HK1161293A - Compositions and methods for controlling disease in animals - Google Patents

Description

Compositions and methods for controlling disease in animals

The present invention claims the benefit of U.S. provisional patent application serial No. 61/187,316, filed 6/3/2009, U.S. provisional patent application serial No. 61/156,902, filed 3/2009, and U.S. provisional patent application serial No. 61/121,258, filed 12/10/2008, under 35 USC § 119 (e).

Background

Pigs and poultry, especially those raised intensively or in large scale operations, have a tendency to suffer from or be at risk of transmitting a variety of diseases and infections, such as mycoplasmosis in pigs and poultry, Lawsonia infection (ileitis) and swine dysentery in pigs and necrotic enteritis in poultry. Drugs have been proposed or used to treat these types of individual diseases or infections.

Lawsonia intracellularis (L.intracellularis) is the causative agent of porcine proliferative enteropathy (PPE; also known as porcine ileitis) and affects virtually all animals, including: rabbits, ferrets, hamsters, foxes, horses, and various other animals such as ostriches and emus. PPE is a common diarrheal dysentery in growing-adult and young breeding pigs, characterized by hyperproliferation and inflammation of the ileum and rectum. It is usually mild and self-limiting, but sometimes causes persistent diarrhea, severe necrotic enteritis, or hemorrhagic enteritis with high mortality.

Necrotic Enteritis (NE) in poultry is caused by the gram-positive anaerobic bacterium Clostridium perfringens (Clostridium perfringens). The disease is acute enterotoxemia, and mainly affects broiler chicks of 2-5 weeks and turkeys of 7-12 weeks. The duration of the disease is short and often the only sign of the disease is a sudden increase in avian mortality.

Clostridium perfringens is an almost ubiquitous bacterium found in soil, dust, feces, feed, and used poultry litter, and is also a resident in the gut of healthy chickens. Enterotoxemia, which causes necrotic enteritis, occurs after changes in the intestinal microbiota or due to disorders that cause damage to the intestinal mucosa (e.g., coccidiosis, mycotoxin toxemia, salmonella infection, ascaris larvae). High dietary levels of animal by-products, wheat, barley, oats or rye predispose birds to the disease. Any event that promotes excessive bacterial growth and toxin production or slows the feed passage rate (feed passage) in the small intestine may contribute to the occurrence of necrotic enteritis.

The diagnosis of necrotic enteritis is based on macroscopic lesions and gram-stained smears showing mucosal scrapings of large gram-positive bacilli (gram-positive rods). Macroscopic lesions are found primarily in the small intestine, which may be ballooned, friable, and contain a brown liquid that smells malodorous. The mucous membranes are usually covered with a false membrane of tan to yellow color, often referred to as a "turkish delinted towel" appearance. Such a pseudomembrane may extend throughout the small intestine or may be located only in a localized area. The disease persists in the population for 5-10 days with a mortality rate of 2-50%. A conventional product for preventing necrotic enteritis in poultry is medicated feed, which contains virginiamycin (20 g/ton feed), bacitracin (50 g/ton) and lincomycin (2 g/ton). Medicated feed containing an anticoccidial compound belonging to the ionophore class also helps to prevent necrotic enteritis. Necrotic enteritis is usually treated by administering bacitracin, penicillin and lincomycin in drinking water for 5-7 days. NE has been identified as a disease condition that can be prevented or controlled by using Direct-Fed microbial (DFM) products, since the products act on the intestinal microbiota.

Coccidiosis causes considerable economic losses in the poultry industry. This disease is caused by several eimeria species, including eimeria tenella (e.tenella), eimeria necatrix (e.necatrix), eimeria acervulina (e.acervulina), eimeria brunetti (e.brunetti), and eimeria maxima (e.maxima). The coccidiosis stage occurs in chickens both inside and outside the host. At a developmental stage in chicks, microscopic eggs (called zygotic sacs) are produced, which are excreted in the faeces. Normally, most birds shed small numbers of zygotic sacs in their feces without adverse effects, but intensive rearing of domesticated chicks provides conditions that allow infectious zygotic sacs to accumulate in the environment, thereby increasing the likelihood of coccidiosis infection. By the third day of infection, coccidiosis infection may become significant. Symptoms include chicken weakness (chicken dropping), stopping eating, and crowding together. By day four blood began to appear in the feces, and by day eight the chicks either died or had begun to recover.

Prevention of coccidiosis involves mixing an anticoccidial drug with the feed. Ionic carriers such as salinomycin are the most commonly used in the United states for the prevention of coccidiosisA medicine is provided. Salinomycin and salts thereof are typically added to animal feed at a concentration of about 40 to about 60 grams per ton of animal feed (0.0044% to 0.0066%). Salinomycin sodium is the preferred form of salinomycin for use in the united states. Narasin is the most commonly reported ion carrier that has been promoted as an adjunct to NE prevention. In particular, coccidiosis vaccines are also available to those who wish to sell antibiotic-free chickensCoccivac-B is a non-attenuated, live sporogenous ascosis vaccine comprising Eimeria acervulina, Eimeria varia (E.mivati), Eimeria maxima and Eimeria tenella.

There is a need for compositions and methods for treating a variety of diseases in animals, particularly diseases of bacterial origin. These compositions should be capable of being used in combination with other treatments and/or compositions for the treatment of other diseases. Optimally, the compositions used to treat the disease have a synergistic effect when used in combination with other compositions or other treatment regimens.

Disclosure of Invention

The present invention relates to compositions and methods for treating diseases in animals, particularly poultry. One embodiment of the present invention relates to a composition for treating a disease in an animal comprising a yeast extract from Saccharomyces cerevisiae (Saccharomyces cerevisiae), Bacillus licheniformis (Bacillus licheniformis) spores and optionally a carrier. Embodiments of the invention also relate to methods for preventing a disease in an animal comprising administering to the animal a composition comprising a yeast extract from saccharomyces cerevisiae, bacillus licheniformis spores, and optionally a carrier.

The present invention also relates to a composition for preventing and/or controlling diseases in animals comprising yeast extract from saccharomyces cerevisiae and QST-713 strain of Bacillus subtilis and optionally a carrier. Embodiments of the present invention also relate to methods for preventing and/or controlling disease in an animal comprising administering to the animal an effective amount of a composition comprising a yeast extract from saccharomyces cerevisiae and QST-713 strain of bacillus subtilis, and optionally a carrier.

The present invention also relates to compositions comprising from about 50% to about 90% by weight of cell wall components from saccharomyces cerevisiae; about 5% to about 50% by weight of Bacillus licheniformis spores containing about 4.5 x 10⁹To about 2.5X 10¹⁰Spores per gram; and an anticoccidial ion carrier. Embodiments of the invention also relate to methods for preventing a disease in an animal comprising vaccinating the animal against coccidiosis and administering to the animal a composition comprising 50% to about 90% by weight of cell wall components from saccharomyces cerevisiae; about 5% to about 50% by weight of Bacillus licheniformis spores containing about 4.5 x 10⁹To about 2.5X 10¹⁰Spores per gram; and about 0 wt% to 45 wt% carrier. Embodiments of the present invention are further directed to methods for reducing mortality in animals challenged with clostridium perfringens comprising vaccinating the animal against coccidiosis and administering to the animal a composition comprising from 50% to about 90% by weight of a cell wall component from saccharomyces cerevisiae; about 5% to about 50% by weight of Bacillus licheniformis spores containing about 4.5 x 10⁹To about 2.5X 10¹⁰Spores per gram; and about 0 wt% to 45 wt% carrier. Embodiments of the invention further relate to methods for reducing necrotic enteritis damage in an animal challenged with clostridium perfringens comprising vaccinating the animal against coccidiosis and administering to the animal a composition comprising from about 50% to about 90% by weight of a cell wall component from saccharomyces cerevisiae; about 5% to about 50% by weight of Bacillus licheniformis spores containing about 4.5 x 10⁹To about 2.5X 10¹⁰Spores per gram; and about 0 wt% to 45 wt% carrier.

Detailed Description

Disclosed herein are unexpected synergistic combinations of probiotics such as bacillus licheniformis and immune enhancing yeast cell wall extracts containing beta-glucan and mannan in combination with anticoccidial or coccidiosis vaccines for the prevention, control and treatment of necrotic enteritis. These compositions, otherwise known as animal feed additives, suitable for treating animal diseases are particularly useful for treating diseases caused by infection with bacteria such as L.intracellularis or Clostridium perfringens.

Direct Fed Microbial (DFM) is a live microorganism that is fed to animals in order to improve yield by regulating the intestinal environment of the animal and by improving digestion. They help to maintain the proper balance of normal intestinal flora in the gastrointestinal tract. While scientists do not fully understand the mechanism of action of DFMs, it is believed that DFMs can act by attaching themselves to the intestinal surface, which reduces or prevents pests from attaching to and colonizing the intestinal surface; stimulating intestinal immunity, which in turn prevents protection from disease; release of enzymes that aid digestion; producing organic acids that alter the pH of the intestine and stimulate beneficial organisms; and the production of nutrients, such as vitamins. AlCare^TM(Alphana Inc. of Bridgewater, N.J.) is a DFM containing a modified strain of Bacillus licheniformis (NCTC 13123). AlCare^TMContaining 1X 10 of calcium carbonate in 70% by weight of a carrier¹⁰Bacillus licheniformis spores per gram. 1 pound AlCrare^TM2.2X 10/ton feed supply⁹Colony forming units (cfu) bacillus licheniformis/pound feed.

Brewer's yeast is a fungus called s.cerevisiae.(Alphana Inc. of Bridgewater, N.J.) is a cross-linked natural polysaccharide fiber produced from the cell walls of food grade brewers yeast.The component (B) is also named Beta(Bri, N.J.)Alphara Inc. of dgewater), BetaIs the cell wall component after lysis of the saccharomyces cerevisiae and extraction of bioactive material from the cells after autolysis.Is a combination of beta-glucan and mannan.The beta-glucans present in (a) have side chains (1, 3-1, 6) which give rise to specific complex structures which cannot be broken down by glucanases. Beta-glucan has a strong immunopotentiating effect in animals, and binds to activated and cytokine-secreting macrophages. Saccharomyces cerevisiae mannan is a polysaccharide-polypeptide complex containing partially phosphorylated D-mannose residues. Mannan has been shown to have an immunostimulatory effect.Typically contains greater than or equal to about 24 wt.% beta-glucan and less than or equal to about 15 wt.% mannan.

Surprisingly, the combination of QST-713 strain of Bacillus subtilis and a yeast extract such as a yeast extract from Saccharomyces cerevisiae, i.e., brewer's yeast, containing beta-glucans and mannans, and compositions thereof, marketed as biological fungicides, can be utilized to prevent and/or control diseases such as infections in animals caused by bacteria such as L.intracellularis or Clostridium perfringens.

As described herein above, the present invention also provides a composition comprising a QST-713 strain of bacillus subtilis and a yeast extract, such as a yeast extract from saccharomyces cerevisiae, and optionally a carrier, for use in the prevention and/or control of diseases in animals, such as infections, in particular necrotic enteritis in poultry and porcine ileitis in pigs.

QST-713 strain of Bacillus subtilis under the trade name of BiofungicideMAX is commercially available (AgraQuest Inc. of Davis, Calif.).MAX is a microbial pesticide based on the naturally occurring strain of Bacillus subtilis QST-713. It produces three groups of lipopeptides: iturin, agrastatins, plipastatins and surfactin act synergistically to inhibit germ tubes, mycelia and bacterial cells.MAX 10-15% by weight of Bacillus subtilis QST-715 in a carrier and provides a minimum of 7.3X 10⁹cfu Bacillus subtilis QST-715 per gram of mixture, wherein the carrier comprises a mixture of inert, inactive ingredients.MAX can be formulated as wettable powders, wettable granules and aqueous suspensions, which are applied like any other foliar fungicide.MAX is not toxic to non-targets and beneficial organisms. Due to the combined action mode, environment friendliness and broad-spectrum control of Serenade, the method is very suitable for comprehensive pest control.

As described herein above, the compositions of the present invention comprise QST-713 strain of bacillus subtilis and a yeast extract, such as a yeast extract from saccharomyces cerevisiae, and optionally a carrier. Preferably, the compositions of the present invention comprise QST-713 strain of bacillus subtilis in an amount ranging from about 5% to about 50% by weight and more preferably from about 10% to about 15% by weight, based on the total weight of the composition. Preferably, the QST-713 strain component of Bacillus subtilis of the present invention comprises about 4.5X 10⁹To about 2.5X 10¹⁰cfu Bacillus subtilis QST-713 strain/g, more preferably at least about 7.3X 10⁹cfu Bacillus subtilis QST-713 strain/g. As described herein above, the present invention also provides an animal feed composition comprising a combination of a QST-713 strain of bacillus subtilis and a yeast extract, such as a yeast extract from saccharomyces cerevisiae, for use in the prevention and/or control of diseases in animals, such as infections, in particular necrotic enteritis in poultry and porcine ileitis in pigs.

One embodiment of the invention includes a composition for preventing disease in an animal comprising a yeast extract from a yeast, such as saccharomyces cerevisiae, a probiotic, and an anti-coccidial ionophore. Coccidiostats include, but are not limited to, saricin, monensin, nigericin, letromycin, renieramycin, griseofulvin, milamycin, calicheamicin, renomycin, rasalomycins ammonium, domimicin and linocillin (lysecelin). Preferred embodiments of the invention include salinomycin (Bio-Alpharma Inc of Bridgewater, new jersey). The anticoccidial ion carrier is present in an amount in the range of about 0.0033 to about 0.0099 weight percent, preferably about 0.0044 to about 0.0066 weight percent.

Embodiments of the invention relate to methods for preventing disease in an animal comprising vaccinating the animal against coccidiosis and administering to the animal a composition comprising yeast, such as saccharomyces cerevisiae, a probiotic, and a carrier. The preferred coccidiosis vaccine is-B (Schering Plough animal health corporation), a non-attenuated, live spore compound ascopathy vaccine containing Eimeria acervuline, Eimeria varia, Eimeria maxima and Eimeria tenella.

The composition optionally comprises a carrier. Many suitable dry carriers can be organic or inorganic. Exemplary inorganic carriers include salts such as calcium carbonate, calcium sulfate, and the like. Suitable organic carriers include lactose and the like. Other flow control agents such as silica may also be used in small amounts. Mixtures comprising one or more of the foregoing may also be used.

The compositions comprise a yeast extract, such as a yeast extract from saccharomyces cerevisiae. In particular, the cell wall component comprises β -glucan and mannan. For example by mechanically disrupting the cells to produce cell wall components. The yeast extract is present in an amount of about 50 wt.% to about 90 wt.%, specifically about 50 wt.% to about 85 wt.%, and more specifically about 50 wt.% to about 55 wt.%, based on the total weight of the composition. In one embodiment, the β -glucan has a (1, 3-1, 6) side chain and is resistant to cleavage by glucanase (cleavage). In another embodiment, the mannan is a polysaccharide-polypeptide complex, partially phosphorylating D-mannose residues. The yeast extract comprises about 40% to about 85% by weight cell walls, more specifically about 40% to about 45% by weight cell walls.

The bacillus licheniformis is present in an amount of about 5 wt.% to about 50 wt.% and more specifically about 7.5 wt.% to about 12.5 wt.%, based on the total weight of the composition. The Bacillus licheniformis component comprises about 4.5 x 10 based on the total weight of the composition⁹To about 2.5X 10¹⁰cfu/g, in particular about 5X 10⁹To about 1.25X 10¹⁰cfu/g.

The carrier is present in an amount of from 0 wt% to 45 wt%, more specifically from about 5 wt% to about 40 wt%, and more specifically from about 37.5 wt% to about 42.5 wt%, based on the total weight of the composition.

In one embodiment, a composition suitable for treating a disease in an animal is administered in a diet of the animal. The manufactured foodstuffs for animals such as cattle, pigs and poultry are usually provided in pellets or similar granular material. Pellets are typically made as follows: mixing a cereal base with ingredients such as oil and protein, steam conditioning the mixture (e.g. for 5 minutes at 70 ℃), extruding the mixture through an annular die (typically between 2mm and 15mm in diameter), cutting to suitable size lengths (e.g. 5-20mm), and drying. The finished pellets are generally cylindrical in shape and have a relatively smooth surface.

In one embodiment, the animal feed composition is prepared by adding an animal feed additive to an animal foodstuff. The animal feed additive composition can be added to food in a variety of ways. An animal feed additive composition containing a specific amount of an active ingredient may be added to a specific amount of feed and mixed or blended to provide a substantially homogeneous drug-containing feed composition. A large batch of feed can be prepared in this way for treating a large number of animals. Alternatively, a feed batch containing feed for a single animal or single meal can be prepared by mixing a predetermined amount of the animal feed additive composition with the animal feed or by adding a predetermined amount of the premix as a topcoating adjuvant (top dressing) to the animal feed.

In another embodiment, the animal feed composition comprises a composition suitable for treating a disease in an animal and an animal foodstuff, wherein the composition suitable for treating a disease in an animal is present in an amount of about 0.5 pounds to about 10 pounds, specifically about 1 pound to about 5 pounds, more specifically about 1.5 pounds to about 2.5 pounds, and most specifically about 2 pounds of the composition suitable for treating a disease in an animal to one ton of the animal foodstuff (in the form of pellets or prills). In one embodiment, the animal feed may be a poultry feed. In another embodiment, the animal feed may be a swine feed.

The invention is further illustrated by the following non-limiting examples.

Examples

Example 1.

One-day-old healthy male broiler chickens (Cobb X Cobb) were obtained from Cobb-Vantress Hatchery, Cleveland, Georgia. After arrival, the chickens are raised in a layered chicken coopThese chickens were used. Eight chickens were placed in each cage. A thermostatically controlled gas furnace/air conditioner is provided to maintain constant temperature and uniform continuous illumination. Water was available ad libitum throughout the experiment. The chicks were divided into groups of 48 chicks. Two control treatment groups were fed a non-medicated commercial chicken feed ration (starter ration) mixed with a usual feed in the united states. One Group (named AlCrare 0.5 Group) is fed with AlCrare^TM(Alpharma, Bridgewater, N.J.) which is a peptide containing 1.0X 10¹⁰Bacillus licheniformis spores/gram DFM product at a ratio of 0.5 lb/ton. Feeding AlCrare 1 Group with another Group^TMThe ratio was 1.0 lb/ton. Feeding two separate groups(Alpharma, Bridgewater, N.J.) which is a brewers yeast (Saccharomyces cerevisiae) extract used in animal feed at a ratio of 1 pound/ton (Alphamune 1 Group) and 2 pounds/ton (Alphamune 2 Group), respectively. Additional groups were fed Alcate and Alphamune at a ratio of Alcate 0.5 lb/ton + Alphamune 1 lb/ton, Alcate 1 lb/ton + Alphamune 1 lb/ton, Alcate 0.5 lb/ton + Alphamune 2 lb/ton, and Alcate 1 lb/ton + Alphamune 2 lb/ton.

The chicks were fed for 28 days. All dead chicks that died during the study were weighed and subjected to necropsy to confirm the presence or absence of Necrotic Enteritis (NE) lesions. On day 14, all groups were challenged with oral E.maxima (5,000 zygotic capsules/chicken). Clostridium perfringens (Clostridium perfringens broth) culture 10 on days 19, 20 and 21⁸cfu/ml) challenge all treatments except control (NC). On day 22, three chicks from each coop were scored for NE damage. The test was terminated on day 28. The test results are shown in table 1.

Table 1.

Treatment of	Necrotic enteritis lesion score	Mortality rate of necrotic enteritis%
			Control	0	0
Attack collation	1.11	20.83
			AlCare 0.5	0.56	6.25
AlCare 1	0.56	6.25
			Alphamune 1	0.72	10.42
Alphamune 2	0.94	12.50
			AlCare 0.5+Alphamune 1	0.44	12.50
AlCare 1+Alphamune 1	0.28	2.08
			AlCare 0.5+Alphamune 2	0.72	10.42
AlCare 1+Alphamune 2	0.56	6.25

Example 2.

One-day-old healthy male broiler chickens (Cobb X Cobb) were obtained from Cobb-Vantress Hatchery, Cleveland, Georgia. Upon arrival, 50 chickens were allocated per treatment pen by zone. For environmental control, ambient humidity and 24 hour lighting were maintained. During the raising period, the gas furnace is the main heat source, and each enclosure is supplemented with heat by heating lamps as required. Ventilation and chicken cooling was achieved using fans and side curtain treatment. Water was available ad libitum throughout the experiment. Two control treatment groups were fed non-medicated commercial chicken diets, medium and large chicken diets (starter and finishing rates) mixed with a commonly used food in the united states. One Group is named as SM Group and is fedMAX, ratio 91 g/ton of feed. The other group is named as SM +(AM) Group with a feed ratio of 91 g/tonMAX and ratio of 1 lb/ton of feedThe process was repeated and randomized in 6 blocks.

The chicks were fed ad libitum from the day they arrived until day 42 of the study. All dead chicks that died during the study were weighed and subjected to necropsy to confirm the presence or absence of Necrotic Enteritis (NE) lesions. On day 14, all groups were challenged with oral E.maxima (5,000 zygotic capsules/chicken). Culture with Clostridium perfringens (CP, Clostridium perfringens broth 10 on days 19, 20, and 21⁸cfu/ml) challenge all treatments except one control group (NC). On day 22, five chickens from each pen were scored for NE damage. The test was terminated on day 42 and the results are shown in table 2.

Table 2.

Example 3.

One day old healthy male broiler chickens (Cobb X Cobb) were obtained from the hatchery, where they were sex identified and received routine inoculation (HVTSB 1). Only healthy-looking chicks were used in the study. None of the chickens were replaced during the study. Upon arrival, the chicks were housed in litter pens (litter pens). 50 chickens were housed per pen. The pens are partitioned by location within the house. Each treatment occurs only once within each zone. The thermostatically controlled gas furnace is kept at a constant temperature. Water was available ad libitum throughout the experiment.

On days 0, 21 and 42, all chickens were weighed into pens. Chick feed, middle chick feed, and large chick feed consumption were measured from day 0 to day 21, from day 21-35, and from day 35-42, respectively. The study was terminated on day 42. On day 22, five chickens from each pen were scored for NE damage. The scores were based on a score of 0 to 3, with 0 indicating normal and 3 indicating the most severe. The scores were as follows: 0 indicates normal bowel, 1 indicates mild mucus covering and loss of health, 2 indicates severe necrotic-causing enteritis, 3 indicates extreme necrotic-causing enteritis, and blood is present in the lumen. Chickens that died during the study were weighed and subjected to necropsy to confirm the presence of NE. All chickens from the study were processed at the study facility. Mortality is summarized as total mortality and mortality due to NE.

A total of eight different protocols were tested. On day 14, all groups were challenged with oral E.maxima (5,000 zygotic capsules/chicken). Clostridium perfringens (Clostridium perfringens broth) culture 10 on days 19, 20 and 21⁸cfu/ml) challenge all treatments except control (NC) and AlCare and Alphamune treated groups (AA-NC groups). Control feed does not contain AlCare^TM(AC)、(AM) or salinomycin (S). At incubation time, the control group does not contain the recommended dose of the marker(Schering Plough Animal health corporation) machine for spray inoculation. Each panel was named as follows:

NC group (non-challenge control): NC groups were fed a pharmaceutical commercial-type free chick ration mixed with a commonly used food in the united states and which was not challenged with clostridium perfringens.

CP group (challenge control): the CP groups were fed a drugless commercial type chick ration mixed with a commonly used food in the united states and challenged with clostridium perfringens as described above.

AA. NC group (feeding AlCrare)^TMAndnon-challenged): the AA, NC groups were fed a commercial chick diet mixed with a commonly used food in the United states plus AlCare in a ratio of 1.0 lb/ton^TMAnd a ratio of 1 lb/tonThe group is not attacked.

AA. CP group (fed AlCrare)^TMAndchallenged) commercial broiler diet mixed with american diet + AlCare ration in a ratio of 1.0 lb/ton^TMAnd a ratio of 1 lb/tonThe group is attacked as described above.

S, CP groups (salinomycin treated, challenged) were fed commercial type chick diets mixed with a usual feed in the united states plus salinomycin (66ppm) and challenged as described above.

Groups S, AA, CP (Serlimycin-treated, AlCrare-fed)^TMAndchallenged) commercial broiler diet mixed with american diet + AlCare ration in a ratio of 1.0 lb/ton^TMAnd a ratio of 1 lb/ton+ salinomycin (66 ppm). The group is attacked as described above.

On the day of incubation, with the recommended dose of the markerMachine (Schering-Ploughanimal Health corporation)Station for CV and CP groups: (-B treated; challenged) were inoculated by jet and fed a commercial type chick ration mixed with a commonly used food in the united states. The group is attacked as described above.

On the day of incubation, with the recommended dose of the markerMachine pair CV, AA, CP group (-B treated; challenged) were inoculated by jet and fed a commercial chick ration mixed with a usual food material in the united states plus AlCare in a ratio of 1.0 lb/ton^TMAnd a ratio of 1 lb/tonThe group is attacked as described above.

In analyzing data, use is made of(SAS Institute, Inc., Cary, NC, USA, 2002). More analyses were performed comparing AA to the anticoccidial regimens with and without AA.

The calculation of fence data used in statistical analysis was performed using SAS. Average chicken weight gain over days 0-21, 0-42, and 21-42 was calculated. With respect to the NE damage score, the mean number of pens was calculated for five chickens selected for NE damage scoring on day 22. Feed Conversion Rates (FCR) were calculated on a pen basis over days 0-21, 0-42 and 21-42 and adjusted for dead and removed chickens. The total percent mortality and those caused by NE were calculated. Any reference to chicken performance refers to the effect on weight gain and/or FCR.

The study was conducted using a randomized complete block design. There were 6 blocks. The chicken pens were the experimental units. The model includes blocks and treatments. Blocks are random effects. The treatment is a fixed effect and thus the model is mixed.

Summary variables (summary variables) include performance data and data related to NE attacks for each fence. MIXED model analysis was performed on the data using the PROC MIXED program of SAS.

The test results are shown in tables 3 to 8. The following abbreviations are used in the tables:

AA＝AlCare+Alphamune

s-sarithromycin

CP is attacked

NC-not attacked

TABLE 3 Effect of dietary Alcate + Alphamune on weight gain in C.perfringens challenge model in litter ground pens

TABLE 4 Effect of dietary Alcate + Alphamune on feed conversion and necrotic enteritis lesions in broiler chicks in Clostridium perfringens challenge model in litter ground pens

TABLE 5 Effect of dietary Alcate + Alphamune on mortality of broiler chicks in Clostridium perfringens challenge model in litter ground pens

TABLE 6 Effect of dietary Alcate + Alphamune on weight gain in C.perfringens challenge model in litter ground pens

TABLE 7 Effect of dietary Alcate + Alphamune on feed conversion and necrotic enteritis lesions in broiler chicks in Clostridium perfringens challenge model in litter ground pens

TABLE 8 Effect of dietary Alcate + Alphamune on mortality of broiler chicks in Clostridium perfringens challenge model in litter ground pens

The data in tables 3, 4 and 5 are a comparison between the AACP treatment and the S and CV treatments with and without AA, respectively. The S + AA combination generally provides the best performance of all treatments. The S + AA combination provides greater weight gain from 0 to 21 days, 0 to 42 days, and 21 to 42 days, and better FCR from 0 to 21 days, 0 to 42 days, and 21 to 42 days, lower NE mortality compared to S and AACP treatment. The S + AA combination also provided a lower NE damage score compared to AACP treatment.

The CV + AA combination provided greater weight gain from 0-21 days, 0-42 days, 21-42 days, and better FCR from 0-21 days, 0-42 days, 21-42 days, compared to CV and AACP treatment (tables 3 and 4). The CV + AA combination provided lower total mortality from day 0 to 42 and lower NE mortality compared to CV and AACP treatment (table 5).

Chickens treated with AA-containing and AA-free salinomycin drugs generally have optimal performance comparable or superior to NC and lower NE damage scores. The addition of AA to the feed of salomycin-fed chickens tended to improve performance but did not further reduce NE damage scores or mortality. In general, the addition of AA to chicken feed treated with CV improved performance and reduced mortality compared to those treated with CV alone. Chickens challenged with NE and treated with a conventional anticoccidial regimen or coccidiosis vaccine regimen may benefit from the addition of AA to their feed. The data in tables 6, 7 and 8 allow direct comparison of coccidiosis control regimens with each other (with and without AA) and with controls (challenged versus non-challenged). During days 0 to 21, the chickens fed S + AA had a greater weight gain compared to other treatments (table 6). During days 0 to 21, chickens fed S had a weight gain comparable to NC but greater than CP. During days 0 to 21, CV-inoculated chickens had different weight gain than NC or CP chickens. Chickens treated with CV had greater weight gain than CP chickens over the period of 0 to 42 days and 21 to 42 days.

During days 0 to 21, chickens fed S + AA had better FCR than NC and CP chickens (table 7). During days 0 to 21, chickens treated with S, CV and CV + AA drugs had a better FCR than CP chickens. Chickens treated with S, S + AA, CV and CV + AA had better FCR than NC and CP chickens during days 0 to 42 and 21 to 42 (table 7). Addition of AA to the diets of chickens treated with S and CV improved FCR compared to S and CV alone, respectively, during days 0 through 42. Addition of AA to the chicken diet inoculated with CV improved FCR compared to CV alone during days 21 to 42.

NE lesion scores of S and CV chickens (with and without AA) were comparable to NC but lower than CP chickens (table 8). Treatment did not affect mortality during the period from day 0 to day 21 (table 8). S, S + AA and CV + AA chickens' mortality (total mortality versus NE mortality) was comparable to NC but lower than CP chickens (Table 6). Mortality (total mortality and NE mortality) was lower in CV chickens than in CP chickens but higher in NC chickens. Mortality (total mortality and NE mortality) in chickens treated with CV + AA mortality in chickens treated with CV alone.

Example 4.

One day old healthy male broiler chickens (Cobb X Cobb) were obtained from the hatchery, where they were sex identified and received routine inoculation (HVTSB 1). Only healthy-looking chicks were used in the study. None of the chickens were replaced during the study. After arrival, the chickens were raised in litter ground pens. 50 chickens were housed per pen. The pens are partitioned by location within the house. Each treatment occurs only once within each zone. The thermostatically controlled gas furnace is kept at a constant temperature. Water was available ad libitum throughout the experiment.

On days 0, 21 and 42, all chickens were weighed into pens. Chick feed, middle chick feed, and large chick feed consumption were measured from day 0 to day 21, from day 21-35, and from day 35-42, respectively. The study was terminated on day 42. On day 22, five chickens from each pen were scored for NE damage. The scores were based on a score of 0 to 3, with 0 indicating normal and 3 indicating the most severe. The scores were as follows: 0 indicates normal bowel, 1 indicates mild mucus covering and loss of health, 2 indicates severe necrotic-causing enteritis, 3 indicates extreme necrotic-causing enteritis, and blood is present in the lumen. Chickens that died during the study were weighed and subjected to necropsy to confirm the presence of NE. All chickens from the study were processed at the study facility. Mortality is summarized as total mortality and mortality due to NE.

A total of nine different protocols were tested. On day 14, all groups were challenged with oral E.maxima (5,000 zygotic capsules/chicken). Clostridium perfringens (Clostridium perfringens broth) culture 10 on days 19, 20 and 21⁸cfu/ml) challenge all treatments except the non-challenge control group (NC). Control feed contained noMAX(QST)、(AM) or salinomycin (S). At incubation time, the control group does not contain the recommended dose of the markerSpraying inoculation is carried out by the machine. Each panel was named as follows:

NC group (non-challenge control): NC groups were fed a pharmaceutical commercial-type free chick ration mixed with a commonly used food in the united states and which was not challenged with clostridium perfringens.

CP group (challenge control): the CP groups were fed a drugless commercial type chick ration mixed with a commonly used food in the united states and challenged with clostridium perfringens as described above.

AM, CP group (feeding)Attacked): the AM and CP groups were fed a commercial chick diet mixed with the usual food from the United states + 1 lb/ton rationThe group is attacked.

QST, CP group (feeding)MAX; attacked) is fed with a mixture of american dietsCommercial type chick diet ration + ratio of 0.25 lb/tonMAX. The group is attacked as described above.

AQST, CP group (feeding)AndMAX, challenged) was fed a commercial type chick ration mixed with american diet plus a ration of 1 lb/tonAnd a ratio of 0.25 lb/tonMAX, and is attacked as described above.

S, CP groups (salinomycin treated; challenged) were fed commercial type chick diets mixed with american chow diet plus salinomycin (66 ppm). The group is attacked as described above.

S, AQST, CP groups (salinomycin-treated, fed)AndMAX; challenged) commercial broiler diet mixed with american diet + 1.25 lb/ton rationAndMAX and treated with salinomycin (66 ppm). As described above to theThe group attacks.

On the day of incubation, with the recommended dose of the markerMachine pair CV, CP group: (-B treated; challenged) were inoculated by jet and fed a commercial type chick ration mixed with a commonly used food in the united states. The group is attacked as described above.

On the day of incubation, with the recommended dose of the markerMachine pair CV, AQST, CP group (-B treated, feedingAndMAX; challenged) were inoculated by jet and fed a commercial type chick ration mixed with a commonly used food in the united states + a ration of 1.25 lbs/tonAndMAX. The group is attacked as described above.

In analyzing data, use is made of(SAS Institute, Inc., Cary, NC, USA, 2002). More analyses were performed comparing AA to the anticoccidial regimens with and without AA.

The calculation of fence data used in statistical analysis was performed using SAS. Average chicken weight gain over days 0-21, 0-42, and 21-42 was calculated. With respect to the NE damage score, the mean number of pens was calculated for five chickens selected for NE damage scoring on day 22. Feed Conversion Rates (FCR) were calculated on a pen basis over days 0-21, 0-42 and 21-42 and adjusted for dead and removed chickens. The total percent mortality and those caused by NE were calculated. Any reference to chicken performance refers to the effect on weight gain and/or FCR.

The study was conducted using a randomized complete block design. There were 6 blocks. The chicken pens were the experimental units. The model includes blocks and treatments. Blocks are random effects. The treatment is a fixed effect and thus the model is mixed.

The summary variables include performance data and data related to NE attacks for each fence. MIXED model analysis was performed on the data using the PROC MIXED program of SAS.

The test results are shown in tables 9 to 17. The following abbreviations are used in the tables:

s-sarithromycin

CP is attacked

NC-not attacked

TABLE 9 Effect of dietary Alphamune + QST on weight gain in C.perfringens challenge model in litter ground pens

TABLE 10 Effect of dietary Alphamune + QST on feed conversion and necrotic enteritis lesions in broiler chicks in Clostridium perfringens challenge model in litter ground pens

TABLE 11 Effect of dietary Alphamune + QST on mortality of broiler chicks in Clostridium perfringens challenge model in litter ground pens

TABLE 12 Effect of dietary Alphamune + QST on weight gain in C.perfringens challenge model in litter ground pens

TABLE 13 Effect of dietary Alphamune + QST on feed conversion and necrotic enteritis lesions in broiler chicks in Clostridium perfringens challenge model in litter ground pens

TABLE 14 Effect of dietary Alphamune + QST on mortality of broiler chicks in Clostridium perfringens challenge model in litter ground pens

TABLE 15 Effect of dietary Alphamune + QST on weight gain in C.perfringens challenge model in litter ground pens

TABLE 16 Effect of dietary Alphamune + QST on feed conversion and necrotic enteritis lesions in broiler chicks in Clostridium perfringens challenge model in litter ground pens

TABLE 17 Effect of dietary Alphamune + QST on mortality of broiler chicks in Clostridium perfringens challenge model in litter ground pens

The addition of AM, QST and AQST to broiler feed improves performance compared to CC. Chickens treated with AQST-containing as well as AQST-free salinomycin drugs generally have optimal performance comparable or superior to NC with low NE damage and mortality. In general, the addition of AQST to the chicks feed treated with Coccivac improved performance and reduced mortality compared to those treated with Coccivac alone.

Claims (48)

1. A composition for treating a disease in an animal comprising:

yeast extract of saccharomyces cerevisiae, bacillus licheniformis spores and optionally a carrier.

2. The composition of claim 1, comprising:

(a) from about 50% to about 90% by weight of cell wall components from Saccharomyces cerevisiae,

(b) about 5% to about 50% by weight of Bacillus licheniformis spores comprising about 4.5×10⁹To about 2.5X 10¹⁰Spores per gram, and

(c) about 0 wt% to about 45 wt% carrier.

3. The composition of claim 1, comprising:

(a) 50% by weight of cell wall components from Saccharomyces cerevisiae,

(b)10 wt.% of Bacillus licheniformis spores comprising about 5 x 10⁹To about 1.25X 10¹⁰Spores per gram, and

(c) 40% by weight of carrier.

4. The composition of claim 1, wherein the carrier comprises calcium carbonate, calcium sulfate, lactose, or a combination thereof.

5. An animal feed composition comprising the composition for treating a disease in an animal of any one of claims 1-4 and an animal feed, wherein the amount of the composition for treating a disease in an animal is from about 0.5 pounds to about 10 pounds per ton of animal feed.

6. The animal feed composition of claim 5, wherein the amount of the composition for treating a disease in an animal is from about 1 pound to about 5 pounds per ton of animal feed.

7. The animal feed composition of claim 6, wherein the amount of the composition for treating a disease in an animal is about 2 pounds per ton of animal feed.

8. The animal feed composition of claim 7, wherein the animal feed is a poultry feed or a swine feed.

9. A method for preventing a disease in an animal comprising administering to the animal the composition for treating a disease in an animal of any one of claims 1-8.

10. The method of claim 9, wherein the animal is poultry and the disease is necrotic enteritis.

11. The method of claim 9, wherein the animal is a pig and the disease is porcine ileitis.

12. A composition for preventing and/or controlling disease in an animal comprising yeast extract from saccharomyces cerevisiae and QST-713 strain of bacillus subtilis and optionally a carrier.

13. The composition of claim 12, comprising from about 75% to about 99% by weight of said yeast extract from saccharomyces cerevisiae; from about 1% to about 3% by weight of the QST-713 strain of bacillus subtilis; and about 0 wt% to about 24 wt% of the carrier.

14. The composition of claim 12, comprising about 80% by weight cell wall components from saccharomyces cerevisiae; 2.5% by weight of QST-713 strain of Bacillus subtilis; and 17.5 wt% of the carrier.

15. The composition of claim 12, wherein the carrier comprises calcium carbonate, calcium sulfate, lactose, or a combination thereof.

16. An animal feed composition comprising the composition of any one of claims 12 to 15 and an animal diet, wherein the composition for preventing and/or controlling animal disease is present in an amount in the range of from about 0.5 pounds to about 10 pounds per ton of animal diet.

17. The animal feed composition of claim 16, wherein the composition for preventing and/or controlling animal disease is present in an amount of about 1.25 pounds per ton of animal feed.

18. The animal feed composition of claim 16, wherein the animal feed is a poultry feed or a swine feed.

19. A method for preventing and/or controlling disease in an animal comprising administering to the animal an effective amount of a composition according to any one of claims 12 to 18.

20. The method of claim 19, wherein the animal is poultry and the disease is necrotic enteritis.

21. The method of claim 19, wherein the animal is a pig and the disease is porcine ileitis.

22. A composition comprising the composition of any one of claims 1 to 8, further comprising an anticoccidial ion carrier.

23. The composition of claim 22, wherein the anticoccidial ion carrier is salinomycin.

24. The composition of claim 23, wherein said salinomycin is present at a concentration of about 66ppm of said composition.

25. An animal feed composition comprising the composition for treating necrotic enteritis in an animal and an animal foodstuff of any of claims 22 to 24, wherein the amount of the composition for treating a disease in an animal is from about 0.5 pounds to about 10 pounds per ton of animal foodstuff.

26. A method for improving the performance of an ionic carrier in treating necrotic enteritis in an animal, comprising administering to the animal the composition of any one of claims 22 to 25.

27. The method of claim 26, wherein the ion carrier is salinomycin.

28. A method for preventing disease in an animal comprising:

inoculating said animal with a coccidiosis vaccine; and

administering to the animal a composition comprising the composition of any one of claims 1 to 8.

29. The method of claim 28, wherein the animal is poultry and the disease is necrotic enteritis.

30. The method of claim 28, wherein the vaccine comprises a non-attenuated, live sporozoite coccidiosis vaccine.

31. The method of claim 28, wherein the coccidiosis vaccine is

32. A method of reducing mortality in an animal challenged with clostridium perfringens comprising vaccinating the animal with a coccidiosis vaccine; and administering to the animal a composition comprising the composition of any one of claims 1 to 8.

33. The method of claim 32, wherein the animal is poultry.

34. The method of claim 33, wherein the vaccine comprises a non-attenuated, live sporozoite coccidiosis vaccine.

35. The method of claim 34, wherein the step of converting the signal into a signal comprises converting the signal into a signalThe coccidiosis vaccine is

36. A method of reducing necrotic enteritis damage in an animal challenged with clostridium perfringens comprising vaccinating the animal with a coccidiosis vaccine; and administering to the animal a composition comprising the composition of any one of claims 1 to 8.

37. The method of claim 36, wherein the animal is poultry.

38. The method of claim 36, wherein the vaccine comprises a non-attenuated, live sporozoite coccidiosis vaccine.

39. A composition comprising the composition of any one of claims 12 to 18, further comprising an anticoccidial ion carrier.

40. The composition of claim 39, wherein the anticoccidial ion carrier is salinomycin.

41. The composition of claim 40, wherein said salinomycin is present at a concentration of about 66ppm of said composition.

42. An animal feed composition comprising the composition for treating necrotic enteritis in an animal and an animal foodstuff of any of claims 39 to 41, wherein the amount of the composition for treating a disease in an animal is from about 0.5 pounds to about 10 pounds per ton of animal foodstuff.

43. A method for improving the performance of an ionic carrier in treating necrotic enteritis in an animal, comprising administering to the animal the composition of any one of claims 39 to 41.

44. The method of claim 43, wherein the ionophore is salinomycin.

45. A method of preventing disease in an animal comprising:

vaccinating the animal with a coccidiosis vaccine; and

administering to the animal a composition comprising the composition of any one of claims 12 to 18.

46. The method of claim 45 wherein the animal is poultry and the disease is necrotic enteritis.

47. The method of claim 45, wherein the vaccine comprises a non-attenuated, live sporozoite coccidiosis vaccine.

48. The method of claim 45, wherein the coccidiosis vaccine is