CA1116520A - Increasing the production of propionic acid - Google Patents

Abstract

26,750 A B S T R A C T

A method for improving the production of propionic acid and thus increasing the propionate:acetate ratio in the course of the microbiological conversion of cellulose and starch containing feedstuffs to short chain volatile fatty acids in the ruminant animal by the use of antibiotic AV290, antibiotic AV290 sulfate, an antibiotic AV290-syntan complex, an antibiotic AV290-alkyl sulfate complex and/or an anti-biotic AV290 alkylated derivative.

Description

This inven-tion relates to a method for improving the production of propionic acid and thus increasing the propionate:
acetate ratio in the course of the microbiological conversion of cellulose and starch containing feedstuffs to short chain volatile fatty acids in the ruminant animal. This invention further relates to the use of antibiotic AV290, antibiotic AV290 sulfate~ an antibiotic AV290-syntan complex, an antibiotic AV290~alkyl sulfate complex, and/or an antibiotic AV290 alkylated derivative for increasing the production of propionic acid and suppressing the production of acetic acid during the micro-biological digestive processes taking place in the animal rumen.
This invention also relates to novel compositions of matter useful in favorably altering the acetate:propionate ratio in the course of the microbiological digestive processes taking place in the rumen of animals such as cattle, sheep and goats. More particularly, it relates to compositions in oral dosage unit form comprising from about 10 mg. to about 1 gram per daily dosage unit of one or more of the following active ingredients tin any proportions) which are useful for enhancing the production of propionates in said animals rumen while the animals feed is digested:
(1) antibiotic A~290 whose preparation and properties are disclosed in United States Patent 3,338,786;

(2) antibiotic AV290 sulfate which is disclosed in United States Patent 3,855,410;

(3) an antibiotic AV290-syntan complex prepared as described in United States Patent 3,832,462;

(4) an antibiotic AV290-alkyl sulfate complex derived by treatment of the antibiotic with an alkali metal alkyl sulfate as set forth in United States Patent 3,856,937;

(5) an antibiotic AV290 alkylated derivative derived by treatment of the antibiotic with a lower alkyl halide as .'', ~

fiS~a~) defined and described in United States Patent 3,954,973.
The invention also rela-tes to a stable premix com position for the treatment and prevention of ketosis in ruminants comprising a feed premix containing from about 1% to about 90% by weight of an active ingredient selected from the group consisting of antibiotic AV290, antibiotic AV290 sulfate, an antibiotic AV290-syntan complex, and antibiotic AV290-alkyl sulfate complex, an antibiotic AV290 alkylated derivative and mixtures thereof in any proportion.
The major nutritive portion of the ruminants diet consists primarily of polysaccharides such as cellulose and starches. These primary sources of said animals energy requirements~ and of basic intermediates for tissue building processes, are hydrolized in said animals rumen by microbio-logical processes to monosaccharides, primarily to glucose.
The thus formed glucose is then further degraded by enzymatic processes to pyruvic acid and derivatives thereof. These in turn are further converted through various enzymatic processes to acetic acid and propionic acid and derivatives thereof.
Simultaneously, and also in the rumen, some of the acetic acid is converted to butyric acid. Although butyric acid is the component most efficiently metabolized by ruminants while acetic acid is the least efficiently utilized product of the above referred-to digestive processes, the formation of butyric acid in the course of said microbiological processes is energetically not very favorable. As stated above, butyric acid is formed from acetic acid, and since one mole of glucose yields two moles of acetic acid and two moles of methane and/or carbon dioxide gas, and since two moles of acetic acid are consumed in the for-mation of one mole of butyric acid, therefore, for every mole ofbutyric acid formed, two moles of methane and/or carbon dioxide gas are generated, representing a considerable energy loss.

Additionally, it is known that production of acetic acid (and/or acetates) in larger than normal amounts may lead to the produc-tion of ketone bodies (acetoacetate, acetone and ~-hydroxy butyrate) which can cause ketosis in ruminants, especially if said animals are under stress. It has been found that the supplementary feeding of propionic acid (and/or propionates) is beneficial for minimizing the effects of ketosis. A further advantage of feeding a diet high in propionic acid to ruminants such as cattle, sheep and goats is to lower the incidence of ketosis.
Thus it would be of advantage both to the animal grower and feedlot operator if the microbiological processes of the rumen could be altered so that the production of propionic acid from carbohydrates is enhanced while that of acetic acid is suppressed.
The novel method of the present invention comprises orally administering to ruminants such as cattle, sheep and goats antibiotic AV290 or the above-identified salts, complexes, and derivatives thereof in amounts sufficient to promote the formation of propionates while suppressing the formation of acetates.
Advantageously, we have found that by orally administer-ing to said ruminants antibiotic AV290 or the above-identified salts, complexes, or derivatives thereof (and/or mixtures thereof) in amounts of from about 10 mg. to about one gram per head per day or from about 0.05 mg. to about 50.0 mg./kg. of body weight per day, the acetate-propionate ratio is altered in favor of the propionates in the course of the microbiological digestive processes taking place in the rumen of said animals. Thus, by promoting the formation of propionates and suppressing the formation of aceta-tes, the incidence of ketosis among said .
rUmlnantS lS mlnlmlZed.

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1rrhe active compounds of this invention, an-tibiotic AV290 and the salts, complexes, and derivatives thereof, can be conveniently adminis-tered to ca-t-tle, s~leep and goats as feed addltives in amounts ranging from about 8 grams to about one kilogram per ton of feed to provide 10 mg. to one gram per head per day or 0.05 mg. to 50.0 mg./kg. of body weight per day of said compound. Should it be desired, antibiotic AV290 and its pharmaceutically acceptable salts, complexes, and derivatives may also be formulated for oral administra-tion as feed premixes~ concentrates, tablets, pills and bol-uses, which may be prepared by accepted and well known meth-ods, using pharmaceutically acceptable carriers, diluents, binding and lubricating agents.
The present invention is further illustrated by the set forth below:
Example 1 Evaluation of the in vitro efficacy of the antibiotic AV2~0 for altering the acetic acid:propionic acid ration during fermentation in rumen fluid~

.
The steers used in the e~periment have surgically installed fistulas that open into the rumen. The steers are offered water and a daily ration of alfalfa hay ad libitum as well as 10 lbs. of a standard 15% protein grain ration of the following composition:
25 Component Percent by Weight Corn32.7 Barley10.0 Oats 7 5 Molasses 10.0 30 Soybean Oil Meal (48~ protein) 13.8 52~

1 seet Pulp 2.5 Corn Gluten Feed 12.5 Distillers Grain 7.5 Trace Mineral Mix 0.05 5 Salt 1.0 Dicalcium Phosphate 2.0 Method A sample of rumen fluid (500 ml.) is aspirated into a flask and strained -through four layers of cheesecloth to remove coarse feed particles. Ten ml. of the clarified rumen fluid is introduced into an incubation flask contain-ing 200 mg. of a substra-te of the following composition:
Component Percent by Weight Corn Starch 68 Soybean Oil Meal 15 Alpha Cellulose 17 The drug to be tested ~AV290) is mixed into this feed substrate at levels of from 40 to 5,000 ppm. These levels are equivalen-t to 8 to 100 ug of drug per flask or 0.4 to 50 ug per ml. of final incubation fluid. To the above mixture of drug/feed substrate and rumen rluid, 10 ml.
of a buffer of the following composition is added:
Component grams/liter Sodium bicarbonate 19.6 Disodium hydrogen phosphate heptahydrate 14.0 Potassium chloride 1.14 Sodium chloride 0.94 Magnesium sulfate heptahydrate 0.24 2~

l Calcium chlori~e 0.08 The pH of the thus prepared fermentation mixture is 7.2. Nex-t, each flask is flushed with carbon dioxide to produce an anaerobic atmosphere and the fermentation mix-tures are incubated for 24 hours in a 37C. constant temper-ature bath while the flasks are shaken. Simultaneously, un-treated control samples are also incubated~ Additionally several samples are taken at the start of the incubation per-iod and acidified with 6N hydrochloric acid to pH 2.0 to serve as zero-time control samples. After 24 hours, the fermenta-tion in the treated and control samples is stopped by the addition of enough 6N hydrochloric acid to lower the pH of the fermentation mixtures to 2Ø A portion of each sample is then centrifuged at 2,000 x g for 15 minutes and the clear supernatant removed for analysis by gas chromatography to determine concentrations of acetic and propionic acid, and butyric acid in the samples. The data obtained are summar-ized in Table I and show the molar percent of each volatile fatty acid (VFA) produced during fermentation in the presence of antibiotic AV290 as well as the ratio, acetic to propionic acid; while in Table II the same data are expressed as per-cent change from the control.

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Example 2 Evaluation of the in vivo efficacy of antibiotic AV290 for .
altering the acetic acid:propionic acid ratio during fermen-tation in the rumen of sheep.

.Twenty-four whether lambs, weighing approximately 40 kg. each, are randomly allotted to four groups of 6 lambs each. One group serves as unmedicated control and receives water and the following diet ad libitum:

Component Percent by Weight Ground corn cob 35.0 Dehydrated alfalfa meal 20~0 Ground corn lg.4 Cane molasses 12.0 Soybean oil meal (48% protein) 12.0 Dicalcium phosphate 1.0 Iodized salt 0.5 Trace mineral mix 0.1 Vitamin A and D3 premix 0.02 Corn oil 0.01 The other three groups receive water and the same diet ad libitum, except that 20, 50 or 100 ppm. of AV290 -lauryl sulfate is incorporated into the diet of the respec-tive groups. After 19 days on the diet, the sheep are sac-rificed and samples of rumen fluid are obtained and analyzed as in Example 1. The data obtained are summarized in Table III and show the molar percent of each volatile fatty acid 1 (VFA) produced in the presence of AV290 as well as the ratio of acetic to propionic acid; while in Table IV the same data are expressed as percent change from the control.

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Th~ above data, especicllly those found in Table IV, clearly show that AV290 lauryl sulfate is very effective in increasing the propionic acid production while suppressing the acetic acid production in vivo, when orally administered to sheep.
Example 3 Evaluation of the efficacy of AV290 for altering the acetic acid:propionic acid ratio during fermentation in the rumen , of cattle.
Eighteen feedlot cattle, weighing approximately 350 kg. each, are randomly allotted to three groups of 6 animals each. Water and the ration described in Example 2 are offered to the animals ad libitum. One group serves as unmedicated control, while groups two and three each receive lS in their daily diet 33 ppm. or 99 ppm. of AV290 lauryl sulfate, respectively.
The feeding trial is conducted for a period of 56 days, with rumen fluid samples being obtained from the animals on days 14, 28 and 56 aft~r the beginning of the 2d trial. The rumen fluid samples are handled and analyzed for volatile fatty acids (VFA) by the method of Example l.
The thus obtained data are summarized in Table V, while in Table VI the same data are expressed as percent change from the controls.

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It can be clearly seen from Table V and especially from Table Vl that the oral adrninistration of AV290 lauryl sulfate to cattle via their diet very effectively alters the VFA production during fe~nentation in the r~nen of said animals in favor of propionic acid.

Claims (8)

1. THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
   PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
   
   l. A therapeutic composition in oral dosage unit form useful for the treatment of ketosis in ruminants comprising from about 10 mg. to about one gram per daily dosage unit of an active ingredient selected from the group consisting of anti-biotic AV290, antibiotic AV290 sulfate, an antibiotic AV290-syntan complex, an antibiotic AV290-alkyl sulfate complex, an antibiotic AV290 alkylated derivative, and mixtures thereof in any proportion, and an edible carrier.
2. 2. A composition according to Claim 1 wherein the active ingredient is antibiotic AV290 sulfate, an antibiotic AV290-syntan complex or antibiotic AV290 alkylated with ethyl bromide.
3. 3. A composition according to Claim l wherein the active ingredient is a 1:1 mixture of antibiotic AV290-cetyl sulfate and antibiotic AV290 alkylated with iso-propyl iodide.
4. 4. A composition according to Claim 1 wherein the active ingredient is AV290 sulfate.
5. 5. A composition according to Claim 1 wherein the active ingredient is AV290 lauryl sulfate.
6. A stable premix composition for the treatment and prevention of ketosis in ruminants comprising a feed premix containing from about 1% to about 90% by weight of an active ingredient selected from the group consisting of antibiotic AV290, antibiotic AV290 sulfate, an antibiotic AV290-syntan complex, and antibiotic AV290-alkyl sulfate complex, an antibiotic AV290 alkylated derivative and mixtures thereof in any proportion.
7. 7. A premix composition according to Claim 6 wherein the active ingredient is AV290 sulfate.
8. 8. A premix composition according to Claim 6 wherein the active ingredient is AV290 lauryl sulfate.