EP3979812A1 - Antimicrobial combination for use in animal feeds - Google Patents
Antimicrobial combination for use in animal feedsInfo
- Publication number
- EP3979812A1 EP3979812A1 EP20730273.8A EP20730273A EP3979812A1 EP 3979812 A1 EP3979812 A1 EP 3979812A1 EP 20730273 A EP20730273 A EP 20730273A EP 3979812 A1 EP3979812 A1 EP 3979812A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- cinnamaldehyde
- animal feed
- terrestrial
- ethylene glycol
- aquatic animals
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
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- 230000000845 anti-microbial effect Effects 0.000 title description 8
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- DHKHKXVYLBGOIT-UHFFFAOYSA-N acetaldehyde Diethyl Acetal Natural products CCOC(C)OCC DHKHKXVYLBGOIT-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
Definitions
- Antimicrobial combination for use in animal feeds
- the present invention relates to the field of animal feed additives. Particularly, it relates to a combination of cinnamaldehyde derived compounds that can be added in animal feeds, and to the combination for use in the prophylactic and/or therapeutic treatment of bacterial infections of terrestrial and aquatic animals. It also relates to a process for the preparation of animal feeds with improved stability and traceability of this antimicrobial combination.
- Cinnamaldehyde (CAS 14371-10-9), which is extracted from cinnamon, is a natural compound with activity against bacteria and an immunomodulatory function. Particularly, it is known the use of cinnamaldehyde as an antimicrobial agent and immunostimulant to protect against bacterial infections in fish and shrimp species in aquaculture, as well as in livestock. Thus, cinnamaldehyde has been used as an additive in feed for aquatic animals including fish, such as salmon, and shrimp for protecting the animals against infections caused by pathogenic or opportunistic microorganisms and, as a consequence reducing mortality as well as to improve the feed conversion ratio through modulation of the gut microbiota.
- cinnamaldehyde has been known to have a low stability in animal feeds, since losses of around 30-50% (or even higher, depending on the thermal treatment of feed) occur in a short period of time such as in 20 to 40 days. Additionally, given the high reactivity of cinnamaldehyde, its analytical determination in animal feeds, particularly in animal feeds comprising a high protein content, provide inaccurate results and with a high variability depending on the storage time of the sample. As a consequence, traceability of this component in animal feeds is a problem, due both to its interaction with proteins and to losses related to its low stability. Besides, the low stability of cinnamaldehyde in animal feeds implies a diminution of its initial effectiveness.
- Document W02004091307 discloses an animal feed additive, for aquatic or terrestrial animals, which has a plant-derived bioactive primary composition and a plant-derived bioactive secondary composition.
- a plant-derived bioactive primary composition and a plant-derived bioactive secondary composition.
- bioactive compounds both of the primary and of the secondary compositions cinnamaldehyde ethylene glycol acetal is mentioned. Nevertheless, no specific composition comprising this compound is disclosed.
- cinnamaldehyde ethylene glycol acetal (CAS 5660-60-6) has a much higher stability than cinnamaldehyde, the inventors have verified that the acetal derivative on their own has a lower antimicrobial and immunostimulant activity than cinnamaldehyde to protect terrestrial and aquatic animals against pathogenic infections.
- the present inventors in an attempt for developing an improved animal feed formulation, have found that by incorporating a combination of cinnamaldehyde and cinnamaldehyde ethylene glycol acetal in an animal feed, particularly in a feed having a high protein content, such us is characteristic for aquatic animals, an appropriate antimicrobial activity is achieved. Additionally, the demonstration of a higher stability of the cinnamaldehyde ethylene glycol acetal, when incorporated in animal feeds, and the higher precision of its analytical determination over time, compared to cinnamaldehyde, advantageously enables its traceability in animal feeds. It has surprisingly been found that the combination of cinnamaldehyde and
- cinnamaldehyde ethylene glycol acetal provides an unexpected effect, since even though cinnamaldehyde is having a higher antimicrobial activity than cinnamaldehyde ethylene glycol acetal, when a part of cinnamaldehyde is substituted by the corresponding amount of this cinnamaldehyde derivative, the activity of the mixture is not reduced.
- This aspect together with the higher stability of cinnamaldehyde ethylene glycol acetal, enables maintaining the efficacy of the feed over time.
- the present invention relates to a combination comprising cinnamaldehyde and cinnamaldehyde ethylene glycol acetal.
- the present invention relates to an animal feed additive for terrestrial and aquatic animals, including livestock and aquaculture species (namely crustaceans, fishes, molluscs, cephalopods, reptiles, amphibious and other invertebrates), comprising cinnamaldehyde and cinnamaldehyde ethylene glycol acetal and food, feed or pharmaceutically acceptable excipients or carriers.
- livestock and aquaculture species namely crustaceans, fishes, molluscs, cephalopods, reptiles, amphibious and other invertebrates
- the present invention relates to an animal feed for terrestrial and aquatic animals, including livestock and aquaculture species, comprising a prophylactically and/or therapeutically effective amount of the animal feed additive as defined herein above and below.
- the present invention relates to a process for the preparation of an animal feed for terrestrial or aquatic animals, including livestock and aquaculture species, as defined herein above and below comprising the steps of:
- step b) mixing the animal feed additive obtained in step a) with a feed in order to obtain an animal feed as defined herein above and below.
- the animal feed additive of the present disclosure can be used as an antimicrobial and/or immunostimulant to protect terrestrial or aquatic animals, including livestock and aquaculture species, against pathogenic infections. Accordingly, another aspect the invention relates to a combination comprising
- cinnamaldehyde and cinnamaldehyde ethylene glycol acetal for use in the prophylactic or therapeutic treatment of bacterial infections in terrestrial and aquatic animals including livestock and aquaculture species such as fish or shrimp.
- This aspect can also be formulated as the use of a combination comprising cinnamaldehyde and cinnamaldehyde ethylene glycol acetal for the manufacture of an animal feed for the prophylactic or therapeutic treatment of bacterial infections in terrestrial and aquatic animals including livestock and aquaculture species such as fish or shrimps.
- It also relates to a method for the prophylactic or therapeutic treatment of bacterial infections in terrestrial and aquatic animals including livestock and aquaculture species such as fish or shrimps susceptible to being or already infected by bacteria comprising administering a prophylactic or therapeutic effective amount of the animal feed additive as defined herein above or below to the terrestrial or aquatic animal in need of the treatment.
- Another aspect of the invention relates to a combination comprising cinnamaldehyde and cinnamaldehyde ethylene glycol acetal for use in improving the feed conversion ratio (FCR) or weight gain of terrestrial and aquatic animals, including livestock and aquaculture species such as fish or shrimps.
- FCR feed conversion ratio
- Fig. 1 shows the amount of cinnamaldehyde (AC) measured over time in samples of an animal feed for aquatic animals containing cinnamaldehyde (AC) stored at freezer temperature (FT) or at room temperature (RT).
- Fig. 2 shows the amount of cinnamaldehyde (AC) and of cinnamaldehyde ethylene glycol acetal (EAAC) measured over time in samples of an animal feed for aquatic animals containing a mixture of AC and EAAC in a molar ratio of 50:50, stored at freezer temperature (FT) or at room temperature (RT).
- AC cinnamaldehyde
- EAAC cinnamaldehyde ethylene glycol acetal
- animal feed additive refers to a composition suitable for or intended for being incorporated in an animal feed.
- animal feed refers to a preparation or mixture suitable for or intended for intake by an animal.
- feed conversion ratio refers to a measure of an animal's efficiency in converting feed mass input into increases of the desired output. In animals raised for meat (such as livestock, shrimp and fish) the output is the body mass gained by the animal.
- FCR is calculated as feed intake divided by weight gain, all over a specified period.
- Improvement in FCR means reduction of the FCR value.
- room temperature refers to about 20 °C to 25 °C and the term“freezer temperature” (FT) refers to about -20 °C to -16 °C.
- an effective amount refers to the minimal amount of a substance and/or agent, which is enough to achieve a desired and/or required effect.
- an effective amount of an "animal feed additive comprising cinnamaldehyde and cinnamaldehyde ethylene glycol acetal" is the minimum amount required in order to obtain a composition such as an animal feed having the desired properties associated therewith.
- the desired or required effects can be the prophylactic or therapeutic treatment of cultured aquatic animals susceptible to being or already infected by bacteria in aquaculture, and/or improving the feed conversion ratio (FCR) or weight gain of terrestrial or aquatic animals including livestock and aquaculture species such as fish or shrimps.
- FCR feed conversion ratio
- prophylactically and/or therapeutically effective amount refers to the amount of a compound that, when administered, is enough to prevent the development of, or alleviate to some extent, one or more of the symptoms of the disorder or condition being treated.
- the particular dose of compound administered according to this invention will of course be determined by the particular circumstances surrounding the case, including the particular condition being treated, the duration of the treatment, the nature of any concurrent treatment, the specific combination of active ingredient used, and any other factor known to the expert.
- total equivalent amount of cinnamaldehyde when having a mixture of cinnamaldehyde and cinnamaldehyde ethylene glycol acetal, refers to the amount of cinnamaldehyde in the form of cinnamaldehyde itself and of cinnamaldehyde ethylene glycol acetal, the latter being expressed as the corresponding amount of cinnamaldehyde.
- percentage by weight or “wt%” refers to the percentage by weight of the ingredient per weight of the overall composition, unless otherwise stated.
- the combination or the animal feed additive for terrestrial or aquatic animals of the invention comprises cinnamaldehyde and cinnamaldehyde ethylene glycol acetal.
- the molar ratio of cinnamaldehyde to cinnamaldehyde ethylene glycol acetal is from 20:80 to 80:20, or from 25:75 to 75:25, or from 30:70 to 70:30, or from 40:60 to 60:40, or 50:50. More particularly, of 50:50 or of 75:25.
- the combination or animal feed additive for terrestrial or aquatic animals of the present disclosure can be incorporated in an animal feed composition in order to obtain an animal feed comprising cinnamaldehyde to cinnamaldehyde ethylene glycol acetal as additional components.
- Molar ratios of the different components in the combination or animal feed additive correspond to the weight ratios of the same components in the animal feed.
- cinnamaldehyde and cinnamaldehyde ethylene glycol acetal are in a total amount equivalent to from 0.01 g to 2.5 g of cinnamaldehyde per each kg of animal feed.
- the total equivalent amount of cinnamaldehyde is from 0.05 g to 1.5 g, or from 0.1 g to 1.0 g per each kg of animal feed.
- the total equivalent amount of cinnamaldehyde is 0.5 g per each kg of animal feed.
- the incorporation of the combination or animal feed additive of the present disclosure in an animal feed composition in order to obtain an animal feed comprising the specific combination of cinnamaldehyde and cinnamaldehyde ethylene glycol acetal as defined above, particularly in the relative amounts also stated above, allows providing a stable animal feed wherein cinnamaldehyde ethylene glycol acetal is traceable.
- the sought efficiency of the prophylactic or therapeutic treatment can be achieved without the activity of the active ingredients being significantly reduced over time, while allowing the traceability of the specific combination of cinnamaldehyde ethylene glycol acetal and cinnamaldehyde.
- the animal feed additive and animal feed of the present disclosure can be used with terrestrial and aquatic animals, including livestock and aquaculture species.
- livestock are cattle, buffaloes, sheep, goats, pigs, chickens, ducks, geese, turkeys, horses, asses, mules, camels and rabbits.
- aquatic animals are crustaceans (e.g. shrimps and crabs), fishes (e.g. salmon, bream, bass, carps, tilapias, and catfishes), molluscs (e.g. clams, oysters, and abalone), cephalopods (e.g. octopus and cuttlefishes) and other invertebrates (e.g. sea cucumbers and sea urchins).
- crustaceans e.g. shrimps and crabs
- fishes e.g. salmon, bream, bass, carps, tilapias, and catfishes
- molluscs e.g.
- the animal feed has a high protein content, i.e. a protein content from 20 to 60 wt%, from 25 to 55 wt%, or from 30 to 50 wt%.
- the animal feed is a fish feed or a shrimp feed.
- animal feed additive and animal feeds of the present disclosure can be prepared by conventional methods known to those skilled in the art, such as the one described in the examples.
- a process for the preparation of an animal feed as defined herein above and below can comprise the steps of mixing cinnamaldehyde,
- cinnamaldehyde ethylene glycol acetal and, optionally, a food, feed or pharmaceutically acceptable excipient or carrier in order to obtain an animal feed additive and, then, mixing the animal feed additive with an animal feed composition.
- ingredients of a feed composition can be mixed for instance in a mixer, then the mixture can be mixed with the feed additive in order to obtain a feed in the form of a powder.
- the feed composition can be transformed in pellets by extrusion or pressing, and then the feed additive of the present disclosure comprising water or an edible oil such as a vegetable oil can be coated onto the pellets, particularly in the amounts defined herein above, in order to obtain the final animal feed.
- Examples of food, feed or pharmaceutically acceptable excipients or carriers include vegetable oils such as soybean oil; emulsifying agents such us lecithins and binders such as colloidal silica.
- Animal feed compositions wherein the combination of the animal feed additive of the present disclosure can be added are commercially available.
- an animal feed composition for terrestrial or aquatic animals can comprise one or more components selected from proteins, carbohydrate, lipids including animal fats, vegetable meals, vegetable oils, gluten, amino acids and other allowed feed additives and feed materials.
- the animal feed used was a commercial fish feed having a 52 wt% of crude protein.
- a sterile plastic bag was spread out on a laboratory tray, and 200 g of the commercial feed was deposited on it. With the help of a liquid diffuser bottle, by means of short pulsations, 3 g of the mixture A, previously diluted to 10 wt% in vegetable oil, was applied to the feed.
- the bag was closed and shaken, and the feed additive mixture was spread by manual friction in order to distribute it homogeneously all over the feed. Then, the feed containing the mixture A was put in a plastic container. The same process was carried out with mixture B instead of mixture A.
- feed samples (200 g) containing mixture A or mixture B were divided into two and put into sterile cans (100 g of feed in each) for the different thermal treatments at room temperature (RT) and freezer temperature (FT).
- RT room temperature
- FT freezer temperature
- Example 2 Stability test in an animal feed
- Carrier gas Helium
- concentration of standard was used to spike the sample.
- Results were calculated by standard addition method, taking into account the weight of the feed samples extracted and the standard concentrations.
- Table 1 shows the amount of cinnamaldehyde (AC) measured over time in the feed samples containing cinnamaldehyde (AC) stored at freezer temperature (FT) and at room temperature (RT).
- Table 2 shows the amount of cinnamaldehyde (AC) and of cinnamaldehyde ethylene glycol acetal (EAAC) detected over time in the feed samples containing a mixture of AC and EAAC in a molar ratio of 50:50, stored at freezer temperature (FT) or at room temperature (RT).
- AC cinnamaldehyde
- EAAC cinnamaldehyde ethylene glycol acetal
- Example 3 In vitro bacteriostatic activity of oil-based feed additives
- the antibacterial activity of the combination of the invention was assessed against some of the most common fish and shrimp bacterial pathogens by finding the Minimum Inhibitory Concentration (MIC). In Table 3 below the bacterial pathogens tested are shown.
- the MIC assay can be carried out following the procedure disclosed in Parasa, L.S. et al. (cf. Parasa, L.S. et al. "In vitro antibacterial activity of culinary spices aniseed, star anise and Cinnamon against bacterial pathogens of fish", International Journal of Pharmacy and Pharmaceutical Sciences,
- Test samples a series of 10 concentrations from 12000 to 23.44 mg/I, in two-fold dilutions Temperature: 27 °C
- the MIC assay was aimed at determining the minimum concentration of experimental product that could completely inhibit the growth of the experimental bacteria in a liquid medium. This intended to simulate a situation where the experimental products interact with pathogens in liquid phases such as digestive fluids or seawater.
- MIC is a direct but inverse measure for the power of the experimental product bacteriostatic activity towards pathogens. The lower the MIC, the higher the inhibiting power of the experimental product.
- the activity of the antibiotic gentamycin against Vibrio campbellii (positive control) showed inhibition at the expected concentration of 4 mg/L.
- the control for bacterial growth (no antibacterial product) showed normal growth.
- the control for bacterial contamination of the culture medium did not show any bacterial growth, as expected.
- pathogen V. anguillarum with a MIC value ranging between 375 to 93.8 mg/L.
- Table 5 shows an overview of the results of MIC values of each experimental product towards each experimental bacterium (mg/L).
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Abstract
Description
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Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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EP19382483.6A EP3747280A1 (en) | 2019-06-07 | 2019-06-07 | Antimicrobial combination for use in animal feeds |
PCT/EP2020/065588 WO2020245333A1 (en) | 2019-06-07 | 2020-06-05 | Antimicrobial combination for use in animal feeds |
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EP3979812A1 true EP3979812A1 (en) | 2022-04-13 |
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EP19382483.6A Withdrawn EP3747280A1 (en) | 2019-06-07 | 2019-06-07 | Antimicrobial combination for use in animal feeds |
EP20730273.8A Pending EP3979812A1 (en) | 2019-06-07 | 2020-06-05 | Antimicrobial combination for use in animal feeds |
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EP19382483.6A Withdrawn EP3747280A1 (en) | 2019-06-07 | 2019-06-07 | Antimicrobial combination for use in animal feeds |
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EP (2) | EP3747280A1 (en) |
CL (1) | CL2021003199A1 (en) |
EC (1) | ECSP21091831A (en) |
WO (1) | WO2020245333A1 (en) |
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US3615597A (en) * | 1967-03-10 | 1971-10-26 | Pillsbury Co | Solid food condiment |
PT762837E (en) * | 1995-03-31 | 2002-07-31 | Schur Jorg Peter | PROCESS FOR IMPROVING THE DURABILITY AND / OR STABILIZATION OF MICROBIOLOGICALLY DEGRADABLE PRODUCTS |
WO2004091307A2 (en) | 2003-04-08 | 2004-10-28 | Advanced Bionutriton Corporation | Feed additives against diseasse infection in terrestrial and aquatic animals |
WO2008155536A1 (en) * | 2007-06-18 | 2008-12-24 | Danisco A/S | Animal feed product |
EP2453758A1 (en) * | 2009-07-17 | 2012-05-23 | DSM IP Assets B.V. | Use of natural substances as feed additives for aquatic animals |
US20160206654A1 (en) * | 2013-09-30 | 2016-07-21 | Furst-Mcness Company | Methods and feed supplements for livestock |
EP3356356B1 (en) * | 2015-10-01 | 2021-05-26 | Firmenich Incorporated | Compounds useful as modulators of trpm8 |
-
2019
- 2019-06-07 EP EP19382483.6A patent/EP3747280A1/en not_active Withdrawn
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2020
- 2020-06-05 EP EP20730273.8A patent/EP3979812A1/en active Pending
- 2020-06-05 WO PCT/EP2020/065588 patent/WO2020245333A1/en active Application Filing
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2021
- 2021-12-01 CL CL2021003199A patent/CL2021003199A1/en unknown
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WO2020245333A1 (en) | 2020-12-10 |
ECSP21091831A (en) | 2022-01-31 |
EP3747280A1 (en) | 2020-12-09 |
CL2021003199A1 (en) | 2022-08-12 |
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