MXPA99003314A - Livestock mucosal competitive exclusion culture to reduce enteropathogenic bacteria - Google Patents

Abstract

A preparation from the scrapings of healthy animals is cultured and administered to animals. This preparation confers a strong measure of protection against the subsequent colonization by enteropathogenic bacteria, including Salmonella species, Campylobacter species and Escherichia coli 0157:H7, which currently cause an unacceptably high incidence of morbidity and mortality in humans and reduce productivity in livestock populations.

Description

CULTURE OF COMPETITIVE EXCLUSION IN LUCKY MUCOSA TO REDUCE ENTEROPATHOGENIC BACTERIA BACKGROUND OF THE INVENTION Field of the Invention This invention relates to the bacteriological culture prepared from the intestinal system of mammalian animals free from pathogenesis. It also refers to subcultures of such crops and to methods for using subcultures in order to protect livestock from the colonization of enteropathogenic bacteria.

Oev í j'fs? Rt tir * 8 - f nol gía R fK tenada The animals, which cause an unacceptably high incidence of oibidity and mortality in humans, can reduce productivity in the livestock population. Gastrointestinal pathogens in humans are typically associated with the intestinal contagion of animals. meats that humans consume. In a symposium on "Locating pathogens in farm-to-table foods" The information needs to be evaluated by the control groups (Tr cking Foodborne Pathogens from Farm to Table: Data Needs to Evaluate Control Groups, Washington, DC, 3-29, 1995 ), held in January 1995, a question was asked: "How important are diseases derived from food?". It was stated that in the United States an estimated 6.5 million to 33 million cases of foodborne illnesses per year have been reported, resulting in a total of up to 9,000 deaths. The Economic Research Service of the United States Department of Agriculture (USDA Economic Research Service) estimates that medical costs in the United States and losses in productivity due to seven foodborne pathogens are from 5 6 billion dollars to 9.4 billion dollars per year. Menning estimates that there are more than 5 million cases of foodborne illnesses derived from red meat and chicken meat in the United States per year; and a large percentage of this is attributed to infections by Salmonella and Ca pylobacter (J Am. Vet Med Assoc, Yol 1 ° 2, 4 ^ - 4f> 7. Q88) Roberts estimates that each case of salt onelosis Does it cost 700 dollars? Americans Lima. J. Agt. Ion, Vol. 71, 468-474, 1989) Based on studies in which the amount of food deposited in other countries is not, it would not be possible for the number of diaircas per year caused by elimenios throughout the world ntribuiblc to the S < tftnnnefla probably exceeds 100 billion with an estimated cost exceeding 25 billion dollars. These pathogens also cause pain, suffering and loss of life. In addition to this, the enteropathogenic bacteria can also tire substantial economic losses through cattle infection. Comparative elimination (CE) techniques are used to decrease the colonization of enteropathogenic bacteria in The chicken. Nurmi et al (Nature [Nature], Vol. 241, 210-211, 1973) found that preparations of healthy, mature chickens gave protection to younger chickens, whose microflora had not yet been established, against colonization of Salmonella . The administration of undefined CE preparations to young chickens accelerates the maturation of intestinal floia in newly incubated or hatched chickens and provides a substrate for the natural process of transmission of microflora by the adult chicken to its young Snoeyenbos et al (U.S. Patent No. 4,335, 107, June 1982) developed a CE microflora technique to prevent colonization of Salmonella by lyophilizing fecal waste and cultivating this preparation anaerobically. Mikola et al (U.S. Patent No. 4,657,762, April 1987) used fecal content and intestine caeca! as a source of EC microlensia to prevent the colonization of the salt? neihíl treatment with this ü "of cultivation that the media would be anaerobic and balanced in PH Stem al (U.S. Patent No. 5,451,400 of September 19, 1995) discloses an EC mucosal compound for protection of chickens against colonization of Salmonella and Campylobacter in which the mucin layer of the prewashed mint is Scraping and scraping, preserved in an oxygen-free site, is anaerobically cultivated.

Nisbct et al (US Pat. No. 5,478,557, Decree 26 of 19%) discloses a probiotic that can be obtained from a variety of domestic animals, including but not limited to poultry and also equine, porcine and bovine animals . Nisbet et al reveal that a stable defined as probiotic, preferably obtained by continuous cultivation of a batch culture produced directly from fecal, caecal and / or large intestinal contents of the adult animal under study. They later reveal that large quantities of the probiotic can be produced either by batch or continuous culture in which the batch culture is continued until the concentration of acetic acid is greater than or equal to about 20 mM, the concentration of the proprionic acid is greater than or equal to about 10 mM and the concentration of butyric acid plus isobutyric acid is greater than or equal to 15 mM. ? splund et a! . { .Journal of Applied Vol. 81, 217-223, 1996) report an intracellular model of the intestine potcipo and its use to show the inhibition of Yersinia enterovoliiica 0: 3 by ileal porcine microfiora and raecaf. Parts of caeca and disdl of the small intestine are collected, grouped and cultured. The caecium and ileal inocula are shown to suppress the growth of cultured Y. enterolltica, with caecal flora, in some way more effective than the ileal flora. No efficacy was reported in vivo.

The present invention provides, for the first time, a compound and method for reducing in vivo and / or preventing the colonization of enteropathogenic bacteria in mammals, especially in cattle.

OBJECTIVE OF THE INVENTION It is, therefore, the object of the present invention to provide a culture derived from animal mucosa for the control of enteropathogenic bacterial colonization in animals.

Another objective of the present invention is to shield a method for treating animals in order to control the colonization of enteropathogenic bacteria in animals by the use of a piece derived from a mucosal culture.

More objectives and advantages will appear from the following description DETAILED DESCRIPTION OF THE INVENTION The importance of enteric infections in humans has been increasingly recognized in the last twelve years. The relationship between livestock contamination and human infection has also become well documented. During the production and processing of animals, such as cattle, fecal material containing pathogens can be transferred into the meat and persist in chickens from food processing. Pork, along with chicken, cattle and shellfish are important carriers of Salmonella (Bean et al, J. Food Protect, Vol 50, 804-817, 1990, Lammerding et al, J. Food Protect. Protection of Foods], Vol. 51, 47-52, 1988). Innocent animals are infected with contaminated food, chronic carriers that have been introduced into the population, infected rodents, or personnel from contaminated farms (Heard, Vet., Rec, Vol 85, 482-484, 1969); Williams et al, J. Hyg. Camb., Vol. 66, 281-293, 1968; Wilcock et al, Piscases in Swine, [Fauna in the Pig] Leman et al, eds., H a State tj tversity Feet. (Press of the State of the Aj A is? A, 570-583, 1992, Duhñtne! Et al, Proc. Í2, Int. Syn. New and Kmerging Infecí Dis. [International Symposium on New and Emerging Infections] , San Diego, C ?, 381, 1992). For the pig, in the iladeio, laurel or slaughterhouse, the initial source of contamination is the pig carried, and it is thought that the tianstpitión occurs in pig-to-pig contact or exposure to the contaminated physical environment (Newell et al, J. A. Vet. Kíed, Assoc, Vol. 158, 89-98, 1971). These infected animals, in turn, contaminate facilities, equipment and personnel, which leads to contamination of the final product (Williams et al, Am. J. Pub. Health, Vol. 60, 926-929, 1970; Newel et al, supra, Morgan et al, Epidemiol, Infecí., Vol. 98, 323-330, 1987). The initial source, however, is still the carrier pig. Recent efforts to identify and eradicate the carrier population that contaminates livestock have been made impossible due to a lack of information regarding the epidemiology and pathogenesis of salmonellosis in cattle. Because the species of Salmonella is widely dispersed and resists the environment well, its elimination and control has been difficult. There is a substantial amount of information regarding the viability of Salmonella relevant to the pathogenesis in man. However, very little information is available regarding the animal sources of this infectious agent. An understanding of the infectious process in animal sources to produce food has assumed a higher priority and the control and elimination of the carrier animal will prevent the zoonotic transmission of the disease. The control of the diseases produced by the consumption of food can be obtained in a better way through the identification and eradication of the carrier population. While the carrier state can occur at any time throughout the life of an animal, the exposure at birth It will be the first opportunity for the animal to be exposed to the pathogen. The application of a mucous culture for the control of pathogenic colonization in the animals with which food is produced has been discovered. The term control means the reduction or prevention of colonization of the enteropathogenic bacteria. The term 'animals with which food is produced' means any animal consumed by humans.

A unique bacterial culture is obtained from the scrapes of the digestive system of animals free of the pathogen. Animals can be of any age, what is most preferred is the use of young animals, to protect newborns and older animals. This initial culture is subcultured and then administered to young animals. The method of this invention is applicable to any animal, whether domesticated or wild and particularly to cattle reared for human consumption, which could serve as a carrier of pathogens object of this study. Cattle include cattle, calves, pigs, piglets, sheep, lambs, buffalos, rams, ras, shellfish and the like. It is preferred to administer the elimination subculture preparation by comparison of cattle mucosa (LMCES) twice within the first 24 hours after calving. However, the preparation could be administered at any time during the life of the animal, either on a continuous basis or at selected times throughout the life of the animal, for example. The term "continuous base" means that a significant source of LMCES is provided by administration through drinking water, food, forced oral feeding or by aerosol application (aerosolization). The LMCES can be administered daily in food or water, weekly, monthly, etc. on a continuous basis. The term "sometimes selected throughout the life of the animal" means administering the LMC'L culture at critical control points, such as occurs during birth, ablation, disease, administration of antibiotics, the excessive heat, the dehydrated !!, the cold, during the transport as it happens when they move to other buildings in the production process and before the transport in the slaughterhouse, etc.

The target pathogens include all human enteropathogenic bacteria capable of colonizing animals, especially livestock bred for human consumption. As referred to herein, "human enteropathogenic bacteria" are bacteria capable of or known to colonize the human alimentary canal or to spread toxins therein and which are capable of causing intestinal diseases in a human host. Examples of human enteropathogenic bacteria include, but are not limited to, the Salmonella species, the Campylobacter species or the Escherichia coli species.

The LMCES can be combined with other cultures or effective treatments for the control of Salmonella in animals, such as, for example, Lactobacillus, fichto-oligosaccharides and yeasts Ott s treatments with entional or known animals and particularly for the inhibition of cntcropatógenos, can be added to the MCES as long as they do not affect the activity of the preparation of the LMCES.

In methods of the present invention, compositions for elimination subcultures by comparison of livestock mucosa (t MCES) are administered to animals. As used herein, "administering" includes any method suitable for orally feeding the compositions. to the animals, as it is known in the art, such as, for example, forced feeding, normal feeding, spraying or application of a paste on the maternal nipples or artificial nipples, through administered milk , etc. The LMCES can also be administered through the opening of the large intestine. The preparation may be made in any known form of the trade, such as, for example, liquid, paste, gelatin capsule or aerosol for administration. Preparations of LMCES are administered to animals at any age including newborn animals in effective amounts to reduce at least the human enteropathogenic bacteria found in the intestine of animals.

As used in this document, 'a reduction of bacteria' or 'at least reducing the human enteropathogenic bacteria' refers to the reduction in numbers of bacteria compared to what would be expected in an animal not receiving the treatment according to the methods of the present invention. Any suitable method of measuring, calculating or comparing bacteria present in the intestinal system of animals can be used for such comparisons, as would be clear and apparent to those skilled in the art. As will be indicated herein, "in effective amounts", "an effective amount", or "an efeetha quantity", refer to the amount of preparation of LMCES administered, in which the effect of the administration acts to at least reduce the human enteropathogenic bacteria found in all ages of the animals.

The amount of preparation will vary depending on the size of the animal being treated and the method of administration. For small animals, including newborn small animals, between 4 and 8 months, from the second or last subculture period of 48 hours, the culture of LMCEs can be administered by feeding now! forced, with approximately mi. of dosage preferably. For large animals, including large newborn animals, the second subculture period of 48 hours or the later of the LMCES may be left undiluted or diluted up to 10 times for oral administration in a liquid suspension in which the diluent may be, for example, milk, water, etc. Additional periods of subculture can be used but will most likely be less effective. Diluted or undiluted liquid LMCES preparations can be administered directly. For newborn animals, the preparation of LMCES ßo applies within approximately 2 to 48 hours after birth, with an approximate, preferably, 2 to 6 hours from birth, followed by a second dose approximately 18 to 24 hours later. The most preferred treatment schedule is the first dose within approximately 6 hours from birth, followed by the second dose, approximately 24 hours after birth. In general, the first dose of the treatment is given approximately between 2 and 48 hours before the ablactation, transportation or moving to another building, with a preferred time of approximately 2 to 6 hours. In these examples, only one treatment may be required, but a second treatment may be administered 18 to 24 hours after the first treatment. Variations in treatment programs will reflect commercial practices of agricultural production. Additional doses can be applied in the ablactation before moving along a production plant or before transportation in H slaughterhouse. If it is ajilica in food or water, daily administration pu * occurred *.

The preparation of LMCES is prepared by aseptically removing the large intestine, including the cecum of an animal and placing it in a sterilized container. As referred to herein, the large intestine is defined as the portion originating at the end of the stomach, up to and including the cecum. This container is maintained in an anaerobic environment throughout the preparation of the crops. The selected length of The intestine is inverted by any means known in the art. The contents of the intestine are eliminated by a combination of washing and scraping. The washing is done with an appropriate anaerobic medium. The washing step can use any effective means for the purpose formulated, including water. A preferable means is an anaerobic medium, and an anaerobic equilibrium medium Eh is particularly preferred in a previously reduced form.

H) Surface scraping is done with a smooth shore medium, as for example a shaving razor worn on the shore. After scraping, the lumen is washed again, followed by another scraping with a sharp edge medium, such as the sharp edge of a scalpel or other suitable instrument. The sharp edge medium and the wall of the lumen are washed with a medium, as described above, to obtain the epithelial cells and the microflora naturally. 1? and her . I am a toieeta e, t a sterilized container F! Fabric can also be cut directly in the culture medium without having to scrape first. It is preferable to keep the tissue in a reduced environment, for example, under a stream of nitrogen when the process is started. F! washing or cuts, with associated epithelial cells and microflora, are suspended and the content inoculated in an anaerobic medium sterilized for the crop As used in e! In this document, the term "microflora" is intended to include bacteria in natural cysts. The culture is incubated apaerobically at approximately 35-40 ° C for approximately 48 hours, then transferred to a fresh anaerobic medium and re-incubated for approximately 48 hours. . This second incubation is the second subculture period of 48 hours and is the most preferred LMCES preparation used. Other periods of subculture can also be used as described above. The culture is then put to test to discover if there is presence of human enteropathogens using some appropriate and conventional isolation technique. The pathogen-free cultures are administered immediately, freeze-dried or frozen using conventional techniques to freeze cultured cells. The blood of donor animals should be examined serologically to discover antibody levels in the corresponding host animal, including human pathogens. Only non-seroconverted animal cultures should be used.

The following examples are intended only to further illustrate the invention and are not intended to limit the scope of the invention as defined in the claims.

EXAMPLE 1 PREPARATION OF EC CROPS OF THE BOVINE CATTLE MUCOSA The intestines of an adult and healthy bovine are obtained in a local establishment in "unapproved" or killed and transported in a plastic bag to the laboratory within one hour in a sterile anaerobic environment. The mucosal culture is prepared from the large intestine system. Before removing the piece of intestine to be used, the surrounding area is tied with rope to prevent runoff of intestinal contents. A length of about 4 to 5 inches is removed aseptically and placed in a sterile container. The vessel is then placed in a tank that is continuously leveled with oxygen-free nitrogen gas. All maneuvers are carried out under the edge of the tank to maintain the crop to be harvested in an anaerobic environment. The length of the intestinal tissue is carefully inverted in a glass measuring rod, sterilized, without touching the inner surface. Once invested, the contents are removed with a combination of washing and scraping. The washing is done by syringe application of a previously reduced heart-brain broth infusion (PR-BHIB). Any other appropriate anaerobic medium is usable for the procedure of the washed. The scraping is done with the blunt edge of a sterilized scalpel blade. The lumen is washed, scraped and then washed again. In this step, the cecal epithelium is scraped gently, preferably removing it with the cutting edge of the scalpel in a scraping procedure. However, the tissue could be cut directly into the culture medium. After scraping the inverted intestine, the scalpel and the tip of the wall of the lunun are again washed with PR-BIUB. This final wash is collected in a sterile container. Washing, with the cor- responding epithelium) and the bacterial cells are aspirated with a sterilized syringe and needle and used to inoculate in a PR-BH1B tube through a rubber tube. These tubes are incubated at 5 ° C for 48 hours, transferred and re-incubated for another 48 hours. The culture is examined to detect if there is presence of important enteropathogens, such as Salmonella, E. Coli or the species Canfyl bacter. If the culture is free of both human and animal pathogens and if it is considered safe, it will be ready to be used immediately or to be stored and used later.

EXAMPLE 2 EXAMINATION OF THE EFFICACY OF BOVIN LMCES Bovine LMCCS compounds, as described in Example 1 above, are administered twice on average to newborn calves within, approximately, the first 24 hours of life. The culture is applied orally through the use of a bottle or bottle. Each treated calf receives around 500 ml. of the LMCES, undiluted or up to 10 times diluted in milk for each application. After being administered, it is allowed to feed the beceps as usual. Approximately twenty-four years after being admired, each calf is given approximately 10 * cells of an E. Coli 0157: 117 resistant to nalidixic acid by forced oral feeding. Following the gone (or exam), calves are allowed to grow in isolated chambers,? T >r rimately seven days, to allow any cell E. Coli 0157: H7 in transition saiga of the intestinal system. The calves are then sacrificed, the mint is removed aseptically and placed in sterilized plastic bags. The mint is then analyzed to discover the presence and level of the / - '. Coli 0157.117. The average registration number of the target pathogen per gram of mint for the whole group is called the colonization factor (CF).

The proportion of CF (untreated control / CF (Treatment Group) is called the protection factor (FP) Comparing the FP of an EC culture of mucosa with the FP of another culture, a relative value is obtained by degree of protection for the crop.

EXAMPLE 3 PREPARATION OF EC CROPS OF THE SWINE MUCOSA A healthy juvenile pig, approximately 1 to 6 months old at full maturity, calms down with a cocktail of medications, as is the routine practice of the trade. A mixture of 8 mg / kg of Ketaset, 7 mg / kg of Telazol and 4 mg / kg of Rompun was used. The pig is then sacrificed by bleeding. A cecum is aseptically removed and placed in a sterilized Petri dish. The plate is then placed in a vat that is continuously leveled with oxygen-free nitrogen gas. All maneuvering must be done below the top edge of the tank to maintain the crop to be harvested in an anaerobic environment. The length of the crop should be carefully invested in a sterilized implement without touching the inner surface. Once inverted, the contents are removed by a combination of washing and scraping. The washing is carried out by applying, with a syringe, the previously reduced infusion. Scraping is done with the blunt edge of a sterilized instrument. The lumen is washed, scraped and washed again at this point, the cecal epithelium is gently scraped with the sharp urine of a scalpel or it can be cut into small sections (approximately 5 cm.). After scraping the inverted cecum, the tip of the lumen wall and the scalpel are again washed with PR-B? HB. This final wash or cecal sections are collected in a sterilized container. The wash, with the related bacterial cells, is aspirated with a sterilized syringe and needle and used to inoculate in a PR-BHIB tube sterilized by a rubber septum.

These tubes are incubated at approximately 35 ° C for about 48 hours, transferred and re-incubated for another 48 hours or so. The culture is examined for the presence of human enteropathogens that are of interest, such as that of the Salmonella, E. Coli or Ca pylobacter species. If the crops are free of pathogens and are considered safe, will be ready for immediate application or to be stored and used later.

EXAMPLE 4 EXAMINATION OF THE EFFECTIVENESS OF THE EC CULTURE (LMCES) OF THE MUCOSA DEL GANADO PORCINO The =; Seeds with known mating dates were kept in breeding cages in isolation units. Each sow was checked every four hours starting one day before the intended mating date to ensure that the first LMCES culture was administered immediately. During mating, the mother fed piglets, in order to make sure that they obtained the colostrum and each piglet was given approximately 5 ml of LMCES for forced oral feeding between 2 and 6 hours after mating. A second dose of approximately 5 ml. was administered in approximately 24 hours. The piglets were examined by applying approximately 103 CFU S. choleraesuis by intronasal instillation approximately 48 hours after mating (24 hours after the last administration of LMCES). After administering the preparation, the piglets are allowed to feed as usual Rectal temperatures and rectal samples were taken daily from each of the piglets for approximately 7 days after the examination and were cultured to examine the presence of Salmonella. Approximately on the seventh day after the examination, all piglets were sacrificed and necropsied. Tissues were collected to perform qualitative bacteriology and included tonsils (ton), mandibular lymph nodes (mln), lungs, nodes lymphatic brachials (bln), liver, spleen (spl), middle ileus, ilcocolic junction (icj), ileocolic lymph nodes, cecum (cec), cecal content (ce), colon (col), the colonic lymph nodes and the stomach wall (sw) Quantitative bacteriology was also performed on the cecal content and the ilcocolic junction to determine the level of Salmonella within the tissues.

In order to assess the impact that sowing may have had on piglets, the faeces of the seed are also collected and cultivated before mating, within the approximate 48 hours after mating and approximately on day 7 after mating. the piglets (the sowings were never directly examined with respect to S Choleraesuis). The control piglets were never administered the LMCES but they were examined approximately 48 hours after birth. The tissues were collected and processed as described above.

The clinical signs were not apparent in all the piglets throughout the experiment The recovery of Salmonella in the rectal samples was variable. However, about 41% of the tissues were positive in the piglets treated with LMCES against approximately 63% positive tissues in the controlled piglets (Table 1, Summary of Tests 1 and 2). In piglets originating in the negative sherds (Test 1 -seed 1, 2,3 and Test 2-sowers 2) 39% [120/310] of the tissues was positive against 78% [47/60] (Test 1 - Negative seed control) the positive tissue of the control seed. The reduction of Salmonella is imparted in those treated with CE compared to untreated piglets An approximate reduction was observed in the 2 to 5 Salmonella registry in cecal content (CC) or ileocolic junction (ICJ) in piglets treated with LMCES when compared with the controls (Salmonella was cleared from ICJ and CC in piglets of 2 and 1 seed, respectively) (Table 2, Tests 1 and 2) In trial 1, all seedlings were free of pathogens. All the sowings were originally throwing a serogroup and then stopped before parking! laugh tito During e! mating, seeding 1 and control semen were housing serogroup B In trial 2, piglets from semen 1 and 2 yielded only Salmonella type B and not S. choleraesuis examined. The controls for tests 1 and 2 did not receive any LMCE, only the body examined. As can be seen, there are not only reduced levels of the examined organism but also reduced levels of native colonization. While reduction was observed in piglets already colonized with Salmonella rather than by the one received with the test, the degree of protection is reduced, suggesting that in the baby piglets, an earlier administration can be guaranteed.

The above detailed description is made for the purpose of illustration. So much detail is done only for that purpose and the experts of the trade can make variants in it without abandoning the spirit and overall aspect of the invention.

TABLE ONE The incidence and levels of Salmonella in piglets treated with LMCE or not treated (Ctrl) TABLE TWO The incidence and levels of Salmonella 2 days after the Test in piglets in the which the LMCE Treatment was applied and in which the treatment was not applied Test 1: STEM or ms No POS% POS. Reg 10 Reg 10 N ° PIGS Pass N ° Sample Fabrics Fabrics CC ICJ 1 - . 1 -? 4/2 0 36/40 90 3.86 3.65 2 - / '/. / 2 1 21/60 35 2.51 3.35 3- / 12 4/5 2 43/120 36 1.5 0 SUMA N * 100/720 45 3.16 314 CONTROL C / 6 0 47 /? O 79 539 5.74 All negative sowings Fabrics Ton bln pul híg spl col 1C1 cec ce sw (4) 3 4 4 3 4 4 4 3 4 3 2 (6) 0 3 3 3 3 0 4 2 2 1 3 (12) 2 12 9 7 6 0 3 1 3 0 c (6 ) 4 4 5 5 5 5 5 5 5 4 Test 2: All red mullet: SEMPLER / o.POS. No POS. % POS. Reg.10 Reg.10 N-LECHONGS Pass N ° Samples Fabrics Fabrics CC ICJ I! p 4/4 50/56 * 37/70 53 2.32 1.38 -2 - / 9 6/7 4/49 * 20/90 22 -0- -0- SUM NA 57 / J60 36 2.02 1.08 C * • / 9 72/72 * 48/90 53 3.33 4.23 * Throwing only B (Cont. Table Two) Fabrics Ton bln pul híg spl col icj cec ce S 1 (7) 2 6 6 5 4 4 4 2 4 0 2 (9) 1 5 3 2 3 1 1 2 2 0 c 2 3 3 3 2 6 9 9 9 2 OnlyCl. STOREY / No IOS No IOS% POS Reg 10 Reg 10 N ° PIGS Pass N ° Samples Fabrics Fabrics CC 1CJ 1 + 7 4/4 na 19/70 27 -0- -0- 2- / 9 6/7 na 18/90 20 -0- -0- SUMA na 37/160 23 -0- -0- C + / 9 na 16/98 18 2.98 4.23 Fabrics Ton bln pul híg spl co! icj cec ce sw 1 (7, I 4 4 0 0 0 0 0 2 (VJ I 2 3 1 I 1 0

Claims (15)

CLAIMS We claim:

1. A method to treat livestock, which includes: providing an elimination culture by comparing the mucosa of cattle derived from a pathogen-free animal, and administering said culture to an animal, in those cases in which said animal has at least a reduced level of enteropathogenic bacteria in comparison with what would be expected to have not been administered said treatment.

2. The method of claim number 1 in which said culture is administered to said animal at least within 48 hours after birth.

3. The method of claim number 1 wherein said culture is administered to said animal twice within the first 24 hours after birth.

4. The method of claim number 1, wherein said culture is administered to said animal within 6 hours after birth and then, again, within 24 hours after birth.

5. The method of claim number 1, wherein said culture is administered to the animal during its moments of stress

6. The method of claim number 1, in which said crop is administered to the animal throughout its life.

7 The method of claim number 1, in which said culture is an anaerobic culture

8. The method of claim 1, wherein said human enteropathogenic bacterium is selected from the group consisting of Salmonella, Campylobacter, E. Coli and mixtures thereof

9. The method of claim 1, wherein said culture is administered in effective amounts to at least reduce the human enteropathogenic bacterial colonization of said animal.

10. A compound derived from animal mucosa consisting of subculture of cattle mucosa for its elimination by comparison derived from a pathogen-free animal.

1 1. The compound of Claim 10 wherein said subculture is an anaerobic culture.

12. The compound of Claim 10 wherein said culture is obtained by scraping the intestinal system of said pathogen-free cattle.

13. The compound of Claim 12 wherein said intestinal system is the large intestine.

14. The compound of claim 10 wherein said compound is prepared by aseptically removing the large intestine of said pathogen-free animal, inverting said intestine, washing said inverted intestine, scraping said inverted intestine, repeating said washing and scraping steps to obtain epithelial cells. and microflora, Inoculating said washes and scrapes in a sterilized anaerobic medium to form in inoculum, Cultivating said inoculum for 48 hours to form a culture and subculturing said culture to obtain said compound.

15. A compound of intestinal mucosa of cattle collected from pathogen-free animals with the property of reducing the colonization of intestinal systems of enteropathogenic bacteria of animals in which said compound is prepared by aseptically removing the large intestine of said pathogen-free animal, inverting said intestine , Washing said inverted intestine, Scraping said inverted intestine, Repeating said steps of washing and scraping to obtain the epithelial cells and microflora, Inoculating said washes and scraping in sterile anaerobic media to form an inoculum, Cultivating said inoculum for 48 hours to form a culture. , and subculturing said culture to obtain said compound.