Management of nutritional programs is also an essential aspect of controlling PWD. Conventional swine production systems rely on the in-feed application of antibiotics, pharmacological levels of zinc, fiber, high digestible protein, spray-dried plasma, dietary acidification, functional feed additives, and others to help the newly weaned pigs to overcome post-weaning challenges. Antibiotics and other in-feed antimicrobial growth promoters such as zinc oxide are phased out more and more globally due to rising concerns regarding the development of bacterial resistances to clinically important antibiotic and accumulation in manure and the environment. Therefore, regulatory provisions, combined with an increasing pressure of consumers demanding more “natural” solutions, necessitates the search for alternatives to traditional antimicrobial growth promoters (AGPs) to maintain efficient farming in the future.Especially, promising in this regard are plant-derived bio-active compounds (hereinafter referred to as phytogenies).SUMMARY

[0006] PWD is a syndrome affecting piglets with a major impact on welfare, growth performance and cost efficacy of swine production. The present description relates to a unique blend of plant-derived components to support post-weaning (PW) piglets. The plant-derived components interact specifically with the intestinal tract and its microbiome, targeting microorganisms to reduce pathogenicity and increasing intestinal barrier integrity. Bacteria such as pathogenic E. coh. an etiological agent most commonly associated with PWD, are influenced in their ability to express virulence factors or invade intestinal epithelia. Successful prevention and / or mitigation of pathogenicity supports growth performance, animal welfare and antibiotic free management of livestock production, both in terms of use as growth promoters and limiting the need for medication to treat disease.

[0007] The present description relates to modem livestock production methods to support piglets during the critical PW period. The methods and compositions described herein therefore are part of modern health and nutrition management strategies to support piglets during the critical weaning period.

[0008] In one aspect, the present disclosure provides a method of feeding a livestock animal. The method includes providing a basal feed supplemented with a phytogenic composition to post-weaning piglets, wherein the phytogenic composition comprises an essential oil core, herbal powders and preferably, a bulking agent. The essential oil core comprises essential oils, wherein the essential oils comprise carvacrol, eugenol and star anise oil, wherein the PW piglets have improved growth performance relative to PW piglets provided the basal feed without the phytogenic composition.

[0009] The phytogenic composition preferably comprises carvacrol at between 20% by weight and 60% by weight of the essential oil mixture. The phytogenic composition more preferably comprises carvacrol at 40% by weight of the essential oils. The phytogenic composition preferably comprises eugenol at between 10% by weight and 50% by weight of the essential oils. The phytogenic composition more preferably comprises eugenol at about 40% of the essential oils. The phytogenic composition preferably comprises star anise oil between 10% by weight and 30% by weight of the essential oils. The phytogenic composition more preferably comprises star anise oil at about 20% of the essential oils. The one or more of the essential oils aremicroencapsulated, wherein the microencapsulated oils comprise essential oils and encapsulation matrix. The bulking agent in the phytogenic composition may be limestone.

[0010] The basal feed is preferably supplemented with the carvacrol at a dose of between 10 ppm and 50 ppm, the eugenol at a dose of between 10 ppm and 50ppm and the star anise oil at a dose of between 5 ppm and 50 ppm. The basal feed is more preferably supplemented with carvacrol at a dose of about 20 ppm, the eugenol at a dose of about 20 ppm and the star anise oil at a dose of about 9 ppm in the basal feed.

[0011] The herbal powders in the phytogenic composition preferably comprise fenugreek seed powder, more preferably the herbal powders comprise fenugreek seed powder, licorice powder and / or turmeric powder. The fenugreek seed powder in the basal feed is preferably at a dose of between 200 ppm and 1000 ppm, more preferably at a dose of about 660 ppm. The turmeric powder in the basal feed is at a dose of between 5 ppm and 50 ppm, more preferably at a dose of about 25 ppm. The licorice powder in the basal feed is preferably at a dose of between 1 ppm and 50 ppm, more preferably at a dose of about 14 ppm.

[0012] The basal feed in the method is preferably supplemented with between 0.25 kilogram / metric ton (kg / t) and 2.5 kg / t of the phytogenic composition, more preferably with about 1 kg / t of the phytogenic composition.

[0013] The method preferably provides the phytogenic composition to the PW piglets from day 18 to day 75, more preferably from day 20 to day 75, more preferably from day 21 to day 70 and any range there between. The method provides the phytogenic composition to healthy PW piglets and / or to PW piglets infected with a pathogenic microorganism. The method preferably improves the growth performance wherein the growth performance preferably comprises improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG) and / or improved fecal scores.

[0014] In another aspect, the present disclosure also provides a PW piglet feed. The PW piglet feed comprises a basal feed and a phytogenic composition, wherein the phytogenic composition comprises an essential oil core and herbal powders, wherein the essential oil core comprises essential oils, wherein the essential oils comprises carvacrol oil, eugenol and star anise oil, wherein the PW piglet feed comprises the phytogenic composition at an inclusion rate of between 0.25 kilogram / ton (kg / t) and 2.5 kg / t and wherein PW piglet feed comprises carvacrol at a dose between 10 ppm and 50 ppm, eugenol at a dose between 10 ppm and 50 ppm and star anise oil at a dose of between 5 ppm and 50 ppm, wherein the PW piglet feed improves growth performance relative to PW piglet feed without the phytogenic composition. The PW piglet feedpreferably comprises the phytogenic composition at an inclusion rate of 1 kg / t and wherein PW piglet feed comprises 20 ppm of carvacrol, 20 ppm of eugenol and 10 ppm of star anise oil. The PW piglet feed preferably further comprises fenugreek seed powder, more preferably fenugreek seed powder, turmeric powder, and licorice powder. The PW piglet feed preferably further comprises fenugreek seed powder at an amount between 200 ppm and 1000 ppm, turmeric powder at an amount between 5 ppm and 50 ppm and licorice powder at an amount between 1 ppm and 50 ppm. The PW piglet feed more preferably comprises about 660 ppm of fenugreek seed powder, 25 ppm of turmeric powder and 14 ppm of licorice powder. The PW piglet feed preferably comprises basal feed supplemented with about 1 kg / t of the phytogenic composition. The PW piglet feed is preferably provided to the PW piglets from day 18 to day 70 and any range there between. The method provides the phytogenic composition to healthy PW piglets and / or to PW piglets infected with a pathogenic microorganism. The PW piglet feed preferably provides improved growth performance comprising improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG) and / or improved fecal scores. The one or more of the essential oils are microencapsulated wherein the microencapsulated oils comprise essential oils and an encapsulation matrix. The phytogenic composition comprises a bulking agent, preferably limestone.

[0015] In a further aspect, the present disclosure further provides a phytogenic composition comprising an essential oil core and herbal powders wherein the essential oil core comprises essential oils, wherein the essential oils comprise carvacrol between 30% by weight and 50% by weight of the essential oils, eugenol between 30% by weight and 50% by weight of the essential oils and star anise oil between 10% by weight and 30% by weight of the essential oils and wherein the herbal powders comprise fenugreek seed powder, turmeric powder and / or licorice powder. The essential oils preferably comprise carvacrol at 40% by weight of the essential oils, more preferably the essential oils comprise carvacrol at 40% by weight and eugenol at 40% by weight of the essential oils, more preferably, the essential oils comprise carvacrol at 40% by weight, eugenol at 40% by weight and star anise oil at 20% by weight of the essential oils. The phytogenic composition preferably comprises the fenugreek seed powder at about 66% by wt. of the phytogenic composition, the turmeric powder between about 2.4% by wt. of the phytogenic composition and about 2.5% by wt. of the composition and the licorice powder between about 1.4% by wt. and about 1.5% by wt. of the composition. The phytogenic composition is preferably used at an inclusion rate in a basal feed at an amount between 0.25 kg / t and 2.5 kg / t, more preferably at an amount of 1 kg / t. The phytogenic composition is preferably in the PW piglet feedat a dosage of carvacrol between 10 ppm and 50 ppm, eugenol between 10 ppm and 50 ppm and star anise oil between 5 ppm and 50ppm, more preferably the dosage of carvacrol is about 20 ppm, the dosage of eugenol is about 20 ppm, and the dosage of star anise oil is about 10 ppm in the feed. The phytogenic composition is preferably added to the PW piglet feed at a fenugreek seed powder dose of between 200 ppm and 1000 ppm, more preferably at a dose of 660 ppm. The phytogenic composition is preferably added to the PW piglet feed at a turmeric powder dose of between 5ppm and 50 ppm, more preferably at a dose of 25 ppm. The phytogenic composition is preferably added to the PW piglet feed at a licorice powder dose between Ippm and 50 ppm, more preferably at a licorice powder dose at 14 ppm. The phytogenic composition is preferably added to the PW piglet feed at a dose of 660 ppm of fenugreek seed powder, 25 ppm of turmeric powder, 14ppm of licorice powder, 20 ppm of carvacrol, 20 ppm of eugenol and 10 ppm of star anise oil. The one or more of the essential oils are microencapsulated oils, wherein the microencapsulated oils comprise essential oils and an encapsulation matrix. The phytogenic composition further comprises a bulking agent, preferably limestone. The phytogenic composition preferably comprises essential oils between about 4% by weight and 5% by weight of the composition, more preferably between about 4.8%% by weight and 5% by weight of the composition. The phytogenic composition preferably comprises the herbal powders in the composition between about 65% by weight and 70% by weight, more preferably at about 67% by weight.

[0100] In yet a further aspect, the present disclosure provides a method of mitigating symptoms from PWD-associated microorganisms in PW piglets comprising providing a basal feed supplemented with a phytogenic composition to the PW piglets, wherein the phytogenic composition comprises an essential oil core, herbal powders and preferably, a bulking agent, wherein the essential oil core comprises essential oils, the essential oils comprise carvacrol, eugenol and star anise oil, wherein the PW piglets have lower incidence of diarrhea and / or reduced fecal shedding relative to PW piglets provided the basal feed without the phytogenic composition. The PWD-associated microorganisms may be pathogenic E. coli. The E. coli may be enterotoxigenic E. coli (ETEC).BRIEF DESCRIPTION OF THE DRAWINGS

[0016] This patent or application contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawings will be provided by the Office upon request and the payment of the necessary fee.

[0017] The drawings illustrate generally, by way of example, but not by way of limitation, various aspects discussed herein.

[0018] FIG. 1A is a plot showing the effect of carvacrol on biofilm formation of E. coli strains O143:H4 and O88:H8 and violacein production of C. violaceum. (A) Results after 18 h incubation are presented as mean ± standard deviation of absorbance values, which were normalized to the positive control (growth without test substance).

[0019] FIG. IB is a plot showing the effect of PFA Core 1 on biofilm formation of E. coli strains O143:H4 and O88:H8 and violacein production of C. violaceum. (A) Results after 18 h incubation are presented as mean ± standard deviation of absorbance values, which were normalized to the positive control (growth without test substance).

[0020] FIG. 1C is a plot showing the effect of PFA Core 2 on biofilm formation of E. coli strains O143:H4 and O88:H8 and violacein production of C. violaceum. (A) Results after 18 h incubation is presented as mean ± standard deviation of absorbance values, which were normalized to the positive control (growth without test substance).

[0021] FIG. ID is a plot showing the effect of tea tree oil on biofilm formation of E. coli strains O143:H4 and O88:H8 and violacein production of C. violaceum. (A) Results after 18 h incubation are presented as mean ± standard deviation of absorbance values, which were normalized to the positive control (growth without test substance).

[0022] FIG. IE is a plot showing the effect of ZnO on biofilm formation of E. coli strains 0143 :H4 and O88:H8 and violacein production of C. violaceum. (A) Results after 18 h incubation are presented as mean ± standard deviation of absorbance values, which were normalized to the positive control (growth without test substance).

[0023] FIG. 2 is a plot of Mucus-bound F4+ fimbriated E. coli presented as percentage of introduced labelled bacteria.

[0024] FIG. 3 is a schematic diagram of one exemplary formulation of PFA1.

[0025] FIG. 4 is a schematic diagram of one exemplary formulation of PFA2.

[0026] FIG.5 is a schematic diagram of another exemplary formulation of PFA2.DETAILED DESCRIPTION

[0027] Reference will now be made in detail to certain aspects of the disclosed subject matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will beunderstood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.

[0028] In this document, the terms “a,” “an,” or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.

[0029] Values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range were explicitly recited. For example, a range of “about 0.1% to about 5%” or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.

[0030] Unless expressly stated, ppm (parts per million), percentage, and ratios are on a by weight basis. Percentage on a by weight basis is also referred to as wt% or % (wt) below.

[0031] The present disclosure relates to a method of feeding piglets during the PW phase by providing basal feed supplemented with phytogenic compositions. The PW piglets are provided the phytogenic compositions during the post-weaning stage to minimize post-weaning diarrhea (PWD) and improve growth performance. The phytogenic compositions are provided to healthy PW piglets and / or to PW piglets infected with a pathogenic microorganism. The basal feed is combined with the phytogenic compositions in described inclusion rates and doses for improved growth performance by PW piglets.

[0032] The phytogenic compositions comprise an essential oil core, herbal powders and preferably, a bulking agent. The essential oil core includes a mixture of essential oils. Preferably, the essential oils include at least three essential oils at the disclosed percentages and / or ratios. The herbal powders include one or more herbal powders. In one aspect, the mixture of essential oilscomprise carvacrol, eugenol and star anise oil. In another aspect, the mixture of essential oils comprise tea tree oil, garlic oil, and citrus oil. The herbal powders included in the phytogenic composition preferably include fenugreek seed powder, licorice powder, turmeric powder and combinations thereof. One or all of the essential oils may be microencapsulated. Microencapsulation may be performed prior to combining the essential oils or after combining the essential oils.

[0033] The term “post-weaning piglets” or “PW piglets” as used herein relates to the period from weaning of piglets until the start of the fattening stage. The PW phase as referred to herein is from about day 25 of life to about day 75 of life. It will be understood that the exact day of weaning may vary and post-weaning refers to any day of life when the weaning of piglets is initiated. The terms “PW phase”, “PW stage” and “PW period” are used interchangeably.

[0034] The term “phytogenic” as used herein relate to plant-derived herbs, spices or extracts and / or bioactive compounds found in plants. Phytogenic includes natural products directly derived from plants. Phytogenic also includes synthetic and / or nature-identical compounds / compositions that can be made artificially but are in their chemical composition identical to molecules found in plants. Phytogenic also includes synthetic and / or nature-identical compositions that are formed by combining or formulating individual components, natural and / or synthetic components, to form a composition that is found naturally. The present description will refer to plant-derived compounds, but it will be understood that synthetic / nature-identical compounds or compositions may also be used.

[0035] The term “phytogenic composition” as used herein relates to a composition comprising an essential oil mixture and herbal powders. The phytogenic compositions described herein are standardized, specific and science-based combinations of bioactive compounds found in plants with proven efficacy and a sustainable impact on animals, people, and / or the environment. The phytogenic composition is combined or included in basal piglet feed to generate PW piglet feed.

[0036] The term “plants” and plant derivatives” as used herein relates to any portion of a growing plant, including the roots, stems, stalks, leaves, branches, seeds, flowers, fruits, and the like.

[0037] The term “essential oil mixture” or “essential oils” as used herein relates to a combination of essential oils in an essential oil core, preferably at least 3 essential oils, that are included in the phytogenic composition. Essential oils and essential oil mixture will be used interchangeably. “Essential oils” as used herein relates to aromatic, volatile liquids extracted fromplant material. Essential oils are often concentrated hydrophobic liquids containing volatile aroma compounds. Essential oil chemical constituents can fall within a wide range of chemical classes such as terpenes, terpenoids, phenols, aldehydes, ketones and phenylpropanoids. Essential oils can be natural (i.e., derived from plants), synthetic or a combination to achieve a nature identical essential oil.

[0038] The term “essential oil core” as used herein relates to the mixture of essential oils that are included and further includes carriers and / or an encapsulation matrix. One or more of the essential oils may be microencapsulated with an encapsulation matrix and / or combined with a carrier to form the essential oil core.

[0039] The term “carvacrol” as used herein relates to 2-methyl-5-propan-2-ylphenol which can be derived from plants, synthetically produced or a combination to achieve a nature identical carvacrol. Carvacrol can be derived from, for example, oregano (Origanum vulgar e), thyme (Thymus vulgaris). Carvacrol derived from other plants can also be used and is also within the scope of this disclosure.

[0040] The term “eugenol” as used herein relates to 2-methoxy-4-prop-2-enylphenol which can be derived from plants, synthetically produced or a combination to achieve a nature identical eugenol. Eugenol can be derived from, for example, clove (Syzygium aromalicum). nutmeg (Myristica fragrans). cinnamon (Cinnamomum verum), basil (Ocimum basilicum . and / or bay leaf (Taurus nobilis). Eugenol derived from other plants can also be used and is also within the scope of this disclosure.

[0041] The term “star anise oil” as used herein relates to an essential oil composed of mainly (>70%) 1 -methoxy -4-[(E)-prop-l-enyl]benzene (trans-anethole) which can be derived from plants, for example from star anise (Illicium verum). “Star anise oil” also can be composed of synthetically produced trans-anethole or a combination of natural and synthetic trans-anethole. “Star anise oil” also includes nature-identical compositions with one or more natural or synthetic components found in star anise oil combined to achieve the function of star anise oil. Essential oils derived from other plants and composed of mainly trans-anethole can also be used and are also within the scope of this disclosure. “Star anise oil” as used herein also includes pure transanethole.

[0042] The term “tea tree oil” as used herein relates to an essential oil composed of mainly terpinene-4-ol ([lS]-4-methyl-l-propan-2-ylcyclohex-3-en-l-ol) and y-terpinene (l-methyl-4- propan-2-ylcyclohexa-l,4-diene) which can be derived from plants, for example from Melaleuca alternifolia. “Tea tree oil” also can be composed of synthetically produced terpinene-4-ol and y-terpinene or a combination of natural and synthetic terpinene-4-ol and y-terpinene. “Tea tree oil” also includes nature-identical compositions with one or more natural or synthetic components found in tea tree oil combined to achieve the function of tea tree oil. Essential oils derived from other plants and composed mainly of terpinene-4-ol and y-terpinene can also be used and are also within the scope of this disclosure. Tea tree oil as used herein includes about 50 to 80% terpinene- 4-ol ([lS]-4-methyl-l-propan-2-ylcyclohex-3-en-l-ol) and y-terpinene (l-methyl-4-propan-2- ylcyclohexa- 1 ,4-diene).

[0043] The term “garlic oil” as used herein relates to an essential oil composed of mainly allyl polysulfides like diallyl disulfide (3-(prop-2-enyldisulfanyl)prop-l-ene) and diallyl trisulfide (3-(prop-2-enyltrisulfanyl)prop-l-ene) which can be derived from plants, for example from garlic (Allium sativum). “Garlic oil” also can be composed of synthetically produced allyl polysulfides or a combination of synthetic and natural derived allyl polysulfides. “Garlic oil” also includes nature-identical compositions with one or more natural or synthetic components found in garlic oil combined to achieve the function of garlic oil. Essential oils derived from other plants and composed mainly of diallyl disulfide and diallyl trisulfide can also be used and are also within the scope of this disclosure.

[0044] The term “citrus oil” as used herein relates to an essential oil composed of mainly D-limonene ([4R]-l-methyl-4-prop-l-en-2-ylcyclohexene) which can be derived from plants, for example from lemon (Citrus x limon) and other members of the Rutaceae. “Citrus oil” can be composed of synthetically produced D-limonene or a combination of natural and synthetic D- Limonene. “Citrus oil” also includes nature-identical compositions with one or more natural or synthetic components found in citrus oil combined to achieve the function of citrus oil. Essential oils derived from other plants and composed mainly of D-limonene can also be used and are also within the scope of this disclosure.

[0045] The term “PFA core 2” or “PFA core 2 essential oil mixture” as used herein relates to essential oils core. The essential oil core may include encapsulation matrix, a carrier and combinations thereof. The essential oils comprise carvacrol, eugenol and star anise oil. These terms will be used interchangeably.

[0046] The term “PFA core 1” or “PFA core 1 essential oil mixture” as used herein relates to essential oils core. The essential oil core may include encapsulation matrix, a carrier and combinations thereof. The essential oils comprise tea tree oil, garlic oil and citrus oil. The two terms will be used interchangeably.

[0047] The term “PFA 2 composition” or “PFA 2” as used herein relates to a phytogenic composition comprising PFA core 2, herbal powders from fenugreek seeds, licorice, and turmeric and bulking agents, e.g., limestone.

[0048] The term “PFA 1 composition” or “PFA 1” as used herein relates to a phytogenic composition comprising PFA core 1, herbal powders from fenugreek seeds, licorice and turmeric and bulking agents, e.g., limestone and wheat bran.

[0049] The term “basal feed” as used herein relates to feed that is fed to the PW piglets during the post-weaning period. It includes the ingredients and nutrients needed for the PW piglets. Basal feed does not include the phytogenic compositions.

[0050] The term “PW piglet feed or “piglet feed” as used herein relates to basal feed supplemented with a phytogenic composition.

[0051] In one aspect, the present description comprises a method of feeding a livestock animal a basal feed combined with a phytogenic composition. The animal is preferably a piglet, more preferably a PW piglet. The PW piglet is preferably at least 21 days old, or more preferably at least 23 days old, or more preferably between 21 days and 25 days old at the initiation of the feeding method described herein. The feeding method may also be initiated when the PW piglets are older than 25 days old.

[0052] The method of feeding is initiated and / or provided to the PW piglets anytime during the PW period. The method of feeding described herein is provided to PW piglets between about day 21 and about day 75 of life. The method of feeding is preferably applied directly in cereal-based diets after weaning (transition from milk feeding to cereal-based diets) and is applied preferably for at least until 14 days PW but preferably at least until 42 days PW. It will be understood that weaning times differ depending on the region and swine production systems, but the method described herein can be initiated as soon as piglets are weaned.

[0053] The method of feeding described herein is provided to PW piglets is between about day 21 and about day 75 of life, preferably between days 21 and 70, more preferably between days 21 and 66, more preferably between days 21 and 60, more preferably between days 23 and 75, more preferably between days 23 and 70, more preferably between days 23 and 66, more preferably between days 23 and 60, more preferably between days 25 and 75, more preferably between days 25 and 70, more preferably between days 25 and 66, more preferably between days 25 and 60, more preferably between days 27 and 70, more preferably between days 27 and 66, more preferably between days 27 and 60.

[0054] In one aspect, the basal feed supplemented with a phytogenic composition is provided to PW piglets that are healthy, e.g. no signs of disease. Preferably, the healthy piglets are not infected by a pathogenic microorganism. The basal feed supplemented with a phytogenic composition may also be provided to PW piglets that are challenged due to a pathogenic infection. The pathogen can be, for example, pathogenic strains of E. coli, e.g. enterotoxigenic E. coli (ETEC). The ETEC can be F4-fimbriated enterotoxigenic E. coli (ETEC). The basal feed supplemented with a phytogenic composition may be provided to PW piglets that develop a pathogenic infection at any time during the PW period. Providing the basal feed supplemented with the phytogenic composition can mitigate the symptoms of a pathogenic infection.

[0055] The method comprises providing a basal feed supplemented with a phytogenic composition / formulation. The basal feed can vary. A variety of basal feeds are known in the art and appropriate for PW piglets. An example of basal feed for PW piglets formulated by Lindenberger Muhle, Germany, is disclosed in the Examples below (Table 1). Basal feeds also include basal feed manufactured by Agropodnik Masovice, a.s. in Masovice, Czech Republic or by Purina, USA. Other basal feeds appropriate for PW piglets are within the scope of this disclosure. The basal feed may be supplemented with the phytogenic compositions described herein at the inclusion rates and doses disclosed.

[0056] The phytogenic compositions comprise an essential oil core comprising a combination of preferably at least two essential oils, more preferably at least three essential oils. The essential oil core may include more than three essential oils or more than four essential oils. One or more of the essential oils on the essential oil core may be microencapsulated. A variety of combinations of essential oils can be used in the essential oil cores. The essential oil cores may comprise, for example, carvacrol, eugenol, trans-anethole, terpinene-4-ol, y-terpinene, diallyl disulfide, diallyl trisulfide, D-limonene, and combinations thereof. Essential oils included in the essential oil core comprise, for example, carvacrol, eugenol and star anise oil. Essential oils included in another exemplary essential oil core comprise for example, tea tree oil, garlic oil and citrus oil.

[0057] The phytogenic compositions described herein further comprise herbal powders in addition to the essential oil core. In one aspect, the phytogenic composition comprises fenugreek seed powder. The phytogenic composition may further comprise turmeric powder, turmeric oleoresin, licorice extracts and / or licorice root powder.

[0058] The phytogenic compositions may comprise an essential core wherein one or more of the essential oils are microencapsulated and / or premixed essential oils. Essential oils may bepremixed with carriers prior to combining to form the essential oil core. Carriers can include sipernate, bentonite, sepiolite and the like. The phytogenic compositions may also comprise one or more of the essential oils that are microencapsulated in an encapsulation matrix. Encapsulation matrix may include silica, proteins such as wheat and yeast protein, citric acid, natural polymers such as starch, maltodextrin, glucose-fructose syrup and the like.

[0059] The phytogenic compositions may further comprise bulking agents. Bulking agents include, for example, wheat bran, limestone, corncob, wheat semolina, phylite, dextrose, wheat starch, com starch, pea starch, potato starch and the like. Preferably, the phytogenic composition comprises wheat bran and / or limestone as the bulking agent(s).

[0060] In one aspect, the phytogenic compositions are combined with a basal feed at a rate that improves the growth performance of the PW piglets. The inclusion rate of the phytogenic composition in the basal feed can be between about 0.25kilogram / metric ton (kg / t) and about 2.5kg / t, preferably between about 0.25kg / t and about 2.0kg / t, more preferably between about 0.25kg / t and about 1.5kg / t, more preferably between about 0.5kg / t and about 2.5kg / t, more preferably between about 0.5kg / t and about 2.0kg / t, more preferably between about 0.5 kg / t and about 1.5 kg / t, more preferably between about 0.8 kg / t and about 1.2 kg / t, more preferably between about 0.9 kg / t and about 1.1 kg / t and more preferably about 1 kg / t.

[0061] In one aspect, the essential oil core comprises carvacrol and / or eugenol. The essential oil core, preferably, further comprises star anise oil. Essential oil core comprising carvacrol, eugenol and star anise oil will be referred to herein as the PFA core 2. In one aspect, the PFA core 2 consists essentially of carvacrol, eugenol and star anise oil. By consists essentially of, it is meant that the PFA core 2 may not include any other essential oils except carvacrol, eugenol and star anise oil.

[0062] The PFA core 2 may comprise microencapsulated essential oils wherein the carvacrol, eugenol and the star anise oil are combined to form an essential oil mixture and then microencapsulated in the presence of an encapsulation matrix as shown in FIG. 5. Alternatively, carvacrol and / or eugenol may be microencapsulated prior to mixing with the star anise oil to form a partially microencapsulated essential oil mix as shown in FIG. 4.

[0063] The PFA core 2 comprises carvacrol at an amount between 20% by weight and 80% by weight of the of the weight of all the essential oils in the PFA core 2 prior to any microencapsulation, preferably between 20% by weight and 60% by weight, preferably between 30% by weight and 70% by weight, more preferably between 30% by weight and 60% by weight, more preferably between 30% by weight and 50% by weight, more preferably between 35% byweight and 45% by weight, more preferably between 38% by weight and 42% by weight, more preferably between 39% by weight and 41% by weight, more preferably about 40% by weight of all the essential oils in the core.

[0064] The PFA core 2 may further comprise eugenol at an amount between 0% by weight and 80% by weight of the of all the essential oils in the PFA core 2 prior to microencapsulation, preferably between 10% by weight and 50% by weight, more preferably between 20% by weight and 50% by weight, more preferably between 30% by weight and 50% by weight preferably, more preferably between 35% by weight and 45% by weight, more preferably between 38% by weight and 42% by weight, more preferably between 39% by weight and 41% by weight, more preferably about 40% by weight.

[0065] The PFA core 2 comprises the combination of carvacrol and eugenol at an amount between about 70% by weight and about 90% by weight of all the essential oils in the PFA core 2 prior to microencapsulation, preferably about 75% by weight and about 85% by weight, more preferably about 80% by weight of all the essential oils in the PFA core 2 prior to mi croencap sul ati on .

[0066] The PFA core 2 further comprises star anise oil at an amount between 10% by weight and 30% by weight of the of all the essential oils in the PFA core 2 prior to microencapsulation, preferably between 15% by weight and 25% by weight, preferably between 18% by weight and 22% by weight, more preferably between 19% by weight and 21% by weight, more preferably about 20% by weight.

[0067] In one aspect, the PFA core 2 comprises the combination of carvacrol and eugenol at an amount between about 70% by weight and about 90% by weight of the, preferably about 75% by weight and about 85% by weight, more preferably about 80% by weight of all the essential oils in the PFA core 2. The PFA core 2 further comprises star anise oil at an amount between 10% by weight and 30% by weight of the of all the essential oils in the PFA core 2, preferably between 15% by weight and 25% by weight, preferably between 18% by weight and 22% by weight, more preferably between 19% by weight and 21% by weight, more preferably about 20% by weight all the essential oils in the PFA core 2.

[0068] The PFA core 2 may include essential oils or essential oil mixtures that are microencapsulated. The matrix for encapsulation may comprise modified starch and maltodextrins. The essential oils may be microencapsulated as described, for example, in EP patent 1419811 Al, incorporated herein by reference. Other methods of encapsulation may also be used and are within the scope of this description.

[0069] In one aspect, the present description includes PFA 2 composition comprising the PF A core 2, as described herein, herbal powders and a bulking agent. A variety of herbal powders may be used, and all are within the scope of this description. 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, more preferably the turmeric powder is organic turmeric powder.

[0070] The PFA 2 composition comprises the PFA core 2 at an amount between about 5% by weight and about 30% by weight of the PFA 2 composition, preferably between about 10% by weight and about 20% by weight, more preferably between about 12 % by weight and about 18% by weight, more preferably between about 14% by weight and about 15% by weight. [

[0071] The PFA core 2 in the PFA 2 composition may comprise an encapsulation matrix if the essential oils are microencapsulated. The PFA 2 composition may comprise an encapsulation matrix at a weight between about 5% by weight and about 20% by weight of the PFA 2 composition, preferably between about 5% by weight and about 15% by weight, more preferably between about 8% by weight and about 12% by weight, more preferably between about 9% and about 10% by weight of the PFA 2 composition.

[0072] The PFA 2 composition may further comprise a bulking agent. A variety of bulking agents are known in the art and all are within the scope of this description. Bulking agents can be, for example, limestone, wheat bran and the like. Preferably, the bulking agent is limestone. The PFA 2 composition may include a bulking agent at an amount between about 5% by weight and about 40% by weight of the PFA 2 composition, preferably about 10% by eight and about 30% by weight, more preferably between about 10% by weight and about 20% by weight, more preferably between about 14% by weight and about 16% by weight, more preferably between about 15% by weight and about 16% by weight of the PFA 2 composition.

[0073] The PFA 2 composition further comprises one or more herbal powders. PFA 2 composition may comprise fenugreek powder, turmeric powder, and licorice powder. PFA 2 composition further comprises fenugreek powder at an amount between about 50% by weight and 80% by weight of the of the PFA 2 composition, preferably between 60% by weight and 75% by weight, preferably between 60% by weight and 70% by weight, more preferably between 62% by weight and 70% by weight, more preferably about 66% by weight of the PFA 2 composition.

[0074] PFA 2 composition further comprises turmeric powder at an amount between 2% by weight and 3% by weight of the of the PFA 2 composition, preferably between 2.2% by weightand 2.8 % by weight, more preferably between 2.4% by weight and 2.6 % by weight, more preferably between about 2.5 % by weight and 2.6% by weight, more preferably 2.5% by weight.

[0075] PFA 2 composition further comprises licorice powder at an amount between 0.1% by weight and 5% by weight of the of the PFA 2 composition, preferably between 0.5% by weight and 3.5 % by weight, more preferably between 1% by weight and 2.5% by weight, more preferably between about 1.2 % by weight and 1.8% by weight, more preferably 1.4% by weight.

[0076] The PW piglet feed comprises PFA 2 composition at an inclusion rate of between about 0.25 kg / t and about 2.5kg / t, preferably between 0.25kg / t and 2.0kg / t, more preferably between about 0.25kg / t and about 1.5kg / t, more preferably between about 0.5kg / t and about 2.5kg / t, more preferably between 0.5kg / t and 2.0kg / t, more preferably between about 0.5 kg / t and about 1.5 kg / t, more preferably between about 0.8 kg / t and about 1.2 kg / t, more preferably between 0.9 kg / t and 1.1 kg / t and more preferably about 1 kg / t.

[0077] The amounts of the PFA 2 components, e.g., the essential oils in the essential oil mixture and the herbal powders, are suitable for improvement of the growth performance of the PW piglets. In one aspect, the fenugreek seed powder is included at an amount between about 200ppm and about 1500 ppm in the PW piglet feed, preferably between about 500ppm and about 1000 ppm in the PW piglet feed, more preferably between about 600ppm and about 750ppm, more preferably between about 640ppm and about 680ppm, more preferably about 660 ppm.

[0078] The turmeric powder is included at an amount between about 5 ppm and about 50 ppm in the PW piglet feed, preferably between about 15 ppm and about 35 ppm, more preferably between about 20 ppm and about 30 ppm, more preferably between 22 ppm and about 28 ppm and more preferably about 25 ppm.

[0079] The licorice root powder is included at an amount between about 1 ppm and about 50 ppm in the PW piglet feed, preferably between about 5 ppm and about 35 ppm, more preferably between about 10 ppm and about 25 ppm, more preferably between 12 ppm and about 18 ppm and more preferably about 14 ppm.

[0080] The carvacrol is included at an amount between about 10 ppm and about 50 ppm in the PW piglet feed, preferably between about 15 ppm and about 50 ppm, more preferably between about 15 ppm and about 40 ppm, more preferably between about 15 ppm and about 35 ppm, more preferably between about 15 ppm and about 30 ppm, more preferably between about 15 ppm and about 25 ppm, more preferably between about 10 ppm and about 30 ppm, more preferably between about 10 ppm and about 25 ppm, more preferably about 20 ppm.

[0081] The eugenol is included at an amount between about 10 ppm and about 50 ppm in the PW piglet feed, preferably between about 15 ppm and about 50 ppm, more preferably between about 15 ppm and about 40 ppm, more preferably between about 15 ppm and about 35 ppm, more preferably between about 15 ppm and about 30 ppm, more preferably between about 15 ppm and about 25 ppm, more preferably between about 10 ppm and about 30 ppm, more preferably between about 10 ppm and about 25 ppm, more preferably about 20 ppm.

[0082] The star anise oil is included at an amount between about 5 ppm and about 50 ppm in the PW piglet feed, preferably between about 5 ppm and about 40 ppm, more preferably between about 5 ppm and about 30 ppm, more preferably between about 5 ppm and about 25 ppm, more preferably between about 5 ppm and about 20 ppm, more preferably between about 5 ppm and about 15 ppm, more preferably between about 7 ppm and about 12 ppm, more preferably about 10 ppm.

[0083] In one aspect, the essential oil core comprises tea tree oil, garlic oil and citrus oil and will be referred to herein as the PFA core 1. In one aspect, the PFA core 1 consists essentially of tea tree oil, garlic oil and citrus oil. By consists essentially of, it is meant that the PFA core 1 may not include any other essential oils except tea tree oil, garlic oil and citrus oil.

[0084] The PFA core 1 comprises tea tree oil at an amount between about 50% by weight and about 80% by weight of the of the all the essential oils in PFA core 1, preferably between about 55% by weight and about 75% by weight, more preferably between about 62% by weight and about 68% by weight, more preferably about 65% by weight of all the essential oils in PFA core 1. The PFA core 1 comprises garlic oil at an amount between about 20% by weight and about 50% by weight of the weight of all the essential oils in PFA core 1, preferably about 32% by weight and about 38 % by weight and more preferably about 34% by weight and about 36% by weight. The PFA core 1 comprises citrus oil at an amount between about 1% by weight and about 2% by weight of the weight of all the essential oils in PFA core 1, preferably about 1.2% by weight and about 1.8 % by weight and more preferably about 1.4% by weight and about 1.6% by weight.

[0085] The PFA 1 composition comprises the PFA core 1, as described above, herbal powders, and one or more bulking agents. The herbal powders are preferably fenugreek seed powder and turmeric powder, more preferably the turmeric powder is organic turmeric powder. The bulking agents are preferably wheat bran and limestone.

[0086] The PFA 1 composition comprises the PFA core 1 at an amount between about 5% by weight and about 30% by weight of the PFA 1 composition, preferably between about 10% by weight and about 20% by weight, more preferably between about 12% by weight and about 18%by weight, more preferably between about 15% by weight and about 17% by weight, more preferably between about 16% by weight and about 17% by weight.

[0087] PFA 1 composition further comprises fenugreek powder at an amount between about 40% by weight and 60% by weight of the PFA 1 composition, preferably between 45% by weight and 55% by weight, more preferably between 48% by weight and 52% by weight, more preferably about 50% by weight of the PFA 1 composition.

[0088] PFA 1 composition further comprises turmeric powder at an amount between 2% by weight and 8% by weight of the PFA 1 composition, preferably between 3% by weight and 7% by weight, more preferably between 4% by weight and 6 % by weight, more preferably about 5 % by weight of the PFA 1 composition.

[0089] PFA 1 composition further comprises a bulking agent at an amount between about 15% by weight to about 40% by weight of the PFA 1 composition, preferably at an amount between about 20% by weight and about 30% by weight, more preferably between about 25% by weight and about 30% by weight, more preferably between about 28% by weight and about 29% by weight. The PFA 1 composition can comprise limestone as a bulking agent at an amount between about 10% by weight to about 20% by weight of the PFA 1 composition, preferably at an amount between about 13% by weight and about 18% by weight, more preferably between about 15% by weight and about 16% by weight. The PFA 1 composition can comprise wheat bran as a bulking agent at an amount between about 10% by weight to about 20% by weight of the PFA 1 composition, preferably at an amount between about 10% by weight and about 15% by weight, more preferably between about 13% by weight.

[0090] The PW piglet feed comprises PFA 1 composition at an inclusion rate of between about 0.25 kg / t and about 2.5kg / t, preferably between 0.25kg / t and 2.0kg / t, more preferably between about 0.25kg / t and about 1.5kg / t, more preferably between about 0.5kg / t and about 2.5kg / t, more preferably between 0.5kg / t and 2.0kg / t, more preferably between about 0.5 kg / t and about 1.5 kg / t, more preferably between about 0.8 kg / t and about 1.2 kg / t, more preferably between 0.9 kg / t and 1.1 kg / t and more preferably about 1 kg / t.

[0091] The amounts of the PFA 1 components, e.g., the essential oils in the essential oil mixture and the herbal powders, are suitable for improvement of the growth performance of the PW piglets. In one aspect, the fenugreek seed powder is included at an amount between about 400 ppm and about 600 ppm in the PW piglet feed, preferably between about 450 ppm and about 550 ppm, more preferably about 500 ppm. The turmeric powder is included at an amount betweenabout 30 ppm and about 70 ppm in the PW piglet feed, preferably between about 40 ppm and about 60 ppm and more preferably about 50 ppm.

[0092] The tea tree oil is included in the PW piglet feed at an amount between about 20 ppm and about 40 ppm in the PW piglet feed, preferably between about 25 ppm and about 35 ppm, more preferably about 30 ppm. The garlic oil is included at an amount between about 5 ppm and about 25 ppm in the PW piglet feed, preferably between about 10 ppm and about 20 ppm, more preferably about 15 ppm. The citrus oil is included at an amount between about 0.4 ppm and about 1 ppm in the PW piglet feed, preferably between about 0.5 ppm and about 0.8 ppm, more preferably about 0.6 ppm.

[0093] In one aspect, the present description includes a method for feeding an animal, preferably PW piglets, a basal feed supplemented with the phytogenic compositions described herein. Preferably, the method includes a basal feed supplemented with the PFA 1 composition or the PFA 2 composition. The method may include providing the PW piglets ad libitum access to the feed. The method may include providing the PW piglets defined amounts of feed, e.g., not ad libitum access. Preferably, the method includes providing the PW piglets ad libitum access to the feed in a mash feed and / or pellet feed. Preferably, the method includes providing PW piglets feed in a mash feed. More preferably, the method includes providing the PW piglets ad libitum access to the feed in a mash form. Piglets are always provided access to water.

[0094] The method of feeding the PW piglets as described herein results in improved growth performance. The method includes providing the PW piglet feed, e.g., the basal feed supplemented with a phytogenic composition, described herein, to the PW piglets, preferably starting between day 21 and day 26. The PW piglets may be provided the piglet feed anytime during the PW phase. Improved growth performance results in improved average daily gain (ADG), improved feed conversion rate (FCR), increased average daily feed intake (ADFI) increased body weight (BW) and / or improved fecal scores compared to PW piglets fed only the basal feed, e.g., without the inclusion of the phytogenic composition. Improved growth performance also includes reduced incidence of PWD. Other improved growth performance indicators may also be improved and are within the scope of this description.

[0095] In the methods described herein, PWD-associated microorganisms such as pathogenic E. coli, are reduced in the intestinal microbiome of the PW piglets. This leads to reduced severity of PWD symptoms, i.e., diarrhea, dehydration, mortality, and growth retardation in surviving pigs. In PW piglets infected with PWD-associated microorganisms, the method can decrease the fecal shedding of the PWD-associated microorganisms, decrease the diarrhealoccurence and improve the fecal scores of the infected PW piglets. Improved growth performance, animal welfare and reduced need for medication are the direct effects when providing the phytogenic compositions described herein to piglets during the post-weaning period, especially under conditions of antibiotic free production. Advantageously, an additional benefit by using this unique blend of phytogenic composition as replacement for traditional antimicrobial growth promoters is a prevention of antibiotic resistance development through antibiotic use in livestock production.

[0096] Improvement of growth parameters will be described with reference to basal feed supplemented with PFA 2 composition, but it will be understood that other phytogenic compositions described herein, e.g., PFA 1 composition, also result in PW piglets having one or more of the improved growth parameters. The PW stage is generally about 45 days in length between about day 25 and about day 70 of piglet’s age. The PW period may vary by about + / - 7 days, e.g., from day 18 to about day 77.

[0097] Without being bound by any theory, the phytogenic compositions improve palatability and welfare of the animals. Sufficient feed intake early in life helps piglets develop a normal and functional gut, supporting their resilience against stress factors.

[0098] PW piglets supplemented with the PFA 2 composition described herein can have improved ADG relative to the PW piglets that are not provided the PFA 2 composition. The ADG is improved in PW piglets provided the PFA 2 composition by at least 1%, preferably at least 2%, more preferably by at least 4%, more preferably by at least 6%, relative to PW piglets that have not been provided feed supplemented with the PFA 2 composition.

[0099] PW piglets supplemented with the PFA 2 composition described herein can have improved FCR relative to the PW piglets that are not provided the PFA 2 composition. The FCR is improved in PW piglets provided the PFA 2 composition by at least 2%, preferably by at least 3%, more preferably by at least 4%, relative to PW piglets that have not been provided feed supplemented with the PFA 2 composition.

[0100] PW piglets supplemented with the PFA 2 composition described herein can have increased BW relative to the PW piglets that are not provided the PFA 2 composition. The BW is improved in PW piglets provided the PFA 2 composition by at least 2%, preferably by at least 4%, more preferably by at least 5%, relative to PW piglets that have not been provided feed supplemented with the PFA 2 composition.

[0100] PW piglets supplemented with the PFA 2 composition described herein can have improved ADFI relative to the PW piglets that are not provided the PFA 2 composition. The ADFIis improved in PW piglets provided the PFA 2 composition by at least 2%, preferably by at least 4%, more preferably by at least 6%, relative to PW piglets that have not been provided feed supplemented with the PFA 2 composition.

[0101] PW piglets supplemented with the PFA 2 composition described herein can maintain normal fecal consistency and / or improve the fecal consistency of the PW piglets. PW piglets supplemented with the PFA 2 composition described herein can have improved fecal scores relative to the PW piglets that are not provided the PFA 2 composition. Determination of fecal scores are known in the art and are described, for example, in Jensen et al., 2006 or Fairbrother et al., 2017.

[0102] PW piglets supplemented with the PFA 2 composition described herein can have reduced propensity for acquiring PWD during the PW stage. The incidence of PWD in PW piglets provided the PFA 2 composition is reduced by at least 10%, preferably by at least 25%, more preferably by at least 50%, more preferably by at least 75% relative to PW piglets that have not been supplemented with the PFA 2 composition.

[0103] PW piglets supplemented with the PFA 2 composition described herein can have reduced mortality by PWD during the PW stage. The incidence of mortality caused by PWD in PW piglets provided the PFA 2 composition is reduced by at least 10%, preferably by at least 25%, more preferably by at least 50%, more preferably by at least 75% relative to PW piglets that have not been supplemented with the PFA 2 composition.

[0104] PW piglets that have acquired a PWD-associated microorganism infection and are supplemented with the PFA 2 composition described herein can have reduction in or elimination of PWD symptoms. The PWD symptoms that can be affected include a reduction in the amount and / or the duration of fecal shedding of the PWD-associated microorganisms. The PWD symptoms that can also be affected include an increase in the fecal consistency scores. The incidence of PWD in PW piglets that have acquired an PWD-associated microorganism infection and provided the PFA 2 composition is reduced by at least 10%, preferably by at least 25%, more preferably by at least 50%, more preferably by at least 75% relative to PW piglets that have not been supplemented with the PFA 2 composition. The incidence of fecal shedding of the infectious pathogen in PW piglets that have acquired an PWD-associated microorganism infection and provided the PFA 2 composition is reduced by at least 10%, preferably by at least 25%, more preferably by at least 30%, more preferably by at least 40% relative to PW piglets that have not been supplemented with the PFA 2 composition.

[0105] In one aspect, the present description includes a PW piglet feed. The PW piglet feed comprises a basal feed as described above and a phytogenic composition as described herein, preferably the PW piglet feed comprises PF A core 2 core and / or PF A core 1, more preferably the PW piglet feed comprises PFA core 2. Preferably, the PW piglet feed is supplemented with PFA 2 composition and / or PFA 1 composition, more preferably the PW piglet feed is supplemented with PFA 2 composition. The PW piglet feed is supplemented with the PFA 2 composition or the PFA 1 composition at an inclusion rate of between about 0.25 kg of the composition / ton of the feed and about 2.5kg / t, preferably between 0.25kg / t and 2.0kg / t, more preferably between about 0.25kg / t and about 1.5kg / t, more preferably between about 0.5kg / t and about 2.5kg / t, more preferably between 0.5kg / t and 2.0kg / t, more preferably between about 0.5 kg / t and about 1.5 kg / t, more preferably between about 0.8 kg / t and about 1.2 kg / t, more preferably between 0.9 kg / t and 1.1 kg / t and more preferably about 1 kg / t.

[0106] In one aspect, the PW piglet feed comprises the fenugreek seed powder at an amount between about 200 ppm and about 1000 ppm in the PW piglet feed, preferably between about 500 ppm and about 800 ppm, more preferably between about 600 ppm and about 700 ppm, more preferably between about 640 ppm and about 680 ppm, more preferably about 660 ppm.

[0107] The PW piglet feed comprises turmeric powder at an amount between about 5 ppm and about 50 ppm in the PW piglet feed, preferably between about 15 ppm and about 35 ppm, more preferably between about 20 ppm and about 30 ppm, more preferably between 22 ppm and about 28 ppm and more preferably about 25 ppm.

[0108] The PW piglet feed comprises licorice root powder at an amount between about 1 ppm and about 50 ppm in the PW piglet feed, preferably between about 5 ppm and about 35 ppm, more preferably between about 10 ppm and about 25 ppm, more preferably between 12 ppm and about 18 ppm and more preferably about 14 ppm.

[0109] The PW piglet feed comprises carvacrol at an amount between about 10 ppm and about 50 ppm, preferably between about 15 ppm and about 50 ppm, more preferably between about 15 ppm and about 40 ppm, more preferably between about 15 ppm and about 35 ppm, more preferably between about 15 ppm and about 30 ppm, more preferably between about 15 ppm and about 25 ppm, more preferably between about 10 ppm and about 30 ppm, more preferably between about 10 ppm and about 25 ppm, more preferably about 20 ppm.

[0110] The PW piglet feed comprises eugenol at an amount between about 10 ppm and about 50 ppm, preferably between about 15 ppm and about 50 ppm, more preferably between about 15 ppm and about 40 ppm, more preferably between about 15 ppm and about 35 ppm, morepreferably between about 15 ppm and about 30 ppm, more preferably between about 15 ppm and about 25 ppm, more preferably between about 10 ppm and about 30 ppm, more preferably between about 10 ppm and about 25 ppm, more preferably about 20 ppm.

[0111] The PW piglet feed comprises star anise oil at an amount between about 5 ppm and about 50 ppm in the PW piglet feed, preferably between about 5 ppm and about 40 ppm, more preferably between about 5 ppm and about 30 ppm, more preferably between about 5 ppm and about 25 ppm, more preferably between about 5 ppm and about 20 ppm, more preferably between about 5 ppm and about 15 ppm, more preferably between about 7 ppm and about 12 ppm, more preferably about 10 ppm.

[0112] In one aspect, the PW piglet feed comprises the fenugreek seed powder at an amount between about 400 ppm and about 600 ppm, preferably between about 450 ppm and about 550 ppm, more preferably about 500 ppm. The PW piglet feed comprises turmeric powder at an amount between about 30 ppm and about 70 ppm, preferably between about 40 ppm and about 60 ppm and more preferably about 50 ppm. The PW piglet feed comprises tea tree oil at an amount between about 20 ppm and about 40 ppm, preferably between about 25 ppm and about 35 ppm, more preferably about 30 ppm. The PW piglet feed comprises garlic oil at an amount between about 5 ppm and about 25 ppm, preferably between about 10 ppm and about 20 ppm, more preferably about 15 ppm. The citrus oil is included at an amount between about 0.4 ppm and about 1 ppm in the PW piglet feed, preferably between about 0.5 ppm and about 0.8 ppm, more preferably about 0.6 ppm.EXAMPLES

[0113] The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.Example 1Materials and MethodsBacterial strains and test compounds

[0114] Chromobacterium violaceum DSM 30191 was purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany) and culturedin trypto-casein soy broth (Biokar Diagnostics, Allone, France; TSB) at 30°C under aerobic conditions.

[0115] Escherichia coli field isolates O88:H8 and O143:H4 (both isolated from pig) were provided by the veterinarian pathology department of AGES GmbH Linz (Austria) and serotyped by the national reference laboratory for Escherichia coli of AGES GmbH Graz (Austria). Both strains were classified as pathogenic due to their high-level growth in pure culture in various organs and their mucoid or hemolytic properties.

[0116] For the mucus adhesion assay, a F4+ fimbriated E. coli strain isolated in Finland was obtained as field isolate by Alimetrics Ltd (Espoo, Finland). The E. coli field isolates were cultured under aerobic conditions at 37°C in trypto-casein soy broth (Biokar Diagnostic), that was supplemented with 2 g / 1 glucose (Carl Roth GmbH + Co. KG, Karlsruhe, Germany; TSB+) for biofilm assays.

[0117] Zinc oxide for the bacterial assays was purchased by Auhof Apotheke (Linz, Austria). Carvacrol and tea tree oil as well as “PFA Core 1” and “PFA Core 2” were provided by Delacon Biotechnik GmbH (Engerwitzdorf, Austria)

[0118] For PFA Core 1, the essential oils are mixed at a ratio of 32.571 (tea tree oil): 16.236 (garlic oil):0.693 (citrus oil). In the PFA Core 1, the essential oil mix is the tea tree oil at 65.8% by weight, the garlic oil was at 32.8% by weight and the citrus oil was at 1.4% by weight. The PFA Core 1 inclusion rate in the feed was 49.5 ppm.

[0119] The PFA 1 composition is used at doses of 500 ppm fenugreek seed powder, 50ppm turmeric powder, 33 ppm tea tree oil, 16 ppm garlic oil, 0.7 ppm citrus oil in the basal feed.

[0120] For PFA Core 2, the essential oils are mixed at a ratio of 20 (carvacrol): 20 (eugenol):9 (star anise oil). The PFA 2 composition is used at doses of 660 ppm fenugreek seed powder, 25 ppm turmeric powder, 14 ppm licorice root powder, 20 ppm carvacrol, 20 eugenol, and 9 ppm star anise oil in the basal feed.Determination of Minimum Inhibitory Concentrations

[0121] The minimal inhibitory concentration (MIC) in each assay was optically determined as the lowest concentration of the test compound that completely inhibit visible growth (no obvious cell pellet or turbidity) after the incubation period (18 h at 30°C for C. violaceum, 18 h at 37°C for A. coli O88:H8 and 0143 :H4 and 24 h at 37°C for the F4+ fimbriated A. coli strain). The median value of all replicates was defined as the MIC value of the respective test compound.

[0122] For MIC evaluation of the F4+ fimbriated E. coli strain, carvacrol and tea tree oil were diluted in TSB containing 2% Tween 80 in a 2-fold dilution series.Quorum sensing inhibition assay (Chromobacterium violaceum)

[0123] The influence of the phytogenic test compounds on C. violaceum was assessed in concentrations ranging from 10000 ppm to 5 ppm. Violacein production was quantified based on the protocol of Skogman et al. (2016). Serial dilutions ranging from 20000 ppm to 10 ppm were prepared from stock solutions (200000 ppm) of the phytogenic test compounds in >99.8% ethanol (VWR International GmbH, Vienna, Austria). Four technical replicates of each concentration were arranged horizontally in Nunc® polystyrene 96-well microtiter plates (Thermo Scientific, Waltham, USA). An overnight culture of C. violaceum in 10 ml TSB was diluted to approximately 1-2 x io6CFU-ml'1in fresh TSB and aliquots of 50 pl were added to each test well and four wells containing only medium, serving as positive growth control without test compounds. An additional four wells containing only TSB were used as negative (sterility) controls. The starting bacterial concentration was verified using the colony count method. The microtiter plates as well as the colony count plates were incubated for 18 h at 30°C. After the incubation period, MIC values were determined before the cells and violacein pigment were pelleted by centrifugation at 3000 rpm for 10 min. The supernatant was removed from the wells by pipetting and 200 pl >99.8% ethanol (VWR International GmbH) were added to each well. The plates were sealed with Microseal 'B' PCR Plate Sealing Film (Bio-Rad, Hercules, USA) and incubated overnight in the dark to allow the pelleted violacein pigment to dissolve. The next day, the plates were centrifuged at 3000 rpm for 10 min to sediment the decolored cells. Subsequently, 100 pl of the violacein- stained supernatant were transferred to a new microtiter plate and the absorbance was measured at 570 nm using a PHOmo microplate reader (Anthos Mikrosysteme GmbH, Friesoythe, Germany).Biofilm inhibition assay

[0124] The effect of the phytogenic test compounds on biofilm formation of E. coli O88:H8 and 0143 :H4 was assessed in a broth micro-dilution assay followed by crystal violet staining according to Axmann et al. (2021) with minor modifications. Briefly, phytogenic test compounds were serially diluted in TSB+and arranged vertically in 96-well microtiter plates as described above in “Quorum sensing inhibition assay (Chromobacterium violaceum)'’'’ . The outer columns of the microtiter plates were filled with sterile TSB+to prevent evaporation from the central wells. Overnight cultures of the respective A. coli in TSB+were adjusted to approximately 2 x 106CFU-ml'1in fresh TSB+and 50 pl of this dilution were placed in each test well. Four wellscontaining either only bacterial culture without phytogenic test compounds or sterile TSB+medium served as positive or negative control, respectively. The plates were incubated under aerobic conditions for 18 h at 37 °C.

[0125] The antimicrobial activity of the phytogenic test compounds was evaluated as described above in section “Determination of Minimum Inhibitory Concentrations”. After removal of the culture medium, the microtiter plates were washed twice with deionized H2O (difLO) to remove the unattached cells and media components and air-dried for 30 min. The attached biomass was stained with 130 pl of a 0.1% (w / v) crystal violet solution (Alfa Aesar by Thermo Fisher GmbH, Kandel, Germany) in diH2O for 20 min at room temperature. Afterwards, the microtiter plates were washed 3 times with diH2O and left to air-dry completely. The adherent dye was solubilized with 130 pl 30% (v / v) acetic acid (Merck KgaA, Darmstadt, Germany) and transferred to a new microtiter plate. Absorbance was measured at 570 nm with a PHOmo microplate reader (Anthos Mikrosysteme GmbH). The mean of the four replicates was calculated followed by subtraction of the negative control measurements and the results were expressed as a percentage biofilm relative to the positive control. Each test compound was assayed three to four times. In this study, the entirety of adherent cell mass which was stained by the crystal violet solution is referred to as biofilm without distinction of the maturation stage.In vitro mucus adhesion assay

[0126] In preparation of the mucus adhesion assay, an overnight culture of F4+ fimbriated E. coli was grown in TSB without test compounds. From this seed culture, two successive overnight cultivations were performed for each treatment using 10% inoculum. All treatments were cultured in three replicate vials. The turbidity of the cultures was followed by measuring the absorbance at 600 nm to ensure that the cell densities were close to those of the control culture without a test product. On the third day, each culture was further diluted tenfold in growth medium containing the respective test product at its specific concentration ( 1 / 8thof the MIC, and in addition for tea tree oil a second concentration with l / 80thof the MIC value to reach a comparable concentration level to the tested carvacrol concentration). Tritium labelled thymidine was introduced in each vial to radioactively label the E. coli cells.

[0127] To prepare the in vitro mucus adhesion assay, authentic intestinal mucus was recovered from sacrificed piglets und used to coat the wells of microtiter plates. Mucus coated microtiter plates were washed with HEPES-Hank’s buffer before labelled E. coli cells were introduced (nine replicate wells per treatment) and incubated for 1 hour at 37°C. The unbound cells were removed by washing the wells twice with HEPES-Hank’s buffer. Scintillation liquidwas added prior to measuring the radioactivity of each well. Additionally, the total radioactivity of each labelled E. coli culture used in the adherence study was measured to be able to determine the proportion of adhered cells.Animals, diets, and in vivo trial design

[0128] A total of one thousand healthy post-weaning piglets (500 ; 500DanBred x Duroc), with an age at weaning of 25 ± 2 days, were allocated to four treatments and 100 pens in total (5 & 5 $ per pen, 25 pens per treatment). The average housing temperature was kept at about 30°C during the first week after weaning and was gradually reduced by 1.8°C per week down to 22°C from d 53 of age onwards. Post-weaning piglets had ad libitum access to feed provided in mash form and water supplied by drinking bowls throughout the experiment.

[0129] The four dietary treatments included a negative control group receiving an unsupplemented basal diet (negative control; NC), a positive control group whose diet was supplemented with zinc oxide at pharmacological level (3 kg / t of feed; ZnO) and the treatment groups PFA 1 and PFA 2 that were fed basal diets supplemented with PFA 1 composition or PFA 2 composition, respectively, at 1 kg / t of feed each.

[0130] Phytogenic prototypes PFA1 (FIG.3) included of 50% fenugreek seed powder, 5% turmeric root powder, 15.5% limestone, and 13% wheat bran. Microencapsulated PFA Core 1 was added to PFA1 at 16.5% (4.6% essential oil).

[0131] Phytogenic prototypes PF A2 composition (FIG.4) included of 66% fenugreek seed powder, 2.5% turmeric root powder, 1.4% licorice root powder and 15.4% limestone. PFA Core 2 was added to PFA2 composition at 14.7% (4.9% total amount of essential oil and the remaining encapsulation matrix and carrier. Carvacrol and eugenol were microencapsulated using an encapsulation matrix at concentrations as shown in FIG. 4. Star anise oil was premixed with a carrier (sipemate, 40.5%) as shown in FIG. 4. The final concentration in the PFA2 composition is 1.99% of each carvacrol and eugenol and 0.9%-star anise oil.

[0132] The 42-day observation period was divided into two feeding periods: a starter period of two weeks (from 25 to 38 days of age) and a subsequent grower period of four weeks (from 39 to 66 days of age). The basal diets for each period were formulated as recommended by the Society of Nutrition Physiology (2006) with exception of zinc. Diet composition and calculated nutrients of starter and grower diets are presented in Table 1 below. Delacon Biotechnik GmbH (Austria) supplied the phytogenic prototypes. Zinc oxide (Spezialfutter Neuruppin GmbH & Co. KG, Neuruppin, Germany) was supplemented at expense of Tixosil (> 97% silicon dioxide), while PFA 1 and PFA 2 were included at expense of limestone.Table 1

[0133] The formulation for basal feed / PFA 1 composition includes: a. 500 ppm Fenugreek seed powder b. 50 ppm turmeric powder c. 30 ppm Tea tree oil d. 15 ppm Garlic oil e. 0.625 ppm Citrus oil

[0134] The formulation for basal feed / PFA 2 composition includes: f. 660 ppm Fenugreek seed powder g. 25 ppm turmeric powder (organic) h. 14 ppm licorice root powder i. 20 ppm Carvacrol j . 20 ppm Eugenol k. 9 ppm Star anise oil*

[0135] Table 2 provides the feed ingredients and calculated analysis of the starter diets from 25 to 38 d of age (as is). Table 3 provides the feed ingredients and calculated analysis of the grower diets from 39 to 66 d of age. Treatment diet T1 is the NC, T2 includes the zinc oxide, T3 includes PFA1 composition and T4 includes PFA2 composition.Table 2Table 3Piglet growth performance and determination of diarrhea score

[0136] All piglets were observed twice daily for any abnormalities, abnormal behaviour, and clinical signs of sickness throughout the 42-d experimental period.

[0137] Pen body weights (BW) were recorded weekly, as was the amount of feed supplied to each pen during the preceding week. Piglet body weight gain was calculated by dividing the mean BW per pen at the end of each period by the mean BW per pen at the start of each period and number of piglets per pen. Feed consumption per piglet was estimated as the total feed supplied per pen and period corrected for dispersed / leftover feed and number of piglets per pen. Feed conversion ratio (FCR) was calculated from the relationship of weekly corrected feed intake and body weight gain per piglet for this period. Diarrhea scores were determined on pen-basis using a scale from 0 to 3 (0 = normal; 1 = soft feces; 2 = mild diarrhea; 3 = severe diarrhea).Statistical analysis

[0138] C. violaceum violacein production and E. coli biofilm formation was analyzed using the glimmix procedure of SAS (SAS 9.4, SAS Institute Inc., Cary, NC, USA) with substance concentration as fixed effect and day of measurement (biological replicate) as random effect.

[0139] The in vivo study design was a random complete block design, with pen as the experimental unit for statistical purposes for recoded parameters. Growth performance was analyzed using the glimmix procedure of SAS (SAS 9.4, SAS Institute Inc.). Feces scores were analyzed using a one-way ANOVA via software package SPSS (IBM SPSS Version 21). Treatment was used as fix effect and results were presented as least square means with standard error of the mean. Body weight day 1 was used as co-factor for analysis of BW at day 15, BW at day 42, ADG, and ADFI.ResultsEffect of selected phytogenies on bacterial growth, violacein production and biofilm formation

[0140] In the present study, the influence of ZnO, two phytogenic prototype essential oil cores (PF A Core 1 and PFA Core 2) and their respective main compounds tea tree oil and carvacrol was initially investigated on bacterial susceptibility, biofilm formation of E. coli strains 088 :H8 and 0143 :H4 and violacein production of C. violaceum. Minimum inhibitory concentrations oftested substances [ppm] against three E. coli strains and C. violaceum are shown in Table 4 below. Treatment with each substance exerted a similar effect on the growth of all bacterial strains. In particular, carvacrol showed very strong bactericidal effects (MIC of 150 - 600 ppm) whereas tea tree oil exhibited substantial lower antimicrobial activities against all three E. coli strains (MIC of 2560 - 10000 ppm) and C. violaceum (MIC of 2500 ppm). ZnO did not inhibit growth of the E. coli and C. violaceum strains up to the highest tested concentration of 10000 ppm. The MIC values of substance combinations PFA Core 1 and PFA Core 2 were similar to those of the respective individual substances.Table 4nt = not tested

[0141] The observed values for biofilm formation and violacein production at each treatment concentration were compared to an untreated positive control (PC) to identify potential sub-MIC effects. Both parameters decreased in a dose-dependent manner in the sub-MIC range in response to all tested substances). In particular, carvacrol reduced the biofilm formation of both E. coli strains down to 80 ppm. (See Fig. 1 A and Table 3) Interestingly, although carvacrol showed identical effects on the biofilm formation of both E. coli strains, the responses of the two strains to the carvacrol -based PFA Core 2 differed notably. In fact, a minimum concentration of 150 ppm PFA Core 2 was required to significantly reduce the biofilm formation of E. coli 088 :H8 (-69.2%) in contrast to 40 ppm for A. coli 0143 :H4 (-27.1%). (See Fig. IB and Table 4)

[0142] Tea tree oil significantly reduced the biofilm formation of both E. coli strains in three to four sub-MICs by -96.6% to -48.5%, despite its weak antimicrobial potential. (See Fig. 1C and Table 5) Likewise, PFA Core 1 significantly decreased (-87.7% to -44%) biofilm formation of E. coli O88:H8 and 0143 :H4 in two and three sub-MICs, respectively. (See Fig. ID and Table 6) While ZnO did not inhibit the growth of either A. coli strain, the substance did showa strong impact on their biofilm formation capabilities with the lowest effective concentration ranging from 150 ppm (-81.4%) forE. coli O88:H8 to 80 ppm (-41.7%) forE. coli 0143 :H4. (See Fig. IE and Table 7).Table 5Table 6Table 7Table 8Table 9

[0143] Significant differences ( <0.05) between treatment concentrations within each bacterial model are indicated by different letters in Tables 3-7. Statistical analysis was performed using the glimmix procedure of SAS.

[0144] Violacein production of C. violaceum was most efficiently reduced with carvacrol and PFA Core 2 at a dosage of 20 ppm as lowest effective concentration. Tea tree oil and PFA Core 1 exhibited similar potentials to inhibit violacein production in the first three sub-MICs (- 96.5% to -32.4% at 1250 - 300 ppm). Interestingly, 10 ppm tea tree oil slightly increased the violacein production of C. violaceum. Comparable to the observations on biofilm formation, ZnO decreased the violacein signal down to a concentration of 40 ppm, despite having no effect on the growth of C. violaceum. (See Fig. 1 A-Fig. IE)Effect of carvacrol and tea tree oil on adhesion of F4+ fimbriated coli to piglets’ small intestinal mucus in vitro

[0145] The effect of carvacrol and tea tree oil on mucus binding properties of an F4+ fimbriated E. coli strain was evaluated in an in vitro mucus adhesion assay. Close to 20% of the radioactive labelled F4+ fimbriated E. coli introduced into the microtiter wells adhered to the mucus coating (Fig. 2). This suggests that all receptor sites in the mucus became occupied by the introduced bacterial cells grown in the absence of the test products. Treatment with tea tree oil and carvacrol showed opposite effects on the adhesion of F4-fimbriated A", coli cells: Exposure to tea tree oil reduced the proportion of adhered cells in both tested concentrations (-52.6% at 320 ppm and -15.8% at 32 ppm) while carvacrol enhanced bacterial adhesion by 24.9%.Effect of phytogenic prototypes on piglet growth performance and health parameters

[0146] The in vivo study was run without any adverse technical events. Effects of ZnO and phytogenic prototypes PFA 1 and PFA 2 on growth performance parameters of post-weaning piglets during the 42-day feeding trial is shown in Table 10 Piglets receiving a pharmacological dosage of 3 kg ZnO per ton of feed showed the best growth performance with a 12.56% improved ADG during the 42-day trial period, resulting in a 9.19% difference on BW at the end of the trial compared to the NC. The two phytogenic prototypes did not differ significantly in their effect on the piglets. However, in comparison to the NC, PFA 2 significantly improved ADG (7.05% from day 1 to day 42) and FCR (-4.4% from day 1 to day 42) which lead to a 5.25% increased final BW at the end of the study. Additionally, dietary supplementation with PFA 2 caused ADFI to increase significantly during the first two weeks of the study compared to the negative control. Supplementation with PFA 1, on the other hand, only improved FCR to a significant extent.Table 10

[0147] BW = body weight [kg]; ADG = average daily gain [g]; ADG = average daily feed intake [g]; FCR = feed conversion ratio [g feed / g body weight gain]

[0148] Means bearing different superscript letters differ significantly with P<0.05

[0149] The mortality observed throughout the whole study period of 42 days was NC:1.6%, ZnO: 0.4%, PFA 1 : 0.8% and PFA 2: 0.4%. The medication rate amounted to NC: 6.4%, ZnO 5.6%, PFA 1 : 7.2% and PFA 2: 5.5%. Clinical signs of PWD were found in 2.4% (6 animals) of piglets fed the unsupplemented basal diet (NC) and 1.2% (3 animals) or 0.8% (2 animals) of piglets fed the diet supplemented with PFA 1 or PFA 2, respectively. No piglets receiving ZnO were affected by clinical signs of PWD. Remaining piglets showed normal activity and alertness, normal coat, and eyes as well as normal feces and urine.

[0150] Fecal scores were recorded based on a scale from 0 to 3 (0 = normal; 1 = soft feces; 2 = mild diarrhea; 3 = severe diarrhea). Effects of ZnO and phytogenic prototypes PFA 1 and PFA 2 on fecal scores of post-weaning piglets during the 42-day feeding trial are shown in Table 11. Signs of severe diarrhea (scale 3) were not observed during the in vivo study. ZnO supplementation and to a lower extend PFA 2 improved fecal consistency during the starter period (0.18 ± 0.16 and 0.32 ± 0.21, respectively) in comparison with the unsupplemented treatment (0.65 ± 0.35; NC). In the subsequent grower period from day 15 to day 42, ZnO reduced fecal scores compared to NC and PFA 1.Table 11

[0151] Means bearing different superscript letters differ significantly with <0.05 Discussion

[0152] Many challenges in livestock production are associated with bacterial infections. This is especially the case for critical phases early in the life of an animal. In this regard, piglets are among the most susceptible livestock species, particularly during the post-weaning period with massive environmental, dietary, and social changes for these young animals. One of the most common strategies to manage post-weaning diarrhea is the application of zinc oxide at pharmacological concentrations.

[0153] The microbroth dilution assay of the individual substances revealed a substantially stronger antibacterial efficacy of carvacrol compared to tea tree oil in all tested strains. The application of potentially lethal concentrations of a substance could, however, be a strong inducer of resistance development. Consequently, we explored the potential of phytogenies to prevent the expression of pathogenic traits without exerting selection pressure towards resistance development or compromising the “healthy” functions of the microbiome. Several bacterial models related to quorum sensing were applied in this study to investigate the potential of phytogenic compositions.

[0154] The first tested model was the biosensor strain C. violaceum which produces the purple pigment violacein in response to QS activity (McCleanl997). Treatment of C. violaceum with both individual substances, carvacrol and tea tree oil, at sub-MIC concentrations inhibited or reduced production of the violacein pigment (See Figs. 1A-1E.). These results suggest that both tested substances can interfere with quorum sensing of this model strain at different concentrations.

[0155] Quorum sensing has also been shown to be involved in the formation of biofilms. The protective environment of biofilms plays an important role in bacterial infections by conferring resistance towards antibiotics and host defense systems. In this study, carvacrol and teatree oil showed a significant impact on biofilm formation of two E. coli field isolates at sub-MIC concentrations although carvacrol exhibited these effects at much lower concentrations than tea tree oil (See Figs. 1 A-1E). Despite the slightly stronger antimicrobial effects against C. violaceum compared with the two pathogenic E. coli field isolates, the phytogenic substances decreased the violacein production, and hence the quorum sensing, of C. violaceum in a similar sub-MIC range than the biofilm formation of the E. coli field isolates. Therefore, it was hypothesized that the observed influence on biofilm formation of carvacrol and tea tree oil is related to their disturbing influence on quorum sensing.

[0156] The third model applied in this study evaluated the impact of tea tree oil and carvacrol on the ability of an F4-fimbriated E. coli strain to adhere to intestinal mucus of piglets. This is especially relevant in the livestock industry as F4-fimbriated E. coli are one of the major pathogens associated with post-weaning diarrhea and production of F4-fimbriae is regulated by quorum sensing. Treatment with carvacrol resulted in a higher number of adherent cells which may indicate increased expression of adhesion factors. It is likely that the chemical stress induced by carvacrol led to the occurrence of autoaggregation events. This type of bacterial behavior serves to protect the involved bacteria from external stresses.

[0157] Based on the results of the examination of the individual phytogenic substances (not shown), two prototypes were formulated, PFA Core 1 and PFA Core 2. PFA Core 1 was based on tea tree essential oil while the second prototype, PFA Core 2, was based on carvacrol. The subsequent study of the MIC values as well as QS and biofilm inhibitory properties of these two prototypes showed that the effects of both PFA cores correlated with those of their respective individual main component (Table 2).

[0158] The in vitro test of ZnO, which served as positive control in the in vivo feeding trial, revealed no bactericidal activity in the applied concentration range. Evidence from literature suggests that particle size has a great influence on the antibacterial activity of ZnO with particularly Zinc oxide nanoparticles apparently showing strong effects. Possibly a high average particle size in combination with low water solubility was responsible for the lack of bactericidal activity with the pharmaceutical grad ZnO applied in the present study. Despite this, a strong inhibition of E. coli biofilm formation and C. violaceum violacein production was observed in a wide range of applied ZnO concentrations. These findings indicate that ZnO may not only function through bactericidal effects, but also inhibit bacterial traits in the sub-MIC range.

[0159] Side effects like the onset of toxic effects during prolonged administration and the accumulation of the heavy metal zinc in the environment, prompted the European Union to banthe use of medicinal doses of ZnO, starting from June 2022. Therefore, this study evaluated the in vivo potential of the formulated phytogenic feed additive prototypes as ZnO alternatives in piglets during the nursery period.

[0160] In a similar study, benefits of ZnO on piglet growth parameters were reported by Molist et al. (2011). There an 8.2% increase in body weight was observed on day 12 post-weaning in piglets receiving 3000 ppm ZnO compared to an unsupplemented control, which correlates with the 9.1% increase on day 15 post-weaning in the present study. This result of the present study indicate that the pharmacological dosage of ZnO showed the expected improvement of the animal’ s growth performance accompanied by reduced occurrence of post-weaning diarrhea. This effect did not rely on high incidence of pathogens as overall mortality and medication rates as well as growth performance suggest a good health status of the whole group of animals observed in this study. The incidence of diarrhea, with a total of only 1.1% of all animals requiring medication due to PWD, was lower than stated averages in literature. Reported PWD rates in piglets receiving no ZnO or antibiotic supplementation are in the range of 34% to 51.1% for individual groups in Denmark (Eriksen2021, Carstensen2015), 24% in a survey of commercial Australian pig herds (Van Breda2017), and between 3.6% and 14.3% in organized Indian farms (VinodhKumar2019). The variations in the incidence of diarrhea and PWD might be the result of differing housing conditions, since, for example, low hygiene standards can lead to increased susceptibility of piglets to stressors and diarrhea (Bonetti2021).

[0161] Due to low overall morbidity of the piglets significant differences are difficult to achieve with the experimental setup and difference could be discussed if they are medically relevant. Still both ZnO and PFA were able to improve the conditions of piglets in terms of fecal scores between day 1 and 14 of the trial. This ranking of this result was found also in growth performance, with PFA 2 performing closer to the group receiving ZnO. PFA 1 supplementation improved FCR in a significant manner. These results reflect the in vitro observations (data not shown), with PFA Core 2 showing effects at much lower concentrations compared to PFA Core 1 in the A. coli biofilm inhibition assay and the C. violaceum quorum sensing inhibition assay.Conclusion

[0162] The results of this study showed that dietary inclusion of a carvacrol -based phytogenic feed additive improves growth parameters and fecal scores in weaned piglets similar to ZnO. The applied in vitro assays suggest this effect is probably associated but not limited to carvacrol's ability to influence bacterial behavior, potentially interfering with virulence factor expression. Therefore, phytogenic feed additives based on carvacrol provide an acceptablealternative to traditional antimicrobial growth promoters in livestock feed, especially in pig farming, to improve animal welfare and increase productivity.Example 2 -Exploring dietary strategies in weaned piglets challenged with ETECObjective: To evaluate the effects of feed supplementation against a F4-fimbriae bearing enterotoxigenic Escherichia coli (F4-ETEC) challenge in weaned piglets.

[0163] Materials and Methods:

[0164] Thirty piglets, 9.4±1.28 kg BW, weaned at 30 days of age. The piglets of mixed gender in 1 : 1 ratio. The piglets were F4-susceptible based on SNP2 gene. 3.0xl010CFU F4-ETEC inoculation at day 9 post- weaning. Three treatment groups with 10 piglets / treatment, and 5 piglets / pen were used. Diets were fed as pellet, ad lib from weaning until 21 days post- weaning. One pen contained two feeding troughs and drinking nipples.

[0165] The four dietary treatments included a negative control group receiving an unsupplemented basal diet (negative control; NC), a positive control group that was provided colistine via drinking water on Day 8-14 in addition to Day 0-3. Treatment group #3 were fed basal diets supplemented with PFA 2 composition at 1 g / kg of feed.

[0166] Phytogenic prototypes PFA2 composition (FIG.4) included of 66% fenugreek seed powder, 2.5% turmeric root powder, 1.4% licorice root powder and 15.4% limestone. PFA Core 2 was added to PFA2 composition at 14.7% (4.9% total amount of essential oil and the remaining encapsulation matrix and carrier. Carvacrol and eugenol were microencapsulated using an encapsulation matrix at concentrations as shown in FIG. 4. Star anise oil was premixed with a carrier (sipemate) as shown in FIG. 4. The final concentration in the PFA2 composition is 1.99% of each carvacrol and eugenol and 0.9%-star anise oil. The contents of the basal feed are as described in Tables 1-3 above.

[0167] The treatment groups are summarized in Table 12.Table 12

[0168] Between Day 0-3 post-weaning, all 30 piglets received colistin orally via drinking water. Piglets were inoculated with F4-ETEC inoculation on Day 9. Fecal samples were collected on day 8, 10, 11, 12, 13, 14, 16 and 18. Fecal consistency was scored on day 0, 8, 10, 11, 12, 13, 14, 16, 18 and 21.

[0169] Data Processing: Data missing due to two piglets reaching a humane end point were handled as missing values in the dataset. The performance of these animals was taken into account until the days they were excluded from the experiment.

[0170] Fecal sample qPCR: Each sample was analysed in triplicate in the F4-ETEC qPCR. The average cycle threshold (CT) was used to determine F4-ETEC concentrations. For some samples, if one CT was undetermined (>40 cycles), the average of the other two CTs was used. If two were undetermined, the third CT (<40 cycles) quantified the F4-ETEC. The limit of detection (LOD) was set at 200 CFU F4-ETEC per gram of feces. Samples with an undetermined CT (>40 cycles) were given a value of 0.5 times the LOD (100 CFU). Finally, a loglO transformation was performed on the F4-ETEC concentrations before statistical analyses.

[0171] Statistical Analyses: The experimental data were analyzed using GenStat Version 23 for Windows(VSN International Ltd, Hemel Hempstead, UK). Missing values were estimated. Analyses were run with and without excluding outliers. Observations were marked as outliers if the residual (fitted - observed value) exceeded 2.5 x standard error of the residuals of the dataset. F4-ETEC concentration (on each day separately) was analysed using one-way analysis of variance (ANOVA) with replicate as random effect. Chi-square tests were used to analyse the prevalence of F4-ETEC diarrhea and shedding. Significant differences are declared at <0.05, with near significant trends at 0.05< <0.10.

[0172] Proximate diet (crude protein and fat) analyses are shown in Table 13.Table 13

[0100] RESULTS:

[0101] Table 14 shows the percentage of pigs with diarrhea (feces score < 4) during the experimental period (n = 10 animals / treatment).Table 14Different superscripts (AB) within a row indicate a tendency between treatments (0.05<P<0.10).

[0101] Table 15 shows the percentage of pigs with F4-ETEC shedding during the experimental period (n = 10 animals / treatment).Table 15

[0102] Table 16 shows the results of F4-ETEC shedding from piglets in logio CFU / g feces during the experimental period. Inoculation took place at day 9 post-weaning.Table 16

[0103] After inoculation of pigs at day 9 post-weaning with the F4-ETEC strain overall percentage of diarrheic piglets was below 50% throughout the trial (Table 14). No diarrhea was observed in the PC group. At day 13 post-weaning the pigs in the NC group showed a tendency for higher proportion of diarrhea compared to the PC group. Piglets receiving PFA2 show numerically a similar or lower proportion of diarrheic animals’ post inoculation compared to the NC, but numerically higher proportion compared to the PC group. The percentage of pigs with F4-ETEC shedding during the experimental period was generally lowest in the PC group (Table 15). This aligns well with the observation that no pigs within the PC group showed signs of diarrhea post-inoculation with the F4 ETEC strain at day 9 post-weaning (Table 14). Animalsshedding F4 ETEC in the PC group showed on average lower logio CFU / g feces values compared to both the NC and PFA2 groups (Table 16). Together these results demonstrate the clear effect of colistin treatment protecting the piglets for the artificial F4 ETEC challenge. While differences are less pronounced between the NC and PFA2 group, results suggest a beneficial effect of the phytogenic additive. While not statistically significant on the individual day on average 24.3% piglets showed signs of diarrhea in the NC group compared to 13.5% of animals in the PFA2 group (Table 14). During the time after inoculation with F4 ETEC at day 9 post-weaning until the end of the study at day 18. This is further supported by data in Table 15 and Table 16, with less animals in the PFA2 group shedding the F4 ETEC strain on days 13, 14, and 16 and the logio CFU / g feces values for the F4 ETEC strain are numerically lower throughout the study except for day 10 post- weaning. The tendency of more pigs in the PFA2 group shedding F4 ETEC while also having logio CFU / g feces values on day 10 post- weaning may be speculated to an interference of the product to the E. coli strains ability of establishing a larger population in the intestinal tract. This is based on the information generated with different assays (biofilm formation, in vitro mucus adhesion, C. violaceum quorum sensing assay) showing a potential to influence bacterial behavior with the active compounds of PFA2. It is hypothesized that compared to the NC group the F4 ETEC has more difficulties to establish a population in PFA2 supplemented animals, leading to reduced adhesion on intestinal surfaces and excreting a larger proportion of the inoculum on day 10 post-weaning compared to the NC group. This would in turn result in a reduced pathogen pressure responsible for the benefits throughout the study period.

[0104] Representative features of the present invention are set out in the following clauses, which stand alone or may be combined, in any combination, with one or more features disclosed in the text of the Specification.

[0105] The present invention is as set out in the following clauses:

[0106] Clause 1 : A method of feeding a livestock animal comprising: providing a basal feed supplemented with a phytogenic composition to post-weaning (PW) piglets, wherein the phytogenic composition comprises an essential oil core, herbal powders and preferably, a bulking agent, wherein the essential oil core comprises essential oils, the essential oils comprise carvacrol, eugenol and star anise oil, wherein the PW piglets have improved growth performance relative to PW piglets provided the basal feed without the phytogenic composition.

[0107] Clause 2: The method of clause 1, wherein the carvacrol comprises between 20% by weight and 60% by weight of the essential oils.

[0108] Clause 3: The method of clauses 1-2, wherein the carvacrol comprises 40% by weight of the essential oils.

[0109] Clause 4: The method of any one of the preceding clauses, wherein the eugenol comprises between 10% by weight and 50% by weight of the essential oils.

[0110] Clause 5: The method of any one of the preceding clauses, wherein the eugenol comprises about 40% by weight of the essential oils.

[0111] Clause 6: The method of any one of the preceding clauses, wherein the star anise oil comprises between 10% by weight and 30% by weight of the essential oils.

[0112] Clause 7: The method of any one of the preceding clauses, wherein the star anise oil comprises about 20% of the essential oils.

[0113] Clause 8: The method of any one of the preceding clauses, wherein the carvacrol in the basal feed is at a dose of between 10 ppm and 50 ppm, the eugenol is at a dose of between 10 ppm and 50ppm and the star anise oil is at a dose of between 5 ppm and 50 ppm.

[0114] Clause 9: The method of any one of the preceding clauses, wherein the carvacrol is at a dose of about 20 ppm, the eugenol is at a dose of about 20 ppm and the star anise oil is at a dose of about 9 ppm in the basal feed.

[0115] Clause 10: The method of any one of the preceding clauses, wherein the basal feed comprises fenugreek seed powder.

[0116] Clause 11 : The method of any one of the preceding clauses, wherein the basal feed comprises fenugreek seed powder, licorice powder and / or turmeric powder.

[0117] Clause 12: The method of any one of the preceding clauses, wherein the basal feed comprises fenugreek seed powder at a dose of between 200 ppm and 1000 ppm.

[0118] Clause 13: The method of clauses any one of the preceding clauses, wherein the basal feed comprises fenugreek seed powder at a dose of about 660 ppm.

[0119] Clause 14: The method of any one of the preceding clauses, wherein the basal feed comprises turmeric powder at a dose of between 5 ppm and 50 ppm.

[0120] Clause 15: The method of any one of the preceding clauses, wherein the basal feed comprises turmeric powder at a dose of about 25 ppm.

[0121] Clause 16: The method of any one of the preceding clauses, wherein the basal feed comprises licorice powder at a dose of between 1 ppm and 50 ppm.

[0122] Clause 17: The method of any one of the preceding clauses, wherein the basal feed comprises licorice powder at a dose of about 14 ppm.

[0123] Clause 18: The method of any one of the preceding clauses, wherein the basal feed is supplemented with between 0.25 kilogram / metric ton (kg / t) and 2.5 kg / t of the phytogenic composition in the basal feed.

[0124] Clause 19: The method of any one of the preceding clauses, wherein the basal feed is supplemented with about 1 kg / t of the phytogenic composition.

[0125] Clause 20: The method of any one of the preceding clauses, wherein one or more of the essential oils are microencapsulated, wherein the microencapsulated oils comprise essential oils and encapsulation matrix and wherein the bulking agent is limestone.

[0126] Clause 21 : The method of any one of the preceding clauses, wherein the phytogenic composition is provided to the PW piglets from day 18 to day 70 and any range there between.

[0127] Clause 22: The method of any one of the preceding clauses, wherein the improved growth performance comprises improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG), reduced fecal shedding of pathogenic microorganisms and / or improved fecal scores.

[0128] Clause 23: A PW piglet feed comprising: a basal feed; and a phytogenic composition, wherein the phytogenic composition comprises an essential oil core and herbal powders, wherein the essential oil core comprises essential oils, wherein the essential oils comprise carvacrol, eugenol and star anise oil, wherein the PW piglet feed comprises the phytogenic composition at an inclusion rate of between 0.25 kilogram / ton (kg / t) and 2.5 kg / t and wherein PW piglet feed comprises carvacrol at a dose between 10 ppm and 50 ppm, eugenol at a dose between 10 ppm and 50 ppm and star anise oil at a dose of between 5 ppm and 50 ppm, wherein the PW piglet feed improves growth performance relative to PW piglet feed without the phytogenic composition.

[0129] Clause 24: The PW piglet feed of clause 23, wherein the PW piglet feed comprises 20 ppm of carvacrol, 20 ppm of eugenol and 10 ppm of star anise oil.

[0130] Clause 25: The PW piglet feed of any one of clauses 23-24, wherein the feed further comprises fenugreek seed powder.

[0131] Clause 26: The PW piglet feed of any one of clauses 23-25, wherein the feed further comprises turmeric powder.

[0132] Clause 27: The PW piglet feed of any one of clauses 23-26, wherein the feed further comprises licorice powder.

[0133] Clause 28: The PW piglet feed of any one of clauses 23-27, wherein the feed further comprises fenugreek seed powder between 200 ppm and 1000 ppm, turmeric powder between 5 ppm and 50 ppm and licorice powder between 1 ppm and 50 ppm.

[0134] Clause 29: The PW piglet feed of any one of clauses 23-28, wherein the feed further comprises about 660 ppm of fenugreek seed powder, 25 ppm of turmeric powder and 14 ppm of licorice powder.

[0135] Clause 30: The PW piglet feed of any one of clauses 23-29, wherein the basal feed is supplemented with about 1 kg / t of the phytogenic composition.

[0136] Clause 31 : The PW piglet feed of any one of clauses 23-30, wherein the phytogenic composition is provided to the PW piglets from day 18 to day 70 and any range there between.

[0137] Clause 32: The PW piglet feed of any one of clauses 23-31, wherein the improved growth performance comprises improved average daily gain (ADG), improved feed conversion ratio (FCR), improved body weight gain (BWG) and / or improved fecal scores.

[0138] Clause 33: The PW piglet feed of any one of clauses 23-32, wherein one or more of the essential oils are microencapsulated, wherein the microencapsulated oils comprise essential oils and encapsulation matrix.

[0139] Clause 34: A phytogenic composition comprising an essential oil core and herbal powders, wherein the essential oil core comprises essential oils, wherein the essential oils comprise carvacrol between 30% by weight and 50% by weight of the essential oils, eugenol between 30% by weight and 50% by weight of the essential oils and star anise oil between 10% by weight and 30% by weight of the essential oils and wherein the herbal powders comprise fenugreek seed powder, turmeric powder and / or licorice powder.

[0140] Clause 35: The composition of clause 34, wherein the essential oils comprise carvacrol at 40% by weight of the essential oils.

[0141] Clause 36: The composition of any one of clauses 34-35, wherein the essential oils comprise eugenol at 40% by weight of the essential oils.

[0142] Clause 37: The composition of any one of clauses 34-36, wherein the essential oils comprise star anise oil at 20% by weight of the essential oils.

[0143] Clause 38: The composition of any one of clauses 34-37, wherein the fenugreek seed powder is at about 66% by wt., the turmeric powder is between about 2.4% by wt. and about 2.5% by wt. and the licorice powder is at about 1.4% by wt. and about 1.5% by wt. of the composition.

[0144] Clause 39: The composition of any one of clauses 34-38, wherein the composition is used at an inclusion rate in a basal feed at an amount between 0.25 kg / t and 2.5 kg / t.

[0145] Clause 40: The composition of any one of clauses 34-39, wherein the composition is used at an inclusion rate in a basal feed at an amount of 1 kg / t.

[0146] Clause 41 : The composition of any one of clauses 34-40, wherein the composition is used in the basal feed at a dosage of carvacrol between 10 ppm and 50 ppm, eugenol between 10 ppm and 50 ppm and star anise oil between 5 ppm and 50ppm.

[0147] Clause 42: The composition of any one of clauses 34-41, wherein the dosage of carvacrol is about 20 ppm, the dosage of eugenol is about 20 ppm, and the dosage of star anise oil is about 10 ppm in the feed.

[0148] Clause 43: The composition of any one of clauses 34-42, wherein the composition is used in the feed with fenugreek seed powder dose between 200 ppm and 1000 ppm.

[0149] Clause 44: The composition of any one of clauses 34-43, wherein the composition is used in the feed with fenugreek seed powder dose at 660 ppm.

[0150] Clause 45: The composition of any one of clauses 34-44, wherein the composition is used in the feed with turmeric powder dose of between 5ppm and 50 ppm.

[0151] Clause 46: The composition of any one of clauses 34-45, wherein the composition is used in the feed with turmeric powder dose at 25 ppm.

[0152] Clause 47: The composition of any one of clauses 34-46, wherein the composition is used in the feed with licorice powder dose between Ippm and 50 ppm.

[0153] Clause 48: The composition of any one of clauses 34-47, wherein the composition is used in the feed with licorice powder dose at 14 ppm.

[0154] Clause 49: The composition of any one of clauses 34-48, wherein the composition is used in the feed with 660 ppm of fenugreek seed powder, 25 ppm of turmeric powder, 14ppm of licorice powder, 20 ppm of carvacrol, 20 ppm of eugenol and 10 ppm of star anise oil.

[0155] Clause 50: The composition of any one of clauses 34-49, wherein the one or more of the essential oils are microencapsulated oils, wherein the microencapsulated oils comprise essential oils and an encapsulation matrix.

[0156] Clause 51 : The composition of any one of clauses 34-50, wherein the composition further comprises a bulking agent.

[0157] Clause 52: The composition of any one of clauses 34-51, wherein the composition further comprises limestone.

[0158] Clause 53: The composition of any one of clauses 34-52, wherein the amount of the essential oils in the composition is between about 4% by weight and 5% by weight of the composition.

[0159] Clause 54: The composition of any one of clauses 34-53, wherein the amount of the essential oils in the composition is between about 4.8%% by weight and 5% by weight of the composition.

[0160] Clause 55: The composition of any one of clauses 34-54, wherein the amount of the amount of the herbal powders in the composition is about 55% by weight and about 75% by weight.

[0161] Clause 56: The composition of any one of clauses 34-55, wherein the amount of the herbal powders in the composition is about 70% by weight.

[0162] Clause 57: The method of any one of the clauses 1-22, wherein the PW piglets are harboring PWD-associated microorganisms.

[0163] Clause 58: A method of mitigating symptoms from PWD-associated microorganisms in PW piglets comprising providing a basal feed supplemented with a phytogenic composition to the PW piglets, wherein the phytogenic composition comprises an essential oil core, herbal powders and preferably, a bulking agent, wherein the essential oil core comprises essential oils, the essential oils comprise carvacrol, eugenol and star anise oil, wherein the PW piglets have lower incidence of diarrhea and / or reduced fecal shedding relative to PW piglets provided the basal feed without the phytogenic composition.

[0164] Clause 59: The method clause 58, wherein the microorganisms are pathogenic E. coli.

[0165] Clause 60, The method according to any one of clauses 58-59, wherein the microorganism is enterotoxigenic / ■ / coli (ETEC).

Claims

CLAIMSWhat is claimed is:

1. A method of feeding a livestock animal comprising: providing a basal feed supplemented with a phytogenic composition to postweaning (PW) piglets, wherein the phytogenic composition comprises an essential oil core, herbal powders and preferably, a bulking agent, wherein the essential oil core comprises essential oils, the essential oils comprise carvacrol, eugenol and star anise oil, wherein the PW piglets have improved growth performance relative to PW piglets provided the basal feed without the phytogenic composition.

2. The method of claim 1, wherein the carvacrol comprises between 20% by weight and 60% by weight of the essential oils, preferably the carvacrol comprises 40% by weight of the essential oils.

3. The method of any one of the preceding claims, wherein the eugenol comprises between 10% by weight and 50% by weight of the essential oils, preferably the eugenol comprises about 40% by weight of the essential oils.

4. The method of any one of the preceding claims, wherein the star anise oil comprises between 10% by weight and 30% by weight of the essential oils, preferably the star anise oil comprises about 20% of the essential oils.

5. The method of any one of the preceding claims, wherein the carvacrol in the basal feed is at a dose of between 10 ppm and 50 ppm, the eugenol is at a dose of between 10 ppm and 50ppm and the star anise oil is at a dose of between 5 ppm and 50 ppm.

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

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

8. The method of any one of the preceding claims, wherein the basal feed is supplemented with between 0.25 kilogram / metric ton (kg / t) and 2.5 kg / t, preferably about 1 kg / t of the phytogenic composition in the basal feed.

9. The method of any one of the preceding claims, wherein one or more of the essential oils are microencapsulated, wherein the microencapsulated oils comprise essential oils and encapsulation matrix and wherein the bulking agent is limestone.

10. The method of any one of the preceding claims, wherein the phytogenic composition is provided to the PW piglets from day 18 to day 70 and any range there between.

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

12. The method of any one of the preceding claims, wherein the PW piglets are harboring PWD-associated microorganisms.

13. A PW piglet feed compri sing : a basal feed; and a phytogenic composition, wherein the phytogenic composition comprises an essential oil core and herbal powders, wherein the essential oil core comprises essential oils, wherein the essential oils comprise carvacrol, eugenol and star anise oil, wherein the PW piglet feed comprises the phytogenic composition at an inclusion rate of between 0.25 kilogram / ton (kg / t) and 2.5 kg / t, preferably at an inclusion rate of about 1 kg / t of the phytogenic composition, and wherein PW piglet feed comprises carvacrol at a dose between 10 ppm and 50 ppm, eugenol at a dose between 10 ppm and 50 ppm and star anise oil at a dose of between 5 ppm and 50 ppm, wherein the PW piglet feed improves growth performance relative to PW piglet feed without the phytogenic composition.

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

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

16. The PW piglet feed of any one of claims 13-15, wherein one or more of the essential oils are microencapsulated, wherein the microencapsulated oils comprise essential oils and encapsulation matrix.

17. A phytogenic composition comprising an essential oil core and herbal powders, preferably the amount of herbal powders in the composition is about 55% by weight and about 75% by weight, wherein the essential oil core comprises essential oils, wherein the essential oilscomprise carvacrol between 30% by weight and 50% by weight of the essential oils, preferably the essential oils comprise carvacrol at 40% by weight of the essential oils, eugenol between 30% by weight and 50% by weight of the essential oils, preferably the essential oils comprise eugenol at 40% by weight of the essential oils and star anise oil between 10% by weight and 30% by weight of the essential oils, preferably the essential oils comprise star anise oil at 20% by weight of the essential oils and wherein the herbal powders comprise fenugreek seed powder, turmeric powder and / or licorice powder.

18. The composition of any one of claims 17, wherein the composition is used at an inclusion rate in a basal feed at an amount between 0.25 kg / t and 2.5 kg / t, preferably at an inclusion rate in a basal feed at an amount of 1 kg / t.

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

20. The composition of any one of claims 17-19, wherein the composition is used in the feed with fenugreek seed powder dose between 200 ppm and 1000 ppm, with turmeric powder dose of between 5ppm and 50 ppm and / or with licorice powder dose between Ippm and 50 ppm.

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

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

23. A method of mitigating symptoms from PWD-associated microorganisms in PW piglets comprising providing a basal feed supplemented with a phytogenic composition to the PW piglets, wherein the phytogenic composition comprises an essential oil core, herbal powders and preferably, a bulking agent, wherein the essential oil core comprises essential oils, the essential oils comprise carvacrol, eugenol and star anise oil, wherein the PW piglets have lower incidence of diarrhea and / or reduced fecal shedding relative to PW piglets provided the basal feed without the phytogenic composition.

24. The method claim 23, wherein the microorganisms are pathogenic E.coli.

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

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