Patent Description:
Vitamin B<NUM> is unique among the vitamins in that it contains not only an organic molecule, but also the essential trace element cobalt. Vitamin B<NUM> is not made by either plants or animals and can be synthesized by only a few species of microorganisms. Bacteria in the human intestinal tract can make enough Vitamin B<NUM> for normal daily requirements from inorganic cobalt salts in the diet. Vitamin B<NUM> is also made in large amounts by rich populations of bacteria in the rumen of ruminant animals and in the cecum of other herbivorous species.

Vitamin B<NUM> participates in many biochemical processes that are essential for life. It acts as a co-enzyme for several enzymes which catalyze the shift of a hydrogen atom from one carbon atom to an adjacent one in exchange for an alkyl, carboxyl, hydroxyl or amino groups. Deficiency of Vitamin B<NUM> results in the development of the serious disease pernicious anemia. Pernicious anemia, as the name implies, involves a low concentration of hemoglobin resulting from the condition, but the effects also include serious disturbances of the central nervous system that may result in abnormal sensations, motion, and in humans, thought.

Vitamin B<NUM> is known to have a positive effect on lactation performance of dairy cows, see for example the <NPL>et al. which shows that in early lactation the supply of Vitamin B<NUM>, if deficient, limits the lactation performance of cows. Since cobalt is needed in animal nutrition as a key component to Vitamin B<NUM> production it therefore follows that ruminants need adequate dietary supply of cobalt for effective animal nutrition and efficient lactation performance. In ruminants, the soluble portion of the ruminal material (solids and liquids) is turned over faster than the solids. This means that highly soluble sources of cobalt will remain in the rumen less time than insoluble forms. This disclosure relates to the ability of a ruminant's bacteria to produce Vitamin B<NUM>. The need for increased B<NUM> production can be seen by the increased performance when B<NUM> is injected, see Journal Dairy Science article previously cited.

<CIT> discloses a pellet for administration to ruminants for providing a biologically active substance over an extended period of time. The pellet can contain <NUM>% cobaltic oxide and <NUM>% clay.

<NPL> discloses a mixed cobalt source composition for feeding to cows comprising <NUM>/kg Co-glucoheptonate and <NUM>/kg Co from an inorganic Co source.

<NPL> discloses a mixed cobalt source composition for feeding to cows comprising <NUM>% DM of a trace mineral source comprising Co-sulfate and <NUM>% DM of a complexed mineral source comprising Co-glucoheptonate.

Horizons No. <NUM>, May <NUM>, p. <NUM>-<NUM>, www. org/content/organic-trace-minerals-improving-livestock-production discloses the utilization of Co by sheep fed dietary levels of <NUM>-<NUM>/kg from cobalt-sulphate and cobalt-glucoheptonate. <CIT> discloses a feed pellet for increasing ruminal microbial activity. The feed pellet is formed of a complex of polysaccharides of low ruminal solubility suspended within the complex and a combination of chemicals which increase ruminal microbial activity over extended periods of time as the chemicals are slowly released into the ruminal fluids.

In one aspect of the present invention there is provided a mixed cobalt source composition for feeding to ruminants to control rumen release rate of cobalt for conversion by rumen bacteria to vitamin B<NUM> according to claim <NUM>.

The composition of the present invention provides a unique source of cobalt for enhanced production of Vitamin B<NUM> in the rumen. It has the advantage of a slow release source of cobalt combined with a fast release source of cobalt. The fast cobalt ion release source which is a soluble source of cobalt is turned over faster in the rumen than the solid cobalt sources are turned over. Thus slower release , for example longer polymer strands of cobalt ligands, are digested over a longer period of time, giving slower release of the cobalt. In this way, a portion of the cobalt mixture is soluble and another portion substantially insoluble; the insoluble portion will remain in the rumen longer. As the bacteria consume, for example, a polysaccharide of cobalt, it will release the cobalt for B<NUM> production. Alternatively, non-rumen degradable polymers like polyacrylic acid would just slowly release cobalt. Surprisingly such control or slow release of cobalt to the rumen bacteria in combination with a faster release is shown by data to have greater B<NUM> production than an all soluble cobalt source, or an all insoluble source.

This invention involves a mixture of a quick release cobalt source and a slow release cobalt source as defined in claim <NUM>, which when fed to a ruminant animal, allows some cobalt ions (the quick release) to immediately be used by the rumen bacteria in making Vitamin B<NUM>, with the balance to slowly release additional cobalt ions as it is more slowly metabolized by the microorganisms of the rumen. Studies have surprisingly demonstrated this combination of quick release and slow release cobalt sources results in more efficient and effective production of Vitamin B<NUM> than either source alone. Not wishing to be bound by any theory, it is believed this occurs because the quick release passes through the rumen faster than the bacteria can metabolize it to Vitamin B<NUM> thus much of it "wasted". In contrast the slow release source of cobalt occurs as the rumen microorganisms gradually metabolize larger molecules of the more insoluble polymers releasing the cobalt to make Vitamin B<NUM>. In the case of all slow release time is "wasted" with limited cobalt available to result in less B<NUM>.

Quick release sources of cobalt include soluble inorganic and organic sources of cobalt such as cobalt chloride, cobalt sulfate, cobalt acetate and other fast release (soluble) cobalt monomer sugar complexes like for example cobalt glucoheptonate or cobalt gluconate, as shown in the assignees earlier <CIT>.

Slow release cobalt sources include polymer complexes with pendant carboxylic acid groups, all of which are largely insoluble.

The quick release cobalt source allows the cobalt to immediately be available to the bacteria from the liquid in the rumen, although much of it passes through so quickly that it cannot all be used. The slow release cobalt source remains in the rumen and passes through only when the microorganisms metabolize the polymer or the salt slowly releases the cobalt which results in a gradually available cobalt source for the making of Vitamin B<NUM> or a gradual release of cobalt from the insoluble form.

The key is to have some polymer bound soluble cobalt and some portion of that being insoluble. This creates a situation where the soluble fraction can immediately start being utilized by the bacteria. As the fluid is turned over in the rumen the cobalt available to the bacteria in an all soluble (inorganic or low molecular weight ligands) will be removed from the rumen. By this time the bacteria will have been degrading the polymer from the polymer bound cobalt releasing this cobalt fraction to the rumen fluid. In essence this "insoluble" fraction is a controlled release source of cobalt. Surprisingly it is best to have both types of cobalt present to optimize vitamin B12 production. The polymer bound insoluble portion can be derived from any acid containing polymer. The prime examples are pectin (part of the present invention) and alginic acid (not part of the present invention).

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 338mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

<FIG> compares the milk production where the only difference in the animal feed is the cobalt source. The CoPro source is cobalt glucoheptonate. This is a soluble monomer sugar ligand to the cobalt. When we used an example of the new cobalt source being <NUM>% cobalt chloride and <NUM>% cobalt pectin an increase in milk production was noted.

This milk production increase was statistically significant.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 170mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 112mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 085mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution alginic acid (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark yellow brown suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 338mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution alginic acid (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark yellow brown suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 168mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution alginic acid (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark yellow brown suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 112mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution alginic acid (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark yellow brown suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 085mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution <NUM>% polyacrylic acid (<NUM>, <NUM> mols of carboxylic acid subunits) is added in one portion. The clear solution is allowed to stir for <NUM> minutes at which point CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 102mols) is added in one portion. The pink suspension is stirred for an additional one hour at room temperature and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution <NUM>% polyacrylic acid (<NUM>, <NUM> mols of carboxylic acid subunits) is added in one portion. The clear solution is allowed to stir for <NUM> minutes at which point CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 051mols) is added in one portion. The pink suspension is stirred for an additional one hour at room temperature and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution <NUM>% polyacrylic acid (<NUM>, <NUM> mols of carboxylic acid subunits) is added in one portion. The clear solution is allowed to stir for <NUM> minutes at which point CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 034mols) is added in one portion. The pink suspension is stirred for an additional one hour at room temperature and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. At this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 169mols) is added to the reaction in one portion and the pH of <NUM> is consequently reduced to <NUM>. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels and molecular weight.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl (<NUM>, 126mmols) acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 170mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. At this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 112mols) is added to the reaction in one portion and the pH of <NUM> is consequently reduced to <NUM>. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels and molecular weight.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. At this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 085mols) is added to the reaction in one portion and the pH of <NUM> is consequently reduced to <NUM>. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels and molecular weight.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl (<NUM>, 258mmols) acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 169mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl (<NUM>, 516mmols) acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 169mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

To <NUM> of dI water is added sodium hydroxide (<NUM>, <NUM>. To this alkaline solution pectin (<NUM>, <NUM> mols of carboxylic acid subunits) is added slowly so as to disperse finely upon the surface of the solution. The fine dispersion quickly becomes a dark orange suspension which is allowed to stir for <NUM> hours at <NUM>. The pH is adjusted to <NUM> with <NUM> HCl (<NUM>, 130mmols) acid at this point solid CoCl<NUM> <NUM><NUM>O (<NUM>, <NUM>. 26mols) is added to the reaction in one portion. The resulting pink suspension is stirred for an additional <NUM> hour and then dried at <NUM> for <NUM> hours. The final blue powder is homogenized and analyzed for cobalt levels.

The pre and post treatment data after receiving the given cobalt source by sheep is shown in <FIG>. In <FIG> three cobalt sources were studied. CSK16254 was <NUM>% CoCl2 and <NUM>% cobalt pectin (Example <NUM>) (not part of the present invention), CSK16255 was <NUM>% CoCl2 and <NUM>% cobalt pectin (Example <NUM>) according to the present invention, CSK16256 was <NUM>% cobalt pectin (Example <NUM>) (not part of the present invention). This study demonstrates that having a combination of soluble and monomeric ligand with a polymer bound cobalt source is advantageous over only the cobalt polymer bound form.

<FIG> and the data shown reflect the change in B<NUM> levels in sheep. The pretreatment is before the addition of the cobalt treatments and the post is after the cobalt treatments. CSK17049 is <NUM>% cobalt glucoheptonate and <NUM>% cobalt pectin from example <NUM> (not forming part of the present invention), CSK17050 is <NUM>% cobalt chloride and <NUM>% cobalt pectin from example <NUM> (not forming part of the present invention), and CSK17051 is <NUM>% cobalt chloride and <NUM>% cobalt alginic acid from example <NUM> (not forming part of the present invention). The data demonstrates that all of these combinations give a strong increase of vitamin B<NUM> in the sheep.

<FIG> and the data shown reflect the average change in B<NUM> levels from pre and post cobalt treatments in sheep. The pretreatment is before the addition of the cobalt treatments and the post is after the cobalt treatments. CSK17057 is <NUM>% cobalt chloride and <NUM>% cobalt pectin from example <NUM> (forming part of the present invention), CSK17058 is <NUM>% cobalt chloride and <NUM>% cobalt pectin from example <NUM> (forming part of the present invention), and CSK17059 is <NUM>% cobalt chloride and <NUM>% cobalt pectin from example <NUM> (not forming part of the present invention). The data demonstrates that all of these combinations give a strong increase of Vitamin B<NUM> in the sheep.

<FIG> and the data shown reflects the average change in B12 levels from pre and post cobalt treatment in sheep. The pretreatment is before the addition of the cobalt treatments and the post is after the treatment. CSK17139 is <NUM>% cobalt chloride and <NUM> % cobalt polyacrylic acid from example <NUM> (not forming part of the present invention). CSK17140 is <NUM>% cobalt chloride and <NUM>% cobalt polyacrylic acid from example <NUM> (not forming part of the present invention). CSK17141 is <NUM>% cobalt chloride and <NUM>% cobalt alginic acid from example <NUM> (not forming part of the present invention). The data demonstrates that all of these combinations give a strong increase of Vitamin B12 in the sheep.

Claim 1:
A mixed cobalt source composition for feeding to ruminant animals to control rumen release rate of cobalt for conversion by rumen bacteria to vitamin B<NUM> comprising:
a quick release source of cobalt and a slow release source of cobalt;
wherein the mixed cobalt source is <NUM>% by weight of the quick release source of cobalt and <NUM>% by weight of the slow release source of cobalt;
and wherein the slow release source of cobalt is cobalt pectin and the quick release source of cobalt is cobalt chloride.