| WO2010106488A2 | 2010-09-23 | |||
| WO2010106488A2 | 2010-09-23 |
| FR2937507A1 | 2010-04-30 | |||
| EP1342419A1 | 2003-09-10 | |||
| EP0519458A1 | 1992-12-23 | |||
| US5434182A | 1995-07-18 | |||
| IS2005000026W | 2005-12-16 |
S DONKIN ET AL: "Glycerol as a feed ingredient in dairy rations", PROCEEDINGS OF THE TRI-STATE DAIRY NUTRITION CONFERENCE, 2007, pages 97 - 103, XP055082900, Retrieved from the Internet
J. KABARA; DENNIS M. KABARA; DENNIS M. SWIECZKOWSKI; ANTHONY J, CONLEY; JOSEPH P. SWIECZKOWSKI; ANTHONY J, CONLEY; JOSEPH P. TRUAN, FATTY ACIDS AND DERIVATES AS ANTIMICROBIAL AGENTS, 1972
G. BERGSSON; J: ARNFINNSSON; S. KARLSSON; O. STEINGRIMSSON; H. THOMAR, IN VITRO INACTIVATION OF CHLAMYDIA TRACHOMATIS BY FATTY ACIDS AND MONOGLICERIDES, 1998
G. BERGSSON; J. ARNFINNSON; O. STEINGRIMSSON; H. THORMAR, IN VITRO KILLING OF CANDIDAALBICANS BY FATTYACIDS AND MONOGLICERIDES
| CLAIMS 1. Compositions for zootechnical use having antibacterial activity, comprising C8- C-H organic acid monoglyceride esters in an amount within the range from 10% - 95% and glycerol from 5% to 90% by weight calculated on the total weight of the composition. 2. Compositions according to claim 1 , wherein said Cs - Ci4 organic acid monoglycerides are lauric acid monoglycerides. 3. Compositions according to claim 2, comprising: lauric acid monoglyceride ester 48 - 55% - lauric acid diglyceride ester 25 - 35% lauric acid triglyceride ester 3 - 7% glycerol 8 - 15% 4. Compositions according to claim 3, further comprising C8- C-|4 organic acids in percentages within the range from 1-25% calculated by weight with respect to the other components, and/or essential oils in percentages within the range from 1-5%. 5. Compositions according to claim 3, consisting of: lauric acid monoglyceride ester 50% lauric acid diglyceride ester 30% - lauric acid triglyceride ester 5% glycerol 15% 6. Liquid or solid animal feeds, supplemented with compositions according to claims 1 - 5. 7. Feeds according to claim 6, wherein the compositions are added in amounts within the range from 0.1 to 0.8%, preferably from 0.3-0.6%. 8. Feeds according to claims 6 and 7, wherein said feeds are for feeding pigs, chickens, fish, cattle, sheep, and household pets. |
ORGANIC ACIDS AND GLYCEROL, AND ANIMAL FEEDS COMPRISING TH
Field of the invention
The present invention relates to the field of compositions for animal feeds.
Prior art
As is known, the most frequent problems to affect farm animals and household pets relate to the damage caused by bacterial infections.
These bacterial infections, intestinal infections in particular, often present themselves in the form of severe diarrhoea, which may seriously jeopardise the health of the animal and therefore the profitability of the farm as a whole.
Normally attempts are made to avoid these inconveniences by using medium- chain organic acids, alone or in admixture, such as: caprylic acid (C8H 16 O2), capric acid (CioH2o0 2 ), lauric acid (C12H24O2), myristic acid (C 4 H 2 80 2 ).
There are a number of physiological limits to the action of organic acids as such or in the form of the salt, when used as antibacterial agents in the diets of animals; for example, it is known that in order to express a really antibacterial action organic acids require an acid pH in which under these conditions they are present in the undissociated RCOOH form. This undissociated RCOOH form passes through the cell wall of the micro-organism, once inside which it is dissociated as a function of the intracellular pH. To maintain its intracellular pH, the micro-organism expels H + ; there is therefore an increase in the dissociated amount of the organic acid which, because it remains within the microorganism, kills it.
In the intestines, however, the pH is slightly basic (approximately 7) and there is therefore a fairly limited presence of the undissociated form of the organic acid and consequently a fairly limited antibacterial function.
Patent application WO2010/106488 in the name of the same Applicants discloses that compositions containing organic acid monoglycerides, with up to a maximum of 7 carbon atoms, can be used as antibacterial agents in animal breeding, but nothing is said in the application regarding the possibility of using, for the same purpose, analogous acid monoglycerides, having a number of carbon atoms greater than 7. The technology of micro-encapsulation of organic acids with lipids was developed as a method for transporting the organic acid within the intestine and lowering the pH thereof, but the results thus obtained are fairly unsatisfactory due to the buffering action of the sodium bicarbonate produced by the pancreas as a regulator of the intestinal pH. Furthermore, as well as being ineffective, this technology is also very expensive.
The antibacterial action of some fatty acid monoglycerides has also been investigated in numerous studies, for example:
- J. Kabara, Dennis M. Kabara, Dennis M. Swieczkowski, Anthony J, Conley, Joseph P. Swieczkowski, Anthony J, Conley, Joseph P. Truant 1972 FATTY ACIDS
AND DERIVATES AS ANTIMICROBIAL AGENTS;
- G. Bergsson, J: Arnfinnsson S. Karlsson, O. Steingrimsson, H. Thomar 1998
IN VITRO INACTIVATION OF CHLAMYDIA TRACHOMATIS BY FATTY ACIDS AND MONOGLICERIDES;
- G. Bergsson, J. Arnfinnson, O. Steingrimsson, H. Thormar 201. IN VITRO KILLING OF CANDIDA ALBICANS BY FATTY ACIDS AND MONOGLICERIDES;
- H. Thorma, H. Hilmarsson, G. Bergsson 205. STABLE CONCENTRATED EMULSIONS OF THE 1-MONOGLYCERIDE OF CAPRIC ACID (MONOCAPRIN) WITH MICROBICIDAL ACTIVITIES AGAINST THE FOOD-BORNE BACTERIA CAMPYLOBACTER JEJUNI, SALMONELLA SPP AND ESHERICHIA COLI PCT/IS2005/000026 "STABLE CONCENTRATED ANTI-BACTERIAL EMULSIONS OF MONOCAPRIN IN WATER" INFECTION;
- Hilmarsson, H. Thormar, J.H. Thrainsson, E. Gunnarsson 2006 EFFECT OF GLYCEROL MONOCAPRATE (MONOCAPRIN) ON BROILER CHICKENS: AN ATTEMPT AT REDUCING INTESTINAL CAMPYLOBACTER.
Although all of these studies have produced evidence of a promising, albeit non- exhaustive, line of research there is currently no knowledge of studies proving a specifically antibacterial action of medium-chain fatty acid monoglyceride compositions in combination with glycerol.
Brief description of the drawings
Fig. 1 is an electron microscope view of an aqueous suspension of a composition according to the invention. Summary of the invention
The present invention relates to compositions containing C 8 to C M organic acid monoglycerides in percentage within the range from 10% - 95% and glycerol from 5 to 90% by weight (calculated on the basis of the whole composition) for addition to cereals, feeds, to feedstuffs in general and to drinking water intended for animal consumption.
DETAILED DESCRIPTION OF THE INVENTION
It has now surprisingly been found that compositions containing monoglyceride esters of C 8 to Ci 4 organic acids in combination with glycerol have a strong antibacterial capacity both at an acid (4.5) and at a neutral pH, such as is present in the intestines of animals (pH 7).
In the compositions according to the invention, the monoglyceride esters of organic acids as defined above are present in quantities within the range 0% - 95% and glycerol from 5% to 90% by weight (calculated on the basis of the whole composition); the said quantities are preferably within the range 40% - 95% and 5% - 60%, respectively.
By Cs to Ci4 organic acids is preferably meant the acids: caprylic, capric, lauric, myristic.
Examples of compositions according to the invention are compositions consisting of:
(a)
caprylic acid monoglyceride ester 73 - 83%
caprylic acid diglyceride ester 7 - 10%
caprylic acid triglyceride ester 0.5 - 2%
glycerol 5 -15%
(b)
capric acid monoglyceride ester 45 - 50%
capric acid diglyceride ester 25 - 30%
capric acid triglyceride ester 1 - 5%
glycerol 10 -20%
Particular examples of compositions according to the invention are compositions consisting of: (C)
caprylic acid monoglyceride ester 75%
caprylic acid diglyceride ester 8%
caprylic acid triglyceride ester 1 %
glycerol 16%
(d)
Capric acid monoglycerides 48%
Capric acid diglycerides 28%
Capric acid triglycerides 3%
glycerol 21 %
(e)
lauric acid monoglyceride ester 48 - 55%
lauric acid diglyceride ester 25 - 35%
lauric acid triglyceride ester 3 - 7%
glycerol 8 -15%
(f)
lauric acid monoglycerides 50%
lauric acid diglycerides 30%
lauric acid triglycerides 5%
glycerol 15%
In table 1 below are indicated the antibacterial capacity of the organic acids alone by comparison with that of the compositions according to the invention.
TABLE 1
SUBSTANCE CONC. pH E.coli Salmonella Campylobacter
USED (cfu/ml) typhimurium Jejuni
(cfu/ml) (cfu/ml)
Positive 7 102x10 b 95x10 b 410x10 b control
Positive 4.5 74x10 ¾ 90x10 b 171x10 ¾ control
Caprylic 1 :100 7 12x10 4 80x10 a 33x10 acid
NB: Monocaprylin 75 is composed of: NB: Monocaprin 48 is composed of: 75% caprylic acid monoglycerides 48% capric acid monoglycerides 8% caprylic acid diglycerides 28% capric acid diglycerides 1 % caprylic acid triglycerides 3% capric acid triglycerides
16% glycerol 21 % glycerol
Table 2 compares the in vitro antibacterial action against Clostridium perfrigens of caprylic acid, caprylic acid monoglycerides without free glycerol, and of a mixture of caprylic acid monoglycerides with free glycerol.
Whereas the mixture of caprylic acid monoglycerides with glycerol already shows, in all three
replications, an inhibitory capacity ("no growth") at the 1000 ppm concentration, the caprylic acid
monoglycerides show no inhibitory capacity against the bacterium, while the caprylic acid
shows inhibitory activity only from 3000 ppm.
TABLE 2
Bacteria Chlostridium perfringes CP27
Inoculation concentration; 10 5
Medium; Brain Heart Infusion
Incubation time and growth that had been appeared; + at 24hr, ++ at 37hr and +++ at 96hr - 3 replications for each concentration Negative Positive
Control Control Ppm Caprylic Monocaprylin Caprylic Acid
(PC) acid 75 Monoglycerides
+ 500 + + +
+ + + +
++ + + +
Mean
1.000 + no growth +
+ no growth +
+ no growth +
Mean
1 ,500 + no growth +
+ no growth +
+ no growth +
Mean
2,000 ++ no growth +
++ no growth +
++ no growth +
Mean
2,500 ++ no growth +
++ no growth +
++ no growth +
Mean
3,000 no growth no growth +
no growth no growth +
no growth no growth +
Mean
4,000 no growth no growth +
no growth no growth +
no growth no growth +
Mean Table 3 compares the in vitro antibacterial action against Porcine Salmonella Typhimurium, of pure capric acid, capric acid monoglycerides without free glycerol, and of a mixture of capric acid monoglycerides with free glycerol. Whereas the mixture of capric acid monoglycerides with glycerol (Monocaprin 48) shows, in all three replications, an inhibitory capacity ("no growth") at the 5,000 ppm concentration, the capric acid monoglycerides show an inhibitory capacity against the bacteria from 20,000 ppm, while the capric acid shows inhibitory activity from 15,000 ppm.
TABLE 3
Bacteria Porcine Salmonella Typhimurium,
Inoculation concentration; 10 5
Medium; Brain Heart Infusion
Incubation time and growth that had been appeared; + at 24hr, ++ at 37hr and +++ at 96hr - 3 replications for each concentration
pH 6
Negative Positive
Control Control Ppm Capric Monocaprin Capric Acid
(PC) acid 48 Monoglycerides
+ 500 + + +
+ + + +
++ + + +
Mean
5,000 + no growth +
+ no growth +
+ no growth +
Mean
10,000 + no growth +
+ no growth +
+ no growth +
Mean
15,000 no growth no growth +
no growth no growth + no growth no growth +
Mean
20,000 no growth no growth no growth
no growth no growth no growth no growth no growth no growth
Mean
25,000 no growth no growth no growth
no growth no growth no growth no growth no growth no growth
Mean
30,000 no growth no growth no growth
no growth no growth no growth no growth no growth no growth
Mean
NB: Monocaprin 48 is composed of:
48% capric acid monoglycerides
28% capric acid diglycerides
3% capric acid triglycerides
21 % glycerol
Table 4 compares the in vitro antibacterial action against Porcine Salmonella Typhimurium, of pure lauric acid, lauric acid monoglycerides without free glycerol, and of a mixture of lauric acid monoglycerides with free glycerol. Whereas the mixture of lauric acid monoglycerides with glycerol (Monolaurin 50) shows, in all three replications, an inhibitory capacity ("no growth") at the 500 ppm concentration, the lauric acid monoglycerides show an inhibitory capacity against the bacteria from 25,000 ppm, while the lauric acid shows inhibitory activity from 10,000 ppm. TABLE 4
Bacteria Porcine Salmonella typhimurium,
Inoculation concentration; 10 5
Medium; Brain Heart Infusion
Incubation time and growth that had been appeared; + at 24hr, ++ at 37hr and +++ at 96hr - 3 replications for each concentration
pH 6
Negative Positive
Control Control Ppm Laurie Monolaurin Laurie Acid
(PC) acid 50 Monoglycerides
+ 500 + no growth +
+ + no growth +
++ + no growth +
Mean
5,000 + no growth +
+ no growth +
+ no growth +
Mean
10,000 no growth no growth +
no growth no growth +
no growth no growth +
Mean
15,000 no growth no growth +
no growth no growth +
no growth no growth +
Mean
20,000 no growth no growth +
no growth no growth +
no growth no growth +
Mean
25,000 no growth no growth no growth no growth no growth no growth
no growth no growth no growth
Mean
30,000 no growth no growth no growth
no growth no growth no growth no growth no growth no growth
Mean
NB: Monolaurin 50 is composed of:
50% lauric acid monoglycerides
30% lauric acid diglycerides
5% lauric acid triglycerides
15% glycerol
If preferred, the compositions according to the invention may also contain active principles of essential oils (cinnamic aldehyde, timolol, carvacrol) in percentages within the range 1-20% (calculated by weight based on the total weight of the composition) as conventionally envisaged with these products for feeding to animals because these active principles are soluble in lipids and insoluble in glycerol.
It is known that, when the composition according to the invention is dispersed in water, the glycerol surrounds the actual monoglyceride, forming droplets which enclose the monoglyceride and remain suspended in water (the other active principles which may be added dissolve in the monoglyceride and therefore they too are enclosed in the droplet of glycerol) (see Fig. 1 ).
The compositions according to the invention may be prepared according to the normal processes of esterification of fatty acids that are widely described in the literature, but using a great excess of glycerol (however never less than 200% by weight relative to the weight of the fatty acids used) so as to obtain an elevated quantity of monoglycerides with elevated quantities of free glycerol.
To compositions according to the invention may be added to the animal feeds and/or drinking water in quantities within the range 0.1 to 1.5%, preferably 0.3- 0.6% calculated by weight based on the weight of the feed or drink. The compositions according to the invention are particularly indicated in the feed of pigs, chickens, fish, cattle, sheep, and domestic pets.
EXAMPLE 1
Sixty Dandread race piglets at the age of 5 weeks were assigned to two groups A) - Control group and B) Treated group of thirty piglets each, divided into 6 sties of ten animals each. After the first 10 days of habituation to the accommodation, all the animals were inoculated by mouth with Salmonella Typhimurium, which had been isolated at the Forli Institute of Zooprophylaxis from faecal samples from infected pigs, at a dose equal to 7x10 7 cfu.
The following day, some of the animals in each sty had diarrhoea.
Faecal samples were obtained on the fifth day post-infection, and the bacterial count was found to be equal to 220,000 cfu in control group A) and 210,000 cfu in treated group B). Group B) was then treated with a mixture composed of:
Caprylic acid monoglycerides = 75%
- Caprylic acid diglycerides = 8%
Caprylic acid triglycerides = 1 %
Glycerol = 16%
administered in the drinking water at a dose of 0.5% for three days. On the third day following the treatment, new faecal samples were obtained for analysis of the bacterial count. The control group A) had a mean cfu number equal to 200,000, whereas in the treated group B), the cfu number was equal to 300. The use of the mixture "caprylic acid esters and glycerol" in the percentages indicated resulted in a decline in the salmonella cfu's 3 logarithms to the base 10 with administration for 3 days. This fact confirms the bactericidal effect of the mixture.
EXAMPLE 2
This field experiment was conducted on an Italian farm which had health problems such as Brachispira spp. enteritis. 1 ,530 Dandread pigs weighing approximately 25 kg (68 days old) were divided into two groups: control group A) and treatment group B), consisting of 760 and 770 animals respectively.
The two groups were given feed of identical formulation, with the exception of the following constituents: the feed of the control group was supplemented with Lincomycin 250 ppm and Doxycycline 250 ppm for the first 14 days of the experiment, and with lincomycin alone for the remaining period. Treatment group B) receive no antibiotics in the feed, but a mixture of "lauric acid esters and glycerol" of the following composition: - Lauric acid monoglycerides = 50%
Lauric acid diglycerides = 30%
Lauric acid triglycerides = 5%
Glycerol = 15%
administered in the feed in a proportion of 0.5% in place of the 0.5% soya oil. The experiment lasted 63 days. The growth and feed efficiency results are summarised in Table 5 below.
TABLE 5
Group A) Group B) - Delta
Control Lauric acid
esters and
glycerol
No. of animals 760 770
Age at the start of the 68 68
experiment (days)
Age at the end of the 131 131
experiment (days)
Mean weight at the 24 24
start of the experiment
(kg)
Mean weight at the 61 .10 62.51
end of the experiment
(kg)
No. of dead animals 4 2 2
No. rejected 4 2 2
Mean daily weight 0.588 0.610 + 0.022
gain (kg) Feed consumed, total 53,520 53,220 - 300 in kg
Meat produced in kg 19,400 20,220 + 820
Feed conversion 2.75 2.63 0.12 index
The diarrhoeal episodes were much more limited in the treatment group B). The growth parameters, the feed conversion index of group B) are comparable, and tend to be improved by comparison with those of control group A), the diet of which contained the antibiotics mentioned above. The mixture of "lauric acid esters and glycerol" allowed the demonstrated enteric pathogens to be controlled without the use of antibiotics. The experiment demonstrated an antibacterial effect of the "lauric acid esters and glycerol" mixture with a consequent improvement in intestinal health.