Methods and compositions for the control of coccidiosis
OwnerCharles S. Schasteen, Jackie Green +4inventors
InventorsCharles S. Schasteen, Jackie Green, Farooq Uraizee, Lance Bull, Mary Ann Pfannenstiel, Tony Allington
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Abstract
Methods are provided for the sporulation, sterilization and storage of coccidial oocyst which are characterized by an absence of the highly toxic chemical potassium dichromate. Also provided are compositions containing sporulated oocysts which are free of potassium dichromate.
Claims143
A composition for the prevention or control of coccidiosis comprising viable wild type sporulated oocysts of at least one species of protozoa known to cause coccidiosis, wherein said composition is sterile and contains at least about 10,000 oocysts per milliliter and less than about 0.8% by weight of alkali metal dichromate.
A composition for the prevention or control of coccidiosis comprising viable wild type sporulated oocysts of at least one species of protozoa known to cause coccidiosis, wherein said composition is sterile and contains at least about 300 oocysts per milliliter and less than about 0.002% by weight of alkali metal dichromate.
A composition for the prevention or control of coccidiosis comprising viable wild type sporulated oocysts of at least one species of protozoa known to cause coccidiosis, wherein said composition is sterile and contains less than about 5.0×10−3 μg of alkali metal dichromate per oocyst.
A method for producing a composition for the prevention or control of coccidiosis comprising:
- collecting manure from host animals wherein said manure contains oocysts known to cause coccidiosis;
- diluting said manure in an aqueous medium to create a slurry;
- separating unwanted fecal matter from said slurry and collecting the aqueous fraction containing oocysts;
- subjecting said aqueous fraction to solid/liquid phase centrifugal-based separation and collecting the solid phase;
- combining a dense aqueous liquid with said collected solid phase wherein said dense liquid has a density greater than about 1.09 g/ml and wherein the oocysts are buoyant;
- subjecting the combination of said dense aqueous liquid and collected solid phase to centrifugation and collecting the dense liquid fraction containing oocysts;
- diluting said dense liquid fraction to a specific gravity wherein the oocysts are no longer buoyant;
- separating oocyst solids from said diluted liquid fraction by centrifugal-based separation and re-collecting the solid phase.
A method of separating oocysts from a liquid suspension by the use of a hydrocyclone.
A method for isolating oocysts comprising:
- collecting manure from host animals wherein said manure contains oocysts known to cause coccidiosis;
- diluting said manure in an aqueous medium to create a slurry;
- separating unwanted fecal matter from said slurry and collecting the aqueous fraction containing oocysts;
- subjecting said aqueous fraction to solid/liquid phase centrifugal-based separation by means of a hydrocyclone.
A method for isolating oocysts comprising:
- collecting manure from host animals wherein said manure contains oocysts known to cause coccidiosis;
- diluting said manure in an aqueous medium to create a slurry;
- separating unwanted fecal matter from said slurry and collecting the aqueous fraction containing oocysts;
- subjecting said aqueous fraction to solid/liquid phase centrifugal-based separation and collecting the solid phase;
- combining a dense aqueous liquid with said collected solid phase wherein said dense liquid has a density greater than about 1.09 g/ml and wherein the oocysts are buoyant;
- subjecting the combination of said dense aqueous liquid and collected solid phase to centrifugation and collecting the dense liquid fraction containing oocysts;
- diluting said dense liquid fraction to a specific gravity wherein the oocysts are no longer buoyant;
- separating oocyst solids from said liquid phase by means of a hydrocyclone and re-collecting the solid phase.
A method for inducing sporulation of oocysts comprising:
- introducing into an aqueous sporulation medium oocysts of at least one species of protozoa known to cause coccidiosis;
- incubating said oocysts in said aqueous sporulation medium, thereby causing sporulation of oocysts;
- and
- introducing an oxidizing agent into said medium at a rate sufficient to maintain the average dissolved oxygen content during sporulation at at least 30%.
A method as set forth in 92 further comprising substantially separating said sodium hypochlorite from the sporulated oocysts by filtration.
A method for inducing sporulation of oocysts comprising:
- introducing into an aqueous sporulation medium oocysts of at least one species of protozoa known to cause coccidiosis;
- incubating said oocysts in said aqueous sporulation medium, thereby causing sporulation of oocysts;
- and
- introducing an oxidizing agent having a standard reduction potential of at least about 0.5 V into said medium at a rate sufficient to maintain the oxidation potential of said medium equivalent to the oxidation potential of a medium containing dissolved molecular oxygen in concentration of at least 30% of saturation during sporulation;
- said medium containing less than about 0.8% by weight alkali metal dichromate during incubation of said oocysts.
A method for monitoring sporulation of oocysts comprising:
- incubating viable oocysts in an aqueous sporulation medium;
- and
- during incubation, monitoring said medium to detect a change in at least one of the following parameters:
- (i) dissolved oxygen content;
- (ii) pH;
- (iii) rate of introduction of oxidizing agent into said medium;
- (iv) flow rate of acid or base into said medium.
A kit for the prevention or control of coccidiosis comprising,
- a composition containing, sterile, viable, sporulated oocysts of at least one species of protozoa known to cause coccidiosis, said composition containing 0.8% by weight of alkali metal dichromate;
- and
- instructions for administration of said composition to an animal.
A composition for the storage of sporulated oocysts comprising an aqueous diluent and a bactericide, said composition characterized as substantially free of alkali metal dichromate wherein said composition is characterized as having:
- a diluent comprising 0.5× phosphate buffered saline;
- a pH from about 5.0 to about 8.0;
- and
- wherein said bactericide is selected from the group consisting of an alkali metal perchlorate, an alkali metal hypochlorite, hydrochlorous acid, sodium hydroxide and antibiotics.
A composition for the storage of sporulated oocysts comprising:
- 5× PBS;
- and
- about 30 μg/ml gentamicin,
- said composition characterized as substantially free of alkali metal dichromate, and wherein said composition is characterized in that oocysts in contact with said composition decrease in viability no more than about 20% over a period of at least about 26 weeks at about 5° C.
Dependencies are inferred from the claim text.