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WO1996010420A1 - Composition et traitement d'hyperimmunisation a l'aide de bacteries non tuees thermiquement - Google Patents

Composition et traitement d'hyperimmunisation a l'aide de bacteries non tuees thermiquement Download PDF

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Publication number
WO1996010420A1
WO1996010420A1 PCT/US1995/013427 US9513427W WO9610420A1 WO 1996010420 A1 WO1996010420 A1 WO 1996010420A1 US 9513427 W US9513427 W US 9513427W WO 9610420 A1 WO9610420 A1 WO 9610420A1
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Prior art keywords
animal
composition
human
bacteria
animals
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PCT/US1995/013427
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English (en)
Inventor
Konky Altur Sotomayor Hermosilla
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Sotomayor, Tevic, Emar
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Application filed by Sotomayor, Tevic, Emar filed Critical Sotomayor, Tevic, Emar
Priority to AU39614/95A priority Critical patent/AU3961495A/en
Publication of WO1996010420A1 publication Critical patent/WO1996010420A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/40—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum bacterial
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/04—Mycobacterium, e.g. Mycobacterium tuberculosis

Definitions

  • the present invention relates to a composition and method for providing a therapeutic treatment for humans and animals by vaccination with enteric non heat-killed bacteria for the treatment of several conditions, including but not limited to the following; intestinal diseases such as diarrhea, bacterial infections, toxic infections, for reducing the bacterial content of the gastrointestinal system, for stimulation of the immune system, for accelerating growth and for increasing the protein to fat ratio.
  • enteric non heat-killed bacteria for the treatment of several conditions, including but not limited to the following; intestinal diseases such as diarrhea, bacterial infections, toxic infections, for reducing the bacterial content of the gastrointestinal system, for stimulation of the immune system, for accelerating growth and for increasing the protein to fat ratio.
  • This invention also provides a composition and method for the treatment of intestinal diseases such as diarrhea and includes administration of serum from an animal, such as a cow, immunized by means of specific, non heat-killed bacteria that are pathogenic to humans or animals.
  • Hormones that have been used to regulate the carcass ratio of protein to fat include anabolic steroids and growth hormone, and also neurotransmitter adrenergic agonists.
  • the immune system of the host will produce antibodies that will neutralize the effects of the antigen. Exposure to such foreign antigens can occur either naturally, or deliberately by administration of the antigen in vaccine form. The latter is generally referred to as active immunization of the host species exposed to the antigen. In general, merely inducing an immune state in an animal does not alter the levels of carcass fat or protein. Williamson, E.D., et al., Livestock Prod. Sc. 12:251-264 (1985).
  • Beck has also described a milk having anti-arthritic properties, U.S. Patent No. 4,732,757, and has patented a method of treating inflammation using milk from hyperimmunized cows.
  • Beck U.S. Patent No. 4,284,623.
  • Stolle, et al. have disclosed a method of using milk from a hyperimmunized cow for the treatment of diseases of the vascular and pulmonary systems.
  • Patent No. 3,911,108 discloses a patent that uses Peterson to produce a signal.
  • Peterson U.S. Patent No. 3,376,198 and Canadian Patent No. 587,849
  • Holm U.S. Application (published) Serial No. 628,987)
  • Tannah et al. British Patent No. 1,442,283
  • the present invention provides an composition and method for the treatment of developing humans to promote growth.
  • the present invention also provides a composition and method to increase the ratio of protein to fat in the human body.
  • the present invention also provides a composition and method for the reduction of the gasttointestinal content of bacteria in humans.
  • Intestinal infections such as diarrheal infections
  • Many bacterial infections are caused by bacteria such as Escherichia sp., Pseudomonas sp., Salmonella sp. and Aerobacter sp.
  • Diarrhea is a major intestinal disease annually affecting billions of humans and animals.
  • Diarrhea is a major cause of mortality in neonates, infants and the elderly, accounting for about 25% of the annual infant mortality. Diarrhea is also a major cause of illness and mortality in livestock and other farm animals, resulting in a tremendous loss of life, food sources and financial resources. Most treatments involve antibiotics which sometimes have undesirable side effects such as reducing immunological resistance to other diseases and decreasing food absorption due to exfoliation of the intestinal epithelium. In addition, many strains of bacteria that cause intestinal diseases such as diarrhea develop resistance to antibiotics, rendering conventional treatment ineffective.
  • Enterobacteria is defined generally as any of various gram-negative, rod-shaped bacteria of the family Enterobacteriaceae that includes some pathogens of humans and animals, such as the Escherichia coli and Salmonella sp.
  • Vaccination with compositions consisting of killed bacterial may provide protection against bacterial infections and also facilitate the immune response to a bacterial infection.
  • such vaccinations may provide a general protective effect through stimulation of the immune system.
  • compositions and methods that treats and prevents diarrhea in humans and animals.
  • This composition should produce fewer side effects than previous compositions and methods.
  • the composition should be capable of being administered through intraperitoneal, intravenous, intramuscular or subcutaneous injection to facilitate treatment of humans and animals, especially herd animals. These modes of administration would be much faster and more economical for treatment of large numbers of animals than previous methods of oral administration.
  • compositions and methodology that is relatively inexpensive and applicable to large numbers of humans and animals.
  • This composition and method should also promote growth in humans, especially developing humans, and increase the protein to fat ratio in humans.
  • This composition and method should promote growth and increase the protein to fat ratio in the offspring of pregnant humans that received this composition before becoming pregnant, and in the offspring of pregnant animals that received this composition before or during pregnancy.
  • This composition and treatment should be effective in increasing growth rates of animals and in producing lean animals that are acceptable to consumers.
  • This composition and method should also effectively reduce the intestinal content of bacteria in animals before slaughter, thereby producing meat products with reduced levels of bacterial contamination.
  • This composition should avoid the use of hormones such as anabolic steroids, expensive genetically engineered products, and expensive anti-hormone antisera.
  • a method and composition which provides a treatment for bacterial infections, intestinal diseases such as diarrhea, and for conditions resulting from affliction with toxins, such as toxic bacterial products.
  • This method and composition should also provide for stimulation of the immune system.
  • This composition should be a serum obtained from animals hyperimmunized with selected bacteria.
  • the present invention provides a therapeutic treatment for humans and animals comprising administering to the human or animal an amount of a composition comprising a mixture of non heat-killed bacteria, wherein the bacteria are Escherichia sp., Pseudomonas sp., Salmonella sp., and Aerobacter sp., effective for therapeutic treatment.
  • the present invention provides a new vaccine composition and method of administering that vaccine composition to provoke an immune response that increases growth in animals and humans.
  • the increased growth is accompanied by an increase in the protein content and relative decrease in fat content of the animal or human, thereby producing a higher protein to fat ratio.
  • the present invention provides a vaccine composition
  • a vaccine composition comprising a mixture of non heat-killed bacteria wherein the bacteria are selected from the group consisting of Escherichia sp., Pseudomonas sp., Salmonella sp., and Aerobacter sp. Chemical means are preferred for killing bacteria. Use of formalin is a preferred method of killing the bacteria.
  • the vaccine composition of the present invention contains an adjuvant such as an oil, lanolin or liquid Vaseline, although other adjuvants including, but not limited to, aluminum hydroxide or aluminum potassium sulfate could be used.
  • the vaccine composition may be administered in the form of microspheres, including but not limited to polylactide microspheres.
  • the vaccine composition comprising a mixture of four, formalin- killed bacteria from Escherichia sp., Pseudomonas sp., Salmonella sp., and Aerobacter sp., is administered to humans to provide a therapeutic treatment for a variety of conditions including but not limited to the following; infections, bacterial infections, complications due to toxins, especially bacterial toxins, and poor growth rate. Treatment of animals with this composition is effective in reducing the bacterial content of the gastrointestinal lumen. This treatment of an animal before slaughter decreases the bacterial contamination of animal products such as meat.
  • the present invention also provides a method of increasing the protein content and ratio of protein to fat in an animal or human by hyperimmunizing the animal by administering to the animal a composition comprising a mixture of non heat-killed bacteria wherein the bacteria are selected from
  • Escherichia sp. Pseudomonas sp., Salmonella sp., and Aerobacter sp.
  • the animal's growth is increased substantially.
  • growth increase has been shown to be 20% or more over non-vaccinated cattie.
  • the vaccine composition can be administered to birds, such as chickens, to substantially increase the size of eggs from the adult female bird.
  • a composition comprising a mixture of the following non heat-killed bacteria; Escherichia sp., Pseudomonas sp., Salmonella sp., and Aerobacter sp.
  • the present invention also provides a method of treating diarrhea by administering to an animal or human with diarrhea antidiarrheal serum obtained from an animal, desirably a cow, that has been immunized with a composition comprising a mixture of non heat-killed bacteria selected from Escherichia sp,
  • the antidiarrheal serum and method of the present invention is especially useful for treating diarrhea in newborns and in the elderly.
  • the antidiarrheal serum of the present invention can be administered by injection. This composition is effective in both treating and preventing diarrhea caused by infection with enteric bacteria.
  • the antidiarrheal serum is highly effective after only one administration, and rarely requires a second administration.
  • antidiarrheal serum derived from cows to which has been administered the composition comprised of a combination of the following non heat-killed bacteria; Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Aerobacter aerogenes.
  • Still another object of the present invention is to provide antidiarrheal serum from animals other than cows, including but not limited to goats, sheep, and pigs, to which has been administered the composition described above wherein the selected bacteria are pathogenic to the species receiving the antidiarrheal serum.
  • Another object of the present invention is to provide a therapeutic treatment for diarrhea in humans and animals by administration of a composition comprising a Combination of the following non heat-killed bacteria; Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Aerobacter aerogenes
  • a further object of the present invention is to provide a method of treating diarrhea in humans especially neonates, infants and the elderly.
  • Another object of the present invention is to provide a method of treating diarrhea in non-human animals, including, but not limited to, livestock, pigs, sheep, birds, rodents, and rabbits.
  • a further object of the present invention is to provide a method of preventing diarrhea in humans especially neonates, infants and the elderly.
  • Another object of the present invention is to provide a method of preventing diarrhea in non-human animals, including but not limited to livestock, pigs, sheep, birds, rodents, and rabbits.
  • Still another object of the present invention is to increase growth and increase the protein to fat ratio in animals and humans.
  • Another object of the present invention is to increase growth of developing humans.
  • Yet another object of the present invention is to preferentially increase the growth of the offspring of pregnant animals that received this composition before or during pregnancy.
  • Still another object of the present invention is to preferentially increase the growth of the offspring of pregnant humans that received this composition before pregnancy.
  • a further object of the present invention is to increase growth by administering to animals and humans one injection of a composition that induces an immune response.
  • Still another object of the present invention is to provide a method to stimulate an immune response in animals and humans by administering the composition.
  • Another object of the present invention is to preferentially increase the protein content and the protein to fat ratio of animals and humans receiving this composition.
  • Another object of the present invention is to mix the non heat-killed bacteria with an adjuvant for administration to animals and humans.
  • Yet another object of the present invention is to administer the vaccine composition of the present invention in a single injection.
  • Yet another object of the present invention is to increase the size of eggs from birds.
  • Another object of the present invention is to decrease the cost of raising livestock and birds by increasing growth rate.
  • Still another object of the present invention is to decrease the time required for livestock and birds to grow to a desired size before sacrifice.
  • Yet another object of the present invention is to provide meat with higher protein content and higher protein to fat ratio from livestock and birds.
  • Another object of the present invention is to decrease the bacterial content of the gastrointestinal tract of animals.
  • Still another object of the present invention is to decrease the bacterial contamination of animal products, such as meat, obtained at slaughter of the animal.
  • administered means oral administration and intramuscular, intravascular, intraperitoneal or subcutaneous injection of an animal or human with the composition of the present invention in a pharmaceutically acceptable vehicle.
  • non heat-killed bacteria refers to bacteria killed through means other than heat.
  • therapeutic treatment means administration of the combination of the following non heat-killed bacteria; Escherichia sp., Pseudomonas sp., Salmonella sp., and Aerobacter sp., optionally combined with one or more adjuvants in order to stimulate growth, reduce bacterial levels, reduce the gastrointestinal bacterial content, decrease diarrhea, reduce bacterial toxin levels, stimulate the immune system, or increase the protein to fat ratio.
  • the term “therapeutic treatment” also means administration of serum obtained from an animal immunized with a combination of these non heat-killed bacteria in order to decrease diarrhea.
  • growth or “increased growth” means an increase in the length of the axial skeleton, an increase in the length of the long bones such as the femur or humerus, an increase in the muscle mass of the body, or an increase in the girth of the body.
  • diarrhea means excessive and frequent evacuation of watery feces, usually indicating gastrointestinal distress or disorder or intestinal disease.
  • the term “food animal” means any animal that is consumed in the diet of humans or other animals.
  • Typical food animals include bovine animals, for example, cattle; ovine animals, for example, sheep; swine, for example, pigs; fowl, for example, chickens and turkeys; ratites, for example emus and ostriches; fish; rabbits and the like.
  • the present invention encompasses a vaccine composition
  • a vaccine composition comprising a mixture of non heat-killed bacteria wherein the bacteria are selected from Escherichia sp. , Pseudomonas sp., Salmonella sp., and Aerobacter sp. and a method for using the composition to increase growth, to increase the protein to fat ratio, to stimulate the immune system, to treat and prevent diarrhea, to reduce bacterial levels, to decrease the bacterial gastrointestinal content, to reduce the level of bacterial toxins, to increase the growth of the offspring of animals receiving this composition during or before pregnancy, and to increase the growth of the offspring of humans receiving this composition before conception.
  • the bacteria are Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Aerobacter aerogenes.
  • Various means may be used to kill bacteria including chemicals, radiation and other methods. Radiation sources may include the use of x-ray, gamma and ultraviolet irradiation. Chemicals which may be used to kill bacteria include, but are not limited to, formalin, phenol, glutaraldehyde, acrolein, alcohol, paraformaldehyde, sodium periodate, lysine, other cross-linking agents and mixtures thereof.
  • the bacteria are killed with formalin.
  • the bacteria can be killed by other means such phenol, and the like.
  • the composition of the present invention comprises bacteria that are selected from the geographical area in which the vaccine composition is to be used. Although it has been found that including at least the four species listed above provides excellent growth increase, it is contemplated that the vaccine composition of the present invention can include additional bacteria.
  • the amounts of bacteria used in a typical preparation of the present invention are between approximately 10 8 and 10 12 bacteria per ml with the desired concentration of bacteria of between 10 10 to approximately 10 11 bacteria per ml.
  • the amounts of bacterial species per 10 ml are: 7 x 10 10 for Escherichia sp., and 1 x 10 for each of Pseudomona sp., Salmonella sp., and Aerobacter sp. An adult cow will receive
  • Preferred methods of administration to cows include intramuscular and subcutaneous administration. Dosages for different species are approximately 1.5 to 1.7 x 10 bacteria per pound of body weight. In poultry, a sufficient response can be obtained with half the standard concentration of bacteria and 10% of the above dosage as inoculum.
  • the optimal volume of the vaccine for each administration will vary depending upon the species of animal to be vaccinated. For example, for cattle, pigs, sheep, and goats, the optimal volume of the vaccine for administration is between approximately 1 ml and 15 mis with the desired volume of approximately 10 mis. For chickens the optimal volume of the vaccine for administration to the chick is between approximately 0.05 mis and 2 mis at half the concentration of bacteria.
  • the optimal volume of the vaccine for administration to the human is between approximately 0.05 to 0.2 ml per 10 kg of body weight at a desired concentration of non heat-killed bacteria of approximately 10 9 bacteria per ml.
  • the preferred method of administration to humans is intramuscular administration. Neonates, infants, and children under 40 kg body weight are not to be treated with the vaccine. Pregnant women should also not be treated with the vaccine. It is to be understood that the volume of the vaccine to be administered to the human or animal is at least partially dependent upon the individual being treated, the concentration of the bacteria in the vaccine, on the type of adjuvant being used, the condition being treated and the method of administration. Thus, the desired volume may be outside the stated ranges depending upon the final formulation and method of administration of the vaccine.
  • the composition of the present invention contains an adjuvant to increase the immune response such as an oil or Vaseline, although aluminum hydroxide or aluminum potassium sulfate may be used.
  • an adjuvant to increase the immune response such as an oil or Vaseline, although aluminum hydroxide or aluminum potassium sulfate may be used.
  • hyperimmunization of food animals is achieved by a single administration of a composition of the present invention with the adjuvant.
  • Many different compositions for the slow release of vaccines have been described which would be applicable to the method of the invention, for example, as described in Sanders, H.J., Chem. & Engineering News, Apr. 1, 1985, pp. 30-48, which is incorporated herein by reference. It is to be understood that the composition of the present invention is desirably injected only once, but can be injected more than once over a period of time.
  • a booster injection may be given several weeks or up to a year after the first injection.
  • Other methods of administration including but not limited to administration of microspheres, such as polylactide microspheres and polyglycolide microspheres, may be used in the present invention.
  • microspheres are described in Cleland, J.L., Design and Production of Single-Immunization Vaccines Using Polylactide Polyglycolide Microsphere Systems, pp. 439-462 in Vaccine Design: The Subunit and Adjuvant Approach, Powell, M.F. and
  • the present invention provides a method for increasing the protein content and increasing the protein to fat ratio in an animal or human by vaccinating the animal or human through administration of a composition comprising non heat- killed bacteria wherein the bacteria are selected from Escherichia sp., Pseudomonas sp., Salmonella sp., and Aerobacter sp.
  • the vaccine of the present invention can also be administered to birds, such as chickens, turkeys, ducks and the like, in order to increase the size of eggs laid by the birds.
  • the method of the present invention is also applicable to any animal used as a source of food and especially as a source of meat such as mutton and lamb from sheep; beef and veal from cattle; pork from pigs; poultry meat including meat from chickens and turkeys, and rabbit meat.
  • animals and humans are vaccinated while still growing and maturing into the adult state.
  • the first vaccination may be administered as soon as the animal is competent to immunologically respond to the antigen.
  • Animals and humans immunologically competent to respond to the antigen are also capable of being placed in a hyperimmune state.
  • pregnant animals are injected with the composition.
  • pregnant animals are injected with the composition and their offspring are injected as soon as the offspring animal is competent to immunologically respond to the antigen.
  • An advantage of the present invention is that only a single injection is required to accomplish increased growth and increased ratios of protein to fat. Accordingly, there is usually no need to perform booster injections, resulting in savings in labor and materials, especially in the application to large numbers of farm animals such as cattle, chickens and turkeys. A single injection is also less stressful to the animals and decreases the chances of infection, thereby increasing the overall livestock survival rate. Additional savings are provided by the nature of the composition itself because all components (bacteria and optionally adjuvant) are inexpensive, especially when compared to prior art methods of increasing growth such as beta receptor agonists, somatostatin antisera, growth hormone and synthetic steroids as taught in the prior art. The financial benefits provided by the present invention also decrease the cost of food production thereby cutting the cost of food to the consumer.
  • the vaccine composition of the present invention does not involve administration of hormones such as anabolic steroids or growth hormone, neurotransmitter agonists such as beta adrenergic agonists, or genetic alteration of animals, so the consumer concerns regarding the effects of these treatments on lean meat are avoided.
  • hormones such as anabolic steroids or growth hormone
  • neurotransmitter agonists such as beta adrenergic agonists
  • genetic alteration of animals so the consumer concerns regarding the effects of these treatments on lean meat are avoided.
  • Another aspect of the present invention is the reduction or elimination of a species of microorganism from the gut of an animal before slaughter.
  • a variation of the vaccine composition of the present invention could be used to achieve this purpose.
  • Cows destined for the slaughterhouse are vaccinated 4 to 6 months before sacrifice with a variation of the vaccine composition containing higher amounts E. coli at a dose of approximately 10 mis at approximately 10 11 bacteria per ml.
  • the relative proportions and components of the vaccine composition may be selected based on the animal selected or the microorganisms prevalent in a particular geographical area or animal.
  • Additional selection of the amounts of other bacteria may occur to decrease the relative percentage of a particular microorganism in the gastrointestinal tract of a selected animal.
  • the present invention is particularly useful for reducing the gastrointestinal content of microorganisms in livestock, especially cattle, cows, sheep, goats, buffalo and pigs; and birds such as chickens, turkeys, hens, doves, and quail.
  • vaccinations of cows may occur annually starting at 6 months of age.
  • Pigs, sheep, and goats could be injected annually after attaining approximately 9 to 11 kg of body weight.
  • Chickens and turkeys could be injected 3 weeks after birth and twice a year thereafter. It is contemplated that other administration schedules may be used.
  • This treatment would greatly reduce or eliminate a selected species or group of species of microorganisms from the gut of an animal before slaughter, thereby decreasing bacterial contamination of the animal products, especially meat, reducing the costs of producing products low in bacterial contamination and decreasing the risks of food poisoning.
  • a species of microorganism from the gastrointestinal tract of humans especially if imbalances in the relative proportions of enteric bacteria produce distress, upset or infection.
  • a variation of the vaccine composition of the present invention could be used to achieve this purpose.
  • Such a treatment might include a vaccine containing an enrichment of formalin-killed E. coli relative to the other antigenic bacteria.
  • the present invention provides a method of treating diarrhea by administering to an animal or human, preferably in a single injection, an antidiarrheal serum obtained from cows or other animals previously injected with a composition comprising a mixture of non heat-killed bacteria selected from Escherichia sp, Pseudomonas sp, Salmonella sp and Aerobacter sp.
  • the non heat-killed bacteria are Escherichia coli, Pseudomonas aeruginosa, Salmonella enteritidis and Aerobacter aerogenes.
  • the bacteria are killed with formalin.
  • the antidiarrheal composition of the present invention comprises bacteria that are selected from the geographical area in which the vaccine composition is to be used.
  • the composition is optionally mixed with an adjuvant and administered to the animal.
  • the adjuvant can be liquid Vaseline, lanolin, and/or vegetable oil although other adjuvants are within the scope of the present invention.
  • the vaccine composition may be administered in the from of microspheres, such as polylactide and polyglycolide microspheres.
  • the amount of bacteria used in immunizing the animal from which the serum is to be extracted generally is between approximately 10 8 and 10 12 per ml, although higher or lower concentrations may be used.
  • the optional volume of the vaccine for each injection will vary defending upon the species of animal to be vaccinated. For example, for cattie the optimal volume of the vaccine for administration to the cow is between approximately 5 mis and 15 mis of inoculum having 10 8 and 10 12 bacteria per ml. For chickens the optimal volume of the vaccine for administration to the chick is between approximately 0.05 mis and 2 mis of inoculum having 10 8 and 10 12 bacteria per ml.
  • the preferred animal for immunization and subsequent collection of serum is the cow. It has been determined that serum from a cow causes fewer side effects in recipients when administered to other species including humans.
  • the present invention includes an antidiarrheal composition
  • serum from an animal immunized with formalin-killed bacterial cultures pathogenic to an infant.
  • This serum contains antibodies useful for treating the intestinal infections induced by these bacteria in infants, young children and trn* elderly and is administered by injection.
  • This invention is a m dicament enabling successful treatment of enterobacterial infection without antibiotics, thus avoiding the cost drawbacks and risks associated with antibiotic administration.
  • the animal is immunized with an antigen mixture of non heat-killed bacteria, such as formalin-killed bacteria, selected from Escherichia coli, Pseudomona aeruginosa,
  • the antigen mixture is typically injected 3 times into an adult animal over a 30 day period. The number of injections and the timing of the injections can vary.
  • blood is collected from the animal and serum is prepared from the blood using standard methods, well known to those of ordinary skill in the art. Typically, the blood is allowed to clot and serum is collected either by compression of the clot or by centrifugation. The serum is collected, extracted, mixed with 0.5% phenol, and stored.
  • the vaccine composition may be administered in the from of microspheres, such as polylactide and polyglycolide microspheres, and blood collection may occur at later dates.
  • the bacteria selected from Escherichia sp., Pseudomonas sp., Salmonella sp., and Aerobacter sp. desirably Escherichia coli, Pseudomona aeruginosa, Salmonella enteritidis and Aerobacter aerogenes responsible for intestinal infections of infants and young children.
  • the composition is a serum preferably obtained from a cow immunized against a mixture of non heat-killed bacteria Escherichia sp., Pseudomonas sp., Salmonella sp., and Aerobacter sp., preferably formalin- killed Escherichia coli, Pseudomona aeruginosa, Salmonella enteritidis and Aerobacter aerogenes, optionally mixed with an adjuvant. Immunization of a cow can be effected in accordance with the general techniques of immunization well known to those of ordinary skill in the art.
  • the new serum composition of the invention can be administered orally or through a single subcutaneous, intravascular or intramuscular injection.
  • a desirable average dose is approximately 0.1-1.0 ml/kg.
  • a desirable average dose is between approximately 0.1-0.2 ml/kg.
  • the doses to administer to neonates and infants, as well as other humans such as adults and the elderly will vary from individual to individual. Consequently, a medical provider may elect to administer a low dosage to a particular patient initially, followed by one or more additional administrations.
  • the dose to be provided to animals through subcutaneous, intramuscular, or intravascular injection may vary depending upon the type of animal, its individual characteristics and its condition of health.
  • the dose of the serum from hyperimmunized cows to be injected into animals subcutaneously, intravascularly, or intramuscularly is tailored to the animal's species, age and body weight. A single injection is the preferred mode of treatment, although some types of infections and some animals may require additional injections.
  • the use of the serum from cows is the preferred embodiment of the invention, some recipient species may respond better to serum produced in the same species. Accordingly, for these cases, serum is produced in the same species as the recipient animal using a composition of the four bacteria selected for maximal compatibility with the animal to be immunized.
  • the preparation of the formulation is as follows: 10 Roux flasks, 1,000 ml capacity each, with 1% dextrose nutritive agar are autoclaved at about 121 °C for 30 minutes. Seven test tubes containing nutritive agar are seeded with a selected bacteria from a strain of Escherichia coli, one test tube with Pseudomonas aeruginosa (pyocyanic bacillus), one test tube with Salmonella enteritidis, and one test tube with Aerobacter aerogenes. The tubes are incubated at approximately 37°C for 24 hours. Each test tube of culture is washed with 10 ml of sterile physiological solution until all colonies are washed loose.
  • Each Roux flask is inoculated with the wash of each test tube of E. coli, P. aeruginosa, S. enteritidis, and A. aerogenes.
  • the Roux flasks are incubated at approximately 37°C for 36 hours, with the agar in the internal superior surface of the flasks. The agar surface is washed using the liquid contained in the bottom part of each Roux flask and these mixtures are saved in a sterile container.
  • each Roux flask is washed with 45 ml of sterile physiological solution until all the colonies are washed loose. These mixtures are collected in the same sterile container. Formalin is added to the sterile container in a proportion of 1% of the total collected volume. Flasks are incubated at approximately 37°C for 24 hours or at room temperature for 72 hours.
  • the components of the adjuvant are 400 ml of liquid Vaseline, 400 ml of vegetable oil and 200 gm of anhydrous lanolin (Sigma Chemical Co., St. Louis MO) (ratio of 2:2:1, respectively). The mixture is melted in a water bath, boiled for 30 minutes, and slowly cooled at room temperature. Ratios of adjuvant components may be adjusted for application to different animals.
  • Equal parts of the antigen and the adjuvant are admixed so that the bacterial count is approximately 1 x 10 11 per 10 ml.
  • the mixture remains undisturbed at 22 to 26°C for 72 hours. This formulation tends to sediment in layers especially at temperatures lower that 22 to 26 ⁇ C.
  • 10 mis of the vaccine are injected subcutaneously.
  • the antidiarrheal immune serum of the invention is obtained after hyperimmunizing a cow with an antigen and adjuvant.
  • the antigen composition comprises a mixture of non heat-killed, preferably formalin-killed bacteria, selected from Escherichia coli, Pseudomona aeruginosa, Salmonella enteritidis and Aerobacter aerogenes, combined with an adjuvant containing Vaseline, lanolin, and vegetable oil.
  • the antigen is administered in 3 injections at 1, 15 and 30 days in 10, 15, and 20 ml, respectively, to adult cows no less than 18 months old. Approximately 45 days after the third administration, 1,000 ml of blood per 100 kg body weight is removed from the jugular vein. Booster injections are performed approximately three times a year with blood removal 35 to 45 days after each injection at 1,000 ml/100 kg body weight.
  • the serum can be obtained following formation of a clot at 4°C - 24°C and compression, or by centrifugation at 1,000 x g. The serum is extracted with techniques familiar to those skilled in the art. The serum may be stored with 0.5% phenol and bottled in 30 ml flasks and stored at -90°C to 4°C.
  • the serum described in Example 1 is injected intramuscularly, intravenously, or subcutaneously into neonates, infants, and elderly persons with diarrhea based on their body weight and the titer of the serum.
  • a 70 kg man with diarrhea is injected with 5.0 ml of the serum intramuscularly in the gluteal region and with 9.0 ml of the serum intravenously.
  • a new born calf was infected at two days after birth with a 5 ml volume of a live bacterial culture containing 10 7 bacteria/ml, where the type and number of bacteria in culture were in the same proportion as was used to prepare the antigen, 27
  • the infected calf subsequently developed diarrhea by day five.
  • Five day-old calves suffering from diarrhea were treated by administering a single injection of 50 mis of the antidiarrheal serum of Example IV.
  • the calves were cured of diarrhea in approximately 6 to 12 hours.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Mycology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Communicable Diseases (AREA)
  • Pulmonology (AREA)
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  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

Selon la présente invention, une composition et un procédé thérapeutique sont utilisés pour traiter l'animal et l'homme. Cette invention porte sur une composition et un procédé visant à stimuler la croissance, augmenter le rapport protéines/graisses, à traiter et prévenir la diarrhée et à stimuler le système immun chez l'animal et chez l'homme. L'invention porte également sur la diminution des coûts de production alimentaire se traduisant par l'accélération du développement des animaux de ferme et la réduction de la population de bactéries dans les voies gastro-intestinales au moment de l'abattage. La composition comprend un mélange de bactéries non tuées thermiquement, ces bactéries étant Escherichia sp., Pseudomonas sp., Salmonella sp. et Aerobacter sp.
PCT/US1995/013427 1994-10-04 1995-10-04 Composition et traitement d'hyperimmunisation a l'aide de bacteries non tuees thermiquement WO1996010420A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU39614/95A AU3961495A (en) 1994-10-04 1995-10-04 Composition and treatment for hyperimmunization with non heat-killed bacteria

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US31803094A 1994-10-04 1994-10-04
US08/318,030 1994-10-04
US31850294A 1994-10-05 1994-10-05
US08/318,502 1994-10-05

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999058133A3 (fr) * 1998-05-08 2000-01-06 Dcv Inc Antidiarrheique et procede d'utilisation dudit produit
US6420183B1 (en) 1996-05-28 2002-07-16 Bayer Aktiengesellschaft Masking background fluorescence and luminescence in optical analysis of biomedical assays
NL1019143C2 (nl) * 2001-10-08 2003-04-09 Nutricia Nv Farmaceutisch product of voedingssupplement en daarbij te gebruiken tussenproduct.
RU2523389C1 (ru) * 2013-05-27 2014-07-20 Федеральное государственное бюджетное учреждение "Федеральный Центр токсикологической, радиационной и биологической безопасности" (ФГБУ "ФЦТРБ-ВНИВИ") Способ получения гипериммунной сыворотки против анаэробной энтеротоксемии и эшерихиозной диареи телят
CN109414461A (zh) * 2016-04-26 2019-03-01 曲生物制品公司 治疗性地触发靶组织中的先天性免疫应答

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3127318A (en) * 1964-03-31 Swine treatment
US3139382A (en) * 1961-10-31 1964-06-30 American Home Prod Bacterins and process of producing the same
US3992521A (en) * 1972-07-07 1976-11-16 Agence Nationale De Valorisation De La Recherche (Anvar) Immunological product for the treatment of intestinal infections due to pathogenic enterobacteria of the infant
US4237115A (en) * 1977-11-23 1980-12-02 Bactex, Inc. Method of immunization against enterotoxogenic infection by Escherichia coli

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3127318A (en) * 1964-03-31 Swine treatment
US3139382A (en) * 1961-10-31 1964-06-30 American Home Prod Bacterins and process of producing the same
US3992521A (en) * 1972-07-07 1976-11-16 Agence Nationale De Valorisation De La Recherche (Anvar) Immunological product for the treatment of intestinal infections due to pathogenic enterobacteria of the infant
US4237115A (en) * 1977-11-23 1980-12-02 Bactex, Inc. Method of immunization against enterotoxogenic infection by Escherichia coli

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6420183B1 (en) 1996-05-28 2002-07-16 Bayer Aktiengesellschaft Masking background fluorescence and luminescence in optical analysis of biomedical assays
US7063952B2 (en) 1996-05-28 2006-06-20 Bayer Healthcare Ag Masking background flourescence and luminescence in optical analysis of biomedical assays
US7138280B2 (en) 1996-05-28 2006-11-21 Bayer Healthcare Ag Masking of the background fluorescence and luminescence in the optical analysis of biomedical assays
US8178359B2 (en) 1996-05-28 2012-05-15 Bayer Healthcare Ag Masking of the background fluorescence and luminescence in the optical analysis of biomedical assays
WO1999058133A3 (fr) * 1998-05-08 2000-01-06 Dcv Inc Antidiarrheique et procede d'utilisation dudit produit
AU772785B2 (en) * 1998-05-08 2004-05-06 Arkion Life Sciences Llc Anti-diarrheal and method for using the same
NL1019143C2 (nl) * 2001-10-08 2003-04-09 Nutricia Nv Farmaceutisch product of voedingssupplement en daarbij te gebruiken tussenproduct.
WO2003030918A1 (fr) * 2001-10-08 2003-04-17 Nutricia N.V. Produit pharmaceutique ou complement alimentaire, et produit intermediaire a utiliser avec ces produits
RU2523389C1 (ru) * 2013-05-27 2014-07-20 Федеральное государственное бюджетное учреждение "Федеральный Центр токсикологической, радиационной и биологической безопасности" (ФГБУ "ФЦТРБ-ВНИВИ") Способ получения гипериммунной сыворотки против анаэробной энтеротоксемии и эшерихиозной диареи телят
CN109414461A (zh) * 2016-04-26 2019-03-01 曲生物制品公司 治疗性地触发靶组织中的先天性免疫应答

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