Compositions and methods for improved utilization of non-digestible fiber in animals

Abstract

The present disclosure provides compositions and methods for improving utilization and reducing antinutritional effects of non-digestible fiber (NDF) in animal feed. The methods comprise orally administering to the animal a composition comprising lysozyme. More specifically, the methods comprise orally administering to the animal a composition comprising lysozyme a type C lysozyme or a GH-22 lysozyme. The methods and compositions can be used to improve utilization of NDF, growth performance, livability, or any combination thereof in the non-human animal.

Description

COMPOSITIONS AND METHODS FOR IMPROVED UTILIZATION OF NON-DIGESTIBLE FIBER IN ANIMALSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 USC § 119(e) of U.S. Provisional Application Serial No. 63 / 477,548, filed on December 28, 2022, the entire disclosure of which is incorporated herein by reference.BACKGROUND AND SUMMARY

[0002] The insoluble fiber components in feedstuffs have negative impacts on animal gut health. Previous studies have shown that feeding insoluble fiber increased incidence and severity of swine dysentery by (1) increasing unfavorable type of mucin production, and (2) reducing the abundance of lactic acid producing bacteria, i.e., lactobacillus, which plays a significantly beneficial role in the gut under enteric infection in young pigs (see, e.g., Li et al., 2019; Wilberts et al., 2014). Additionally, insoluble dietary fiber decreases digesta transit time and also negatively affect ileal digestibility of amino acids and other nutrients. Improving the digestion and utilization of insoluble fiber in feed ingredients may also benefit the digestibility of other nutrients of the diet.

[0003] The fibrous component in corn and corn DDGS is primarily insoluble and poorly fermentable, with arabinoxylan and cellulose being the major non-starch polysaccharides (Ranathunga et al., 2018). Previous studies showed feeding insoluble fiber from DDGS exacerbated the E. coli infection in young pigs as measured by increased fecal score and E coli. shedding (Li et al., 2019). One of the nutritional interventions to improve digestibility of fiber, especially the insoluble fiber components, is adding carbohydrase (e.g., xylanase and [)- glucanase) to the diet of nonruminant animals. However, the responses on livability and performance have been highly inconsistent. For example, Petry and Patience (2022) reviewed a total of 19 publications from 2000 to 2020 and reported that in 33% and 26% of the studies, respectively, xylanase improved ADG and G:F, but no improvements in average daily feed intake were reported. The responses of xylanase on apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of “fiber” (NSP, TDF, or NDF) were also quite variable. Xylanase improved the AID and ATTD of fiber by 61% and 50% of the time, respectively. Thus, there exists a need for a feed additive to improve utilization and reduce the antinutritional effects of non-digestible fiber in animal feed.

[0004] Accordingly, the present disclosure provides such compositions and methods. The methods of the present disclosure can comprise orally administering a compositioncomprising a lysozyme such as a type C lysozyme or a GH-22 lysozyme to the animal. As described herein, the methods and compositions can be used to improve utilization of NDF, growth performance, livability, or any combination thereof in the non-human animal.

[0005] Other objects, features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTIONS OF THE DRAWINGS

[0006] The detailed description particularly refers to the accompanying figures in which:

[0007] FIG. 1 shows a plot showing effects of dietary supplementation of lysozyme on apparent total tract digestibility (ATTD) of dry matter in nursery pigs.

[0008] FIG. 2 shows a plot showing the effects of dietary supplementation of lysozyme on apparent total tract digestibility (ATTD) of gross energy (GE) in nursery pigs.

[0009] FIG. 3 shows a plot showing the effects of dietary supplementation of lysozyme on apparent total tract digestibility (ATTD) of nitrogen in nursery pigs

[0010] FIG. 4 shows a plot showing the effects of dietary supplementation of lysozyme on ADG of nursery pigs.

[0011] FIG. 5 shows a plot of meta-analysis of feeding lysozyme to nursery pigs on ADG during early nursery phase.

[0012] FIG. 6 shows a plot of meta-analysis of feeding lysozyme to nursery pigs on ADFI during early nursery phase.

[0013] FIG. 7 shows a plot of meta-analysis of feeding lysozyme to nursery pigs on F:G during early nursery phase.

[0014] FIG. 8 shows a plot of meta-analysis of feeding lysozyme to nursery pigs on BW during early nursery phase.

[0015] FIG. 9 shows a plot of meta-analysis of feeding lysozyme to nursery pigs on total removal rate during early nursery phase.

[0016] FIG. 10 shows a plot of meta-analysis of feeding lysozyme to nursery pigs on mortality during early nursery phase.

[0017] FIG. 11 shows a plot of meta-analysis of feeding lysozyme to nursery pigs on closeout nursery performance during entire nursery phase (49-51-d post- weaning).

[0018] FIG. 12 shows a plot of meta-analysis of feeding lysozyme to nursery pigs on closeout nursery performance during entire nursery phase (49-51-d post- weaning).DETAILED DESCRIPTIONFeed Compositions

[0019] One aspect of the present disclosure encompasses feed compositions for nonhuman animals. The compositions comprise a basal animal diet supplemented with lysozyme. Lysozymes are bacteriolytic enzymes that hydrolyze the beta(l->4) bonds between N- acetylglucos amine and N-acetylmuramic acid in the peptidoglycan of prokaryotic cell walls. It has also been recruited for a digestive role in certain ruminants and colobine monkeys. There are at least five different classes of lysozymes: C (chicken type; Type C lysozymes), G (goose type), phage-type (T4), fungi (Chalaropsis', CH type lysozymes), and bacterial (Bacillus subtilis). There are few similarities in the sequences of the different types of lysozymes.

[0020] In some aspects, the lysozyme of the present disclosure is a type C lysozyme. Type C lysozymes belong to the glycoside hydrolase family 22 (GH-22 lysozymes).Accordingly, in some aspects, the lysozyme of the present disclosure is a GH-22 lysozyme. In some aspects, the lysozyme of the present disclosure is a type C lysozyme. In some aspects, the lysozyme of the present disclosure is egg white lysozyme. In some aspects, the concentration of egg white lysozyme in the feed composition ranges from about 0.1 Ib / ton to about 0.5 Ib / ton. In some aspects, the concentration of egg white lysozyme in the feed composition ranges from about 0.2 Ib / ton to about 0.4 Ib / ton. In some aspects, the concentration of egg white lysozyme ranges from about 0.001% w / w to about 0.5% w / w. In some aspects, the concentration of egg white lysozyme ranges from about 0.005% w / w to about 0.250% w / w.

[0021] In some aspects, the lysozyme of the present disclosure is not a CH type lysozyme. CH type lysozymes belong to the glycoside hydrolase family 25 (GH-25 lysozymes). Accordingly, the lysozyme of the present disclosure is not a GH-25 lysozyme.Feed Additive Compositions

[0022] One aspect of the present disclosure encompasses feed additive compositions for non-human animals. The feed additive compositions comprise a lysozyme. The lysozyme can be as described in Section I herein above. In addition to lysozyme, a feed additive composition of the present disclosure can further comprise at least one additional ingredient such as vitamins, minerals, amino acids, antioxidants, probiotics, essential fatty acids, and pharmaceutically acceptable excipients. The feed additive composition can be added to basalanimal diet for administration to the non-human animals. The feed additive composition can be formulated with basal animal diets to prepare the feed compositions described previously.

[0023] The amount of lysozyme in a feed additive composition can and will vary depending on the lysozyme, the type of non-human animal that will be administered the feed additive composition comprising the lysozyme, the body weight, sex, and medical condition of the non-human animal that will be administered the feed additive composition. Generally, a feed additive composition can comprise between about 20% to about 95% lysozyme.

[0024] In various aspects, a feed additive composition can be introduced to a basal animal diet by way of various methods, depending on whether the feed additive composition is in a liquid or solid form. Non-limiting examples of introducing the feed additive composition to a basal animal diet can be formulating the feed additive composition into the basal animal diet, top-dressing the solid composition of a basal animal diet, spraying a liquid feed additive composition onto a basal animal diet, or combinations thereof. It will be recognized that, when the feed additive is introduced to a basal animal diet, the amount of the feed additive introduced to a basal animal diet is sufficient to provide the therapeutically effective amount of the combination of formulated yeast and capsicum product in the diet of the animal.Methods of Using

[0025] Another aspect of the disclosure encompasses methods of nutritional intervention that uses dietary supplementation with lysozyme. The methods comprise orally administering the animal a feed composition or a feed additive of the present disclosure to non- human animals. A feed composition as described previously can be utilized and a feed additive composition as described previously can be utilized.

[0026] Non-human animals, in broad term, can be defined as any animal which exhibits improved growth, improved health, improved intestinal health, and reduced microbial pathogen counts after administration of the feed additive composition. In various aspects, the non-human animal can be a livestock mammal varying in age and health. Non-limiting example of suitable livestock mammals can be beef cattle, horses, dairy cattle, veal, pigs, goats, sheep, bison, llama, or alpaca. In other aspects, the non-human animal can be an avian species varying in age and health. Non- limiting examples of suitable avian species or poultry can be chickens, including broilers, layers, and breeders, ducks, game hens, geese, pheasants, guinea fowl / hens, quail, turkeys, and ratites, such as emus and ostrich and aquaculture. In another aspect, the non- human animal can be a companion animal varying in age and health. Non-limiting examples of companion animals can be a dog, a cat, a bird, a hamster, or a Guinee pig. In some aspects, thenon-human animal is selected from a group comprising growing pigs, calves, foals, kids (goats), lambs, cria, chicks, poults, ducklings, puppies, kittens, or combinations thereof.

[0027] In some aspects, the non-human animal is a non-ruminant animal. In some aspects, the non-human animal is a pig. In some aspects, the non-human animal is a nursery pig. In some aspects, the non-human animal is a healthy nursery pig.

[0028] The methods can be used to improve utilization of NDF in animal feed. The methods of the present disclosure can also be used to reduce the antinutritional effects of non- digestible fiber in animal feed. In some aspects, a method of the present disclosure comprises reducing the incidence and severity of swine dysentery, reducing unfavorable type of mucin production, increasing the abundance of lactic acid producing bacteria, i.e., Lactobacillus, increasing digesta transit time, positively affecting ileal digestibility of amino acids and other nutrients, or any combination thereof. The methods can improve nutrient digestibility, fiber digestion, and growth performance of the animal without affecting E. coli proliferation.

[0029] In some aspects, the method improves NDF digestibility by at least 36%. In some aspects, the method reduces undigested NDF of DDGS by at least 13%. The method can improve utilization of NDF in the presence of pathogens or absence of pathogens.

[0030] In some aspects, the method improves nutrient digestibility, growth performance of the non-human animal, survivability of the non-human animal, or any combination thereof. For instance, the method can improve ADG, ADFI, feed conversion, increases body weight, reduces mortality, reduces total removal rate, or any combination thereof. The method can also improve apparent total tract digestibility (ATTD) of dry matter, improves ATTD of gross energy, improves ATTD of nitrogen. In some aspects, the method reduces undigestible carbohydrate fractions in a feed composition.

[0031] In some aspects, a method of the present disclosure comprises improving growth performance, livability, or growth performance and livability of a non-human animal. The method comprises orally administering to the animal a feed composition or a feed additive composition comprising a lysozyme. The lysozyme, the feed composition and the feed additive composition can be as described previously.

[0032] In some aspects, the method improves growth performance, livability, or growth performance and livability of an animal irrespective of the diseases state of the animal. In other aspects, the method improves nutrient digestibility, growth performance of the non-human animal, survivability of the non-human animal, or any combination thereof. In some aspects, the method improves ADG, ADFI, feed conversion, increases body weight, reduces mortality, reduces total removal rate, or any combination thereof. In other aspects, the method improvesapparent total tract digestibility (ATTD) of dry matter, improves ATTD of gross energy, improves ATTD of nitrogen. In additional aspects, the method improves growth performance, livability, or growth performance and livability without affecting proliferation of a pathogen in the animal. The method can also reduce undigestible carbohydrate fractions in a feed composition.

[0033] The timing and duration of administration of a composition of the present disclosure to an animal can and will vary. The feed or feed additive composition can be administered throughout the period of feeding the animal. Alternatively, a feed or feed additive composition of the present disclosure can be administered at specific periods during the growth and development of the animal. A feed or feed additive composition of the present disclosure can also be administered at various intervals. For instance, a composition can be administered daily, weekly, monthly, or over a number of months. In some aspects, a composition is administered weekly. In other aspects, a composition is administered monthly. In some aspects, a composition is administered daily. As it will be recognized in the art, the duration of treatment can and will vary depending on the growth and health of the animal.

[0034] The following numbered embodiments are contemplated and are non-limiting:1. A composition for improving utilization and reducing antinutritional effects of non-digestible fiber (NDF) in feed of a non-human animal, the composition comprising a basal animal diet supplemented with lysozyme.2. The composition of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the composition is a feed composition or a feed additive composition.3. The composition of clause 1 or clause 2, any other suitable clause, or any combination of suitable clauses, wherein the lysozyme is not a CH type lysozyme.4. The composition of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the lysozyme of the present disclosure is not a GH- 25 lysozyme.5. The feed composition of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the lysozyme is a type C lysozyme.6. The feed composition of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the lysozyme is a GH-22 lysozyme.7. The feed composition of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the lysozyme is an egg white lysozyme.The feed composition of clause 7, any other suitable clause, or any combination of suitable clauses, wherein the concentration of egg white lysozyme in the feed composition ranges from about 0.1 Ib / ton to about 0.5 Ib / ton. The feed composition of clause 7, any other suitable clause, or any combination of suitable clauses, wherein the concentration of egg white lysozyme in the feed composition ranges from about 0.001% w / w to about 0.5% w / w. The feed composition of clause 8 or clause 9, any other suitable clause, or any combination of suitable clauses, wherein the animal is a non-ruminant animal. The feed composition of any of clauses 8-10, any other suitable clause, or any combination of suitable clauses, wherein the animal is a pig. The feed composition of any of clauses 8-11, any other suitable clause, or any combination of suitable clauses, wherein the animal is a nursery pig. A method of improving utilization and reducing antinutritional effects of NDF in a nonhuman animal, the method comprising orally administering to the animal a composition of any one of clauses 1-12 any other suitable clause, or any combination of suitable clauses,. The method of clause 13, any other suitable clause, or any combination of suitable clauses, wherein the method improves NDF digestibility by at least 36%. The method of clause 13 or clause 14, any other suitable clause, or any combination of suitable clauses, wherein the method reduces undigested NDF of DDGS by at least 13%. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method improves utilization of NDF in the presence of pathogens or absence of pathogens. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method improves nutrient digestibility, growth performance of the non-human animal, survivability of the non-human animal, or any combination thereof. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method improves ADG, ADFI, feed conversion, increases body weight, reduces mortality, reduces total removal rate, or any combination thereof. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method improves apparent total tract digestibility (ATTD) of dry matter, improves ATTD of gross energy, improves ATTD of nitrogen. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method reduces undigestible carbohydrate fractions in a feed composition.21. A method of improving growth performance, livability, or growth performance and livability of a non-human animal, the method comprising orally administering to the animal a composition of any one of clauses 1-12, any other suitable clause, or any combination of suitable clauses,.22. The method of clause 21, any other suitable clause, or any combination of suitable clauses, wherein the method improves growth performance, livability, or growth performance and livability of an animal irrespective of the diseases state of the animal.23. The method of clause 21 or clause 22, any other suitable clause, or any combination of suitable clauses, wherein the method improves nutrient digestibility, growth performance of the non-human animal, survivability of the non-human animal, or any combination thereof.24. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method improves ADG, ADFI, feed conversion, increases body weight, reduces mortality, reduces total removal rate, or any combination thereof.25. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method improves apparent total tract digestibility (ATTD) of dry matter, improves ATTD of gross energy, improves ATTD of nitrogen.26. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method improves growth performance, livability, or growth performance and livability without affecting proliferation of a pathogen in the animal.27. The method of any one of the preceding clauses, any other suitable clause, or any combination of suitable clauses, wherein the method reduces undigestible carbohydrate fractions in a feed composition.

[0035] All patents and publications mentioned in the specification are indicative of the levels of those skilled in the art to which the present disclosure pertains. All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference.

[0036] The publications discussed throughout are provided solely for their disclosure before the filing date of the present application. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.EXAMPLES

[0037] The following examples are included to demonstrate the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the following examples represent techniques discovered by the inventors to function well in the practice of the disclosure. Those of skill in the art should, however, in light of the present disclosure, appreciate that many changes could be made in the disclosure and still obtain a like or similar result without departing from the spirit and scope of the disclosure, therefore all matter set forth is to be interpreted as illustrative and not in a limiting sense.EXAMPLE 1Feeding egg white lysozyme (GH-22) did not demonstrate beneficial responses in nursery pigs challenged with pathogenic E. coli

[0038] A study was conducted to evaluate the effects of feeding egg white lysozyme to weanling pigs on growth performance and signs of diarrhea under E. coli Fl 8 challenge. A total of 36 weanling barrows with weaning age at 19-21 days old between 10 to 16 lb were randomly allotted to one of 3 dietary treatments (Table 1). Pigs were fed respective experimental diets from day 6 to 11 dpi with a 6-day adaptation period and a ll -day post-challenge period. On day 0 and 1-dpi, respectively, each pig was orally inoculated with 5 ml (approximately 2.0 x 108 / mL with a total challenge of 109CFU) of the freshly grown E coli Fl 8 inoculants via a polyethylene tube attached to a syringe placed in the back of the oral cavity. Pigs were continued to be fed their experimental diets and monitored for 11 days post-challenge.Table 1. Experimental dietary treatments

[0039] This E. coli. challenge model resulted in over 75% diarrhea frequency and severe signs of diarrhea in pigs fed Control diet and diet containing egg white lysozyme (Table 2). Feeding pharmacological level of Zn in the diet resulted in 0% mortality, best growth performance, minimal signs of diarrhea and lowest diarrhea frequency in this study. However, feeding egg white lysozyme did not demonstrate any beneficial responses on growth performance, livability or severity of diarrhea compared to Control and ZnO. The lack of responses of egg white lysozyme in this study was different when compared to previous findings that microbial fermented lysozyme (GH-25) improved growth performance in pigs and chickens because of its antimicrobial efficacy (see U.S. Patent No. 10,568,344).Table 2. Effects of feeding egg white lysozyme in weanling pigs challenged with pathogenic E. coli.a bMeans without a common superscript differ (P < 0.05)x yMeans without a common superscript tend to differ (P < 0. 10)1Start BW was used as covariate for all analysesSubjective fecal score was graded on 0 - 3 scale: 0 = no signs of diarrhea; 3 = severe signs of diarrhea.EXAMPLE 2Egg white lysozyme improved in vitro fiber digestibility of feedstuffs

[0040] To determine the fiber digestibility of egg white lysozyme on feedstuff, corn, corn DDGS, soyhulls, and swine grow-finish complete feed were mixed with lysozyme at various concentrations from 0 to 0.25%. Each mixture was prepared in duplicate. Each duplicated sample was digested with a standardized in vitro rumen procedure (Sawyer and Goeser, 2013) and analyzed in triplicate for neutral detergent fiber (NDF) content and digestibility at Rock River Laboratory (Watertown, Wisconsin).

[0041] Neutral detergent fiber concentration was similar within each ingredient regardless of lysozyme concentrations (Table 3). Inclusion of lysozyme as low as 0.005% was able to improve NDF digestibility by at least 36% and reduced undigested NDF of DDGS by at least 13%. In other ingredients, including lysozyme as low as 0.05% NDF digestibility and reduced undigested NDF. These results suggest that egg white lysozyme (GH-22) can improve fiber digestion and utilization in feedstuff that contain NDF.Table 3. Inclusion of lysozyme from 0 to 0.25% improved digestibility of neutral detergent fiber (NDF) of DDGS.Weans differ than Control within each ingredient (P < 0.10) **Means differ than Control within each ingredient (P < 0.05)EXAMPLE 3Feeding egg white lysozyme improved nutrient digestibility and growth performance without affecting E. coli proliferation in nursery pigs fed diets containing NDF

[0042] To evaluate the effects of feeding egg white lysozyme to nursery pigs on nutrient digestibility and growth performance, a total of 60 weanling barrows with weaning age at 19-21 days old between 10 to 16 lb were randomly allotted to one of 3 dietary treatments with lysozyme concentrations of 0, 0.2, and 0.4 Ib / ton of feed (Table 4).Table 4. Dietary treatments

[0043] Pigs were fed respective experimental diets for 10-d post-weaning. The basal diet was formulated to contain 1 .98% crude fiber and 5.60% NDF (Table 5). Each pig received an oral gavage of E. coli. Fl 8 inoculant on day 7 to mimic the enteric challenge that is usually seen in nursery pigs under commercial production condition. On day 10, pigs were euthanized, and ileal mucosa tissue was collected for total coliform and enterotoxigenic E. coli (ETEC) plate count.Table 5. Composition of basal diet

[0044] Dietary supplementation of lysozyme up to 0.4 Ib / ton to nursery pigs linearly improved apparent total tract digestibility (ATTD) of dry matter (P = 0.05; FIG. 1), gross energy (P = 0.03; FIG. 2) and nitrogen (FIG. 3; P = 0.01) in the presence of pathogenic E coli. The improved nutrient digestibility was accompanied with linearly improved pig ADG (P < 0.001; FIG. 4), reduced fecal score (P = 0.01) and diarrhea frequency (Table 6). However, bacteria proliferation, as measured by total coliform and ETEC count in ileum tissue, remained unchanged across treatments (Table 6), suggesting that the antimicrobial efficacy of egg white lysozyme was very limited. Furthermore, the linear correlation analysis suggested ATTD of dry matter, gross energy, and nitrogen all showed positive correlation with pig ADG (P < 0.06) and negative correlation with fecal score (P < 0.05), but no significant correlations with coliform count or ETEC count (Table 7). These results, together with the findings in Example 1 and 2, suggest that the beneficial effects of feeding egg white lysozyme to nursery pigs were related to improved digestibility and utilization of nutrients, particularly NDF, of the diet, and this mechanism was not related to antimicrobial efficacy.Table 6. Effects of dietary supplementation of lysozyme on signs of diarrhea and ETEC colonization in ileum tissue of nursery pigsa bMeans without a common superscript differ (P < 0.05)2Subjective fecal score was graded on 0 - 3 scale: 0 = no signs of diarrhea; 3 = severe signs of diarrhea.Table 7. Linear correlation between apparent total tract digestibility (ATTD) of dry matter (DM), gross energy (GE) and nitrogen (N) with pig ADG, fecal score and bacterial count of ileum tissueEXAMPLE 4Feeding lysozyme improved growth performance and livability in commercial nursery pigs. A meta-analysis of pigs fed from 0 -24 days post-weaning

[0045] Seven studies were conducted following the same procedure to evaluate feeding lysozyme for 21-24 days post- weaning on growth performance and livability of commercial nursery pigs. In each experiment, weanling pigs (initial BW = 12.0 ± 0.1 lb) were housed in pens with 27 pigs per pen and randomly assigned to either a control diet (Control) or a dietcontaining lysozyme with an inclusion level ranging from 0.1 to 0.4 Ib / ton of feed. This resulted in a total of 1,944 pigs in 72 pens for Control treatment and 1,917 pigs in 71 pens for lysozyme treatment over 7 studies. Feed-grade medications were included at standard use in each study.

[0046] Pharmacological levels of Zn (3,000 ppm) and Cu (150 ppm) were included in all studies except for Study #5. An example of the basal diets of early nursery phases (Phase 1 and 2) is shown in Table 8. The NDF concentration was between 4.43 to 6.43% of the complete diet. Data from the 7 experiments were compiled for meta-analysis using the MIXED procedure of SAS.

[0047] Figures 5-8 suggest that feeding weanling pigs a diet containing lysozyme from 12-24 lbs of BW at an inclusion level of 0.1-0.4 Ib / ton of feed consistently and repeatably improved pig growth performance. Specifically, ADG of pigs fed lysozyme was improved in each of the 7 studies by an average of 8.1% (P < 0.05) compared to Control in meta-analysis (FIG. 5). Pig ADFI was improved in 6 out of 7 studies by an average of 2.7% (P < 0.05) in lysozyme treatment compared to Control treatment (FIG. 6). Feed conversion was improved in each of the 7 studies by an average of 4.6% (P < 0.05) compared to Control (FIG. 7). The greater ADG led to an average of 0.8 lb heavier BW (P < 0.05) in pigs fed lysozyme compared to Control (FIG. 8).

[0048] Along with improved growth performance, livability was also improved in response to dietary supplementation of lysozyme. Specifically, total removal rate, which includes mortality and poor performing pigs, was reduced by 1.2 percentage points in pigs fed lysozyme compared to Control (FIG. 9). Data of mortality alone was recorded in Study #5, 6 and 7. Each of these three studies showed improved livability in lysozyme treatment with an average of 2.1 percentage points higher than Control (FIG. 10). The meta-analysis of the early nursery performance in 7 studies suggest that feeding lysozyme consistently and repeatably improved livability and growth performance in nursery pigs fed diets containing 4 to 6% of NDF.Table 8. Example of early nursery phase diets in studies included in meta-analysis.EXAMPLE 5Feeding lysozyme improved growth performance and livability in commercial nursery pigs. A meta-analysis of pigs fed from 0 to 51 days post- weaning

[0049] Three out of the 7 studies described in Example 4 were continued to late nursery phase for evaluation of feeding lysozyme for 49 to 51-day post- weaning with approximate BWfrom 12 of 52 lb on growth performance and livability of commercial nursery pigs. An example of the basal diets of late nursery phases (Phase 3 and 4) is shown in Table 9. The NDF concentration in late nursery diets was between 9.48 to 10.43%. Data from the 3 experiments were compiled for meta-analysis using the MIXED procedure of SAS.

[0050] Data in FIG. 11 suggest that feeding nursery pigs a diet containing lysozyme from 12 - 52 lbs of BW at an inclusion level of 0.1 - 0.4 Ib / ton of feed consistently and repeatably improved pig growth performance. Specifically, closeout nursery ADG of pigs fed lysozyme was improved in each of the 3 studies by an average of 4.9% (P < 0. 10) compared to Control in meta-analysis. Closeout nursery feed conversion was improved in each of the 3 studies by an average of 3.1% (P < 0. 10) compared to Control. The greater ADG led to an average of 1.4 lb heavier closeout nursery BW (P < 0.05) in pigs fed lysozyme compared to Control.

[0051] Along with improved growth performance, nursery mortality, total removal and percentage of medically treated pigs were also reduced in each of the studies (FIG. 12). This meta-analysis of the entire nursery performance in 3 studies suggest that feeding lysozyme consistently and repeatably improved livability and growth performance in nursery pigs from fed diets containing 4 to 10% NDF from 12 to 52 lb of BW.TABLE 9

Claims

WHAT IS CLAIMED IS:

1. A composition for improving utilization and reducing antinutritional effects of non-digestible fiber (NDF) in feed of a non-human animal, the composition comprising a basal animal diet supplemented with lysozyme.

2. The composition of claim 1, wherein the composition is a feed composition or a feed additive composition.

3. The composition of claim 1, wherein the lysozyme is not a CH type lysozyme.

4. The composition of claim 1, wherein the lysozyme is not a GH-25 lysozyme.

5. The feed composition of claim 1, wherein the lysozyme is a type C lysozyme.

6. The feed composition of claim 1, wherein the lysozyme is a GH-22 lysozyme.

7. The feed composition of claim 1, wherein the lysozyme is an egg white lysozyme.

8. The feed composition of claim 7, wherein the concentration of egg white lysozyme in the feed composition ranges from about 0. 1 Ib / ton to about 0.5 Ib / ton.

9. The feed composition of claim 7, wherein the concentration of egg white lysozyme in the feed composition ranges from about 0.001% w / w to about 0.5% w / w.

10. The feed composition of claim 8, wherein the animal is a non-ruminant animal.

11. The feed composition of claim 8, wherein the animal is a pig.

12. The feed composition of claim 8, wherein the animal is a nursery pig.

13. A method of improving utilization and reducing antinutritional effects of NDF in a non-human animal, the method comprising orally administering to the animal a composition of any one of claims 1-12.

14. The method of claim 13, wherein the method improves NDF digestibility by at least 36%.

15. The method of claim 13, wherein the method reduces undigested NDF of DDGS by at least 13%.

16. The method of claim 13, wherein the method improves utilization of NDF in the presence of pathogens or absence of pathogens.

17. The method of claim 13, wherein the method improves nutrient digestibility, growth performance of the non-human animal, survivability of the non-human animal, or any combination thereof.

18. The method of claim 13, wherein the method improves ADG, AD FI, feed conversion, increases body weight, reduces mortality, reduces total removal rate, or any combination thereof.

19. The method of claim 13, wherein the method improves apparent total tract digestibility (ATTD) of dry matter, improves ATTD of gross energy, improves ATTD of nitrogen.

20. The method of claim 13, wherein the method reduces undigestible carbohydrate fractions in a feed composition.

21. A method of improving growth performance, livability, or growth performance and livability of a non-human animal, the method comprising orally administering to the animal a composition of any one of claims 1-12.

22. The method of claim 21, wherein the method improves growth performance, livability, or growth performance and livability of an animal irrespective of the diseases state of the animal.

23. The method of claim 21, wherein the method improves nutrient digestibility, growth performance of the non-human animal, survivability of the non-human animal, or any combination thereof.

24. The method of claim 21, wherein the method improves ADG, AD FI, feed conversion, increases body weight, reduces mortality, reduces total removal rate, or any combination thereof.

25. The method of claim 21, wherein the method improves apparent total tract digestibility (ATTD) of dry matter, improves ATTD of gross energy, improves ATTD of nitrogen.

26. The method of claim 21, wherein the method improves growth performance, livability, or growth performance and livability without affecting proliferation of a pathogen in the animal.

27. The method of claim 21, wherein the method reduces undigestible carbohydrate fractions in a feed composition.

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