WO2018049243A1 - Feed additive comprising allicin - Google Patents

Abstract

Disclosed herein are embodiments of a combination for administrating to an animal. The combination comprises allicin, alliin, allinase, or algae, mineral clay, silica, glucan, mannans, endoglucanohydrolase, yeast, a vitamin, yucca, quillaja, a probiotic, an antimicrobial, a vaccine, sorbic acid or a salt thereof, polyphenol, or a growth promotant, or any combination thereof. Certain embodiments comprise allicin, alliin, allinase, algae or a combination thereof and may also comprise mineral clay, silica, glucan, and mannans. The sorbic acid salt may be potassium, sodium or ammonium sorbate, and the vitamin may include vitamin C. The combination may also comprise an endoglucanohydrolase, a carrier, an adhesive agent and/or a feedstuff. Embodiments of a method for administering the combination to an animal are also disclosed.

Description

FEED ADDITIVE COMPRISING ALLICIN

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 62/385,724, filed September 9, 2016, which is herein incorporated by reference in its entirety.

FIELD

The present disclosure concerns a combination for use as a supplement, alone or in combination with a feedstuff, for administration to an animal. Also disclosed herein is a method of administering the combination to the animal.

SUMMARY

Disclosed herein are embodiments of a combination for administration to an animal. The combination may comprise allicin, alliin, allinase, algae, mineral clay, silica, glucan, mannans, endoglucanohydrolase, yeast, a vitamin, yucca or an extract thereof, quillaja or an extract thereof, a probiotic, an antimicrobial, a vaccine, sorbic acid or a salt thereof, polyphenol or plant material comprising polyphenol, a growth promotant, or any combination thereof. In some embodiments, the combination comprises one or more of allicin, alliin, allinase or algae, and one or more of silica, glucan, mannans, endoglucanohydrolase, yeast, a vitamin, yucca, quillaja, a probiotic, an antimicrobial, a vaccine, sorbic acid or a salt thereof, polyphenol, or a growth promotant. In some embodiments, the combination comprises from 1% to 50% allicin, such as from 15% to 40%. And in certain embodiments, the combination is not 10-25% allicin, 1-5% β-glucanase, 20-40% diatomaceous earth, 5-15% potassium sorbate, 25-50% brewer's yeast cell wall, and 0.5-1% vitamin C.

The combination may be a composition, and/or may comprise a composition I comprising mineral clay, silica, glucan, mannans, or a combination thereof. In any embodiments, the composition or composition I may further comprise an endoglucanohydrolase, such as β-1,3 (4)- endoglucanohydrolase.

In any of the disclosed embodiments, the yeast may be yeast culture, a live yeast, a dead yeast, yeast extract, or a combination thereof. The sorbic acid or salt thereof may be potassium sorbate, sodium sorbate, ammonium sorbate, or a combination thereof. The vitamin may be vitamin A, vitamin Bi, vitamin B2, vitamin B3, vitamin B5, vitamin B₆, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, or a combination thereof. The algae may be a blue-green algae (cyanobacteria), a diatom (bacillariophyta), a stonewort algae (charophyta), a green algae

(chlorophyta), a golden algae (chrysophyta), a dinoflagellate (dinophyta), a brown algae

(phaeophyta) or a red algae (rhodophyta). The yucca may be Yucca schidigera, and/or the quillaja may be Quillaja saponaria, and the probiotic may be a Bacillus species, such as Bacillus coagulans. In some embodiments, the combination comprises an antimicrobial, such as an antibiotic, an antifungal, an antiparasitic, an antiviral, or a combination thereof, and the antiparasitic may be an anticoccidial. Additionally or alternatively, the combination may comprise a vaccine and/or a growth promotant. Also in any embodiments, the combination might further comprise an adhesive agent, such as a syrup, an oil, or a combination thereof, and/or a feedstuff, such as a solid animal feed, a liquid animal feed, a supplement, a premix, water, a feed additive carrier, or a combination thereof.

In any of the embodiments, glucan and mannans may be provided by yeast cell wall, or an extract thereof. In certain disclosed embodiments, the combination comprises silica, mineral clay, glucan, and mannans, and allicin, potassium sorbate, vitamin C, or a combination thereof. And the combination may consist essentially of mineral clay, silica, glucan, mannans, and allicin; allicin, endoglucanohydrolase, silica, mineral clay, potassium sorbate, glucan, mannans, and vitamin C; allicin, endoglucanohydrolase, silica, mineral clay, potassium sorbate, yeast cell wall extract, and vitamin C; allicin, yucca, such as Yucca schidigera, and quillaja, such as Quillaja saponaria; allicin, mineral clay, silica, glucan, mannans, yucca, and quillaja; or allicin, mineral clay, silica, glucan, mannans, endoglucanohydrolase, yucca, and quillaja.

Any of the disclosed embodiments may be formulated for administration to an animal, including a non-human animal. The animal may be a land animal; a mammal, including, but not limited to, a ruminant or ungulate; an aquatic animal, including, but not limited to, a fish, a crustacean, or a mollusk; an avian, particularly chickens or turkeys; or a companion animal.

Also disclosed herein are embodiments of a method of administering the combination to an animal, such as a non-human animal; a mammal, such as a ruminant or ungulate; an avian; an aquatic species, such as a fish, crustacean or mollusk; or a companion animal. The combination may be administered to provide a beneficial result in the animal compared to an animal not administered the combination. The beneficial result may be prevention of a disease, such as an infectious or a non-infectious disease; stress; a stress-related condition or disease; treatment of a disease, stress, a stress-related condition or disease; amelioration of symptoms associated with a disease or stress-related condition; a beneficial effect on the animal's immune system; an increase in the longevity of the animal; or a combination thereof. The animal may be an animal that has or is at risk of developing a disease, stress, a stress-related disease or condition, or a combination thereof. Additionally, or alternatively, administering the combination to the animal may improve a feed conversion ratio of the animal compared to an animal not administered the combination.

The foregoing and other objects, features, and advantages of the invention will become more apparent from the following detailed description, which proceeds with reference to the accompanying figures.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a table illustrating exemplary dose ranges of exemplary embodiments of the combination comprising silica, mineral clay, glucan and mannans for various growth stages.

FIG. 2 provides Western blot results demonstrating the effect of exemplary embodiments of the combination on the expression of neutrophil L-selectin as described in Example 1.

FIG. 3 provides Western blot results demonstrating the effects of a disclosed embodiment of the combination in unheated and heated (pelleted) forms on the expression of neutrophil L-selectin as described in Example 2.

FIG. 4 is a graph summarizing the effects of a disclosed embodiment of the combination on the expression of mRNA encoding L-selectin in rat neutrophils as described in Example 3.

FIG. 5 provides Western blot results demonstrating the effects of a disclosed embodiment of the combination on the expression of neutrophil interleukin-ΐβ (II- 1β) as described in Example 4.

FIG. 6 is a graph summarizing the effects of different compositions on the ability of rat neutrophils to affect the viability of Staphylococcus aureus bacteria as described in Example 8.

FIG. 7 is a graph summarizing the effects of different embodiments of the combination on the expression of mRNA encoding interkeukin-8 receptor in rat neutrophils as described in

Example 8.

FIG. 8 is a graph summarizing the effects of different embodiments of the combination on the expression of mRNA encoding L-selectin in rat neutrophils as described in Example 8.

FIG. 9 is a table comparing the average weight (g) of sea bream being fed two different doses of exemplary embodiments of the combination and a control group.

FIG. 10 is a table showing the feed conversion rate (FCR) of sea bream in the trial at day

158.

FIG. 11 is a table showing the survival rate in the trial at day 128.

FIG. 12 is a table showing the expected growth rate and feeding chart of tilapia.

FIG. 13 is a table comparing the average weight (g) of hybrid tilapia being fed two different doses of the combination and a control group. FIG. 14 is a table of the food conversion ratio results for each group from the hybrid tilapia trial.

FIG. 15 is a table providing the feeding chart of carp by temperature during the common carp trial.

FIG. 16 is a table comparing the average weight (g) of common carp being fed two different doses of the combination and a control group.

FIG. 17 is a table of the food conversion ratio results for each group during the common carp trial.

FIG. 18 is a bar graph illustrating the total mortality of tilapia separated by treatment.

FIG. 19 is a graph of level (ppm) versus time (days), illustrating ammonia and nitrite levels in the water.

FIG. 20 is a graph of level versus time (days), illustrating pH and oxygen (ppm) levels in the water.

FIG. 21 is a table comparing the average survival (percent from 4 replicates) of shrimp using 100 and 200 mg/kg body weight per day doses, and a control group.

FIG. 22 is a bar graph illustrating results obtained from a 48-day trial wherein feed conversion in chickens in pens was measured after 48 days, wherein the data is provided in descending order (highest feed conversion to lowest feed conversion).

FIG. 23 is a table of the antibiotic free (ABF) and treatment diets used in the test.

FIG. 24 is a bar graph illustrating feed conversion based on the type of treatment used

(regular ABF diet versus a treatment embodiment disclosed herein).

FIG. 25 is a bar graph illustrating growth rate based on the type of treatment used (regular ABF diet versus a treatment embodiment disclosed herein).

FIG. 26 is a graph of IL-6 concentration versus treatment illustrating the concentration of IL-6 in rat plasma after being fed a diet comprising each supplement for 28 days.

FIG. 27 is a graph of IL-10 concentration versus treatment illustrating the concentration of IL-6 in rat plasma after being fed a diet comprising each supplement for 28 days.

FIG. 28 is a graph of average daily gain (ADG) versus treatment illustrating the effect of the combination on the ADG of turkeys.

FIG. 29 is a graph of weight versus treatment, illustrating the effect of the combination on the net weight of turkeys.

FIG. 30 is a graph of feed conversion ratio versus treatment, illustrating the effect of the combination on the feed conversion ratio of turkeys. DETAILED DESCRIPTION

I. Definitions

The following explanations of terms and abbreviations are provided to better describe the present disclosure and to guide those of ordinary skill in the art in the practice of the present disclosure. As used herein, "comprising" means "including" and the singular forms "a" or "an" or "the" include plural references unless the context clearly dictates otherwise. The term "or" refers to a single element of stated alternative elements or a combination of two or more elements, unless the context clearly indicates otherwise.

Unless explained otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. The materials, methods, and examples are illustrative only and not intended to be limiting. Other features of the disclosure are apparent from the following detailed description and the claims.

Unless otherwise indicated, all numbers expressing quantities of components, molecular weights, percentages, temperatures, times, and so forth, as used in the specification or claims are to be understood as being modified by the term "about." Accordingly, unless otherwise indicated, implicitly or explicitly, the numerical parameters set forth are approximations that may depend on the desired properties sought and/or limits of detection under standard test conditions/methods.

When directly and explicitly distinguishing embodiments from discussed prior art, the embodiment numbers are not approximates unless the word "about" is recited.

Administering: Administration by any route to a subject. As used herein, administration typically but not necessarily refers to oral administration.

Antimicrobial: An agent that kills and/or inhibits the growth of microorganisms. As used herein, antimicrobials include antibiotics, antifungals, antivirals, and antiparasitics, including without limitation anticoccidials, or combinations thereof.

Binding agent or binder: A material or substance that is used to hold or draw together other materials to form a cohesive unit.

Co-administration: Administering two or more agents simultaneously or sequentially in any order to a subject to provide overlapping periods of time in which the subject is experiencing effects, beneficial and/or deleterious, from each agent. For example, if administration of a first agent results in deleterious side effects, as second agent may be administered to reduce and/or substantially prevent or inhibit those side effects. One or both of the agents may be a therapeutic agent. The agents may be combined into a single composition or dosage form, or they may be administered simultaneously or sequentially in any order as separate agents.

Combination: A combination includes two or more components that are administered such that the effective time period of at least one component overlaps with the effective time period of at least one other component. A combination and/or component thereof may be a composition. In some embodiments, the effective time periods of all components administered overlap with each other. In an exemplary embodiment of a combination comprising four components, the effective time period of the first component administered may overlap with the effective time periods of the second, third and fourth components, but the effective time periods of the second, third and fourth components independently may or may not overlap with one another. In another exemplary embodiment of a combination comprising four components, the effective time period of the first component administered overlaps with the effective time period of the second component, but not that of the third or fourth; the effective time period of the second component overlaps with those of the first and third components; and the effective time period of the fourth component overlaps with that of the third component only. A combination may be a composition comprising all the combination components, a composition comprising one or more components and another separate component (or components) or composition(s) comprising the remaining component(s), or the combination may be two or more individual components. In some embodiments, the two or more components may comprise the same component administered at two or more different times, two or more different components administered substantially simultaneously or sequentially in any order, or a combination of sequential and simultaneous administration.

Excipient or carrier: A physiologically inert substance that is used as an additive in (or with) a combination, composition, or component as disclosed herein. As used herein, an excipient or carrier may be incorporated within particles of a combination, composition, or component, or it may be physically mixed with particles of a combination, composition, or component. An excipient or carrier can be used, for example, to dilute an active agent and/or to modify properties of a combination or composition. Examples of excipients and carriers include, but are not limited to, calcium carbonate, polyvinylpyrrolidone (PVP), tocopheryl polyethylene glycol 1000 succinate (also known as vitamin E TPGS, or TPGS), dipalmitoyl phosphatidyl choline (DPPC), trehalose, sodium bicarbonate, glycine, sodium citrate, and lactose.

Feed efficiency: A measure of an animal's efficiency in converting feed mass into the desired output, e.g. , weight gain, milk production. Feed efficiency also may be referred to as feed conversion ratio, feed conversion rate, or feed conversion efficiency. Feedstuff: As used herein, the term "feedstuff refers to anything that may be consumed by an animal. The term "feedstuff encompasses solid and liquid animal feeds (e.g. , a feed ration), supplements (e.g. , a mineral supplement, a protein supplement), a premix, water, feed additive carriers (e.g. , molasses), and combinations thereof.

Glucocorticoid: A class of steroid hormones that bind to the glucocorticoid receptors in vertebrate animal cells. Exemplary endogenous glucocorticoids include Cortisol (hydrocortisone) and corticosterone.

Mannans: A class of polysaccharides including the sugar mannose. The mannans family includes pure mannans (i.e. , the polymer backbone consists of mannose monomers), glucomannan (the polymer backbone comprises mannose and glucose), and galactomannan (mannans or glucomannan in which single galactose residues are linked to the polymer backbone). Mannans are found in cell walls of some plant species and yeasts.

Mineral Clay: According to the AIPEA (Association Internationale pour l'Etude des Argiles (International Association for the Study of Clays)) and CMS (Clay Minerals Study) nomenclature committees, the term "mineral clay" refers to a mineral that imparts plasticity to a clay and hardens upon drying or firing. Mineral clays include aluminum silicates, such as aluminum phyllosilicates. Mineral clays usually include minor amounts of impurities, such as potassium, sodium, calcium, magnesium, and/or iron.

Pharmaceutically acceptable: The term "pharmaceutically acceptable" refers to a substance that can be taken into a subject without significant adverse toxicological effects on the subject, including a non-human animal subject.

Polyphenols: A class of natural, synthetic, or semisynthetic organic chemicals characterized by the presence of plural phenolic

structural units.

Saponin: A class of chemical compounds, one of many secondary metabolites found in natural sources, with saponins found in particular abundance in various plant species. More specifically, they are amphipathic glycosides grouped, in terms of structure, by their composition.

In certain embodiments, saponin comprises one or more hydrophilic glycoside moieties combined with a lipophilic triterpene and/or steroidal derivative.

Therapeutic agent: An agent that is capable of providing a therapeutic effect, e.g. , preventing a disorder, inhibiting a disorder, such as by arresting the development of the disorder or its clinical symptoms, or relieving a disorder by causing regression of the disorder or ameliorating its clinical symptoms. Therapeutically effective amount: A quantity or concentration of a specified compound, composition or combination sufficient to achieve an effect in a subject.

Additional disclosure is provided by U.S. Patent Application No. 14/699,740, U.S. Patent Application No. 13/566,433, U.S. Patent Application No. 13/872,935, U.S. Patent Publication No. 2013/0017211, U.S. Patent Publication No. 2012/0156248, U.S. Patent Publication No.

2007/0253983, U.S. Patent Publication No. 2007/0202092, U.S. Patent Publication No.

20070238120, U.S. Patent Publication No. 2006/0239992, U.S. Patent Publication No.

2005/0220846, U.S. Patent Publication No. 2005/0180964, and Australian Patent Application No. 2011201420, each of which is incorporated herein by reference in its entirety.

II. The Combination

This disclosure concerns embodiments of a combination comprising silica, mineral clay, glucan, mannans, endoglucanohydrolase, allicin, alliin, allinase, yeast, polyphenol or a polyphenol- containing plant material, one or more vitamins, algae, yucca or an extract thereof, quillaja or an extract thereof, one or more probiotics, an antimicrobial, a vaccine, sorbic acid or a salt thereof, a growth promotant, or any combination thereof, for administering to an animal. The

endoglucanohydrolase may be β-1,3 (4)-endoglucanohydrolase. The glucan and mannans may be provided by yeast cell wall or an extract thereof. The probiotic may be a direct-fed microbial. Additionally, the combination may comprise an adhesive agent, a feedstuff, or a combination thereof. The components of the combination are prepared by methods commonly known in the art and/or can be obtained from commercial sources. Also disclosed are embodiments of a method for administering the combination to an animal, such as a land animal, an avian and/or an aquatic animal.

In some embodiments, one or more of the following provisos apply: the combination is not, or does not consist essentially of, silica, mineral clay, glucan and mannans; the combination is not, or does not consist essentially of, silica, mineral clay, glucan, mannans and endoglucanohydrolase; the combination is not, or does not consist essentially of, yucca and quillaja; the combination is not, or does not consist essentially of, yucca, quillaja and a Bacillus species; the combination is not, or does not consist essentially of, yucca, quillaja and Bacillus coagulans; the combination is not, or does not consist essentially of, Yucca schidigera and Quillaja saponaria; the combination is not, or does not consist essentially of, Yucca schidigera, Quillaja saponaria and Bacillus coagulans; the combination is not, or does not consist essentially of, 10-25% of allicin, 1-5% of β-glucanase, 20- 40% of diatomaceous earth, 5-15% of potassium sorbate, 25-50% of brewer's yeast cell wall, and 0.5-1% of vitamin C; the combination is not, or does not consist essentially of, silica, mineral clay, glucan, mannans, potassium sorbate, and endoglucanohydrolase; the combination is not, or does not consist essentially of, silica, mineral clay, glucan, mannans, and an antibiotic; the combination is not, or does not consist essentially of, silica, mineral clay, glucan, mannans, endoglucanohydrolase, and an antibiotic; the combination is not, or does not consist essentially of, silica, mineral clay, glucan, mannans, and a vaccine; the combination is not silica, mineral clay, glucan, mannans, endoglucanohydrolase, and a vaccine; the combination is not, or does not consist essentially of, mineral clay, silica, β-glucan, or a combination thereof, any one or more of yucca, quillaja, a direct- fed microbial, a vitamin D species, or a plant extract, and optionally including mannans and/or endoglucanohydrolase.

If the composition and/or combination is, consists of, or consists essentially of yucca and/or quillaja; and an antibiotic, an antimicrobial, an anticoccidial agent, or a combination thereof, or if the composition and/or combination is, consists of, or consists essentially of yucca and/or quillaja; an antibiotic, an antimicrobial, an anticoccidial agent, or a combination thereof; and a vaccine, then the fish is not salmon, trout, cod, halibut, snapper, herring, or catfish; the crustacean is not lobster, shrimp, prawns, crabs, krill, crayfish, barnacles, or copepods; and the mollusk is not abalone, conchs, rock snails, whelk, clams, oysters, mussels, or cockles.

If the composition and/or combination is, consists of, or consists essentially of silica, mineral clay, glucan and mannans, and one or more of yucca, quillaja, a direct-fed microbial, a vitamin D species, or a plant extract, then the fish is not salmon, trout or tilapia.

In some embodiments, the antibiotic is not, or does not comprise, hydrogen peroxide.

In some embodiments, the composition and/or combination does not comprise a peroxide compound.

In some embodiments, the composition and/or combination does not comprise hydrogen peroxide.

In some embodiments, the composition and/or combination does not comprise carbamide peroxide.

In some embodiments, the composition and/or combination does not comprise urea.

In some embodiments, the composition and/or combination does not comprise hydrogen peroxide and urea.

A. Silica, mineral clay, glucan and/or mannans

The silica may be obtained from or provided by quartz, sand, diatomaceous earth, synthetic silica, or a combination thereof. Diatomaceous earth is available as a commercially-available, acid- washed product comprising 95% silica (S1O2) and remaining components not assayed but primarily ash (minerals) as defined by the Association of Analytical Chemists (AOAC, 2002). The mineral clays (e.g. , aluminosilicates, such as calcium, potassium and/or sodium aluminosilicates) used in this composition and/or combination may be any of a variety of clays, including commercially- available clays including, but not limited to, montmorillonite clay, bentonite, kaolinitic, or zeolite.

The mannans may be, or comprise, glucomannan, and/or the glucans may be β-glucans, such as those obtained from yeast, or other materials, such as fungi, algae, or the like, β-1,3 (4)- endoglucanohydrolase may be obtained commercially and is produced from submerged fermentation of a strain of Trichoderma longibrachiatum. In some embodiments, the glucans include soluble and/or insoluble β-glucans, such as (1,3/1,4) β-glucan (β-1,3 (4) glucan), (1,3/1,6) β-glucan, or a combination thereof. Commercial sources of glucan and mannans (e.g. , β-1,3 (4) glucan and glucomannan), and/or endoglucanohydrolase include plant cell walls, yeast (e.g. ,

Saccharomyces cerevisiae, Candida utilis, brewer's yeast), certain fungi (e.g. , mushrooms), and bacteria. The yeast and/or yeast extract, such as a yeast cell wall extract, can be administered affirmatively to provide glucan, mannans and endoglucanohydrolase endogenously. The glucan and mannans, optionally in the form of a yeast cell wall extract, can be prepared by a method known to a person of ordinary skill in the art. Yeast cell wall extract may have a composition comprising 0-8% moisture and 92-100% dry matter. The dry matter may comprise 10-55 % protein, 0-25 % fats, 0-2% phosphorus, 10-30% β-glucan, 0-25% mannans or from greater than 0 to 25% mannans, and 0-5% ash. For example, a commercial source of β-1,3 (4) glucan and glucomannan derived from primary inactivated yeast (Saccharomyces cerevisiae) with the following chemical composition can be used: moisture 3.5-6.5%; proteins 1-6%; fats 0-0.5%; phosphorus 0-0.2%; mannans 9-20%; β-1, 3-(4) glucan 9-18%; and ash 75-85%. Alternatively, the yeast cell wall extract may comprise moisture 2-3% and dry matter 97-98%, where the dry matter comprises proteins 14-17%, fats 20-22%, phosphorus 1-2%, mannans 22-24%, β-1, 3-(4) glucan 24- 26%, and ash 3-5%.

B. Allicin, alliin and/or alliinase

Allicin (diallyl thiosulfate; 2-Propene-l-sulfinothioic acid S-2-propenyl ester)

is a compound found in garlic, such as raw garlic.

When extracted, it may be an oily, yellowish liquid. Allicin may have medicinal and/or health benefits when consumed by animals. Benefits of allicin include, but are not limited to, an immunity booster; a blood thinner; an anti-oxidant; an anti-bacterial agent, such as against E. coli; an anti-inflammatory; an anti- viral; an anti-fungal; or may alleviate symptoms of bacterial, viral or fungal infections. Allicin is typically produced from alliin ((2R)-2-amino-3-[(5)-prop-2- enylsulfinyljpropanoic acid) in damaged garlic cells by the action of the enzyme alliinase.

When the garlic cells are damaged, such as by chopping, crushing, or cooking the garlic, the alliinase enzyme converts the alliin into allicin. Allicin, alliin, and/or alliinase may be provided as whole garlic cloves or bulbs; crushed, mashed, or chopped garlic; a garlic extract; and/or as a synthesized or isolated compound.

C. Vitamins, yeast and/or sorbic acid or sorbic acid salt

The one or more vitamins may include vitamin A; vitamin Bi, such as thiamine

mononitrate; vitamin B2, such as riboflavin-5-phosphate; vitamin B3, such as niacin or niacinamide; vitamin B5, such as pantothenic acid or d-calcium pantothenate; vitamin B₆, such as pyridoxine or pyridoxine hydrochloride; vitamin B12; vitamin C, such as ascorbic acid, sodium ascorbate, or calcium sorbate; vitamin D; vitamin E; vitamin K, or a combination thereof. Vitamin D may comprise vitamin Di, vitamin D2, vitamin D3, vitamin D₄, vitamin D5, 25 -hydroxy vitamin D3, 25- dihydroxy vitamin D3, or combinations thereof.

The yeast may be a yeast culture, a live yeast, a dead yeast, yeast extract, or a combination thereof. The yeast may be a baker's yeast, a brewer's yeast, a distiller's yeast, a probiotic yeast or a combination thereof. Exemplary yeast's include, but are not limited to, Saccharomyces cerevisiae, Saccharomyces boulardii, Saccharomyces pastorianus, Brettanomyces bruxellensis, Brettanomyces anomalus, Brettanomyces custersianus, Brettanomyces naardenensis, and Brettanomyces nanus, Candida stellata, Schizosaccharomyces pombe, Torulaspora delbrueckii, or Zygosaccharomyces bailii.

In some embodiments, the combination comprises sorbic acid or a salt thereof. Sorbic acid, or a salt thereof, may act as a preservative, such as by inhibiting mold and/or yeast growth. The salt may be any suitable salt of sorbic acid, and in some embodiments, is a group I, group II, or organic salt of sorbic acid. Suitable salts include, but are not limited to, potassium sorbate, sodium sorbate, or ammonium sorbate.

D. Yucca, quillaja, and/or probiotic

Additionally, or alternatively, the combination may comprise yucca, quillaja or both.

Examples of yucca include, but are not limited to, Yucca aloifolia, Yucca angustissima, Yucca arkansana, Yucca baccata, Yucca baileyi, Yucca brevifolia, Yucca campestris, Yucca capensis, Yucca carnerosana, Yucca cernua, Yucca coahuilensis, Yucca constricta, Yucca decipiens, Yucca declinata, Yucca de-smetiana, Yucca elata, Yucca endlichiana, Yucca faxoniana, Yucca

filamentosa, Yucca filif era, Yucca flaccida, Yucca gigante an, Yucca glauca, Yucca gloriosa, Yucca grandiflora, Yucca harrimaniae, Yucca intermedia, Yucca jaliscensis, Yucca lacandonica, Yucca linearifolia, Yucca luminosa, Yucca madrensis, Yucca mixtecana, Yucca necopina, Yucca neomexicana, Yucca pallida, Yucca periculosa, Yucca potosina, Yucca queretaroensis, Yucca reverchonii, Yucca rostrata, Yucca rupicola, Yucca schidigera, Yucca schottii, Yucca sterilis, Yucca tenuistyla, Yucca thompsoniana, Yucca treculeana, Yucca utahensis, Yucca valida, or a combination thereof. In certain embodiments, the yucca is Yucca schidigera.

Examples of quillaja include, but are not limited to, Quillaja brasiliensis, Quillaja lanceolata, Quillaja lancifolia, Quillaja molinae, Quillaja petiolaris, Quillaja poeppigii, Quillaja saponaria, Quillaja sellowiana, Quillaja smegmadermos, or a combination thereof. In certain embodiments, the quillaja is Quillaja saponaria.

A person of ordinary skill in the art will appreciate that, as used herein, a plant name may refer to the plant as a whole, or to any part of the plant, such as the roots, stem or trunk, bark, leaves, flower, flower stems, or seeds or a combination thereof. These plant parts may be used fresh, or dried, and may be whole, pulverized, or comminuted. The name may also refer to extracts from any part or parts of the plant, such as chemical extracts, or extracts, including the residues, obtained by pressing, or any other methods of concentrating or extracting oils or other extracts known to those in the art or that are hereafter discovered. Plant extracts may include compounds that are saponins, tri terpenoids, polyphenols, antioxidants or resveratrol, or combinations thereof.

The combination may comprise a composition comprising yucca and/or quillaja that may also include carriers and binding agents suitable to formulate the yucca and/or quillaja for administration to an animal. In certain embodiments, such a composition can be a commercially available product, such as a composition comprising Yucca schidigera and Quillaja saponaria, sold under the trademark NUTRAFITO PLUS by Desert King International and/or MAGNI-PHI by Phibro Animal Health Corporation. Such compositions may comprise 85% Quillaja saponaria and 15% Yucca schidigera or 90% Quillaja saponaria and 10% Yucca schidigera.

The combination may also comprise a probiotic, such as a direct-fed microbial. The probiotic may be a Bacillus species. Bacillus is a genus of Gram-positive, rod-shaped bacteria. Examples of Bacillus include, but are not limited to, B. alcalophilus, B. alvei, B. aminovorans, B. amyloliquejaciens, B. aneurinolyticus, B. anthracis, B. aquaemaris, B. atrophaeus, B. boroniphilus, B. brevis, B. caldolyticus, B. centrosporus, B. cereus, B. circulans, B. coagulans, B. firmus, B. flavothermus, B. fusiformis, B. galliciensis, B. globigii, B. infernus, B. larvae, B. laterosporus, B. lentus, B. licheniformis, B. megaterium, B. mesentericus, B. mucilaginosus, B. mycoides, B. natto, B. pantothenticus, B. polymyxa, B. pseudoanthracis, B. pumilus, B. schlegelii, B. sphaericus, B. sporothermodurans, B. stearothermophilus, B. subtilis, B. thermoglucosidasius, B. thuringiensis, B. vulgatis, or B. weihenstephanensis. In some embodiments, the Bacillus is not Bacillus subtilis. In particular embodiments the Bacillus is Bacillus coagulans. A person of ordinary skill in the art will appreciate that, as used herein, the bacterial name may refer to the bacteria, or to a compound or compounds obtained from that bacteria. Methods of obtaining compounds from bacteria are well known in the art. A composition comprising Bacillus may also include additional materials, such as carriers or binding agents, suitable to formulate the Bacillus for administration to an animal. In certain embodiments, Bacillus coagulans is provided by Ganpro^®, a commercial product available from Ganeden Biotech, Ohio. In other embodiments, Bacillus coagulans is Provia 6086^®, available from Prince Agri Products, Inc.

E. Algae

The algae may be a blue-green algae (cyanobacteria), a diatom (bacillariophyta), a stonewort algae (charophyta), a green algae (chlorophyta), a golden algae (chrysophyta), a dinoflagellate (dinophyta), a brown algae (phaeophyta) or a red algae (rhodophyta). In some embodiments, the algae is a chlorophyta, and may be an algae from the genus Chlorella, including, but not limited to, Chlorella vulgaris, Chlorella angustoellipsoidea, Chlorella botryoides,

Chlorella capsulata, Chlorella ellipsoidea, Chlorella emersonii, Chlorella fusca, Chlorella homosphaera, Chlorella luteo-v iridis, Chlorella marina, Chlorella miniata, Chlorella minutissima, Chlorella mirabilis, Chlorella ovalis, Chlorella parasitica, Chlorella peruviana, Chlorella rugosa, Chlorella saccharophila, Chlorella salina, Chlorella spaerckii, Chlorella sphaerica, Chlorella stigmatophora, Chlorella subsphaerica, Chlorella trebouxioides, or a combination thereof. In other embodiments, the algae is a cyanobacteria, such as Arthrospira platensis or Arthrospira maxima (spirulina). Other algae include, but are not limited to, algae of the genus Pediastrum, such as Pediastrum dupl, Pediastrum boryanum, or a combination thereof; algae of the genus

Botryococcus, such as Botryococcus braunii; algae of the genus Porphyra, such as Porphyra dioica, Porphyra linearis, Porphyra lucasii, Porphyra mumfordii, Porphyra purpurea, Porphyra umbilicalis, or a combination thereof.

F. Antimicrobial, vaccine and/or growth promotant

The antimicrobial may be an antibiotic, an antifungal, an antiparasitic, an antiviral, or a combination thereof. An antibiotic may be a tetracycline, a penicillin, a cephalosporin, a polyether antibiotic, a glycopeptide, an orthosomycin, or a combination thereof. The antibiotic may be selected from, by way of example, and without limitation, virginiamycin, Bacitracin MD, Zinc Bacitracin, Tylosin, Lincomycin, Flavomycin, bambermycins, Terramycin, Neo-Terramycin, florfenicol, oxolinic acid, oxytetracycline, hydrogen peroxide (Perox-Aid® 35%), bronopol (2- bromo-2-nitro-l,3-propanediol, Pyceze®), sulfadimethozine, ormetoprim, Sulfadiazine,

Trimethoprim, or a combination thereof. In some embodiments, the antibiotic is not, or does not comprise, hydrogen peroxide. In some embodiments, the antibiotic is virginiamycin, Bacitracin MD, Zinc Bacitracin, Tylosin, Lincomycin, Flavomycin, bambermycins, Terramycin, Neo- Terramycin, florfenicol, oxolinic acid, oxytetracycline, bronopol (2-bromo-2-nitro-l,3-propanediol, Pyceze®), sulfadimethozine, ormetoprim, Sulfadiazine, Trimethoprim, or a combination thereof.

An antifungal may be selected from, by way of example, formalin, formalin-F, bronopol (2- bromo-2-nitro-l,3-propanediol, Pyceze®), or a combination thereof. Exemplary antiparasitics may be selected from an anticoccidal, copper sulfate, fenbendazole, formalin, formalin-F, hyposalinity, hadaclean A, praziquantel, emamectin benzoate (SLICE®), or a combination thereof.

Suitable anticoccidial agents include, but are not limited to, ionophores and chemical anticoccidial products. Ionophores can include, but are not limited to, Monensin, Salinomycin, Lasalocid, Narasin, Maduramicin, Semduramicin, or combinations thereof.

Chemical anticoccidial products can include, but are not limited to, Nicarbazin, Maxiban, Diclazuril, Toltrazuril, Robenidine, Stenorol, Clopidol, Decoquinate, DOT (zoalene), Amprolium, or combinations thereof.

Suitable vaccines can be selected from live coccidiosis vaccines, such as COCCIVAC (e.g. , a composition comprising live oocysts of Eimeria acervulina, Eimeria mivati, Eimeria maxima, Eimeria mitis, Eimeria tenella, Eimeria necatrix, Eimeria praecox, Eimeria brunetti, Eimeria hagani, or combinations thereof), LivaCox (a composition comprising 300 - 500 live sporulated oocysts of each attenuated line of Eimeria acervulina, E. maxima and E. tenella in a 1 % w/v aqueous solution of Chloramine B); ParaCox (a composition comprising live sporulated oocysts derived from E. acervulina HP, E. brunetti HP, E. maxima CP, E. maxima MFP, E mitis HP, E. necatrix HP, E. praecox HP, E. tenella HP, and combinations thereof); Hatch Pack Cocci III (a composition comprising oocysts derived from Eimeria acervulina, Eimeria maxima, Eimeria tenella, or combinations thereof); INOVOCOX (a composition comprising oocysts derived from Eimeria acervulina, Eimeria maxima, Eimeria tenella, and a sodium chloride solution);

IMMUCOX (a composition comprising live oocysts derived from Eimeria acervulina, Eimeria maxima, Eimeria necatrix, Eimeria tenella, and combinations thereof), Advent, or combinations thereof. Vaccines may also comprise live oocysts of the Eimeria genus, for example, Eimeria aurati, Eimeria baueri, Eimeria lepidosirenis, Eimeria leucisci, Eimeria rutile, Eimeria carpelli, Eimeria subepithelialis, Eimeria funduli and/or Eimeria vanasi. Vaccines may also comprise oocysts from the genus Epeimeria, a new genus of coccidia infecting fishes.

Other suitable vaccines include, but are not limited to, ALPHA DIP® 2000, ALPHA DIP® Vibrio, ALPHA MARINE® Vibrio, ALPHA DIP® ERM Salar, ALPHA JECT micro® 1 ILA, ALPHA JECT micro® 7ILA, ALPHA JECT® Panga, ALPHA JECT® 1000, ALHPA JECT® 2000, ALPHA JECT® 3000, ALPHA JECT® 3-3, ALPHA JECT® 4000, ALPHA JECT® 4-1, ALPHA JECT® 5-1, ALPHA JECT® 5-3, ALPHA JECT® 6-2, ALPHA JECT® micro 1 ISA, ALPHA JECT® micro 2, ALPHA JECT® micro 4, Apex®-IHN, AQUA VAC® ERM Oral, AQUA VAC® ERM immersion, AQUA VAC® FNM Injectable, AQUA VAC® IPN Oral,

AQUA VAC® RELERA™, AQUA VAC® Vibrio Oral, AQUA VAC® Vibrio Pasteurella injection, AQUA VAC® Vibrio immersion and injectable, AQUA VAC-COL™ immersion, AQUAVAC- ESC™ immersion, Birnagen Forte 2, Ermogen, Forte Micro, Forte V II, Forte VI, Fry Vacc 1, Furogen Dip, ICTHIOVAC JG injection, ICTHIOVAC® PD immersion, Lipogen DUO, Lipogen Forte, Microvib, Norvax® Compact PD injection, Norvax® Minova 4WD, Norvax® Minova 6 injection, Norvax® STREP Si immersion and injection, Premium Forte Plus, Premium Forte Plus ILA, Renogen, Vibrogen 2, or a combination thereof.

Growth promotants are used to help increase the efficiency of animal production, such as by increasing the rate of weight gain, improved feed efficiency and/or product output. A growth promotant may also increase the quality of a product, such as increase the quality of meat produced. Growth promotants can include, but are not limited to, β-agonists, antibiotics, antimicrobials, steroids and hormones. In some embodiments, a growth promotant may be a compound that has one or more other uses and is used as a growth promotant at a lower dose than the dose for the primary application. For example, an antibiotic or antimicrobial compound may also be useful as a growth promotant when used at a sub-therapeutic dose. Exemplary growth promotants include, but are not limited to, β-agonists such as ractopamine and zilpaterol; somatotropin such as bovine somatotropin (bST) and recombinant bovine somatotropin (rbST); ionophores such as monesin, lasalocid, laidlomycin, salinomycin and narasin; hormones such as oestrogen, progesterone, testosterone and analogs thereof; estradiol benzoate; tetracyclines, such as oxy tetracycline, chlortetracycline, tetracycline, demeclocycline, doxycycline, lymecycline, meclocycline, methacycline, minocycline, rolitetracycline, and salts thereof, for example, hydrochloride, hydrobromide, hydroiodide, calcium, sodium, potassium, magnesium, or lithium salts; arsanilic acid; 4-hydroxy-3-nitrobenzenearsonic acid, erythromycin thiocyanate, tylosin phosphate, melengestrol acetate, iodinated casein, ethopabate, oleandomycin, penicillin G procaine, chlortetracycline, sulfathiazole, bambermycins, bacitracin, virginiamycin, chlortetracycline calcium complex, or salt and/or combinations thereof.

G. Polyphenol

In some embodiments, the polyphenol is provided by a plant extract from a polyphenol- containing plant material. The plant material also may include non-polyphenol compounds, including polyphenol degradation products, such as gallic acid and trans-caftaric acid. Degradation can occur, for example, through oxidative and/or biological processes. Both the polyphenols and the non-polyphenol compounds may have biological activity. The plant extract may be prepared from a single plant material or from a combination of plant materials. Suitable plant materials from which a plant extract can be obtained include, but are not limited to, apples, blackberries, black chokeberries, black currants, black elderberries, blueberries, cherries, cranberries, grapes, green tea, hops, onions, quillaja, plums, pomegranates, raspberries, strawberries, and yucca.

In some embodiments, the plant extract is prepared from a pressed plant material, such as grape pomace, a dried plant material, such as tea, or a combination thereof. Pomace may be obtained substantially immediately post-pressing or as an ensiled product, i.e. , pomace collected and stored for up to several months post-pressing. Suitable plants have a plurality of polyphenols and/or other non-polyphenolic compounds including, but not limited to, non-polyphenolic organic acids (such as gallic acid and/or trans-caftaric acid), flavanols, gallate esters, flavanodiols, phloroglucinol, pyrogallol, and catechol. In some embodiments, the plant extract is prepared from Pinot noir pomace, Pinot gris pomace, or green tea.

In some embodiments, pressed or dried plant material is ground to a fine powder prior to, or during, extraction. Pressed plant materials may be frozen to facilitate grinding. Polyphenols and other non-polyphenolic compounds may be extracted for administration. For example, polyphenols and other non-polyphenolic compounds may be extracted from the powder using a solution comprising a polar solvent, such as water, an alcohol, an ester, or a combination thereof. In some embodiments, the solution comprises a water-miscible alcohol, ester, or combination thereof, such as a lower alkyl alcohol, lower alkyl ester, or a combination thereof. In some embodiments, the solution is water or an aqueous solution comprising 25-99% solvent, such as 25-95% solvent, SO- 80% solvent, or 50-75% solvent, and water. In certain embodiments, the solution is an aqueous solution comprising methanol, ethanol, isopropanol, ethyl acetate, or a combination thereof. The solution may be acidified by addition of an acid. The acid may prevent or minimize oxidative degradation of biologically-active polyphenols and other non-polyphenolic compounds in the extract. The acid may be any suitable acid, such as a mineral acid (e.g. , hydrochloric acid), or an organic acid such as citric acid or acetic acid. In some embodiments, the solution comprises from 0.01% to 1% acid, such as 0.02-0.5%, 0.025-0.25%, or 0.05-0.15%. In some examples, the solution includes 0.1% hydrochloric acid.

Extraction may be performed at a temperature ranging from 0-100 °C. In some embodiments, extraction is performed at a temperature ranging from 20-70 °C, or at ambient temperature. Extraction may be performed for a duration ranging from several minutes to several days. To increase extraction efficiency, the plant material and solution may be mixed or agitated during extraction, such as by grinding the plant material during extraction, stirring the mixture, shaking the mixture, or homogenizing the mixture. In some embodiments, the extraction may be repeated one or more times with fresh solution to increase recovery of polyphenols and other non- polyphenolic compounds from the plant material. The liquid phases from each extraction cycle are then combined for further processing.

The liquid phase can recovered, and the residual solids, or pulp, are discarded. Recovering the liquid phase may comprise decanting the liquid from the remaining solids and/or filtering the liquid phase to remove residual solids. The solvent (alcohol, ester, or combination thereof) can be removed from the liquid solution by any suitable means, such as evaporation (e.g. , roto- evaporation), to produce an aqueous extract containing the biologically-active components in a mildly acidic solution.

In certain embodiments where the plant material includes a significant amount of oils, or lipids, an initial extraction of nonpolar components may be performed before extracting the polyphenols and other polar, non-polyphenolic compounds. Nonpolar components may be extracted by homogenizing the plant material in a nonpolar solvent, e.g., hexanes, heptanes, or a combination thereof. The solvent layer including the extracted nonpolar components is separated from the plant material and discarded.

The aqueous plant extract may be further purified by suitable means, e.g., extraction, chromatographic methods, distillation, etc., to remove non-polyphenolic compounds and/or to increase the concentration of polyphenols relative to other compounds in the extract.

The aqueous plant extract may be dried, for example by freeze-drying or other low- temperature drying methods, and ground to a powder to provide a dried plant extract. In some embodiments, the dried plant extract comprises 0.01 wt% to 25 wt% total polyphenols, such as 0.01 wt% to 10 wt%, 0.01 wt% to 5 wt%, 0.01 wt% to 2.5 wt%, 0.01 wt% to 1 wt%, 0.01 wt% to 0.5 wt%, 0.02 to 0.25 wt%, or 0.03-0.1 wt% total polyphenols. In certain embodiments, the dried plant extract further comprises non-polyphenolic compounds. For example, the dried plant extract may comprise 0.01-1 mg/g gallic acid, such as 0.05-0.5 mg/g or 0.09-0.25 mg/g gallic acid, and/or 0.001-0.1 mg/g trans-caftaric acid, such as 0.005-0.05 mg/g or 0.01-0.025 mg/g trans-caftaric acid.

The aqueous plant extract may be concentrated to a smaller volume, e.g. , by evaporation, and used as an aqueous plant extract. In other embodiments, the aqueous plant extract is mixed with a carrier before drying and grinding. Suitable carriers include, for example, diatomaceous earth, silica, maltodextrin, ground grain (e.g. , corn), meals (e.g. , soybean or cottonseed meal) byproducts (e.g. , distiller's dried grains, rice hulls, wheat mill run), clays (e.g. , bentonite), and combination thereof. The plant extract may be combined with a carrier in a ratio ranging from 10: 1 to 1: 10 by weight, such as from 5: 1 to 1:5. For example, the plant extract may be mixed with diatomaceous earth in a ratio of 3 : 1 by weight.

H. Embodiments of a combination

In some embodiments, the combination comprises one or more of allicin, alliin, allinase, or algae, and one or more of silica, mineral clay, glucan, mannans, endoglucanohydrolase, yeast, one or more vitamins, yucca, quillaja, polyphenol, one or more probiotics, an antimicrobial, a vaccine, sorbic acid or a salt thereof, or a growth promotant. In certain embodiments, the combination comprises one or more of allicin, alliin, allinase, or algae, and silica, mineral clay, glucan, and mannans, and optionally may further comprise endoglucanohydrolase and/or yeast. In other embodiments, the combination comprises one or more of allicin, alliin, allinase, or algae, and yucca and quillaja, and optionally comprises a probiotic, such as Bacillus coagulans. In certain embodiments, the combination comprises allicin, silica, mineral clay, glucan, and mannans; allicin, silica, mineral clay, glucan, mannans, and endoglucanohydrolase; allicin, yucca, such as Yucca schidigera, and quillaja, such as Quillaja saponaria; allicin, silica, mineral clay, glucan, mannans, yucca, such as Yucca schidigera, and quillaja, such as Quillaja saponaria; or allicin, silica, mineral clay, glucan, mannans, endoglucanohydrolase, yucca, such as Yucca schidigera, and quillaja, such as Quillaja saponaria. The combination may comprise from 0.1% to 99.9% allicin, alliin, allinase, algae, or a combination thereof, relative to the amounts of other components in the combination, such as from 1% to 99%, from 1% to 90%, from 1% to 80%, from 1 % to 70%, from 1% to 60%, from 1% to 50%, from 1% to 40%, from 1% to 30%, from 1% to 25%, from 1% to 20%, or from 1% to 10% allicin, alliin, allinase, algae, or a combination thereof.

In some embodiments, the combination comprises silica, glucan, mannans and mineral clay, and may comprise silica, glucan, mannans and mineral clay in the following relative amounts: 1-40 wt% silica, 0.5-25 wt% glucan and mannans, and 40-92 wt% mineral clay, such as 1-40 wt% silica, 1-25 wt% glucan and mannans, and 40-92 wt% mineral clay; 5-40 wt% silica, 0.5-15 wt% glucan and mannans, and 40-80 wt% mineral clay, such as 5-40 wt% silica, 2-15 wt% glucan and mannans, 40-80 wt% mineral clay; 20-40 wt% silica, 0.5-10 wt% glucan and mannans, and 50-70 wt% mineral clay, such as 20-40 wt% silica, 4-10 wt% glucan and mannans, and 50-70 wt% mineral clay; 15-40 wt% silica, greater than zero to 15 wt% glucans such as 1-15 wt% glucans, 0- 10 wt% mannans or greater than 0 to 10 wt% mannans, 50-81 wt% mineral clay; 15-40 wt% silica, 0.5-5.0 wt% glucans, 0.5-8.0 wt% mannans, and 50-81 wt% mineral clay, such as 15-40 wt% silica, 1.0-5.0 wt% glucans, 1.0-8.0 wt% mannans, 50-81 wt% mineral clay; 20-30 wt% silica, 0.5-3.5 wt% glucans, 0.5-6.0 wt% mannans, and 60-70 wt% mineral clay; or 20-30 wt% silica, 1.0-3.5 wt% glucans, 1.0-6.0 wt% mannans, 60-75 wt% mineral clay.

Since β-glucans and mannans may be obtained from yeast, such as from a yeast cell wall or an extract thereof, the combination may comprise silica, mineral clay, and yeast, yeast cell wall or an extract thereof in the following relative amounts: 1-40 wt% silica, 1-30 wt% yeast cell wall extract, and 40-92 wt% mineral clay; 10-40 wt% silica, 5-20 wt% yeast cell wall extract, 40-80 wt% mineral clay; or 15-30 wt% silica, 5-15 wt% yeast cell wall extract, 55-70 wt% mineral clay.

The combination may further comprise an endoglucanohydrolase, such as β-1,3 (4)- endoglucanohydrolase. The combination may include from 0.05 wt% endoglucanohydrolase to 5 wt% endoglucanohydrolase or more, relative to an amount of silica, mineral clay, glucan and mannans, or yeast, yeast cell wall or an extract thereof, such as from 0.05 wt% to 3 wt% β-1,3 (4)- endoglucanohydrolase. In some examples, the combination comprises silica, mineral clay, glucan and mannans, or yeast, yeast cell wall or an extract thereof, and endoglucanohydrolase in the following relative amounts: 0.1-3 wt% β-1,3 (4)-endoglucanohydrolase, 20-40 wt% silica, 2-20 wt% glucan and mannans, 50-70 wt% mineral clay; 0.2-3 wt%, β-1,3 (4)-endoglucanohydrolase, 20-40 wt% silica, 4-10 wt% glucan and mannans, 50-70 wt% mineral clay; 0.05-0.3% β-1,3 (4)- endoglucanohydrolase, 1-40 wt% diatomaceous earth, 1-20 wt% yeast cell wall extract, 40- 92% wt% mineral clay; 0.1-3 wt% β-1,3 (4)-endoglucanohydrolase, 5-40 wt% diatomaceous earth, 2-10 wt% yeast cell wall extract, 40-80 wt% mineral clay; 0.2-3 wt%, β-1,3 (4)- endoglucanohydrolase, 30-40 wt% diatomaceous earth, 4-6 wt% yeast cell wall extract, 50-65 wt% mineral clay; 0.1-3 wt%, β-1,3 (4)-endoglucanohydrolase, 20-40 wt% diatomaceous earth, 2-20 wt% β-glucan and glucomannan, and 50-70 wt% mineral clay; 0.1-3 wt% β-1,3 (4)- endoglucanohydrolase, 20-40 wt% silica, 0.5-20 wt% glucan and mannans, and 50-70 wt% mineral clay; 0.1-3 wt%, β-1,3 (4)-endoglucanohydrolase, 20-40 wt% silica, 0.5-10 wt% glucan and mannans, and 50-70 wt% mineral clay; 0.1-3 wt% β-1,3 (4)-endoglucanohydrolase, 1-40 wt% silica, 5-30 wt% yeast cell wall or an extract thereof, and 40-92 wt% mineral clay; 0.1-3 wt% β-1,3 (4)-endoglucanohydrolase, 10-40 wt% silica, 5-20 wt% yeast cell wall or an extract thereof, and 40- 80 wt% mineral clay; or 0.1-3 wt% β-1,3 (4)-endoglucanohydrolase, 15-30 wt% silica, 5-15 wt% yeast cell wall or an extract thereof, and 50-70 wt% mineral clay.

In some embodiments, the combination comprises a component 1 selected from: 1A) silica; IB) mineral clay; 1C) glucan; ID) mannans; IE) yeast cell wall or an extract thereof; IF) endoglucanohydrolase; 1G) silica and mineral clay; 1H) silica and glucan; II) silica and mannans; 1J) silica and yeast cell wall or an extract thereof; IK) silica and endoglucanohydrolase; 1L) mineral clay and glucan; 1M) mineral clay and mannans; IN) mineral clay and yeast cell wall or an extract thereof; 10) mineral clay and endoglucanohydrolase; IP) glucan and mannans; 1Q) glucan and yeast cell wall or an extract thereof; 1R) glucan and endoglucanohydrolase; IS) mannans and yeast cell wall or an extract thereof; IT) mannans and endoglucanohydrolase; 1U) silica, mineral clay and glucan; IV) silica, mineral clay and mannans; 1W) silica, mineral clay and yeast cell wall or an extract thereof; IX) silica, mineral clay and endoglucanohydrolase; 1Y) silica, glucan and mannans; 1Z) silica, glucan and yeast cell wall extract or an extract thereof; 1AA) silica, glucan and endoglucanohydrolase; 1AB) silica, mannans and yeast cell wall or an extract thereof; 1AC) silica, mannans and endoglucanohydrolase; IAD) silica, yeast cell wall or an extract thereof, and endoglucanohydrolase; 1AE) mineral clay, glucan and mannans; 1AF) mineral clay, glucan and yeast cell wall or an extract thereof; 1AG) mineral clay, glucan and endoglucanohydrolase; 1AH) glucan, mannans, and yeast cell wall or an extract thereof; 1AI) glucan, mannans and

endoglucanohydrolase; 1AJ) mannans, yeast cell wall or an extract thereof, and

endoglucanohydrolase; 1AK) silica, mineral clay, glucan and mannans; 1AL) silica, mineral clay, glucan and yeast cell wall or an extract thereof; 1AM) silica, mineral clay, glucan and

endoglucanohydrolase; IAN) mineral clay, glucan, mannans, and yeast cell wall or an extract thereof; 1AO) mineral clay, glucan, mannans, and endoglucanohydrolase; 1AP) glucan, mannans, yeast cell wall or an extract thereof, and endoglucanohydrolase; 1AQ) yeast cell wall or an extract thereof, and endoglucanohydrolase; 1AR) silica, mineral clay, glucan, mannans, and yeast cell wall or an extract thereof; IAS) silica, mineral clay, glucan, mannans, and endoglucanohydrolase; 1AT) mineral clay, glucan, mannans, yeast cell wall or an extract thereof, and endoglucanohydrolase; 1AU) silica, mineral clay, glucan, mannans, yeast cell wall or an extract thereof, and

endoglucanohydrolase; 1AV) allicin; 1AW) alliin; 1AX) allinase; 1AY) allicin and alliin; 1AZ) allicin and allinase; 1BA) alliin and allinase; IBB) allicin, alliin, and allinase; 1BC) yeast; 1BD) polyphenol; 1BE) algae; 1BF) yucca; 1BG) quillaja; 1BH) Yucca schidigera; 1BJ) Quillaja saponaria; 1BL) Bacillus coagulans; IBM) Yucca schidigera and Bacillus coagulans; 1BN) Quillaja saponaria and Bacillus coagulans; 1BO) Yucca schidigera, and Quillaja saponaria; IBP) Yucca schidigera, Quillaja saponaria and Bacillus coagulans; 1BQ) a probiotic; 1BR) an antimicrobial; IBS) a vaccine; 1BT) sorbic acid or a salt thereof; 1BU) potassium sorbate; 1BV) one or more vitamins; IBW) vitamin C; IBX) vitamin D; IBY) vitamin C and vitamin D; IBZ) an antibiotic; 1CA) Virginamycin; 1CB) an antifubgal; ICC) an antiparasitic; 1CD) an antiviral; or ICE) an anticoccidial.

The combination also comprises a component 2. With respect to the component 1 embodiments, the component 2 may be, in a combination with 1A to ICE: 2A) allicin; 2B) alliin; 2C) allinase; 2D) allicin and alliin; 2E) allicin and allinase; 2F) alliin and allinase; 2G) allicin, alliin, and allinase; 2H) yeast; 21) polyphenol; 2J) algae; 2K) yucca; 2L) quillaja; 2M) Yucca schidigera; 2N) Quillaja saponaria; 20) Bacillus coagulans; 2P) Yucca schidigera and Bacillus coagulans; 2Q) Quillaja saponaria and Bacillus coagulans; 2R) Yucca schidigera, and Quillaja saponaria; 2S) Yucca schidigera, Quillaja saponaria and Bacillus coagulans; 2T) a probiotic; 2U) an antimicrobial; 2V) a vaccine; 2W) sorbic acid or a salt thereof; 2X) potassium sorbate; 2Y) one or more vitamins; 2Z) vitamin C; 2AA) vitamin D; 2AB) vitamin C and vitamin D; 2AC) an antibiotic; 2AD) Virginamycin; 2AE) an antifungal; 2AD) an antiparasitic; 2AE) an antiviral; or 2AF) an anticoccidial.

A person of ordinary skill in the art will understand that any of 2A to 2AF may be combined with any of 1A to ICE, to form any and all compositions and/or combinations between such components.

The combination may also comprise a component 3. With respect to the component 1 embodiments 1A to ICE and the component 2 embodiments 2A to 2AF, component 3 may be, in combination with 1A to ICE and 2A to 2AF: 3A) yeast; 3B) polyphenol; 3C) algae; 3D) yucca; 3E) quillaja; 3F) Yucca schidigera; 3G) Quillaja saponaria; 3H) Bacillus coagulans; 31) Yucca schidigera and Bacillus coagulans; 3J) Quillaja saponaria and Bacillus coagulans; 3K) Yucca schidigera, and Quillaja saponaria; 3L) Yucca schidigera, Quillaja saponaria and Bacillus coagulans; 3M) a probiotic; 3N) an antimicrobial; 30) a vaccine; 3P) sorbic acid or a salt thereof; 3Q) potassium sorbate; 3R) one or more vitamins; 3S) vitamin C; 3T) vitamin D; 3U) vitamin C and vitamin D; 3V) an antibiotic; 3W) Virginamycin; 3X) an antifungal; 3Y) an antiparasitic; 3Z) an antiviral; or 3 A A) an anticoccidial.

A person of ordinary skill in the art will understand that any of 3A to 3AA may be combined with any of 1A to ICE and any of 2A to 2AF, to form any and all compositions and/or combinations between such components.

The combination may further comprise a component 4. With respect to the component 1 embodiments 1A to ICE, the component 2 embodiments 2A to 2AF, and the component 3 embodiments 3A to 3AA, component 4 may be, in combination with 1A to ICE, 2A to 2AF, and 3A to 3AA: 4A) yeast; 4B) polyphenol; 4C) algae; 4D an antimicrobial; 4E) a vaccine; 4F) sorbic acid or a salt thereof; 4G) potassium sorbate; 4H) one or more vitamins; 41) vitamin C; 4J) vitamin

D; 4K) vitamin C and vitamin D; 4L) an antibiotic; 4M) Virginamycin; 4N) an antifungal; 40) an antiparasitic; 4P) an antiviral; or 4Q) an anticoccidial.

A person of ordinary skill in the art will understand that any of 4A to 4Q may be combined with any of 1A to ICE, any of 2A to 2AF, and any of 3A to 3AA to form any and all compositions and/or combinations between such components.

The combination may further comprise a component 5. With respect to the component 1 embodiments 1A to ICE, the component 2 embodiments 2A to 2AF, the component 3

embodiments 3A to 3AA, and the component 4 embodiments 4A to 4Q, component 5 may be, in combination with 1A to ICE, 2A to 2AF, 3A to 3AA, and 4A to 4Q: 5A) yeast; 5B) polyphenol;

5C) algae; 5D) an antimicrobial; 5E) a vaccine; 5F) sorbic acid or a salt thereof; 5G) potassium sorbate; 5H) an antibiotic; 51) Virginamycin; 5J) an antifungal; 5K) an antiparasitic; 5L) an antiviral; or 5M) an anticoccidial.

A person of ordinary skill in the art will understand that any of 5A to 5M may be combined with any of 1A to ICE, any of 2A to 2AF, any of 3A to 3AA, and any of 4A to 4Q to form any and all compositions and/or combinations between such components.

The combination may further comprise a component 6. With respect to the component 1 embodiments 1A to ICE, the component 2 embodiments 2A to 2AF, the component 3

embodiments 3A to 3AA, the component 4 embodiments 4A to 4Q, and the component 5 embodiments 5A to 5M, component 6 may be, in combination with 1A to ICE, 2A to 2AF, 3A to

3AA, 4A to 4Q, and 5A to 5M: 6A) yeast; 6B) polyphenol; 6C) algae; 6D) a vaccine; 6E) sorbic acid or a salt thereof; or 6F) potassium sorbate.

A person of ordinary skill in the art will understand that any of 6A to 6F may be combined with any of 1A to ICE, any of 2A to 2AF, any of 3A to 3AA, any of 4A to 4Q, and any of 5A to

5M to form any and all compositions and/or combinations between such components.

The combination may further comprise a component 7. With respect to the component 1 embodiments 1A to ICE, the component 2 embodiments 2A to 2AF, the component 3

embodiments 3A to 3AA, the component 4 embodiments 4A to 4Q, the component 5 embodiments 5A to 5M, and the component 6 embodiments 6A to 6F component 7 may be, in combination with

1A to ICE, 2A to 2AF, 3A to 3 A A, 4A to 4Q, 5A to 5M, and 6A to 6F: 7A) yeast; 7B) polyphenol; 7C) algae; 7D) a vaccine; 7E) yeast and polyphenol; 7F) yeast and algae; 7G) yeast and a vaccine; 7H) polyphenol and algae; 71) polyphenol and a vaccine; 7J) algae and a vaccine; 7K) yeast, polyphenol, and algae; 7L) yeast, polyphenol, and a vaccine; 7M) yeast, algae and a vaccine; 7N) polyphenol, algae and a vaccine; or 70) yeast, polyphenol, algae and a vaccine.

A person of ordinary skill in the art will understand that any of 7A to 70 may be combined with any of 1A to ICE, any of 2A to 2AF, any of 3A to 3AA, any of 4A to 4Q, any of 5A to 5M, and any of 6A to 6F to form any and all compositions and/or combinations between such components.

In any of the above embodiments, glucan is β-glucan. In any of the above embodiments, endoglucanohydrolase is β-1,3 (4)-endoglucanohydrolase.

Any combination of any of 1A to ICE, and any of 2A to 2AF, and optionally any of 3A to 3AA, any of 4A to 4Q, any of 5A to 5M, any of 6A to 6F, and/or any of 7A to 70, might further comprise one or more additional components, such as an adhesive agent and/or feedstuff, as disclosed herein.

The disclosed combination may comprise from less than 0.1% to greater than 99.9% component 1, such as from 1% to 99%, from 1% to 75%, from 1% to 60%, from 1% to 50%, component 1, and from less than 0.1% to greater than 99.9% component 2, such as from 1% to 99%, from 1% to 75%, from 1% to 60%, from 1% to 50%, component 2. The combination optionally may further comprise from less than 0.1% to greater than 99.9%, such as from 1% to 99%, from 1% to 75%, from 1% to 60%, from 1% to 50%, of any of component 3, component 4, component 5, component 6, component 7, and/or the one or more additional components, such as an adhesive agent and/or feedstuff, as disclosed herein. However, a person of ordinary skill in the art will appreciate that the total relative amounts of the components in the combination cannot exceed 100%.

In particular embodiments, combination comprises, consists essentially of, or consists of, allicin, glucan (e.g., β-1,3 (4)glucan), silica, mineral clay and mannans; allicin, glucan (e.g. , β-1,3 (4)glucan), silica, mineral clay, mannans and endoglucanohydrolase; allicin, silica, mineral clay and yeast cell wall extract; allicin, yucca (e.g., Yucca schidigera) and quillaja (e.g. , Quillaja saponaria); allicin, glucan (e.g., β-1,3 (4)glucan), silica, mineral clay, mannans, yucca (e.g., Yucca schidigera) and quillaja (e.g. , Quillaja saponaria); allicin, glucan (e.g. , β-1,3 (4)glucan), silica, mineral clay, mannans, endoglucanohydrolase, yucca (e.g. , Yucca schidigera) and quillaja (e.g. , Quillaja saponaria); or allicin, silica, mineral clay, yeast cell wall or an extract thereof, yucca (e.g. , Yucca schidigera) and quillaja (e.g. , Quillaja saponaria). The combination may further comprise a feedstuff, such as an animal feed.

Any of these embodiments may further comprise an endoglucanohydrolase, such as β-1,3 (4)-endoglucanohydrolase; sorbic acid or a salt thereof, such as potassium sorbate, sodium sorbate, or ammonium sorbate; and/or a vitamin, such as vitamin C. In any of the above embodiments, the glucan and mannans may be provided by yeast cell wall extract. In some examples, the combination comprises mineral clay, silica, glucan, mannans, β-1,3 (4)-endoglucanohydrolase, allicin, potassium sorbate and vitamin C.

III. Additional components

In any of the above embodiments, the combination includes additional components.

Additional components may be used for any desired purpose, such as a substantially biologically inert material added, for example, as a filler, or to provide a desired beneficial effect. For example, the combination may include a carbonate (including a metal carbonate such as calcium carbonate); a trace mineral, such as, but not limited to, chloride, fluoride, iodide, chromium, copper, zinc, iron, magnesium, manganese, molybdenum, phosphorus, potassium, sodium, sulfur, selenium, or a combination thereof; a bulking agent; a carrier; a colorant; a taste enhancer; a preservative; or a combination thereof. The preservative may be benzoic acid or a salt thereof, e.g. sodium benzoate; lactic acid or a salt thereof, e.g. sodium lactate, potassium lactate or calcium lactate; propionic acid or a salt thereof, e.g. sodium propionate; ascorbic acid or a salt thereof, e.g. sodium ascorbate; gallic acid or a salt thereof e.g. sodium gallate; sulfur dioxide and/or sulfites; nitrites; nitrates; choline, or a salt thereof, such as an anion salt of choline, e.g. choline halide, such as chloride, bromide, iodide, fluoride, or choline hydroxide; or any combination thereof.

Additionally, or alternatively, the combination may further comprise corn, soybean meal, soybean oil, wheat, barley, rye, rice hulls, canola, corn oil, limestone, salt, distillers dried grains with solubles (DDGS), dicalcium phosphate, sodium sesquicarbonate, methionine source, lysine source, L-threonine, mineral oil, biotin, folic acid, kelp, menadione dimethylpyrimidinol bisulfite, calcium aluminosilicate, or any combination thereof.

In some embodiments, the combination does not comprise a peroxide compound.

In some embodiments, the combination does not comprise hydrogen peroxide.

In some embodiments, the composition and/or combination does not comprise carbamide peroxide.

In some embodiments, the combination does not comprise urea.

In some embodiments, the combination does not comprise hydrogen peroxide and urea.

In particular disclosed embodiments, the combination may further comprise a carrier and/or adhesive agent. The carrier and/or adhesive agent may be any carrier and/or adhesive agent known to a person of ordinary skill in the art as being suitable for combining with the combination. The amount of carrier and/or adhesive agent may be from zero to 10% or more by weight, such as from greater than zero to 10% or from 2% to 10% by weight. The carrier and/or adhesive agent is a material selected to, for example, facilitate adhering some or all of the components of the combination together, to a foodstuff, or both. The carrier and/or adhesive agent also may facilitate maintaining adherence of the combination together or to a foodstuff in an aquatic environment to facilitate administration to aquatic species. The carrier and/or adhesive agent also may be edible by animals, such as land animals, avians and/or aquatic animals.

In some embodiments, the carrier and/or adhesive agent is an oil. For example, the oil may be selected from corn oil, coconut oil, linseed oil, cottonseed oil, olive oil, peanut oil, palm oil, canola oil, safflower oil, soy oil, sunflower oil, Naskole oil, or any combination thereof. In some embodiments, the carrier and/or adhesive agent is a syrup. For example, the syrup may be selected from molasses, sorghum, sugar syrup, honey, or any combination thereof. Combinations of oils and syrups also may be used.

The combination may further comprise a feedstuff, such as an animal feed. The feedstuff may be anything that is consumed by the animal. "Feedstuff encompasses solid and liquid animal feeds (e.g. , a feed ration), supplements (e.g. , a mineral supplement, a protein supplement), a premix, water, and feed additive carriers (e.g. , molasses). The feedstuff may be a commercial feed stuff. For aquatic animals, the feedstuff may be a fish meal or crustacean or mollusk food, and may be formulated as a floating or sinking feedstuff. IV. Formulation

In some embodiments, the combination is a composition. In other embodiments, the combination comprises one or more compositions, and/or one or more individual components. For example, a first composition may comprise silica, mineral clay, glucan, mannans, or a combination thereof, and a second composition may comprise allicin, alliin, alliinase, yeast, polyphenol, algae, yucca, quillaja, probiotic, a vitamin, and/or sorbic acid or a salt thereof. Alternatively, or additionally, allicin, alliin, alliinase, yeast, polyphenol, algae, yucca, quillaja, probiotic, a vitamin, and/or sorbic acid or a salt thereof may be formulated as individual components.

The combination and/or any of its components may be formulated in any suitable form, including a powder, a granule, a pellet, a solution, or a suspension. Certain disclosed embodiments are formulated as a dry, free-flowing powder. This powder is suitable for direct inclusion into a commercially-available feed, food product or as a supplement to a total mixed ration or diet. The powder may be mixed with either solid or liquid feed or with water. In another embodiment, the combination and/or any components are formed into pellets. The combination and/or any of its components may be formulated as a powder that is admixed with a feedstuff using a carrier and/or adhesive agent. The carrier and/or adhesive agent may be any carrier and/or adhesive agent known to a person of ordinary skill in the art as being suitable for combining with a feed composition and/or combination, such as molasses, or an oil, such as corn oil or soy oil.

The combination and/or any of its components may be formulated to be suitable to form a substantially homogeneous mixture with the feedstuff, such as by crushing, crumbling, grinding or otherwise sizing the combination. Alternatively, the combination and/or any of its components may be formulated as a solution, suspension or slurry. When the combination and/or any of its components is a combination comprising two or more components, the components may be formulated separately or substantially together, and one component may be used as a solid and another component as a solution or suspension. The components may also be admixed with the feedstuff sequentially, in any order, or substantially simultaneously. V. Method of Using

A. Animals

Embodiments of the disclosed combination are fed and/or administered to an animal, such as a human or non-human animal. The animal may be a land animal, an aquatic animal, an avian, or an amphibian. The animal may be a mammal, or a non-mammal. The non-human animal can be an animal raised for human consumption or a domesticated animal. Examples of animals that can be fed and/or administered the disclosed combination include, but are not limited to, ruminant species, such as a sheep, goat, cow, deer, bison, buffalo, elk, alpaca, camel or llama; ungulates, such as a horse, donkey, or pig; avians, such as chickens, including laying hens and broilers, turkey, goose, duck, Cornish game hen, quail, partridge, pheasant, guinea-fowl, ostrich, emu, swan, or pigeon; aquatic animals, such as an aquaculture species, such as fish (e.g. , salmon, trout, tilapia, sea bream, carp, cod, halibut, snapper, herring, catfish, flounder, hake, smelt, anchovy, lingcod, moi, perch, orange roughy, bass, tuna, mahi mahi, mackerel, eel, barracuda, marlin, Atlantic ocean perch, Nile perch, Arctic char, haddock, hoki, Alaskan Pollock, turbot, freshwater drum, walleye, skate, sturgeon, Dover sole, common sole, wolfish, sablefish, American shad, John Dory, grouper, monkfish, pompano, lake whitefish, tilefish, wahoo, cusk, bowfin, kingklip, opah, mako shark, swordfish, cobia, croaker, or hybrids thereof, and the like), crustaceans (e.g. , lobster, shrimp, prawns, crab, krill, crayfish, barnacles, copepods, and the like), or molluscs (e.g. , squid, octopus, abalone, conchs, rock snails, whelk, clams, oysters, mussels, cockles, and the like). Additionally, or alternatively, the animal may be a companion animal, such as canines; felines; rabbits; rodents, such as a rat, mouse, hamster, gerbil, guinea pig or chinchilla; birds, such as parrots, canaries, parakeets, finches, cockatoos, macaws, parakeets or cockatiel; reptiles, such as snakes, lizards, tortoises or turtles; fish; crustaceans; and amphibians, such as frogs, toads and newts. B. Uses of the combination

The combination may be used to replace or supplement animal feedstuffs, or it may be administered separately from a feedstuff. In some embodiments, the feedstuff is a commercial feedstuff. The combination and/or any of its components, may be formulated in any form suitable for mixing with a feedstuff and/or replacing a feedstuff, including a powder, a granule, a pellet, a solution, or a suspension. Certain disclosed embodiments are formulated as a dry, free-flowing powder. This powder is suitable for direct inclusion into a commercially-available feed, food product or as a supplement to a total mixed ration or diet. The powder may be mixed with either solid or liquid feed and/or with water. In other embodiments, the combination and/or any components are formed into pellets, and in further embodiments, the combination and/or any components are formulated into granules, such as floating or sinking granules, suitable for feeding to aquatic animals.

In some embodiments, the combination, or one or more components of the combination, has an average particle size selected to be compatible with a feedstuff to which the combination, or the one or more components of the combination, may be admixed. The term "compatible" as used herein means that the particle size is sufficiently similar to reduce or eliminate particle size segregation when the combination, or one or more components of the combination, is admixed with the feedstuff. For example, if the combination, or one or more components of the combination, is admixed with a feedstuff having an average particle size of 50-200 μιη, the combination, or one or more components of the combination, may have a similar average particle size, e.g. , from 80-120% of the feedstuff/component particle size with which the combination, or one or more components of the combination, is admixed.

The disclosed combination can be administered to animals to obtain one or more beneficial results. Such benefits may include, but are not limited to, prevention and/or treatment of certain diseases or conditions, such as infectious diseases, non-infectious diseases, stress and stress-related conditions and diseases; a beneficial effect on the animal's immune system; or helping increase longevity of the animal. An animal may have one or more of an increased expression of genes associated with immune system function, such as L-selectin and interleukin 8 receptor; increased super-oxide dismutase (SOD) activity; higher serum glucose concentrations and/or lower concentrations of acute phase proteins, that may demonstrate improved metabolism and/or immune function; improved oxidative respiratory index in liver tissue; improved milk production; or lower Somatic cell count (SCC). An animal may be affirmatively selected based on one or more factors that include the animal's age, decreased immunity, exposure to stressors or stress events (e.g. , heat stress, crowding, ammonia toxicity, work load, chemotherapy, anti-inflammatory therapy), gastrointestinal disturbances (e.g. , diarrheal diseases), or combinations thereof.

Additionally, or alternatively, the combination may improve the feed conversion rate, and/or the Feed:Gain ratio, of an animal, such as an animal raised for consumption; improve the weight gain of the animal; and/or reduce mortality. A feed conversion rate, also known as a feed conversion ratio, is a measure of an animal's efficiency in converting feed mass into increased body mass. Animals with low feed conversion rates are considered efficient, as they require less feed to reach a desired weight. Feed conversion rates vary from species-to-species.

Embodiments of the disclosed combination can be administered to aquatic animals to obtain one or more beneficial results. For example, embodiments of the combination may be used to prevent and/or treat certain aquatic diseases. Additionally, the combination may improve the feed conversion rate of an aquatic animal. Feed conversion rates for aquatic species vary from species- to-species. For example, tilapia typically have a feed conversion ratio of from 1.6 to 1.8, and farm raised salmon typically have a ratio of around 1.2. In some embodiments, the feed conversion rate may be enhanced by from 0.5% to 20% or more, such as from 1% to 20 %, preferably from 2% to 10%, and in certain embodiments, from 3% to 5%.

In some embodiments, the combination is administered daily to the animal at time intervals believed or determined to be effective for achieving a beneficial result. The combination may be administered in a single dose daily or in divided doses throughout the day. In some instances, one or more components of the combination may be administered to the animal at a first time, and remaining components may be administered individually or in combination at one or more subsequent times during the same day. Typically, a time period over which the combination is administered is sufficient such that the animal received a benefit from the combination of components. In some embodiments, the components of the combination may be administered to the animal in any order over a time period sufficient that an effective time period of a first component, or combination of first components, overlaps with an effective time period of a second component, or combination of second components, and any effective time periods of any subsequent components, or combinations of subsequent components. An "effective time period" is a time period during which the animal received a beneficial result from being administered the particular component, or combination of particular components. C. Immune system benefits

Without wishing to be bound by any particular theory, the combination may enhance the animal's immune system, such as the innate system or the adaptive immune system, or both. When administered to an animal, the combination may produce a concomitant change in a level of, for example, an immune system biomarker or an inflammation biomarker in the animal by at least 5%, at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, at least 100%, at least 200%, or at least 500%, such as from 5-600%, from 10-500%, from 10-200%, or from 10-100%, compared to an average level of the biomarker in an animal that has not received the combination. The change may be an increase or a decrease, depending on the particular biomarker. For example, some embodiments of the combination affect levels of immune biomarkers including, but not limited to, neutrophil L-selectin, IL-Ιβ and/or gene expression of Crp, Mbl2, Apes, 115, Ifnal, Ccll2, Csf2, 1113, 1110, Gata3, Stat3, C3, Tlr3, Ccl5, Mx2, Nfkbl, Nfkbia, Tlr9, CxcllO, Cd4, 116, Ccl3, Ccr6, Cd40, Ddx58, 1118, Jun, Tnf, Traf6, Statl, Ifnbl, Cd80, Tlrl, Tlr6, Mapk8, Nod2, Ccr8, Iraki, Cdldl, Stat4, llrl, Faslg, M3, Ifnarl, Slcl lal, Tlr4, Cd86, Caspl, Ccr5, Icaml, Camp, Tlr7, Irf7, Rorc, Cd401g, Tbx21, Casp8, 1123a, Cdl4, Cd8a, Cxcr3, Foxp3, Lbp, Mapkl, Myd88, Stat6, Agrin and/or IL33. As disclosed in U.S. Patent No. 8,142,798, which is incorporated herein by reference, some embodiments of the combination also augment an animal's adaptive immune system, e.g. , by increasing response to a vaccine; antibody levels, such as IgG levels, may be increased, relative to an animal that has received a vaccine but has not been administered the combination. The combination also may reduce the effects of stress in the animal, potentially by ameliorating the effects of stress (e.g. , heat stress, pregnancy stress, parturition stress, etc.) on the animal's immune system. Some embodiments of the combination affect levels of inflammation biomarkers, e.g. , COX-2, IL-Ιβ, tumor necrosis factor alpha (TNF-a), interleukin-8 receptor (IL8R), and/or L-selectin.

In some embodiments, administration of the combination may produce a concomitant change in a level of innate defense mechanisms of fish prior to exposure to a pathogen, or improve survival following exposure to a specific pathogen. Markers of improved innate immune response may include:

1. Total leucocyte count

Abnormal changes in total and differential blood cell counts in fish, such as anaemia, leukopaenia, leukocytosis and thrombocytopaenia, may result from diseases, but may also indicate stress, toxic exposure, hypoxia and changes in reproductive status.

Due to the nucleated nature of red blood cells (erythrocytes) in fish, white blood cells (leukocytes), which serve as an indicator of health, cannot be distinguished using automated cell counting procedures without lysis of erythrocytes and are usually manually counted using a haemocytometer. Differential leukocyte and haemocyte enumerations, which also serve as health indicators, are generally performed either on stained smears or with a haemocytometer in fish and Crustacea, respectively. The disadvantage of manual enumeration is the statistical limitation associated with counting between 100 to 200 cells, the typical range in differential leukocyte procedures.

Flow cytometry is an instrumental technique in which a stream of suspended particles is interrogated by one or more lasers. Particles are analysed and differentiated on the basis of their light- scattering properties, auto- or labelled fluorescence, or a combination of both.

The major advantages of flow cytometry technology are the ability to differentiate and enumerate several thousands of particles per second, and to physically sort multiple populations simultaneously into collection vessels. In haematological applications, the capability to obtain accurate and precise total and 5 differential blood counts on so many more cells than practically achievable with manual methods, in a fraction of the time, is thus dependent only on the ability to accurately discriminate between cell types.

2. Respiratory burst (release of superoxide anion)

Several reactive oxygen species (ROS) are produced by fish phagocytes during the respiratory burst. Once bacteria or fungi are engulfed by leucocytes, the host's NADPH-oxidase is activated, which in turn increases oxygen consumption and subsequently produces ROS such as superoxide anion (O2 ), hydrogen peroxide (H2O2), hydroxyl radical (OH) and singlet oxygen (^lOi). The release of superoxide anion is known as the respiratory burst, and the ROS released and/or formed may be are bactericidal.

3. Phagocytic index and activity

Phagocytosis is an essential component of the non-specific immune response against infectious agents in teleosts. This process involves the recognition and attachment of foreign particles, including pathogens, engulfment and digestion by the phagocyte. In vitro assays have been used for studying fish macrophage phagocytic activity, thereby providing an avenue for evaluating immunocompetence in fish. In vitro assays have also provided insight for non- specifically enhancing disease resistance in finfish aquaculture, and have served as immunological biomarker tests to assess aquatic environmental health.

4. Lysozyme activity

Lysozyme found in cutaneous mucus, peripheral blood and certain tissues rich in leucocytes, is an enzyme which catalyzes the hydrolysis of N-acetyl muramic acid and N-acetyl glucosamine of peptidoglycan in bacterial cell walls. This protein plays a crucial role in the defense system.

In other embodiments, administration of the combination may produce a concomitant change in a level of innate defense mechanisms of crustaceans prior to exposure to a pathogen, or improve survival following exposure to a specific pathogen. Markers of improved innate immune response in crustaceans may include:

1. Total hemocyte count

Haemocytes play a central role in crustacean immune defense. They remove foreign particles in the hemocoel by phagocytosis, encapsulation and nodular aggregation. Additionally, haemocytes take part in wound healing by cellular clumping and initiation of coagulation processes through the release of factors required for plasma gelation.

The hemogram consists of the total haemocyte count (THC) and the differential haemocyte count (DHC). For the DHC, most researchers agree with the identification of three cell types in penaeid shrimp: large granule haemocytes (LGH), small granule haemocytes (SGH) and agranular haemocytes or hyaline cells (HC).

THC can be easily determined using a hemocytometer, whereas determination of DHC requires a more complex haemocyte identification. DHC can be determined by using

morphological criteria such as size and shape of cells and the difference of haemocyte refractivity using a phase contrast microscope. Although this technique is rapid, it should be mentioned that when using this technique it is easy to obtain large variations in results possibly due to

interpretation errors.

Different haemocyte types can be determined using cytochemical studies of enzyme activity detection or specific stains. The results obtained from cytochemical stains for penaeid shrimp indicate that these specific stainings can differentiate between the types of haemocytes and provide additional information on their functions. An alternative method for cell identification is the use of monoclonal antibodies (mAbs) in order to find antigenic markers of different cell types. Using mAbs against different subpopulations of haemocytes separated by isopycnic centrifugation on a Percoll gradient, it has been found in P. japonicus that HC share epitopes with SGH, and that an antigen was specifically expressed for LGH. Monoclonal antibodies could be considered as powerful tools for the development of haemocyte lineages and haemocyte proliferation studies, as well as for the isolation and study of plasma components.

2. Phagocytic activity

Phagocytosis is the most common reaction of cellular defense. During phagocytosis, particles or microorganisms are internalized into the cell which later forms a digestive vacuole called the phagosome. The elimination of phagocyted particles involves the release of degradative enzymes into the phagosome and the generation of reactive oxygen intermediates (ROIs). This last process is known as the respiratory burst. The first ROI generated during this process is the superoxide anion. Subsequent reactions will produce other ROIs, such as hydrogen peroxide, hydroxyl radicals and singlet oxygen. Hydrogen peroxide can be converted to hypochlorous acid via the myeloperoxidase system, forming a potent antibacterial system.

Despite the limited number of studies focusing on respiratory burst in penaeid shrimp, the actual results have value as biomarkers of environmental disturbances. Furthermore, the importance of respiratory burst as a microbicidal mechanism in penaeid shrimp is strongly suggested by the fact that pathogenic bacteria of shrimp have developed ways of circumventing this mechanism. In P. tannamei, O2 generation is not produced when virulent Vibrio tulnificus is used as elicitor, as opposed to strong stimulation generated by V. Iginolyticus and other bacteria, such as Escherichia coli.

3. Phenoloxidase (PO) and Prophenoloxidase (ProPO) activity The PO is responsible for the melanization process in arthropods. The PO enzyme results from the activation of the ProPO enzyme. The ProPO activating system has been very well studied in crustaceans. Using these different approaches, the function of the ProPO system can be better understood in relation to the health status of shrimp. Some studies have shown that ProPO could be used as health and environmental markers because changes are correlated with infectious state and environmental variations, this issue which has recently been confirmed also at the gene expression level. Phenoloxidase, which has been detected in a wide range of invertebrates, is activated by several microbial polysaccharides, including -l,3-glucan from fungal cell walls and

peptidoglycans or lipopoly saccharides from bacterial cell walls.

4. Antibacterial activity

Antibacterial peptides and proteins have been well studied in arthropods, mainly in insects and chelicerata, where the families of antimicrobial molecules have been isolated and

characterized. In crustacean, some studies have shown the ability of crustacean haemolymph to inhibit bacterial growth. Several antibacterial proteins, active in vitro against Gram-positive and Gram-negative bacteria, were found in C. maenas.

In the literature there are reports showing that antibacterial activity in crustaceans can be considered as an environmental marker. Therefore, many researchers have developed quantitative antibacterial assays based on inhibition of bacterial growth on agar plate (zone inhibition assay and colony-forming units (CFU). inhibition assay), or in liquid medium on microtiter plates

(turbidometric assay), to detect the antibacterial ability in crustacean haemolymph. Using the CFU inhibition technique, antibacterial activity has been found in granular haemocytes of the shore crab C. maenas and in other crustacean species. It has been reported that a potent antibacterial activity in the serum of Cal.sapidus, using the zone inhibition assay and turbidimetric test. Using the CFU inhibition assay, bactericidal activity against Gram negative bacteria have been described in the haemolymph of P. monodon. In P. tannamei, strong antibacterial activity of plasma against different marine bacteria has been observed, using a turbidimetric assay.

5. Plasma protein concentration

In recent years blood metabolites have been investigated as a tool for monitoring physiological condition in wild or cultured crustaceans exposed to different environmental conditions. Hemocyanin is the major hemolymph constituent (>60%); the remaining proteins (in order of concentration) include coagulogen, apohemocyanin, hormones, and lipoproteins. Blood protein levels fluctuate with changes in environmental and physiological conditions and play fundamental roles in the physiology of crustaceans from O2 transport to reproduction up to stress responses. In fact, moulting, reproduction, nutritional state, infection, hypoxia, and salinity variations are the major factors affecting the relative proportions and total quantities of the hemolymph proteins.

The shrimp immune system response is largely based on proteins. These are involved for example in recognizing foreign particles and in trapping foreign invading organisms and prevent blood loss upon wounding. Recently, it has been shown that shrimp are well adapted to use protein as a source of energy and molecules. Blood protein concentration has been found to be related to nutritional condition in a number of crustaceans. The concentration of protein in the blood is a possible index of nutritional condition, which decreases in starved prawns and lobsters. The moult cycle imposes constraints on protein levels, blood-proteins typically drop just before moulting as water is taken up and protein is used to synthesize the new exoskeleton. Protein levels then gradually build up again after ecdysis as water is replaced by tissue. Consequently, measuring the blood protein concentration of a crustacean sample group can provide valuable information to identify its condition. The concentration of protein in the blood is directly proportional to the refractive index of the blood. Measurements of the blood refractive index therefore offer potential as a field method for assessing the nutritional condition of prawns.

Colorimetric procedures are generally the preferred choice to measure serum protein concentration; however, they are expensive, time consuming, and not easily performed in the field. Because of ease, rapid mode of operation, and small amount of material required, measuring serum protein concentration using a refractometer provided a nondestructive field method to assess crustacean's physiological state (stress, immunoresponse, nutrition status, molt, etc.) without any need of laboratory facilities; the refractometer is a simple, small portable instrument that can be used in the field or on crustacean farms.

D. Amount administered

The combination may be administered or fed to an animal in a sufficient amount to provide a desired result. The amount may be from greater than zero to 500 grams per animal per day, such as from 0.5 grams to 250 grams, from 5 grams to 200 grams, or from 10 grams to 70 grams per animal per day. Alternatively, the combination may be fed or administered in an amount of from greater than zero to 1000 mgs or more per kilogram of the animal's body weight, such as from greater than zero to 500 mgs per kilogram body weight. In other embodiments, the combination is fed or administered per weight of animal feed. The combination may be fed or administered in an amount of from greater than zero to 150 kg per ton (2000 pounds) of feed, such as from 0.1 kg to 100 kg per ton of feed. Alternatively, the combination may be fed or administered in an amount of from greater than zero to 20 grams per kilogram of feed, such as from greater than zero to 10 grams of feed.

In some embodiments, the combination comprises a composition 1 comprising silica, mineral clay, glucan and mannans. When incorporated directly into feeds, the combination may be added in amounts sufficient to provide an effective amount of the composition 1. An effective amount of the composition 1 may range from 0.1 to 100 kg per ton of feed. In some embodiments, the combination is added in sufficient amount such that composition 1 is added in amounts of from 0.1 to 50 kg per ton, such as from 0.1 to 20 kg per ton of feed, from 0.5 kg to 10 kg per ton of feed, or from 1 to 5 kg per ton of feed.

When expressed as a percentage of dry matter of feed, the combination is added in sufficient amount such that composition 1 may be added to animal feedstuffs or to foods in amounts ranging from 0.01 to 2.5% by weight, such as from 0.0125% to 2% by weight, from 0.05 to 1.5% by weight, from 0.06% to 1% by weight, from 0.1 to 0.7% by weight, or from 0.125% to 0.5% by weight of feed.

Alternatively, the combination may be administered such that composition 1 may be fed directly to animals as a supplement in amounts of from greater than 0.01 gram to 20 gram per kilogram of live body weight, such as from 0.01 gram to 10 gram per kilogram of live body weight, from 0.01 gram to 1 gram per kilogram of live body weight, from 0.01 gram to 0.5 gram per kilogram of live body weight, or from 0.02 gram to 0.4 gram per kilogram of live body weight per day. In some embodiments, composition 1 may be provided for use with many mammalian species, including non-human mammals, in amounts of from 0.05 grams to 0.20 grams per kilogram of live body weight per day. By way of example, for cattle, the combination may be provided in a sufficient amount such that composition 1 is provided in the range of from 10 grams per head per day to 70 grams per head per day, such as from 45 grams per head per day to 70 grams per head per day, or from 50 grams per head per day to 60 grams per head per day. A person of ordinary skill in the art will appreciate that the amount of composition I fed can vary depending upon a number of factors, including the animal species, size of the animal and type of the feedstuff to which composition 1 is added.

For some embodiments concerning aquatic animals, the combination can be administered based on body weight, such as grams of the combination per pound or kilogram body weight of fish per day, or in milligrams of the combination per pound or kilograms of body weight. In some embodiments, the amount of the combination administered is sufficient to provide a desired amount of one or more of the components of the combination. In a particular example, when administered to fish the combination comprises a composition 1 , comprising silica, mineral clay, glucan and mannans, in an amount of from greater than zero to 500 mg of composition 1 per kilogram of body weight per day, such as from 10 mg to 350 mg per kilogram of body weight per day or from 50 mg to 250 mg per kilogram of body weight per day.

Alternatively, the combination may be administered based on the amount of feed provided to the aquatic animals. In some embodiments, the amount of the combination provides composition 1 to the aquatic animals in an amount of from greater than zero to 10,000 mg composition 1 per kilogram of feed or more, such as from 500 mg to 7,500 mg per kilogram of feed, or from 1,000 mg to 5,000 mg per kilogram of feed.

A person of ordinary skill in the art will appreciate that the amount of composition 1 , and/or a combination comprising composition 1 , administered can vary depending upon a number of factors, including the animal species, size of the animal, the age or growth stage of the animal, and type of the feedstuff to which the combination is added. In some embodiments, 100 mg of composition 1 per kilogram of body weight per day is administered, and in other embodiments, 200 mg of composition 1 per kilogram of body weight per day is administered. In certain embodiments, 1,000 mg, 2,000 mg or 4,000 mg of composition 1 per kilogram of feed is administered to the animals.

FIG. 1 provides exemplary ranges for hatchery, nursery and grow-out stages for fish, based on an administration amount of 100 mg of composition 1 per kilogram of body weight per day.

FIG. 1 illustrates that hatchery stage fish being fed at a feeding rate of 10% of body weight per day and being administered 100 mg of composition 1 per kilogram of body weight per day, provides a composition 1 dose of 1,000 mg per kilogram of feed. This increases to 2,000 mg per kilogram of feed for fish at the nursery stage, and up to 4,000 mg per kilogram of feed for fish at the grow-out stage. FIG. 1 also provides exemplary feed sizes of from greater than zero to 1 mm as a starter feed, and from 1 mm to 2 mm as a small feed for the hatchery stage; from 2 mm to 3 mm for the nursery stage, optionally as small pellets; and 3 mm to the largest size useful for feeding the grow- out stage fish, optionally formulated as pellets. The feed size may vary depending on the species of aquatic animal as well as on the growth stage of the animal. Suitable feed sizes for particular aquatic animals at different growth stages are known to persons of ordinary skill in the art.

The combination may comprise a probiotic, for example a Bacillus species, such as Bacillus coagulans. The amount of the combination administered or fed to the animal may be selected to provide a sufficient amount of the probiotic, such as Bacillus coagulans, to provide a desired and/or beneficial result or enhancement in the animal. In some embodiments, the Bacillus coagulans may be administered as Ganpro^® or Previa 6086^®. In poultry the amount of the combination administered may be sufficient to provide an amount of Bacillus coagulans of from about 0.5 to 2.5 grams per head per day, such as about 1 gram per head per day. In embodiments concerning cattle, the amount of the combination administered or fed to the cattle is sufficient to provide an amount of Bacillus coagulans of from about 10 to about 50 grams per head per day, preferably from about 28 to 36 grams per head per day. And for swine the amount of the combination administered or fed may be sufficient to provide an amount of Bacillus coagulans of from about 2 to about 10 grams per head per day, preferably about 5.5 grams per head per day. In some examples, the Bacillus coagulans may be admixed with feedstuff at from about 0.5 grams per ton (2000 pounds) to about 10 grams per ton or more feed stuff. In certain embodiments, the Bacillus coagulans is admixed with feedstuff at about 7.5 grams per ton of feedstuff. In other embodiments, the amount of Bacillus coagulans, administered in certain embodiments as Ganpro^® or Previa 6086^®, is from about 0.5 grams to less than 7.5 grams per ton of feedstuff, such as from 2 grams to 7.25 grams per ton, or from 5 grams to 7 grams per ton. In other examples, the amount of Bacillus coagulans, administered in certain embodiments as Ganpro^® or Previa 6086^®, is from greater than 7.5 grams to greater than 10 grams per ton of feed stuff, such as from greater than 7.5 grams per ton to 10 grams per ton, or from 7.75 grams per ton to 8 grams per ton.

The combination may comprise yucca and/or quillaja, such as Yucca schidigera and/or Quillaja saponaria. In some embodiments, the combination is administered or fed to an animal in an amount sufficient to provide an amount of yucca to the animal of from greater than 0 to greater than about 10 ounces per ton of feedstuff, preferably from about 1 to about 5 ounces. In other embodiments, the combination is administered or fed to an animal in an amount sufficient to provide an amount of quillaja to the animal of from greater than 0 to greater than about 10 ounces per ton of feedstuff, preferably from about 1 to about 5 ounces. In certain embodiments, the combination comprises both yucca and quillaja, and the combination is administered to an animal in an amount sufficient to provide a combined amount of yucca and quillaja of from greater than 0 to greater than about 10 ounces per ton of feedstuff, preferably from about 2 to about 6 ounces.

In some embodiments, the combination comprises a composition 2 comprising yucca and quillaja, typically Yucca schidigera and/or Quillaja saponaria. Composition 2 may be

administered as Nutrafito^® Plus. The combination may be administered to an animal in a sufficient amount to provide an amount of composition 2 to the animal of from greater than 0 ppm to about 500 ppm, such as from about 50 ppm to about 500 pmm, from about 100 ppm to about 500 ppm, from about 50 ppm to about 400 ppm, or from about 100 ppm to about 300 ppm. In some embodiments, the combination is administered in an amount sufficient to provide an amount of composition 2 of from greater than 0 ppm to less than 125 ppm, such as from greater than 0 ppm to 124 ppm or from greater than 0 ppm to 100 ppm. And the combination may be administered in an amount sufficient to provide an amount of composition 2 of from greater than 125 ppm to 500 ppm, such as from about 126 ppm to 400 ppm, or from 150 ppm to 300 ppm. In certain embodiments, a sufficient amount of the combination is administered or fed to avians to provide an amount of

Yucca schidigera and Quillaja saponaria as Nutrafito^® Plus, of from about 2 to about 6 ounces per ton of feedstuff. In other embodiments, the combination comprising Nutrafito^® Plus is

administered to avians in a sufficient amount to provide about 125 ppm (parts per million) or about 4 ounces of Nutrafito^® Plus per ton of feedstuff.

Additional information concerning embodiments of a combination comprising Yucca, quillaja and Bacillus can be found in U.S. application No. 14/699,740, which is incorporated herein by reference in its entirety.

The combination may comprise an antimicrobial or antibiotic. The amount of the antimicrobial or antibiotic may be selected to be within the amounts stated below but may depend on the particular antimicrobial or antibiotic used as will be understood by a person of ordinary skill in the art. In some embodiments, the amount of the antibiotic or antimicrobial that is used can be a therapeutically effective amount that is at an approved or authorized dosage level for a particular antibiotic. In some embodiments, the amount of antibiotic or antimicrobial used can range from greater than 0 ppm to 100,000 ppm, such as 0.25 ppm to 5,000 ppm, or 0.5 ppm to 2,500 ppm, or 0.75 ppm to 2,000 ppm, or 1 ppm to 1,500 ppm, or 5 ppm to 1,000 ppm, or 10 ppm to 500 ppm, or 25 ppm to 300 ppm. In yet additional embodiments, the amount of antibiotic or antimicrobial used can range from greater than 0 mg/kg of body weight to 100,000 mg/kg of body weight, such as 0.5 mg/kg to 2,500 mg/kg, or 1 mg/kg to 1,500 mg/kg, or 5 mg/kg to 1,000 mg/kg, or 10 mg/kg to 500 mg/kg m, or 25 mg/kg to 300 mg/kg, or 10-20 mg/kg. In some embodiments, the amount of the antimicrobial or antibiotic that is included in the composition can range from at least 1 g/ton of feed to 230 g/ton of feed (or at least 1.1 ppm to 256 ppm), such as at least 1 g/ton of feed to 220 g/ton of feed (or at least 1.1 ppm to 243 ppm), at least 1 g/ton of feed to 100 g/ton of feed (or at least 1.1 ppm to 110 ppm), at least 1 g/ton of feed to 50 g/ton of feed (or at least 1.1 ppm to 55 ppm), or at least 1 g/ton of feed to 10 g/ton of feed (or at least 1.1 ppm to 11 ppm). Particular antimicrobials or antibiotics that can be used, and dosage amounts of such antimicrobials and antibiotics include, but are not limited to, the following:

Virginiamycin in an amount ranging from 5 g/ton of feed to 25 g/ton of feed (or 5 ppm to 27 ppm, such as 22 ppm); Bacitracin MD in an amount ranging from 40 g/ton of feed to 220 g/ton of feed (or 44 ppm to 242 ppm, or 50 ppm to 250 ppm in some other embodiments); Zinc Bacitracin in an amount ranging from 40 g/ton of feed to 220 g/ton of feed (or 44 ppm to 242 ppm); Tylosin in an amount ranging from 1 g/ton of feed to 1000 g/ton of feed (or 1 ppm to 1100 ppm); Lincomycin in an amount ranging from 1 g/ton of feed to 5 g/ton of feed (or 1 ppm to 6 ppm); Flavomycin in an amount ranging from 1 g/ton of feed to 5 g/ton of feed (or 1 ppm to 6 ppm); or combinations thereof.

The amount of an anticoccidial agent, as will be understood by a person of ordinary skill in the art (e.g. , a veterinarian), can be selected depending on the particular anticoccidial agent used. In some embodiments, the amount of anticoccidial agent used can be a therapeutically effective amount for a particular animal species. In some embodiments, the amount of anticoccidial agent used can range from greater than 0 ppm to 100,000 ppm, such as 0.25 ppm to 5,000 ppm, or 0.5 ppm to 2,500 ppm, or 0.75 ppm to 2,000 ppm, or 1 ppm to 1,500 ppm, or 5 ppm to 1,000 ppm, or 10 ppm to 500 ppm, or 25 ppm to 300 ppm. In yet additional embodiments, the amount of antibiotic or antimicrobial used can range from greater than 0 mg/kg of body weight to 100,000 mg/kg of body weight, such as 0.5 mg/kg to 2,500 mg/kg, or 1 mg/kg to 1,500 mg/kg, or 5 mg/kg to 1,000 mg/kg, or 10 mg/kg to 500 mg/kg m, or 25 mg/kg to 300 mg/kg, or 10-20 mg/kg.

VI. Examples

The following examples are provided to illustrate certain beneficial effects of administering to an animal a combination comprising silica, glucan, mannans and mineral clay. A person of ordinary skill in the art will appreciate that the scope of the disclosed embodiments is not limited to the features exemplified by these working embodiments. EXAMPLE 1

An experiment was conducted with sheep with the goal of determining the ability of a combination comprising silica, mineral clay, glucan and mannans to increase expression of neutrophil L-selectin, a marker of the innate immune system, in immunosuppressed animals. Animals (six per group) were divided into two groups: Control and Experimental. The Control group received a high energy ration consisting of chopped hay available ad libitum, one pound of ground corn per head per day and one pound of baked wheat mill run per head per day for a period of 28 days. During this time, they also received twice daily injections of dexamethasone, an immunosuppressive drug. The Experimental group received daily intake of the combination (5 grams per head per day) for 28 days and received the same diet and dexamethasone injection protocol as the Control. This composition of the Experimental group was 65.8 weight percent of mineral clay, 0.20 weight percent of endoglucanohydrolase, 9.0 weight percent of glucans and glucomannan, and 25 weight percent of calcined diatomaceous earth. At the end of the study, blood samples were recovered and neutrophils were purified using Percoll gradient centrifugation. The amounts of L-selectin expression in neutrophils were assessed using Western blotting techniques and antibodies specific for L-selectin.

As shown in FIG. 2, top panel, animals that did not receive component I had low and variable expression of L-selectin. As shown in FIG. 2, lower panel, animals that received the combination demonstrated a consistent increase in L-selectin expression. The top panel represents six Control, immunosuppressed animals. The lower panel represents six Experimental immunosuppressed animals which received the combination in their diet.

EXAMPLE 2

In this study, stimulation of the innate immune system in sheep was examined when the Experimental combination of Example 1 was provided in a pelleted diet. The basal diet consisted of 21.55% barley, 10.0% canola meal, 5% distillers grains, 40% ground corn, 1.50% limestone, 0.01% manganese sulfate, 0.01% microvitamin E, 4.0% molasses, 0.25% mono-cal, 0.25% potassium chloride, 0.60% sodium chloride, 0.03% sodium selenite, 15.79% wheat mill run, 0.01% zinc sulfate, 0.75% ammonium sulfate and 0.2 5% cobalt sulfate. When the Experimental combination was added to this diet, it was included at 0.6% replacing that portion of wheat mill run. Twenty-eight sheep were assigned to four treatments which consisted of a Control group, a group which received the Experimental combination in powdered form, a group which received the Experimental combination in pelleted form where pellets were formed at a temperature of 160° F, and a group which received the Experimental combination in pelleted form where pellets were formed at 180° F. All animals were immunosuppressed via daily injection of Dexamethasone.

The study was conducted using methods identical to Example 1 except the combination was administered in pellets that were manufactured by forming the pellets at high temperatures. The rationale for conducting this study was to determine whether heating of the combination (as is required in pellet formation) might inactivate the ability of the combination to augment innate immunity. As shown in FIG. 3, sheep (Control) which did not receive the combination expressed very low levels of L-selectin in neutrophils. The provision of the Experimental combination even in a pelleted (heated) form still increased expression of neutrophil L-selectin markedly.

In FIG. 3, the uppermost panel represents neutrophil L-selectin expression in

immunosuppressed animals fed a control diet without the combination. The second panel (Powder) represents L-selectin expression in immunosuppressed animals which received the Experimental combination in unheated freely-mixed form as in Example 1 (Experimental group). Panels 3 and 4 represent neutrophil L-selectin expression in immunosuppressed animals which received the Experimental combination in pelleted forms. The pellets used in Panel 3 were formed by heating to 160°F and Panel 4 pellets were heating to 180°F during manufacture of the feeds.

EXAMPLE 3

An experiment was performed with rats to investigate whether the combination had ability to augment innate immunity in a non-ruminant model. In this study, rats were assigned to one of two treatments: a Control group (un- supplemented diet) and an Experimental group where the combination of Example 1 was added to the diet at 1% of dry weight of feed. In this experiment, rats were fed a commercial ground rat chow with or without the Experimental combination.

Immunosuppression using dexamethasone injection protocols were not utilized in this study.

Following 14 days, blood samples were taken from anesthetized rats via cardiac puncture.

Neutrophils were isolated from blood samples using Percoll gradient centrifugation and total RNA was isolated using TriZol^®.

The concentration of the messenger RNA (mRNA) encoding rat L-selectin in the neutrophil RNA samples was then determined by quantitative reverse transcriptase polymerase chain reaction (QRT-PCR) using primers which were specifically developed for assay of rat L-selectin. The amounts of L-selectin mRNA were standardized by showing them as a proportion of β-actin mRNA, which is expressed in all cells at a fairly constant level. As shown in FIG. 4, and in agreement with the results in Examples 1 and 2, the combination increased expression of L-selectin mRNA by greater than 6-fold (P<0.05). This study demonstrated that the increased expression of L-selectin protein as shown in by Western blotting in Examples 1 and 2 may be caused by an increase in the mRNA encoding this protein. This implies that the combination alters the rate of transcription of the gene encoding L- selectin.

EXAMPLE 4

Neutrophils, cells of the innate immune system, are able to signal and thereby up-regulate the production of antibodies by the acquired immune system through the secretion of interleukin-ΐβ (IL-Ιβ). To investigate the ability of the combination to induce neutrophils to increase synthesis of IL-Ιβ, the concentration was assessed of IL-Ιβ in neutrophils taken from the same sheep as described in Example 1. To complete this study, Western blotting and antibodies specific for IL-Ιβ were used.

As shown in FIG. 5, animals which did not receive daily provision of the combination contained virtually undetectable levels of IL-Ιβ; however, provision of the combination to animals caused a marked increase in the expression of IL-Ιβ (P < 0.05). In FIG. 5, the top panel represents six Control-fed immunosuppressed animals. The lower panel represents six Experimental combination- fed immunosuppressed animals which received the combination. Concentrations of IL-Ιβ were determined using Western blot analysis and an antibody specific for IL-Ιβ.

These data indicate that the combination not only increases markers of innate immunity (e.g., L-selectin; Examples 1, 2 and 3) but also increases expression of the key signaling molecule (i.e., IL-Ιβ) that up-regulates the adaptive immune system.

EXAMPLE 5

The goal of this experiment was to determine which genes were differentially-expressed in neutrophils after feeding the combination to peri-parturient dairy cattle. In this study, the mechanism(s) by which the combination increased the expression of IL-Ιβ in neutrophils was examined. Peri-parturient dairy cattle are a good model because the stress of pregnancy leads to immunosuppression, making the cows particularly susceptible to infection.

In this experiment, eight peri-parturient dairy cattle were assigned to a Control diet that did not have the Experimental combination and eight cattle were assigned to an Experimental group that received an embodiment of the combination in their diet (56 grams per day per head). Animals were fed the diets for approximately 28 days until parturition. At 12-15 hours following parturition, 500 ml samples of blood were recovered via jugular puncture and neutrophils were prepared via large-scale Percoll gradient centrifugation. RNA was isolated from neutrophils using the TriZol^® method and then reverse-transcribed into cDNA using reverse transcriptase. During reverse transcription, differently-colored nucleotide-based dyes (Cy3 and Cy5) were employed such that complementary DNAs (cDNAs) synthesized from the two different treatment (Control and Experimental) groups incorporated different colors. The cDNA samples from Experimental and Control groups were then applied to a BoTL-5 microarray slide. This microarray was prepared at the Center for Animal Functional Genomics at Michigan State University and contains 1500 genes (each arrayed in triplicate) upon a glass slide. The cDNAs generated from the Experimental and Control group samples were then allowed to compete for binding to the 1500 genes on the array and the relative expression of the genes was then assessed by comparing relative abundance of Cy3 and Cy5 signals on each spot on the array. Data were then statistically analyzed to identify those genes which were differentially- expressed (those genes where P < 0.05).

The results showed that greater than 20 genes were differentially expressed (P < 0.05) in bovine neutrophils taken from the Experimental group. Interleukin-converting enzyme (ICE) was one such up-regulated gene. This was confirmed using QRT-PCR and primers specific to the bovine ICE sequence. ICE is the rate-limiting enzyme in the conversion of inactive pro- IL-Ιβ to the active, secreted IL-Ιβ. Thus, the combination may up-regulate adaptive immunity (i.e., such as increasing antibody titer) through its ability to increase expression of neutrophil ICE activity and, consequently, secretion of IL-Ιβ.

EXAMPLE 6

A total of 60 cows on a commercial dairy were balanced for DIM, parity and milk production and assigned to 1 of 2 treatment groups fed (1) an embodiment of the combination comprising between 15% and 40% silica, between 50% and 81% mineral clay, between 1.0% and 5.0% β-glucans, between 0.05% and 3.0% β-1,3 (4)-endoglucanohydrolase and between 1% and 8.0% mannans (EX, 30 cows) or (2) control (CON, 30 cows) diets for 52 days post calving. At 52 days of lactation cows were randomly selected (n = 12) from both groups (6 EX and 6 CON) and housed in environmentally controlled modules for 21 days. The combination was top-dressed 2x/day with molasses as the carrier and the CON cows received the molasses carrier 2x/day. Both were mixed into the top one-third of the TMR. During the environmental room phase of the study cows fed the combination (EX) had higher feed intake than CON during heat stress (HS) (46.8 kg vs. 42.9 kg, P < 0.0001) and no difference during thermoneutral (TN). A temperature-humidity index (THI) threshold of 68 or greater was used to achieve HS. Feeding the combination maintained a numerical 1 kg milk yield advantage compared with CON (30.3 kg vs. 31.4 kg, P = 0.26) during HS but not during TN. Cows fed the combination had lower milk fat (%) (4.2% vs. 3.8%, P = 0.02) and milk protein (%) (P = 0.04). There was no difference in 3.5% FCM between treatments. Water consumption was lower (12.4 liter/day in the combination treated cows, P < 0.01) than control cows. Respiration rates were lower in treated cows at 1400 hours and 1700 hours (4.7 and 8.4 less respirations/minute, P = 0.05, < 0.001) and rectal temperatures were also lower (0.15 °Celsius and 0.25 °Celsius lower that CON, P = 0.05, < 0.001) in treated cows.

Feeding the combination reduced physiological responses to heat stress in lactating dairy cows.

EXAMPLE 7

A total of 30 cows on a commercial dairy were balanced for DIM, parity and milk production and assigned to 1 of 2 treatment groups fed the combination (EX, 15 cows) or control (CON, 15 cows) diets for 90 days post calving. At 90 days of lactation, cows were randomly selected (n = 12) from both groups (6 EX and 6 CON) and housed in environmentally controlled modules for 21 days. The combination was top-dressed 2x/day with molasses as the carrier. The CON cows received the molasses carrier 2x/day. Both were mixed into the top one-third of the TMR. During the environmental room phase of the study, cows fed the combination (EX) had higher feed intake than CON during heat stress (HS) (46.8 kg vs. 42.9 kg, P < 0.0001) and no difference during thermoneutral (TN). A temperature-humidity index (THI) threshold of 68 or greater was used to achieve HS. Feeding the combination maintained a numerical 1 kg milk yield advantage compared with CON (30.3 kg vs. 31.4 kg, P = 0.26) during HS but not during TN. Cows fed the combination had lower milk fat (%) (4.2% vs. 3.8%, P = 0.02) and milk protein (%) (P = 0.04). There was no difference in 3.5% FCM between treatments. Water consumption was lower (12.4 liter/day in the combination treated cows, P < 0.01) than control cows. Respiration rates were lower in treated cows at 1400 hours and 1700 hours (4.7 and 8.4 less respirations/minute, P = 0.05, < 0.001) and rectal temperatures were also lower (0.15 °Celsius and 0.25 °Celsius lower that CON, P = 0.05, < 0.001) in treated cows. Feeding the combination reduced physiological responses to heat stress in lactating dairy cows.

Experimental Design: The study consisted of two phases; 1) the commercial dairy, and 2) the controlled environmental chambers. During the commercial dairy phase, multiparous lactating Holstein cows (n=30) were balanced by DIM, milk production and parity (91 + 5.9 DIM, 36.2 + 2.5 kg/day, and 3.1 + 1.4). Cows were separated into one of two groups. The control group received the base TMR with no supplement. The treatment group was fed the base diet plus 56 grams/ head/ day of the combination (EX) mixed into the TMR. Daily milk production was measured. The dairy phase lasted for 45 days. The dairy portion was used to meet the manufacture's recommended 45 days feeding for EX to function.

After the on-dairy portion was complete, 12 cows (6 control and 6 treatment) were housed in environmentally controlled rooms. Cows continued the ARC portion in the same treatment groups from the on-dairy portion.

The ARC portion lasted for 21 days. Cows were subjected to 7 days of TN conditions, 10 days of HS, and 4 days of recovery (TN). Feed intake, milk production, and milk composition were measured daily. Rectal temperatures and respiration rates were recorded 3x/day (600, 1400, and 1800 hours). Blood samples were taken on days 7 (TN), 8 (HS), 10 (HS), 17 (HS) and 18 (TN) during the ARC segment.

Statistical analyses were performed using the PROC MIXED procedure (version 9.3, SAS Institute, Cary, NC). Cow was the experimental unit (ARC portion). Data is presented in least square means with significance declared with a P- value < 0.05. (See Table 1, below).

Feeding the disclosed combination to heat stressed dairy cows maintained feed intake during heat stress. Milk yield had a numeric (1 kg) advantage with the combination treatment but did not differ significantly. Respiration rate and rectal temperatures were lower in treated animals during heat stress. There was also a reduction in SCC with treatment. Serum Cortisol levels were lower in on 8 days (the first day of heat stress) at 2000 hours in the combination supplemented cows (P=0.03).

Table 1

EXAMPLE 8

Thirty six male CD rats (ca. 225 grams) were randomly assigned to six treatment groups for a feeding trial. Animals were fed one of the following six diets ad libitum for 28 days:

A. Control diet (Teklad 8604 powdered diet).

B. Diet supplemented with yeast cell wall preparation (including β-glucans and glucomannan).

C. Diet supplemented with diatomaceous earth and -l,3(4)-endoglucanohydrolase.

D. Diet supplemented with the yeast cell wall preparation of Composition B, diatomaceous earth, and -l,3(4)-endoglucanohydrolase.

E. Diet supplemented with the yeast cell wall preparation of Composition B, diatomaceous earth, -l,3(4)-endoglucanohydrolase, and mineral clay.

F. Diet supplemented with 0.5% w/w of a commercially available supplement, comprising 9 wt% Safmannan^® yeast cell wall material (source of β-glucans and mannans), 25 wt% diatomaceous earth, 0.02 wt% Trichoderma extract (a source of -l,3(4)-endoglucanohydrolase), 65.98 wt% AB20™ bentonite, and a mixture of B-vitamins.

The amounts of yeast cell wall extract, diatomaceous earth, -l,3(4)-endoglucanohydrolase and mineral clay used to supplement the diet in Compositions B-E were selected to reflect the amounts that would be added if the diet were supplemented with the commercial supplement recited in Composition F.

On day 28, rats were anesthetized with a mixture of ketamine and xylazine and blood samples (6-10 mL) were taken via cardiac puncture. Neutrophils were isolated from blood samples via Percoll gradient centrifugation. RNA was isolated from a portion of neutrophils in all animals using the Trizol^® method. This was then used to quantify concentrations of L-selectin, interleukin- 8 receptor (IL-8R) and β-actin mRNAs. Another portion of neutrophils from all animals were used in a phagocytosis (cell killing assay). In this assay, neutrophils isolated from rats were combined with Staphylococcus aureus in a ratio of 30: 1 S. aureus bacteria to neutrophil. Neutrophils were allowed to "react" with bacteria for 3 hours after which S. aureus viability was assessed spectrophotometrically.

The study found that Composition B (yeast cell wall preparation) and Composition C (diatomaceous earth and -l,3(4)-endoglucanohydrolase) had no significant effect on any of the three tested markers of innate immunity, as compared to Composition A (control diet) (FIGS. 6-8).

While neither Composition B nor Composition C produced a significant effect on the ability of neutrophils to phagocytose S. aureus, Composition D, which represents a combination of Compositions B and C, unexpectedly improved phagocytosis by 20%, which is significant (FIG. 6). Furthermore, the addition of a mineral clay (Composition E) resulted in a significant improvement in the IL-8R marker, as compared to the control (Composition A) (FIG. 7). Composition E also caused a further significant reduction in S. aureus viability as compared to Composition D (FIG. 6). Composition F (commercially available supplement) was found to have the ability to regulate the three measured markers in innate immunity and substantially mimicked the results obtained with Composition E, indicating that the B vitamins included in the commercially available supplement do not significantly affect regulation of these markers of innate immunity (FIGS. 6-8).

EXAMPLE 9

A study was conducted to identify genes expressed by circulating immune cells that are regulated by a commercial embodiment of a combination comprising silica, mineral clay, glucan and mannans. Rats (n=6 per group) were randomly assigned to the combination and control groups. The combination was supplemented in the diet at 0.5% in the combination group. Total RNA was purified from whole blood and gene expression was analyzed with the use of the Rat Innate and Adaptive Immune Responses RT2 Profiler Polymerase Chain Reaction (PCR) Array

(SABiosciences, Qiagen). A total of 84 target genes were present on the array. Gene expression of circulating immune cells was analyzed at seven, fourteen, twenty-one and twenty-eight days of the combination supplementation. The expression of 67 genes changed following the combination supplementation across the time points. Table 2 lists the genes with altered gene expression following combination supplementation and includes information indicating stimulation (+) or repression (-) of gene expression.

Table 2

Additional subject matter concerning component I is found in U.S. Patent No. 7,939,066, U.S. Patent No. 8,142,798, U.S. Patent No. 8,236,303, U.S. Patent No. 8,431,133, U.S. Patent No. 8,568,715, U.S. Provisional Application No. 61/856,544, and U.S. Provisional Application No. 61/859,689, each of which is incorporated herein by reference in its entirety.

EXAMPLE 10

A. Methods

In this example, the combination was administered as a composition comprising between 15% and 40% silica, between 50% and 81% mineral clay, between 1.0% and 5.0% β-glucans, between 0.05% and 3.0% β-1,3 (4)-endoglucanohydrolase and between 1% and 8.0% mannans. Embodiments of the combination were used as a feed additive for sea bream. Juveniles of Gilthead sea bream (Sparus aurata) were stocked in 12 tanks in the experimental station. Each tank of 1.0 cubic meters was stocked with 55 juvenile sea bream at an average weight of 26 grams. The water source was from a well at a stable temperature of 21 °C, with a constant total salinity of 18.0 parts per thousand. The duration of the experiment was 158 days.

The experimental protocol included continuous assessment for the presence of diseases causing organisms. Growth performance parameters of the fish were recorded regularly. The daily/weekly assessment of water quality parameters included ammonia, nitrite, pH, temperature and oxygen.

Feeding rate was based on the recommended commercial feeding chart of Phibro Aqua and adjusted according to the size of the fish and the water temperature (FIG. 1). Feeding was performed manually twice a day. The feeding quantity for each tank was adjusted after evaluating the average weight of the fish in each tank every two weeks.

The combination was top-coated on the pellets using 2 wt% of soy oil as an adhesive agent. The control group was given the same feed coated with 2 wt% soy oil. The feed preparation for the trial included mixing the weighted feed in a mixer for 5 minutes with 2 wt% soy oil, and then additional 5 minutes coating with the combination. The experiment was carryout in replicates of 4 tanks per treatment. In the trial 2 different doses of the combination in the feed were compared to a control group B: 100 milligrams per kilogram of bodyweight per day (group A); and 200 milligrams per kilogram of bodyweight per day (group C).

The feed used in this trial was made by Raanan Fish Meal and was based on sinking extruded pellets #4932S0 at sizes of 2-4 millimeters; containing 45.0% protein, 12.0% fat, 3.0% carbohydrates, 9% ash and 9.8% moisture.

B. Results

General health parameters:

1. Survival rates in all the tanks for all the treatments were high (99.1-99.5%).

2. No external or internal parasites were detected in the trial.

3. The general health condition as indicated by the vitality and the response to the feeding was very good for all the treatments for the entire trial.

A significantly higher growth rate of the fish fed with dose (A), using 100 milligrams of the combination per kilogram of bodyweight per day was obtained in this trial. A better growth rate in treatment (A) was observed by day 17. This difference became statistically significant by day 59 (FIG. 9). Without being bound to a particular theory, the better growth rate may be due to an improved nutrition for the fish and/or improved immunostimulant ingredients in the feed. As shown in this trial, the response of the fish to the combination was significantly better compared to a control group without the combination. This conclusion emphasized the efficacy and the advantage of the combination as an effective feed additive in aquatic animals such as fish.

Treatment (A) had the lowest significant feed conversion ratio (FCR) value among the 3 treatments (FIG. 10). This demonstrated the advantage of the combination as an advanced performer, improving the feed intake ability of the fish. This ability to lower the FCR value is a major factor in aquaculture management, because it reduced the feeding cost, which is often the highest cost for fish and shrimp farmers.

The environmental conditions of this example in terms of water temperature, dissolved oxygen levels and water quality were optimal for rearing sea bream. The growth rates of all the 3 groups were according to the expected growth rate of sea bream. The high percentage of survival (99.1- 99.5%) in all 3 groups in this study emphasized the optimum conditions during the trial (FIG. 11). The lower temperature at the end of the trial affected the optimal growth rate of the fish but still the advantages of the combination were evident.

EXAMPLE 11

A. Methods

In this example, the combination was administered as a composition comprising between 15% and 40% silica, between 50% and 81% mineral clay, between 1.0% and 5.0% β-glucans, between 0.05% and 3.0% β-1,3 (4)-endoglucanohydrolase and between 1% and 8.0% mannans. The combination was used as a feed additive for tilapia. Juveniles of hybrid tilapia (Oreochromis niloticus X O. aureus) were stocked in 18 cages in the experimental station. The total volume of the experimental system was 600 cubic meters. Each cage of 1 cubic meter in volume with a 25 millimeter mesh net was stocked with 35 fish at an average weight of 95 grams. The water source was from a well at a stable temperature of 24 °C. The duration of the experiment was 149 days.

The experimental protocol included continuous assessment for the presence of disease causing organisms. Growth performance parameters of the fish were recorded regularly. The daily/weekly assessment of water quality parameters included ammonia, nitrite, pH, temperature and dissolved oxygen.

Feeding rate was based on the recommended commercial feeding chart of Phibro Aqua and adjusted according to the size of the fish and the water temperature (FIG. 12). Feeding was performed manually twice a day. The feeding quantity for each cage was adjusted after evaluating the average weight of the fish in each cage every two weeks. The combination was top-coated on the pellets using 2 wt% soy oil as the adhesive agent. The control group was given the same feed coated with 2 wt% soy oil. The feed for the trial was prepared by mixing the weighted feed in a mixer for 5 minutes with 2 wt% soy oil, and then additional 5 minutes coating with the combination. In this trial 2 different doses of the combination in the feed were compared to a control group B: 100 milligrams AI per kilogram of bodyweight per day (group A); and 200 milligram AI per kilogram of bodyweight per day (group C).

Replicates of 6 cages were used per treatment, which were divided equally in the rearing system. The feed for this trial was manufactured by Zemach Feed Mill. The feed is based on floating extruded pellets #4662 at sizes of 2-4 millimeters; containing 35.0% protein, 3.5% fat, 14.0% carbohydrates, 8.0% ash and 10.0% moisture.

B. Results

General health parameters:

1. Survival rate in all the cages for all the treatments were excellent, without mortality. 2. External parasites (Trichodina and Dactylogyrus) were detected at low incidence.

The fish were treated with formalin 37% and Bromex solution (50% Naled).

3. Low presence of dignea parasite, Centrocestus, was detected. No treatment was required.

4. The general health condition as indicated by the vitality and the response to the feeding was very good for all treatments for the entire trial.

A significantly higher growth rate of the fish fed with dose (A), using 100 mg of the combination per kilogram of bodyweight per day was obtained in this trial. A better growth rate in treatment (A) was observed by day 16. This difference became statistically significant by day 86 (FIG. 13). Without being bound to a particular theory, the better growth rate may be due to an improved nutrition for the fish and/or improved immunostimulant ingredients in the feed.

Administering the composition led to a better growth rate and a better feed intake. 100 and 200 milligrams/kilogram bodyweight per day doses were administered. As shown in this trial, the response of the fish to the combination was significantly better compared to the control group without the combination. This conclusion emphasized the efficacy and the advantage of the combination as an effective feed additive in aquatic animals such as fish.

Treatment (A) had the lowest significant FCR value among the 3 treatments (FIG. 14). This demonstrated the advantage of the combination as an advanced performer, improving the feed intake ability of the fish. This ability to lower the FCR value is a major factor in aquaculture management. The feeding cost is often the highest cost for fish and shrimp farmers. The environmental conditions of the experiment in terms of water temperature, dissolved oxygen levels and water quality were optimal for rearing tilapia. The growth rates of all the 3 groups were better when compared to the expected growth rate of tilapia, emphasizing the optimal conditions of the trial (not shown). The high percentage of survival (100%) in all 3 groups in this study also emphasized the optimum conditions during the trial. The lower temperature at the end of the trial affected the optimal growth rate of the fish but still the advantages of the combination were evident.

EXAMPLE 12

A. Methods

In this example, the combination was administered as a composition comprising between 15% and 40% silica, between 50% and 81% mineral clay, between 1.0% and 5.0% β-glucans, between 0.05% and 3.0% β-1,3 (4)-endoglucanohydrolase and between 1% and 8.0% mannans. The combination was used as a feed additive for carp. Juveniles of Common carp (Cyprinus carpio) were stocked in 18 cages in the experimental station. The total volume of the experimental system was 600 cubic meters. Each cage of 1 cubic meter in volume with a 25 millimeter mesh net was stocked with 35 fish at an average weight of 160 grams. The water source was from a well at a stable temperature of 24 °C. The duration of the experiment was 83 days.

The experimental protocol included continuous assessment for the presence of diseases causing organisms. Growth performance parameters of the fish were recorded regularly. The daily/weekly assessment of water quality parameters included ammonia, nitrite, pH, temperature and dissolved oxygen.

Feeding rate was based on the recommended commercial feeding chart of Phibro Aqua and adjusted according to the size of the fish and the water temperature (FIG. 15). Feeding was performed manually twice a day. The feeding quantity for each cage was adjusted after evaluating the average weight of the fish in each cage every two weeks.

The combination was top-coated on the pellets using 2 wt% soy oil as the adhesive agent. The control group was given the same feed coated with 2 wt% soy oil. The feed for the trial was prepared by mixing the weighted feed in a mixer for 5 minutes with 2 wt% soy oil, and then additional 5 minutes coating with the combination. In this trial 2 different doses of the combination in the feed were compared to a control group C: 100 mg per Kg of body weight per day (group A); and 200 mg per Kg of body weight per day (group B).

Replicates of 6 cages were used per treatment, which were divided equally in the rearing system. The feed for this trial was manufactured by Zemach Feed Mill. The feed is based on floating extruded pellets #4212 at size of 4 millimeters; containing 30.0% protein, 5.0% fat, 4.5% carbohydrates, 8.0% ash and 10.0% moisture.

B. Results

General health parameters:

1. Survival rate in all the cages for all the treatments were excellent, without mortality.

2. External parasites (Gyrodectylus and Dactylogyrus) were detected at low incidence. The fish were treated with formalin 37% and Bromex solution (50% Naled).

3. The general health condition as indicated by the vitality and the response to the feeding was very good for all treatments for the entire trial.

A significant higher growth rate of the fish fed with dose (A), using 100 milligrams of the combination per kilogram of bodyweight per day was obtained in this trial. A better growth rate in treatment (A) was observed by day 41. This difference became statistically significant by day 83 (FIG. 16). Without being bound to a particular theory, the better growth rate may be due to an improved nutrition for the fish and/or improved immunostimulant ingredients in the feed.

Administering the combination led to a better growth rate and a better feed intake. 100 and 200 milligram/kilogram bodyweight per day doses were administered. As shown in this trial, the response of the fish to the combination was significantly better compared to the control group without the combination. This conclusion emphasized the efficacy and the advantage of the combination as an effective feed additive in animals like fish.

Treatment (A) had the lowest (insignificant) FCR value among the 3 treatments (FIG. 17). This demonstrated the advantage of the combination as an advanced performer, improving the feed intake ability of the fish. This ability to lower the FCR value is a major factor in aquaculture management. The feeding cost is often the highest cost for fish and shrimp farmers.

The temperatures of the experiment demonstrated a cold water environment (16-21 °C).

This range of temperatures is common in carp culture worldwide. These low temperatures affected the optimal growth rate of the fish but still the advantages of the combination were evident. The water conditions in terms of dissolved oxygen levels, ammonia, nitrite and pH were optimal for rearing carp. The high percentage of survival (100%) in all 3 groups in this study emphasized the optimum conditions during the trial (not shown). EXAMPLE 13

A. Background

In this example, the combination was administered as a composition comprising between 15% and 40% silica, between 50% and 81% mineral clay, between 1.0% and 5.0% β-glucans, between 0.05% and 3.0% β-1,3 (4)-endoglucanohydrolase and between 1% and 8.0% mannans. The combination was used as an immune modulator for hybrid tilapia. Ammonia is a toxic compound that can adversely affect fish health. The nature and degree of toxicity depends on many factors, including the chemical form of ammonia, the pH and temperature of the water, the length of exposure, and the life stage of the exposed fish. In natural surface waters, ammonia occurs in two forms: ionized ammonia, NH₄ ⁺, and un-ionized ammonia, NH3. In fish, ammonia is a byproduct of protein metabolism and is primarily excreted across the gill membranes, with a small amount excreted in the urine. Ammonia's toxicity is principally due to the un-ionized form, NH3. As pH increases, the toxicity of ammonia rises because the relative proportion of unionized ammonia increases. The toxicity of ammonia may cause convulsions, coma and death. Without being bound to a particular theory, elevated NH₄ ⁺ in the fish body may displace K⁺ and depolarize neurons, causing activation of glutamate receptor, which leads to an influx of excessive Ca²⁺ and subsequent cell death in the central nervous system. In the case of larvae of common carp, acute toxicity of 1.76 parts per million of NH3 caused 50% mortality in the group after 24 hours. Chronic effects of ammonia were studied in three batches of turbot {Scophthalmus maximus) juveniles (14, 23 and 104 grams) exposed for 4—6 weeks to constant ammonium chloride solutions. Under the environmental conditions used (16.5-17.5 °C, pH 7.92-8.03, salinity 34.5 parts per thousand, over 80% oxygen saturation), no mortalities occurred up to 0.4 parts per million unionized ammonia. In adapted small turbot, no major physiological disturbances were observed up to 0.4-0.5 parts per million, while large turbot were more sensitive to ammonia.

The ability to improve the resistance of aquatic species to the toxicity of the ammonia has been investigated. Tiger shrimp (Penaeus monodon), 5-day post larvae, were fed diets

supplemented with 0 and 71.5 parts per million astaxanthin for 8 weeks. Shrimp were then subjected to 72 hours exposure of ammonia at 0.02, 0.2, 2 and 20 parts per million. The survival rates of the astaxanthin-fed shrimp were higher than those of the control shrimp under all levels of ammonia except, 20 ppm, showing that the shrimp's resistance to ammonia stress had been improved by dietary astaxanthin. Other research has investigated the effects of dietary mannans oligosaccharide (MOS) on growth performance, gut morphology, and NH3 stress tolerance of Pacific white shrimp Litopenaeus vannamei. After NH₃ stress for 24 hours, survival rates of shrimp fed 2.0, 4.0, 6.0 and 8.0 grams/kilogram MOS-supplemented diets were significantly higher (P < 0.05) than that of shrimp fed a control diet.

The purpose of this study was to evaluate the effect, of the combination on the fish resistance to the stressful condition of toxic ammonia levels in the water.

B. Methods

Hybrid tilapia (Oreochromis niloticus X O. aureus) were stocked in 12 tanks in the experimental station. Each tank of 230 liter in volume was stocked with 10 fish with an average weight of 350 grams per fish. The water source was from a well with a constant water temperature of 22 °C and constant salinity of 1,300 milligrams chloride. The duration of the experiment was 74 days. During the first phase, 6 tanks were fed 100 milligrams of the combination per kilogram of bodyweight per day, while the other 6 tanks were fed with commercial feed without supplement. After 30 days of feeding in optimal conditions of water, the water inlet was reduced, allowing the water quality to deteriorate for an additional 30 days. In the third phase of 14 days the water inlet was closed completely and ammonium chloride (NH₄C1) was added to each tank on a daily basis. This phase was characterized by a continuous mortality of the fish showing clinical symptoms of ammonia toxicity and bacterial infections associated with poor water quality.

The experimental protocol included continuous assessment for the presence of diseases causing organisms. The daily assessment of water quality parameters included ammonia, nitrite, pH, water temperature and dissolved oxygen.

Feeding rate was 1 % of bodyweight, based on the recommended commercial feeding chart of Phibro Aqua and was adjusted according to the water temperature and the response of the fish (FIG. 12). Feeding was performed manually twice a day. The combination was top-coated on the pellets using 2 wt% soy oil as the adhesive agent. The control group was given the same commercial feed coated with 2 wt% soy oil, but without the combination. The feed for the trial was prepared by mixing the weighted feed in a mixer for 5 minutes with 2 wt% soy oil, and then additional 5 minutes coating with the combination. The feed for this trial was manufactured by Zemach Feed Mill. The feed is based on floating extruded 4 mm pellets, #4662; containing 35.0% protein, 3.5% fat, 14.0% carbohydrates, 8.0% ash and 10.0% moisture. C. Results

General health parameters:

1. At the third phase (14 days) the fish didn't respond to the feed.

2. The moribund and the dead fish that were collected during the trial had typical clinical symptoms of toxicity of ammonia.

The results of this trial showed a significant higher resistant fish fed a diet with the combination at a dose of 100 mg/Kg of body weight per day compared to the control without the combination (FIG. 18). In this trial, the moribund and the dead fish that were collected during the trial had typical clinical symptoms of ammonia toxicity, including convulsions, gill necrosis, coma, and death.

Poor water quality suppresses the immune system of the fish, enabling parasites and bacteria to enter the fish body, causing disease outbreak and consequently mortality. In the experiment, the clean water inlet flow was reduced to cause deterioration of the water quality, which finally resulted in death in the most stressed and frail fish in this trial (FIGS. 19 and 20).

Feeding tilapia with the combination at a dose of 100 milligrams/kilogram of body weight per day for 30 days period significantly improved their resistance and survival under poor water conditions such as high levels of ammonia and nitrite.

EXAMPLE 14

The combination as an immune modulator on the survival and the overall health status of the

Pacific White Shrimp (Litopenaeus vannamei)

Materials and Methods

General design of the clinical field study

2,400 Postlarva-20 days of Litopenaeus vannamei were stocked in 12 tanks in the experimental station. Each tank of 500 liter in volume was stocked with 200 PL- 20, at an estimated weight of 0.15 g per postlarvae. The experimental unit included a central collecting tank and a central biofilter. The water source was from a well. Balance marine salt was added to the water to achieve a total salinity of 10 ppt (parts per thousand). The duration of the experiment was 71 days. The average size of the shrimp at the end of the trial was around 10 grams.

Diet supplemented with the disclosed combination

In this trial, the combination was administered as a composition comprising between 15% and 40% silica, between 50% and 81% mineral clay, between 1.0% and 5.0% β-glucans, between 0.05% and 3.0% β-1,3 (4)-endoglucanohydrolase and between 1% and 8.0% mannans. 2 different doses of the combination were compared to a control. 4 tanks were fed 100 mg of the combination per Kg of BW per day, 4 tanks were fed 200 mg of the combination per Kg of BW per day, while the other 4 tanks were fed with commercial feed without supplement.

Feeding rate based on the recommended feeding chart of Phibro Aqua for Shrimp. Feed quantity was adjusted according to the water temperature, the response of the shrimp and the estimation of their average weight. Feeding was performed manually twice a day. The combination was top-coated on the pellets using 2% of Soy oil as the adhesive agent. The control group was given the same feed without supplement coated with 2% Soy oil. The feed for the trial was prepared by mixing the weighted feed in a cement mixer (maximum load of 50 Kg) for 5 minutes with 2% Soy oil, and then additional 5 minutes coating with the supplement.

General conditions

The experimental protocol included continuous assessment for the presence of diseases causing organisms. The daily assessment of water quality parameters included total salinity, ammonia, nitrite, pH, water temperature and dissolved oxygen.

Results

The results illustrate that a significantly greater percentage of the shrimp that were fed the supplement survived, compared to the control group (FIG. 21). The conditions of the trial were excellent for growing shrimp. Shrimp had good body condition and good coloration. No external parasites were detected. The final average weight of the shrimp was around 10 grams, normal for in-door culture. At this stage (nursery), the survival rates of the control groups (60%) are normal.

In a different trial, 2500 shrimp in a pond were administered a combination formulated as a composition comprising between 15% and 40% silica, between 50% and 81% mineral clay, between 1.0% and 5.0% β-glucans, between 0.05% and 3.0% β-1,3 (4)-endoglucanohydrolase and between 1% and 8.0% mannans. After 6 months, the shrimp were compared to 2500 control shrimp in a separate pond that were not administered the combination. The composition-fed shrimp has an 86% survival rate, compared to 22% for the control shrimp, and also had a greater yield (kg/pond) than the control shrimp.

EXAMPLE 15

96 pens were randomly filled with 20 male R X Ross 708 broiler chicks. The chicks were fed one of six diets: 1) a positive control (PC) diet consisting of a Standard OK Foods Diet without saponins; 2) a negative control (NC) diet consisting of the PC diet without any direct-fed microbial (DFM); 3) a PC diet with added Nutrafito^® Plus; 4) a NC diet with Nutrafito^® Plus; 5) a NC diet with added Ganpro^®; and 6) a NC diet with added Nutrafito^® Plus and Ganpro^®. Nutrafito^® Plus was added at an inclusion rate of 125 ppm, or 0.25 pounds per ton of feed. The inclusion rate of Ganpro^® was 7.5 grams per ton of feed. Feed was fed on a per pound basis as follows:

Starter - 1.5 pounds per bird; Grower - 3.0 pounds per bird; Finisher 1 - 4.0 pounds per bird; Finisher 2 - To market. The birds were weighed on days 0, 7, 14, 28, 42 and 48, the end of the trial.

Results: Table 2 and FIG. 22 show the results from the trial. As can be seen in Table 3, and FIG. 22, the combination of Nutrafito^® Plus and Ganpro^® significantly reduced, by about 3-4%, the feed conversion rate by day 48, relative to both the controls and to the chickens given only feed with Nutrafito^® Plus or only feed with Ganpro^®.

Table 3: Results from trial of Nutrafito Plus^® and Ganpro^® combination

EXAMPLE 16

Study Animals: HubxCobb500 Broiler Chickens from O.K. Farms, Inc., Stigler Hatchery, Stigler, OK. Males and females (straight run as hatched), with initial weight 35-60 grams.

TREATMENT

1 ABF Control Diets 5 ABF Control Diets

2 ABF Control Diets 6 ABF Control Diets

3 Treatment Diets 7 Treatment Diets

4 Treatment Diets 8 Treatment Diets

The treatments diets (FIG. 23) included a combination of Nutrafito^® Plus and a composition comprising Bacillus coagulans (Ganpro^® and/or Previa 6086^®). Nutrafito^® Plus was added at an inclusion rate of 125 ppm, or 0.25 pounds per ton of feed. The inclusion rate of the Bacillus coagulans composition was 7.5 grams per ton of feed.

Study Design:

Housing: Eight (8) commercial broiler houses similar in construction, design, size, compass direction, insulation, heating, ventilation, lighting, watering system, and feed equipment were used for the study. The number of birds placed per house and general house environment were similar as possible so that differences in performance due to housing were minimized.

Feeder space: Two lines of Cumberland Hi Lo pan feeders were providing feed for a minimum of 64 birds per pan throughout the study. Additional ChickMate pan feeders provided additional feeder space during the brooding period.

The Floor space: Stocking density was 0.86 sq ft/bird.

Feed and water: Feed and water was provided ad libitum consumption.

Environment: Each house has insulated, solid sidewalls with insulated endwalls, and ceiling. The minimum ventilation system provides slight negative pressure with two, 36" fans mounted on the endwalls of the house. Tunnel ventilation uses eight 48" fans mounted in the end of the house. A computerized controller using 7 sensors placed throughout the house controls the house

temperature.

Vaccinations: The birds received Marek's, IBD and NC/IBV vaccine at the hatchery. LT vaccine was administered at the hatchery.

Basal diet: Diets were a typical commercial starter, grower, finisher 1 and finisher 2 used by O.K. Farms. The only differences in the diets were as indicated in Treatments. All diets were pelleted. The starter diet was crumbled after pelleting. Grower & finisher feeds were also crumbled. PROCEDURE:

Feed preparation: Feed for the trial was made and delivered under the supervision of the

Technical Services Department. Samples were taken from the truck prior to delivery to the bins on the farm. Samples were retained until the study was complete.

Feed was fed on a pound per bird placed basis as follows:

Starter - 1.5 pounds per bird placed;

Grower - 3.00 pounds per bird placed;

Finisher One - 4.00 pounds per bird placed;

Finisher Two - As needed to market.

Bird placement: The eight houses on the farm were filled with 15,300 day-old broiler chicks from HubbxCobb500 breeder flocks. Chicks from each breeder flock were equally distributed in each house. An investigator was stationed at the hatchery to confirm the distribution of each breeder flock. Chick boxes were labeled by house before they were loaded on the delivery truck. The label on each chick box was verified and the total number of boxes for each house was verified before it was unloaded in the house and recorded. They were placed at 0.86 sq. ft. /bird.

Randomization: Technical Services provided the randomization procedure for assigning the color- coded treatments to four houses according to an allotment of treatment to houses. Houses were identified with study number, house number, and treatment color code as follow: House #1 - White, House #2 - Orange, House #3 - Green, House #4 Yellow, House #5 - Orange, House #6 - Green, House #7 - White, House #8 - Yellow.

Observations: The broiler house caretaker observed the birds in all houses daily. In addition, the Investigator was examining each house at least 3 days per week to look for clinical signs of disease and to assess the environment and litter condition.

Mortality: Mortality and culling records were average for the week these birds were processed. Final Bird Weights: At the end of the grow-out period birds were processed. The gross and tare weights of the trucks used to transport the birds from each house to the processing facility was determined and recorded. Likewise the number of birds placed on the trucks at the grow-out facility was recorded. Authenticated copies of these records were placed in the study file as raw data.

Processing: At the end of the grow-out period, birds were processed at O.K. Farms, Inc. Processing Plant, North 6^th and Reed, Fort Smith, AR 72902 (Establishment number P-165S). A USDA Certificate of Condemnation was prepared and copies were collected as a raw data for each house separately. Disposal of unused feed: Any unused or weigh back feed was recorded and documented when it was returned to the mill.

Raw data: All original raw data were assembled as a part of the Investigator's report and forwarded to the sponsor upon completion of the study. A readable, exact, dated copy of the data were retained in a file and stored in a secured area by OK Foods, Inc.

Completed Data: The results of the performance data are enclosed in tabular (Table 4) and graphic form (FIGS. 24 and 25). As can be seen in Table 4, the feed conversion rates are improved for the animals that had the combination of Nutrafito^® Plus and the Bacillus coagulans composition administered with the feed, relative to the animals that only received the feed.

Table 4.

EXAMPLE 17

Study Animals:

Forty-eight female rats ordered from Charles River Labs were randomly assigned to two rats per cage. Three cages were randomly assigned to each treatment. Rats were allowed ad libitum access to powdered Teklad 2014 prior to initiation of the study. Rats weighed approximately 180 g at the start of the experiment. Environmental Conditions:

Temperature was set to 68 °F and lighting to 12 hours light: 12 hours dark. Bedding was changed as needed throughout the study. Actual temperature ranged from 69.4 °F to 75.4 °F. Humidity ranged from 31% to 73%.

Treatment:

On day 1 of the study, old feed from feeders was replaced with freshly-prepared diets. Rats were weighed on the first day of the trial, after 14 days on feed, after 21 days on feed and before sacrifice at day 28.

The control diet was Teklad 2014. Treatment diets were prepared with inclusion of supplement in Teklad 2014. The treatments diets included Bacillus coagulans (Previa 6086^®), Nutrifito^® Plus, Yucca extract, Quillaja extract, a combination of Previa 6086^® and Nutrafito^® Plus, a combination of Previa 6086^®and Yucca and a combination of Previa 6086^® and Quillaja. The inclusion rate of the Previa 6086^®was 0.000825%, Nutrifito^® Plus was 0.0125%, Yucca was 0.0125% and Quillaja: was 0.0125%. Treatment duration was 28 days.

Plasma Purification:

Whole blood was subjected to centrifugation (5000 x g) for 20 minutes. Plasma was aliquoted into 0.500 mL volumes and stored at -80 °C until analysis.

Plasma Cytokine Analysis:

Plasma from all groups was analyzed for Interleukin-6 (IL-6; Cat No. MBS012805, MyBioSource) and Interleukin-10 (IL-10, Cat No. MBS034393, MyBioSource).

Statistical Analysis of ELISA Data:

Data gathered from ELISA was interpolated via standard curve using Graphpad Prism 6.0.

Subsequent data was evaluated for normality and then analyzed by two-way ANOVA to determine effect of diet supplementation. Post-hoc tests were used to determine differences between groups.

Completed Data:

The concentration of IL-6 and IL-10 in the plasma of rats was determined in this project to evaluate the effect of treatment on immune system function. The results of the data are included in graphic form (FIGS. 26-27). As can be seen in FIG. 26, Nutrifito^® Plus, Yucca extract, the combination of Provia 6086^® and Nutrafito^® Plus and the combination of Previa 6086^® and Yucca extract resulted in a lower plasma concentration of IL-6 when compared to control after 28 days of inclusion in the diet. As can be seen in FIG. 27, Quillaja, the combination of Provia 6086^® and Nutrifito^® Plus, the combination of Provia 6086^® and Yucca extract and the combination of Provia 6086^® and Quillaja resulted in a low plasma concentration of IL-10 when compared to control after 28 days of inclusion in the diet. These results indicate that inclusion of these supplements will alter the production of cytokines that regulate homeostasis and possibly physiological inflammation. IL- 6 is generally considered a pro-inflammatory cytokine and IL-10 is generally considered an anti- inflammatory cytokine. Both of these cytokines are considered myokines, or factors produced by muscle tissue during tissue recovery. Thus, these data indicate that the supplements regulate gut- influenced cytokine production and potentially muscle growth through cytokine suppression. This regulation will result in less energy expenditure to cytokine production, improved nutrient uptake and nutrient utilization. Taken in combination with the improved feed conversion and weight gain observed in poultry projects described in this application, these data indicate that one biological action of these supplements is to improve nutrient uptake through alteration of gut enterocyte function and production of cytokines in the lamina propria and connective tissue of the

gastrointestinal tract. Similarly, alteration of the production of myokines can lead to enhanced muscle growth leading to increased weight gain as less energy is expended on the differentiation and proliferation of myosatellite cells versus the increase in size of muscle filaments.

EXAMPLE 18

In a comparison test, two groups of turkeys, were administered a base diet or a base diet supplemented with a combination of Nutrafito^® Plus and Provia 6086^® for the first 12 weeks of life. The birds were in pens for 21 days at 1.1 sq. ft./bird. Each group was then split into 10 pens, at 3.75 sq. ft./bird. The weights of the birds were recorded at the start and after 141 days, and the average daily weight gain (ADG) was calculated (Table 5). A feed conversion ratio was calculated from the Total Pounds of Feed/Net Pounds of bird. The feed cost per live pound of bird was calculated from the Total cost of the Feed/Net pounds of bird. After factoring in other costs, a Total Cost per Live Pound of Bird (Total Live) was calculated for each group (Table 5).

Table 5.

As can be seen in Table 5 and FIGS. 28-30, turkeys fed the supplemented diet had an improved feed conversion ratio, and a lower live weight cost per pound. Table 6 demonstrated the benefits of the combination when the results from Table 5 are extrapolated to a projected annual turnover of over 5 million birds. The improved feed conversion ratio and subsequent lower cost per pound is projected to result in a saving of over two million dollars.

Table 6.

EXAMPLE 19

One-day old broiler chicks are fed a daily feed supplement comprising 10-25% allicin, 1- 5% β-1,3 (4)-endoglucanohydrolase, 20-40% silica, 5-15% of potassium sorbate, 25-50% yeast cell wall and 0.5-1% vitamin C. The supplement is mixed with the feed in an amount of 0.5 wt%. A control group is not fed the supplement. After 45 days the group fed the supplement are expected to increased weight gain compared to the control group, and also a lower mortality rate.

Additionally, innate immunity markers are measured during the trial period. After 14 days, mRNA expression of macrophage inflammatory protein 1β is expected to be significantly higher than in the control group, indicating that the supplement increases the chickens' innate immune system.

Furthermore, the chickens fed the supplement will show an enhanced immune response when administered a vaccine, compared to the control group.

EXAMPLE 20

The study is designed to determine the effect of feeding of a supplement comprising allicin on the performance of broiler chickens when reared in commercial broiler houses. The test period begins on Trial Day 0 (day of hatch) and the chicks are fed a commercial type crumbled feed. Experimental units are 14 replicate groups of 10 male broilers randomly assigned to treatment or control groups. The total length of the project is 42 days.

The chicks are split into four groups -a control group and three experimental groups as described below:

Group 1 - a control group fed feed with no supplement;

Group 2 - an experimental group fed feed comprising 0.025% to 0.25% allicin, and from 100 ppm to 500 ppm, typically about 125 ppm, of a supplement comprising yucca and quillaja, Yucca schidigera and Quillaja saponaria;

Group 3 - an experimental group fed feed containing a supplement comprising 5-50% allicin, 1-5% β-1,3 (4)-endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans, at an inclusion rate in the feed of about 0.5%; and

Group 4 - an experimental group fed feed containing a first supplement comprising 5-50% allicin, 1-5% β-1,3 (4)-endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans, and a second supplement yucca and quillaja, such as Yucca schidigera and Quillaja saponaria. The inclusion rates are 0.5% for the first supplement in the feed, and 100 ppm to 500 ppm, typically 125 ppm, for the second supplement in the feed.

Optionally, additional control groups may include a group fed a supplement comprising yucca and quillaja without allicin; a group fed a supplement comprising 1-5% β-1,3 (4)- endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans without allicin; and/or a group fed a supplement comprising allicin without yucca, quillaja, silica, mineral clay, glucan and mannans. Chickens are checked daily for signs of unusual growth, disease, health problems and mortality and records updated daily. Body weights and food consumption are measured weekly. At the end of the project, one bird per group is sampled for blood collection.

Measurements include the seven-day mortality, 14-day mortality, total mortality, average weight, average daily gain, feed consumed, feed conversion and growth rate. Blood samples at the end of the project are used for complete blood count analysis, super-oxide dismutase (SOD) analysis activity, and the expression of genes that regulate immune function.

The expected outcomes of this project include improved feed conversion, weight gain and reduced mortality in the experimental groups compared to the control group(s). Expression of genes associated with immune system function, such as L-selectin and interleukin 8 receptor, will be higher in the chickens fed the supplemented feed compared to the controls. In addition, higher SOD activity is expected in chickens fed the supplemented feed.

EXAMPLE 21

The study is designed is to compare the performance of nursery pigs fed diets supplement comprising allicin. Four rooms are used to house the pigs. Each room will consist of 36 pens per room with 10 pigs per pen. Treatments are assigned to pens with a completely randomized design and pigs are assigned to pens based on weight on the day of weaning (about 20 days of age). The pens are mixed sex pens and gender balanced within pen. A total of four treatments with 36 replication with the pen as the experimental unit. Rations are fed in a pellet form. Pig health is assessed daily and mortality, disease or health problems are recorded. The total length of the project is 45 days.

The pigs are split into four groups -a control group and three experimental groups as described below:

Group 1 - a control group fed feed with no supplement;

Group 2 - an experimental group fed feed comprising 0.025% to 0.25% allicin, and from 100 ppm to 500 ppm, typically about 125 ppm, of a supplement comprising yucca and quillaja, Yucca schidigera and Quillaja saponaria;

Group 3 - an experimental group fed feed containing a supplement comprising 5-50% allicin, 1-5% β-1,3 (4)-endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans, at an inclusion rate in the feed of about 0.5%; and

Group 4 - an experimental group fed feed containing a first supplement comprising 5-50% allicin, 1-5% β-1,3 (4)-endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans, and a second supplement yucca and quillaja, such as Yucca schidigera and Quillaja saponaria. The inclusion rates are 0.5% for the first supplement in the feed, and 100 ppm to 500 ppm, typically 125 ppm, for the second supplement in the feed.

Optionally, additional control groups may include a group fed a supplement comprising yucca and quillaja without allicin; a group fed a supplement comprising 1-5% β-1,3 (4)- endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans without allicin; and/or a group fed a supplement comprising allicin without yucca, quillaja, silica, mineral clay, glucan and mannans.

Individual pig body weights are obtained at the beg inning of the trial (Day=0). Pen weights of pigs are collected at days 7, 14, 21, 28, 35 and 42. All feed are weighed and recorded and feed weigh-backs recorded on pig weight collection days. Feed consumption is calculated on a pen basis.

Pig performance is assessed based on average daily gain, average daily feed intake and Feed:Gain ratio. Blood is collected from two pigs per pen on Day=2 and Day = 42 and processed for complete blood counts, super-oxide dismutase (SOD) activity analysis and the expression of genes that regulate immune function.

The expected outcomes of this project include improved average daily gain and improved

Feed: Gain ratio for pigs fed the supplement. In addition, fewer health events are expected in pigs fed the supplement. Comparison of the SOD activity is expected to indicate pigs fed the supplement will have higher SOD activity at Day=42 than controls. Similarly, pigs fed the supplement will have higher expression of gene associated with immune system function than pigs fed the control diet. EXAMPLE 22

The study is to compare the performance of lactating multiparous cows fed a dietary supplement with a supplement comprising allicin. Specifically, 32 multiparous cows are fed a conventional corn silage-based diet behind Calan gate feeders. A total of four treatments with eight cows per treatment, with cow as the experimental unit. Feed intake, milk yield, milk composition, body weight, body condition, ruminal pH, and volatile fatty acids (VFA) are measured.

The cows are split into four groups -a control group and three experimental groups as described below:

Group 1 - a control group fed feed with no supplement; Group 2 - an experimental group fed feed comprising 0.025% to 0.25% allicin, and from 100 ppm to 500 ppm, typically about 125 ppm, of a supplement comprising yucca and quillaja, Yucca schidigera and Quillaja saponaria;

Group 3 - an experimental group fed feed containing a supplement comprising 5-50% allicin, 1-5% β-1,3 (4)-endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans, at an inclusion rate in the feed of about 56.7 g/head/day; and

Group 4 - an experimental group fed feed containing a first supplement comprising 5-50% allicin, 1-5% β-1,3 (4)-endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans, and a second supplement yucca and quillaja, such as Yucca schidigera and Quillaja saponaria. The inclusion rates are 56.7g/head/day for the first supplement in the feed, and 100 ppm to 500 ppm, typically 125 ppm, for the second supplement in the feed.

Optionally, additional control groups may include a group fed a supplement comprising yucca and quillaja without allicin; a group fed a supplement comprising 1-5% β-1,3 (4)- endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans without allicin; and/or a group fed a supplement comprising allicin without yucca, quillaja, silica, mineral clay, glucan and mannans.

Cows are fed a diet free of supplements for at least 28 days before a 14-day covariate period and during the covariate period. After the covariate period, cows are blocked by Days In Milk (DIM) and milk yield and assigned randomly within block to dietary treatments.

Cows are fed a total mixed ration for ad libitum intake (approximately 1.10 x expected intake) once daily. Cows are removed from the pen 3 times daily (0430, 1230, and 2030 hours) for milking in a double-twelve parallel milking parlor. Cows are moved through an animal handling area weekly after the 1230 hour milking for body weight determination. Forages, diets, and orts are collected 3 times per week. Dry matter intake (DMI) is determined by recording feed offered daily and refused daily for each cow.

Milk yields are recorded electronically during the covariate and treatment periods. Milk samples from 3 consecutive milkings for each cow are collected weekly during the covariate and treatment periods. The milk samples are analyzed for fat, true protein, lactose, solids nonfat, urea nitrogen, and de novo, mixed, and preformed fatty acids by mid-infrared procedures. Somatic cell count will be analyzed by flow cytometry and weekly milk samples are mathematically composited after analysis in proportion to milk yield at each sampling. Somatic cell count is transformed and analyzed as somatic cell score (SCS) using the equation: SCS = log2(SCC/100) + 3 where SCC is in units of 1,000 cells/mL. Solids-corrected milk is calculated according to the equation: = [(12.3 x kg of fat yield) + (6.56 x kg of solids non-fat) - (0.0752 x kg of milk yield)]. Feed efficiency (kg/kg) is calculated and expressed as milk/dry matter intake and solids-corrected milk/dry matter intake.

Blood is collected from all cows during the covariate period, at Day=0, 14, 28, 35 and 42 for serum and to purify RNA from circulating immune cells.

The expected outcomes of this project include improved milk production and lower SCC in cows fed the supplement. In addition, fewer health events are expected in cows fed the supplement. Similarly, cows fed the supplement will have higher expression of genes associated with immune system function than cows fed the control diet. Cows fed the supplement are expected to have higher serum glucose concentrations and lower concentrations of acute phase proteins

demonstrating improved metabolism and immune function.

EXAMPLE 23

The study is to compare the performance of beef steers fed a dietary supplement with a supplement comprising allicin. Specifically, 32 steer calves (about 225 kg) are fed a conventional backgrounding diet behind Calan gate feeders. A total of four treatments with eight steers per treatment, with steer as the experimental unit. Feed intake, weight, liver function, liver mineral concentration, serum mineral concentration, acute phase proteins, serum metabolites, growth and feed efficiency are measured. The treatment period is 60 days.

The steers are split into four groups -a control group and three experimental groups as described below:

Group 1 - a control group fed feed with no supplement;

Group 2 - an experimental group fed feed comprising 0.025% to 0.25% allicin, and from 100 ppm to 500 ppm, typically about 125 ppm, of a supplement comprising yucca and quillaja, Yucca schidigera and Quillaja saponaria;

Group 3 - an experimental group fed feed containing a supplement comprising 5-50% allicin, 1-5% β-1,3 (4)-endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans, at an inclusion rate in the feed of about 56.7 g/head/day; and

Group 4 - an experimental group fed feed containing a first supplement comprising 5-50% allicin, 1-5% β-1,3 (4)-endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans, and a second supplement yucca and quillaja, such as Yucca schidigera and Quillaja saponaria. The inclusion rates are 56.7g/head/day for the first supplement in the feed, and 100 ppm to 500 ppm, typically 125 ppm, for the second supplement in the feed.

Optionally, additional control groups may include a group fed a supplement comprising yucca and quillaja without allicin; a group fed a supplement comprising 1-5% β-1,3 (4)- endoglucanohydrolase, 20-40% diatomaceous earth, 25-50% brewers yeast cell wall, 50-81% mineral clay, 1.0%-5.0% β-glucans and 1-8% mannans without allicin; and/or a group fed a supplement comprising allicin without yucca, quillaja, silica, mineral clay, glucan and mannans.

Steers are fed a diet free of supplements for at least 28 days before addition of supplements to the diets. Steers are blocked by weight and liver mineral concentration and assigned randomly within block to dietary treatments.

Steers are fed a total mixed ration for ad libitum intake (approximately 1.10 x expected intake) once daily. Steers are moved through an animal handling area weekly prior to feeding for body weight determination. Forages, diets, and orts are collected 3 times per week. Dry matter intake (DMI) is determined by recording feed offered daily and refused daily for each cow. Liver biopsy is collected from each steer at Day= (-) 28, Day=0, 28 and 60. Blood is collected from steers at Day= (-28), 0, 14, 28, 42 and 60 for serum and RNA purification.

The expected outcomes of this project include improved weight gain and Feed:Gain ratio in steers fed the supplement compared to the control group(s). In addition, fewer health events are expected in steers fed the supplement. Similarly, steers fed the supplement will have higher expression of genes associated with immune system function than cows fed the control diet. Steers fed the supplement are expected to have higher serum glucose concentrations and lower concentrations of acute phase proteins demonstrating improved metabolism and immune function. Last, steers fed the supplement will have improved oxidative respiratory index in liver tissue when compared to liver samples collected from control animals.

VII. Exemplary Embodiments

The following numbered paragraphs illustrate exemplary embodiments of the disclosed technology.

Paragraph 1. A combination, comprising one or more of allicin, alliin, allinase, or algae, and one or more of mineral clay, silica, glucan, mannans, endoglucanohydrolase, yeast, a vitamin, yucca, quillaja, a probiotic, an antimicrobial, a vaccine, sorbic acid or a salt thereof, polyphenol, or a growth promotant;

wherein the combination is not 10-25% allicin, 1-5% β-glucanase, 20-40% diatomaceous earth, 5-15% potassium sorbate, 25-50% brewer's yeast cell wall, and 0.5-1% vitamin C. Paragraph 2. The combination of paragraph 1, wherein the combination is a composition.

Paragraph 3. The combination of paragraph 1, wherein the combination comprises a composition I comprising mineral clay, silica, glucan, mannans, or a combination thereof.

Paragraph 4. The combination of any one of paragraphs 1-3, wherein the combination comprises endoglucanohydrolase.

Paragraph 5. The combination of any one of paragraphs 1-4, wherein the yeast is yeast culture, a live yeast, a dead yeast, yeast extract, or a combination thereof.

Paragraph 6. The combination of any one of paragraphs 1-5, comprising allicin, alliin, alliinase, or a combination thereof.

Paragraph 7. The combination of any one of paragraphs 1-6, comprising allicin.

Paragraph 8. The combination of any one of paragraphs 1-7, comprising sorbic acid, or a salt thereof.

Paragraph 9. The combination of paragraph 8, wherein the sorbic acid salt is potassium sorbate, sodium sorbate, ammonium sorbate, or a combination thereof.

Paragraph 10. The combination of paragraph 9, wherein the sorbic acid salt is potassium sorbate.

Paragraph 11. The combination of any one of paragraphs 1-10, wherein the vitamin is vitamin A, vitamin Bi, vitamin B2, vitamin B3, vitamin B5, vitamin B₆, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, or a combination thereof.

Paragraph 12. The combination of paragraph 11, wherein the vitamin is vitamin C.

Paragraph 13. The combination of any one of paragraphs 1-12, comprising algae.

Paragraph 14. The combination of paragraph 13, wherein the algae is a blue-green algae (cyanobacteria), a diatom (bacillariophyta), a stonewort algae (charophyta), a green algae

(chlorophyta), a golden algae (chrysophyta), a dinoflagellate (dinophyta), a brown algae

(phaeophyta) or a red algae (rhodophyta).

Paragraph 15. The combination of any one of paragraphs 1-14, comprising yucca, quillaja, or a combination thereof.

Paragraph 16. The combination of any one of paragraphs 1-15, comprising Yucca schidigera, Quillaja saponaria, or both Yucca schidigera and Quillaja saponaria.

Paragraph 17. The combination of any one of paragraphs 1-16, comprising a probiotic.

Paragraph 18. The combination of paragraph 17, wherein the probiotic is a Bacillus species.

Paragraph 19. The combination of paragraph 17, wherein the probiotic is Bacillus coagulans. Paragraph 20. The combination of any one of paragraphs 1-19, comprising an antimicrobial.

Paragraph 21. The combination of paragraph 20, wherein the antimicrobial is an antibiotic, an antifungal, an antiparasitic, an antiviral, or a combination thereof.

Paragraph 22. The combination of paragraph 21, wherein the antibiotic is a tetracycline, a penicillin, a cephalosporin, a poly ether antibiotic, a glycopeptide, an orthosomycin, or a combination thereof.

Paragraph 23. The combination of paragraph 21, wherein the antibiotic is virginiamycin, Bacitracin MD, Zinc Bacitracin, Tylosin, Lincomycin, Flavomycin, bambermycins, Terramycin, Neo-Terramycin, florfenicol, oxolinic acid, oxytetracycline, hydrogen peroxide, bronopol, sulfadimethozine, ormetoprim, Sulfadiazine, Trimethoprim, or a combination thereof.

Paragraph 24. The combination of paragraph 21, wherein the antifungal is formalin, formalin-F, bronopol, or a combination thereof.

Paragraph 25. The combination of paragraph 21, wherein the antiparasitic is copper sulfate, fenbendazole, formalin, formalin-F, hyposalinity, hadaclean A, praziquantel, emamectin benzoate (SLICE®), or a combination thereof.

Paragraph 26. The combination of paragraph 21, wherein the antiparasitic is an

anticoccidial.

Paragraph 27. The combination of paragraph 26, wherein the anticoccidial is Monensin, Salinomycin, Lasalocid, Narasin, Maduramicin, Semduramicin, , Nicarbazin, Maxiban, Diclazuril, Toltrazuril, Robenidine, Stenorol, Clopidol, Decoquinate, DOT (zoalene), Amprolium, or combinations thereof.

Paragraph 28. The combination of any one of paragraphs 1-27, comprising a vaccine. Paragraph 29. The combination of any one of paragraphs 1-28, comprising a growth promotant.

Paragraph 30. The combination of paragraph 29, wherein the growth promotant is a β- agonist, an antibiotic, an antimicrobial, a steroid, a hormone, or a combination thereof.

Paragraph 31. The combination of any one of paragraphs 1-30, comprising mineral clay, silica, glucan, mannans, and allicin.

Paragraph 32. The combination of any one of paragraphs 1-31, comprising mineral clay, silica, glucan, mannans, and potassium sorbate.

Paragraph 33. The combination of any one of paragraphs 1-32, comprising mineral clay, silica, glucan, mannans, and vitamin C. Paragraph 34. The combination of any one of paragraphs 1-33, wherein the glucan and mannans is provided by yeast cell wall or an extract thereof.

Paragraph 35. The combination of any one of paragraphs 1-30, consisting essentially of mineral clay, silica, glucan, mannans, and allicin.

Paragraph 36. The combination of any one of paragraphs 1-30, consisting essentially of mineral clay, silica, glucan, mannans, endoglucanohydrolase, and allicin.

Paragraph 37. The combination of paragraph 1, consisting essentially of allicin, endoglucanohydrolase, silica, mineral clay, potassium sorbate, glucan, mannans, and vitamin C.

Paragraph 38. The combination of paragraph 1, consisting essentially of allicin, endoglucanohydrolase, silica, mineral clay, potassium sorbate, yeast cell wall extract, and vitamin C.

Paragraph 39. The combination of any one of paragraphs 1-38, wherein the combination is formulated for administration to an animal.

Paragraph 40. The combination of paragraph 39, wherein the animal is a non-human animal.

Paragraph 41. The combination of paragraph 39, wherein the animal is a land animal.

Paragraph 42. The combination of paragraph 41, wherein the land animal is a mammal. Paragraph 43. The combination of paragraph 42, wherein the mammal is a ruminant or ungulate.

Paragraph 44. The combination of paragraph 42, wherein the mammal is a sheep, a goat, a cow, a deer, a bison, a buffalo, an elk, an alpaca, a camel, a llama a horse, a donkey, or a pig.

Paragraph 45. The combination of paragraph 39, wherein the animal is an aquatic animal.

Paragraph 46. The combination of paragraph 45, wherein the aquatic animal is a fish.

Paragraph 47, The combination of paragraph 46, wherein the fish is a salmon, trout, tilapia, sea bream, carp, cod, halibut, snapper, herring, catfish, flounder, hake, smelt, anchovy, lingcod, moi, perch, orange roughy, bass, tuna, mahi mahi, mackerel, eel, barracuda, marlin, Atlantic ocean perch, Nile perch, Arctic char, haddock, hoki, Alaskan Pollock, turbot, freshwater drum, walleye, skate, sturgeon, Dover sole, common sole, wolfish, sablefish, American shad, John Dory, grouper, monkfish, pompano, lake whitefish, tilefish, wahoo, cusk, bowfin, kingklip, opah, mako shark, swordfish, cobia, croaker, or hybrids thereof.

Paragraph 48. The combination of paragraph 45, wherein the aquatic animal is a crustacean.

Paragraph 49. The combination of paragraph 48, wherein the crustacean is a lobster, shrimp, prawns, crab, krill, crayfish, barnacle, or copepod.

Paragraph 50. The combination of paragraph 45, wherein the aquatic animal is a mollusk. Paragraph 51. The combination of paragraph 50, wherein the mollusk is a squid, octopus, abalone, conch, rock snail, whelk, clam, oyster, mussel, or cockle.

Paragraph 52. The combination of paragraph 39, wherein the animal is an avian.

Paragraph 53. The combination of paragraph 52, wherein the avian is a laying hen, chicken, turkey, goose, duck, Cornish game hen, quail, partridge, pheasant, guinea-fowl, ostrich, emu, swan, or pigeon.

Paragraph 54. The combination of paragraph 39, wherein the animal is a companion animal.

Paragraph 55. The combination of paragraph 54, wherein the companion animal is a canine, feline, rabbit, rodent, bird, reptile, fish, crustacean, or amphibian.

Paragraph 56. The combination of any one of paragraphs 1-55, further comprising an adhesive agent.

Paragraph 57. The combination of paragraph 56, wherein the adhesive agent is a syrup, an oil, or a combination thereof.

Paragraph 58. The combination of paragraph 57, wherein the adhesive agent is corn oil, coconut oil, linseed oil, cottonseed oil, olive oil, peanut oil, palm oil, canola oil, safflower oil, soy oil, sunflower oil, Naskole oil, molasses, sorghum, sugar syrup, honey, or any combination thereof.

Paragraph 59. The combination of any one of paragraphs 1-58, further comprising a feedstuff.

Paragraph 60. The combination of paragraph 59, wherein the feedstuff comprises a solid animal feed, a liquid animal feed, a supplement, a premix, water, a feed additive carrier, or a combination thereof.

Paragraph 61. The combination of any one of paragraphs 1-60, further comprising a trace mineral, bulking agent, carrier, colorant, taste enhancer, preservative, benzoic acid or a salt thereof, propionic acid or a salt thereof, ascorbic acid or a salt thereof, gallic acid or a salt thereof, sulfur dioxide, sulfite, nitrite, nitrate, choline or a salt thereof, corn, soybean meal, wheat, barley, rye, canola, corn oil, limestone, salt, distillers dried grains with solubles (DDGS), dicalcium phosphate, sodium sesquicarbonate, methionine source, lysine source, L-threonine, mineral oil, biotin, folic acid, or any combination thereof.

Paragraph 62. The combination of any one of paragraphs 1-61, wherein the combination comprises silica, glucan, mannans, and mineral clay in amounts relative to each other of 1-40 wt% silica, 1-25 wt% glucan and mannans, and 40-92 wt% mineral clay. Paragraph 63. The combination of any one of paragraphs 1-61, wherein the combination comprises silica, glucan, mannans, and mineral clay in amounts relative to each other of 5-40 wt% silica, 2-15 wt% glucan and mannans and 40-80 wt% mineral clay.

Paragraph 64. The combination of any one of paragraphs 1-61, wherein the combination comprises silica, glucan, mannans, and mineral clay in amounts relative to each other of 20-40 wt% silica, 4-10 wt% glucan and mannans, and 50-70 wt% mineral clay.

Paragraph 65. The combination of any one of paragraphs 1-61, wherein the combination comprises silica, glucan, mannans, and mineral clay in amounts relative to each other of 15-40 wt% silica, 1-15 wt% glucans, 0-10 wt% mannans, 50-81 wt% mineral clay.

Paragraph 66. The combination of any one of paragraphs 62-65, wherein the combination further comprises an amount of endoglucanohydrolase of from 0.05 wt% to 5 wt%, relative to the amounts of silica, mineral clay, glucan and mannans in the combination.

Paragraph 67. The combination of paragraph 66, wherein the combination comprises β-1,3 (4)-endoglucanohydrolase, silica, glucan, mannans, and mineral clay in amounts relative to each other of 0.1-3 wt% β-1,3 (4)-endoglucanohydrolase, 20-40 wt% silica, 2-20 wt% glucan and mannans, and 50-70 wt% mineral clay.

Paragraph 68. The combination of paragraph 66, wherein the combination comprises β-1,3 (4)-endoglucanohydrolase, silica, glucan, mannans, and mineral clay in amounts relative to each other of 0.2-3 wt%, β-1,3 (4)-endoglucanohydrolase, 20-40 wt% silica, 4-10 wt% glucan and mannans, and 50-70 wt% mineral clay.

Paragraph 69. The combination of paragraph 1, wherein the combination comprises from 1% to 50% allicin.

Paragraph 70. The combination of paragraph 69, wherein the combination comprises from 15% to 40% allicin.

Paragraph 71. The combination of any one of paragraphs 1-70, wherein the combination comprises from 15 to 99% of a composition I comprising mineral clay, silica, glucan and mannans.

Paragraph 72. The combination of paragraph 71, wherein the combination comprises from 55% to 95% of the composition I.

Paragraph 73. The combination of paragraph 71, wherein the combination comprises from 45% to 60% of the composition I.

Paragraph 74. The combination of paragraph 71, wherein the combination comprises from 10% to 25% allicin, from 5% to 15% potassium sorbate, from 0.5% to 1 % vitamin C, and from 60% to 85% of the composition I. Paragraph 75. The combination of any one of paragraphs 71-74, wherein the composition I further comprises β-1,3 (4)-endoglucanohydrolase.

Paragraph 76. A method, comprising administering the combination of any one of paragraphs 1-75 to an animal.

Paragraph 77. The method of paragraph 76, wherein the animal is a land animal.

Paragraph 78. The method of paragraph 77, wherein the land animal is a non-human animal.

Paragraph 79. The method of paragraph 77, wherein the non-human animal is a mammal.

Paragraph 80. The method of paragraph 79, wherein the mammal is a ruminant or ungulate.

Paragraph 81. The method of paragraph 80, wherein the ruminant is as a sheep, goat, cow, deer, bison, buffalo, elk, alpaca, camel or llama.

Paragraph 82. The method of paragraph 80, wherein the ungulate is a horse, donkey or pig.

Paragraph 83. The method of paragraph 77, wherein the animal is an avian.

Paragraph 84. The method of paragraph 83, wherein the avian is a laying hen, chicken, turkey, goose, duck, Cornish game hen, quail, partridge, pheasant, guinea-fowl, ostrich, emu, swan, or pigeon.

Paragraph 85. The method of paragraph 77, wherein the animal is an aquatic animal.

Paragraph 86. The method of paragraph 85, wherein the aquatic animal is a fish.

Paragraph 87. The method of paragraph 86, wherein the fish is a salmon, trout, tilapia, sea bream, carp, cod, halibut, snapper, herring, catfish, flounder, hake, smelt, anchovy, lingcod, moi, perch, orange roughy, bass, tuna, mahi mahi, mackerel, eel, barracuda, marlin, Atlantic ocean perch, Nile perch, Arctic char, haddock, hoki, Alaskan Pollock, turbot, freshwater drum, walleye, skate, sturgeon, Dover sole, common sole, wolfish, sablefish, American shad, John Dory, grouper, monkfish, pompano, lake whitefish, tilefish, wahoo, cusk, bowfin, kingklip, opah, mako shark, swordfish, cobia, croaker, or hybrids thereof.

Paragraph 88. The method of paragraph 85, wherein the aquatic animal is a crustacean.

Paragraph 89. The method of paragraph 88, wherein the crustacean is a lobster, shrimp, prawns, crab, krill, crayfish, barnacles, or copepods.

Paragraph 90. The method of paragraph 85, wherein the aquatic animal is a mollusk.

Paragraph 91. The method of paragraph 90, wherein the mollusc is a squid, octopus, abalone, conch, rock snail, whelk, clam, oyster, mussel, or cockle.

Paragraph 92. The method of paragraph 77, wherein the animal is a companion animal.

Paragraph 93. The method of paragraph 92, wherein the companion animal is a canine, feline, rabbit, rodent, bird, reptile, fish, crustacean, or amphibian. Paragraph 94. The method of any one of paragraphs 76-93, wherein the combination comprises a composition I comprising mineral clay, silica, glucan and mannans.

Paragraph 95 The method of paragraph 94, comprising administering the composition I in an amount of from greater than zero to 500 mg of composition I per kilogram of body weight per day.

Paragraph 96. The method of paragraph 94, comprising administering the composition I in an amount of from 10 mg to 350 mg per kilogram of body weight per day.

Paragraph 97. The method of paragraph 94, comprising administering the composition I in an amount of from 0.01 gram to 20 gram per kilogram of live body weight.

Paragraph 98. The method of paragraph 94, wherein the animal is a mammal, and the composition I is administered in an amount of from 0.05 grams to 0.2 grams per kilogram of live body weight per day.

Paragraph 99. The method of any one of paragraphs 76-98, wherein administering the combination to the animal provides a beneficial result in the animal compared to an animal not administered the combination.

Paragraph 100. The method of paragraph 99, wherein the beneficial result comprises: prevention of an infectious disease, a non-infectious disease, stress, a stress-related condition or disease;

treatment of an infectious disease, a non-infectious disease, stress, a stress-related condition or disease;

a beneficial effect on the animal's immune system;

an increase in the longevity of the animal; or

a combination thereof.

Paragraph 101. The method of paragraph 100, wherein the animal is an animal that has or is at risk of developing an infectious disease, a non-infectious disease, stress, a stress-related disease or condition, or a combination thereof.

Paragraph 102. The method of paragraph 100, comprising selecting the animal that has or is at risk of developing an infectious disease, a non-infectious disease, stress, a stress-related disease or condition, or a combination thereof.

Paragraph 103. The method of any one of paragraphs 76-102, wherein administering the combination to the animal improves a feed conversion ratio of the animal compared to an animal not administered the combination. In view of the many possible embodiments to which the principles of the disclosed invention may be applied, it should be recognized that the illustrated embodiments are only preferred examples of the invention and should not be taken as limiting the scope of the invention. Rather, the scope of the invention is defined by the following claims. We therefore claim as our invention all that comes within the scope and spirit of these claims.

Claims

We claim:

1. A combination, comprising allicin, and one or more of mineral clay, silica, glucan, mannans, endoglucanohydrolase, yeast, a vitamin, yucca, quillaja, a probiotic, an antimicrobial, a vaccine, sorbic acid or a salt thereof, polyphenol, or a growth promotant;

wherein the combination is not 10-25% allicin, 1-5% β-glucanase, 20-40% diatomaceous earth, 5-15% potassium sorbate, 25-50% brewer's yeast cell wall, and 0.5-1% vitamin C.

2. The combination of claim 1 , wherein the combination is a composition.

3. The combination of claim 1, wherein the combination comprises a composition I comprising mineral clay, silica, glucan, mannans, or a combination thereof.

4. The combination of claim 3, wherein the composition I comprises mineral clay, silica, glucan, mannans and endoglucanohydrolase.

5. The combination of claim 1, wherein the combination comprises yucca, quillaja, or a combination thereof.

6. The combination of claim 5, wherein the combination comprises Yucca schidigera,

Quillaja saponaria, or both Yucca schidigera and Quillaja saponaria.

7. The combination of claim 1, wherein the combination comprises mineral clay, silica, glucan, mannans, endoglucanohydrolase, Yucca schidigera and Quillaja saponaria.

8. The combination of any one of claims 1-7, wherein the yeast is yeast culture, a live yeast, a dead yeast, yeast extract, or a combination thereof.

9. The combination of any one of claims 1-8, further comprising alliin, alliinase, or a combination thereof.

10. The combination of any one of claims 1-7, comprising sorbic acid, or a salt thereof.

11. The combination of claim 10, wherein the sorbic acid salt is potassium sorbate, sodium sorbate, ammonium sorbate, or a combination thereof.

12. The combination of claim 11, wherein the sorbic acid salt is potassium sorbate.

13. The combination of any one of claims 1-7, wherein the vitamin is vitamin A, vitamin B₁, vitamin B2, vitamin B3, vitamin B5, vitamin B₆, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, or a combination thereof.

14. The combination of claim 13, wherein the vitamin is vitamin C.

15. The combination of any one of claims 1-7, further comprising algae.

16. The combination of claim 15, wherein the algae is a blue-green algae

(cyanobacteria), a diatom (bacillariophyta), a stonewort algae (charophyta), a green algae (chlorophyta), a golden algae (chrysophyta), a dinoflagellate (dinophyta), a brown algae (phaeophyta) or a red algae (rhodophyta).

17. The combination of any one of claims 1-7, comprising a probiotic.

18. The combination of claim 17, wherein the probiotic is a Bacillus species.

19. The combination of claim 17, wherein the probiotic is Bacillus coagulans.

20. The combination of any one of claims 1-7, comprising an antimicrobial.

21. The combination of claim 20, wherein the antimicrobial is an antibiotic, an antifungal, an antiparasitic, an antiviral, or a combination thereof.

22. The combination of claim 21, wherein the antiparasitic is an anticoccidial.

23. The combination of any one of claims 1-7, comprising a vaccine.

24. The combination of any one of claims 1-7, comprising a growth promotant.

25. The combination of claim 1, comprising mineral clay, silica, glucan, mannans, and allicin.

26. The combination of claim 25, further comprising potassium sorbate.

27. The combination of claim 26, further comprising vitamin C.

28. The combination of any one of claims 1-7, wherein the glucan and mannans is provided by yeast cell wall or an extract thereof.

29. The combination of claim 1, consisting essentially of mineral clay, silica, glucan, mannans, and allicin.

30. The combination of claim 1, consisting essentially of mineral clay, silica, glucan, mannans, endoglucanohydrolase, and allicin.

31. The combination of claim 1, consisting essentially of allicin, endoglucanohydrolase, silica, mineral clay, potassium sorbate, glucan, mannans, and vitamin C.

32. The combination of claim 1, consisting essentially of allicin, endoglucanohydrolase, silica, mineral clay, potassium sorbate, yeast cell wall extract, and vitamin C.

33. The combination of any one of claims 1-7, wherein the combination is formulated for administration to an animal.

34. The combination of claim 33, wherein the animal is a non-human animal.

35. The combination of claim 34, wherein the animal is a land animal.

36. The combination of claim 35, wherein the land animal is a mammal.

37. The combination of claim 36, wherein the mammal is a ruminant or ungulate.

38. The combination of claim 36, wherein the mammal is a sheep, a goat, a cow, a deer, a bison, a buffalo, an elk, an alpaca, a camel, a llama a horse, a donkey, or a pig.

39. The combination of claim 34, wherein the animal is an aquatic animal.

40. The combination of claim 39, wherein the aquatic animal is a fish, crustacean, or a mollusk.

41. The combination of claim 34, wherein the animal is an avian.

42. The combination of claim 41, wherein the avian is a laying hen, chicken, turkey, goose, duck, Cornish game hen, quail, partridge, pheasant, guinea-fowl, ostrich, emu, swan, or pigeon.

43. The combination of any one of claims 1-7, further comprising an adhesive agent.

44. The combination of claim 43, wherein the adhesive agent is a syrup, an oil, or a combination thereof.

45. The combination of claim 43, wherein the adhesive agent is corn oil, coconut oil, linseed oil, cottonseed oil, olive oil, peanut oil, palm oil, canola oil, safflower oil, soy oil, sunflower oil, Naskole oil, molasses, sorghum, sugar syrup, honey, or any combination thereof.

46. The combination of any one of claims 1-7, further comprising a feedstuff.

47. The combination of claim 46, wherein the feedstuff comprises a solid animal feed, a liquid animal feed, a supplement, a premix, water, a feed additive carrier, or a combination thereof.

48. The combination of claim 46, further comprising a trace mineral, bulking agent, carrier, colorant, taste enhancer, preservative, benzoic acid or a salt thereof, propionic acid or a salt thereof, ascorbic acid or a salt thereof, gallic acid or a salt thereof, sulfur dioxide, sulfite, nitrite, nitrate, choline or a salt thereof, corn, soybean meal, wheat, barley, rye, canola, corn oil, limestone, salt, distillers dried grains with solubles (DDGS), dicalcium phosphate, sodium sesquicarbonate, methionine source, lysine source, L-threonine, mineral oil, biotin, folic acid, or any combination thereof.

49. The combination of any one of claims 1-7, wherein the combination comprises silica, glucan, mannans, and mineral clay in amounts relative to each other of 1-40 wt% silica, 1-25 wt% glucan and mannans, and 40-92 wt% mineral clay.

50. The combination of any one of claims 1-7, wherein the combination comprises silica, glucan, mannans, and mineral clay in amounts relative to each other of 15-40 wt% silica, 1- 15 wt% glucans, 0-10 wt% mannans, 50-81 wt% mineral clay.

51. The combination of any one of claims 1-7, wherein the combination further comprises an amount of endoglucanohydrolase of from 0.05 wt% to 5 wt%, relative to the amounts of silica, mineral clay, glucan and mannans in the combination.

52. The combination of any one of claims 1-7, wherein the combination comprises β-1,3 (4)-endoglucanohydrolase, silica, glucan, mannans, and mineral clay in amounts relative to each other of 0.1-3 wt% β-1,3 (4)-endoglucanohydrolase, 20-40 wt% silica, 2-20 wt% glucan and mannans, and 50-70 wt% mineral clay.

53. The combination of any one of claims 1-7, wherein the combination comprises β-1,3 (4)-endoglucanohydrolase, silica, glucan, mannans, and mineral clay in amounts relative to each other of 0.2-3 wt%, β-1,3 (4)-endoglucanohydrolase, 20-40 wt% silica, 4-10 wt% glucan and mannans, and 50-70 wt% mineral clay.

54. The combination of any one of claims 1-7, wherein the combination comprises from l% to 50% allicin.

55. The combination of claim 54, wherein the combination comprises from 15% to 40% allicin.

56. The combination of claim 54, wherein the combination comprises from 15 to 99% of a composition I comprising mineral clay, silica, glucan and mannans.

57. The combination of claim 56, wherein the combination comprises from 55% to 95% of the composition I.

58. The combination of claim 56, wherein the combination comprises from 45% to 60% of the composition I.

59. The combination of claim 56, wherein the combination comprises from 10% to 25% allicin, from 5% to 15% potassium sorbate, from 0.5% to 1 % vitamin C, and from 60% to 85% of the composition I.

60. The combination of claim 56, wherein the composition I further comprises β-1,3 (4)- endoglucanohydrolase.

61. A method, comprising administering the combination of claim 1 to an animal.

62. The method of claim 61, wherein the animal is a land animal.

63. The method of claim 61, wherein the land animal is a non-human animal.

64. The method of claim 61, wherein the non-human animal is a mammal.

65. The method of claim 61, wherein the mammal is a ruminant or ungulate.

66. The method of claim 65, wherein the ruminant is as a sheep, goat, cow, deer, bison, buffalo, elk, alpaca, camel or llama.

67. The method of claim 65, wherein the ungulate is a horse, donkey or pig.

68. The method of claim 61, wherein the animal is an avian.

69. The method of claim 68, wherein the avian is a laying hen, chicken, turkey, goose, duck, Cornish game hen, quail, partridge, pheasant, guinea-fowl, ostrich, emu, swan, or pigeon.

70. The method of claim 61, wherein the animal is an aquatic animal.

71. The method of claim 70, wherein the aquatic animal is a fish, a Crustacean, or a mollusk.

72. The method of any one of claims 61-71, wherein the combination comprises a composition I comprising mineral clay, silica, glucan and mannans.

73 The method of claim 72, wherein administering the combination comprises administering the composition I in an amount of from greater than zero to 500 mg of composition I per kilogram of body weight per day.

74. The method of claim 73, comprising administering the composition I in an amount of from 10 mg to 350 mg per kilogram of body weight per day.

75. The method of claim 73, comprising administering the composition I in an amount of from 0.01 gram to 20 gram per kilogram of live body weight.

76. The method of claim 73, wherein the animal is a mammal, and the composition I is administered in an amount of from 0.05 grams to 0.2 grams per kilogram of live body weight per day.

77. The method of any one of claims 61-71, wherein administering the combination to the animal provides a beneficial result in the animal compared to an animal not administered the combination.

78. The method of claim 77, wherein the beneficial result comprises:

prevention of an infectious disease, a non-infectious disease, stress, a stress-related condition or disease;

treatment of an infectious disease, a non-infectious disease, stress, a stress-related condition or disease;

a beneficial effect on the animal's immune system;

an increase in the longevity of the animal; or

a combination thereof.

79. The method of claim 78, wherein the animal is an animal that has or is at risk of developing an infectious disease, a non-infectious disease, stress, a stress-related disease or condition, or a combination thereof.

80. The method of claim 78, comprising selecting the animal that has or is at risk of developing an infectious disease, a non-infectious disease, stress, a stress-related disease or condition, or a combination thereof.

81. The method of any one of claims 61-71, wherein administering the combination to the animal improves a feed conversion ratio of the animal compared to an animal not administered the combination.

82. A combination according to claim 1, for use in improving a feed conversion ratio, increasing expression of genes associated with immune system function, increasing super-oxide dismutase (SOD) activity, increasing serum glucose concentrations, lowering concentrations of acute phase proteins, improving milk production or lowering Somatic cell count, in an animal that is administered the combination, compared to an animal that is not administered the combination.

83. The combination for use according to claim 82, wherein the combination comprises allicin, silica, mineral clay, glucan and mannans.

84. The combination for use accordingly claim 82 or claim 83, wherein the combination comprises yucca and quillaja.