Detoxifying antimicrobial molecular complex

Abstract

The invention relates to a food additive which includes at least one clay interspersed with volatile antimicrobial agents. Such an additive can be added to the feed of certain animals, in particular poultry and ruminants, in order to act as a growth promoter and to reduce the risks of infection, as well as the risks of intoxication by mycotoxins.

Claims

1. A composition combining both antimicrobial and toxin-adsorption activities, characterized in that it is formed by a molecular complex comprising an edible clay interspersed by a volatile antimicrobial agent, characterized in that said edible clay is a mixture of Ghassoul from Morocco and bentonite from Morocco.

2. The composition according to claim 1, characterized in that said volatile antimicrobial agent is an antiseptic aromatic alcohol selected from thymol, cresol, carvacrol, eugenol, menthol, cinnamaldehyde or mixtures or isomers thereof.

3. The composition according to claim 1, characterized in that said volatile antimicrobial agent is formed by a mixture of two or more natural or synthetic antimicrobial agents.

4. The composition according to claim 1, characterized in that said mixture of Ghassoul and bentonite from Morocco contains at least 5% by weight Ghassoul from Morocco.

5. The composition according to claim 1, characterized in that said volatile antimicrobial agent is present in a mass ratio from 0.005 to 0.33 based on said edible clay.

6. A method for manufacturing a composition comprising an edible clay and a volatile antimicrobial agent comprising: i) adding said volatile antimicrobial agent into solution with an organic or mineral solvent in order to obtain an antimicrobial solution; and ii) mixing said antimicrobial solution with said edible clay as a powder or pellets under stirring and temperature conditions to obtain a homogenous and stable composition formed by a molecular complex containing said edible clay interspersed by said volatile antimicrobial agent, characterized in that said edible clay is a mixture of Ghassoul from Morocco and bentonite from Morocco.

7. The manufacturing method according to claim 6, characterized in that said volatile antimicrobial agent is selected from thymol, cresol, carvacrol, menthol, eugenol, cinnamaldehyde, or mixtures or isomers thereof, said organic or mineral solvent is a vegetable oil.

8. The manufacturing method according to claim 6 wherein: crystallized thymol is gradually put into solution in a volume of vegetable oil heated to a temperature allowing it to melt so as to obtain a homogeneous and limpid liquid composition (an antimicrobial solution); mixing said antimicrobial solution with an amount of said edible clay as a powder capable of absorbing it in order to obtain a molecular complex as a homogenous and stable powder; and subjecting said powder to a calibration or sieving operation to obtain the desired grain size.

9. The manufacturing method according to claim 6, characterized in that the volatile antimicrobial agent is a mixture of thymol and cresol.

10. The manufacturing method according to claim 6, characterized in that the volatile antimicrobial agent is a mixture of thymol, cresol and menthol.

11. A method of reducing or suppressing the development of pathogenic or toxinogenic germs in an animal comprising administering a composition according to claim 1 to an animal or its surroundings.

12. The method according to claim 11, wherein said composition is administered to said animal in a feed containing said composition.

13. The method according to claim 11, wherein said composition is added to feed, drinking water, litter, walls of the animal enclosure, barn area, rearing equipment or the vehicles transporting the animal.

14. The method according to claim 11, wherein said animal is a mammal, fish, bee or poultry.

15. The method according to claim 14, wherein said mammal is a bovine animal, ovine animal, rabbit, pig, caprine animal or equine animal.

16. The method according to claim 14, wherein said poultry are selected from chicken broilers, laying hens, breeding roosters and hens, guinea fowl, turkeys, quail, ducks, geese and pigeons.

17. The method according to claim 11, said method reducing or suppressing development of bacteria, fungi, parasites, bacteriotoxins and mycotoxins in the animal.

Description

EXAMPLE 1: X-RAY DIFFRACTION TEST

(1) Preparations of a final weight of 1,000 grams were made with 150 grams of thymol put into solution according to the method of the invention with 20 milliliters of table oil at a temperature of 60 C. The antiseptic solution obtained thereby was mixed with 830 grams of Moroccan ghassoul, 830 grams of Moroccan bentonite, or 830 grams of a 50%/50% ghassoul/bentonite mixture (preparation a).

(2) Control compositions were prepared:

(3) Preparation b: The same pure clays without any additive.

(4) Preparation c: The same clays with 20 ml of table oil alone.

(5) Preparation d: The same clays with thymol alone as a crystallized powder.

(6) Preparation e: The same clays with liquefied thymol by heating to 60 C. without any vegetable oil.

(7) Preparations a, b and c were subjected to an x-ray diffraction test according to the usual standards for treating clays in this analytical method.

(8) The results obtained show a displacement of the specific peaks of bentonite and stevensite in the spectrum, in a surprising way in the case of the preparations according to the method of the invention (preparation a) as compared with the controls of pure clays (preparation b) and clays having been mixed with table oil alone (preparation c) (results in Table 1). At the same time, the results show that for preparation d containing thymol as a crystallized powder mixed with clay, the obtained powder is heterogeneous and the obtained x-ray diffraction spectrum was not able to be utilized. For preparation e, in which thymol was liquified by heating and mixed with clay without any vegetable oil, it is seen that thymol recrystallizes and forms hard lumps. The mixture obtained thereby is heterogeneous and cannot meet the criteria for which the invention was elaborated.

(9) This shows on the one hand that the manufacturing method used in the invention actually gives the possibility of ending up with interspersion of thymol in the interfoliar spaces of the clays, which leads to a stable and homogeneous dispersion of thymol in clay and an increase in the distance of the interfoliar spaces of the clays, therefore improving hydrophobicity and the capability of the clays of adsorbing the toxins.

(10) These results show on the other hand that the mixtures with thymol and clay by obvious methods using crystallized thymol or liquified thymol by heating do not end up with the result expected by the invention.

(11) TABLE-US-00001 TABLE 1 Interfoliar distance of the clays according to the preparation modes a, b and c. Bentonite Stevensite With With With table oil and With table oil and Crude table oil essential oil Crude table oil essential oil Clays (prep. b) (prep.c) (prep. a) (prep. b) (prep. c) (prep. a) Distance 15.23 15.23 15.69 15.75 16.15 16.51 between sheets (in angstroms)

EXAMPLE 2: IN VIVO TEST

(12) Batches of 25 young broilers and 12 turkey poults were fed with a feed containing the preparation of the invention in an amount of 100 grams, 500 grams, 1 kilogram or 2 kilograms per metric ton of feed. Control batches of the same size were fed with a feed not containing any antibiotic or antiparasitic agents. Other control batches of the same size were fed with a feed containing Flavomycin in an amount of 200 grams per metric ton of feed and Salinomycin in an amount of 70 grams per metric ton of feed for the broilers and 100 grams per metric ton of monensin for the turkey poults. During five weeks of treatment, feces samples of each subject in each batch were analyzed by microscopy and by suitable microbiological analysis for evaluating the numbers of bacteria, yeast, fungi and oocytes of protozoa, in particular of the species Eimeria. sp., involved in coccidiosis in chickens and turkeys. At the end of the five weeks of the experiment, the subjects were sacrificed and autopsies were carried out by professional veterinarians, specialists in avian pathology. The goal of the autopsies was to evaluate the lesional scores of the intestine and the integrity condition of the liver, kidneys, lungs, spleen and intestine.

(13) The results obtained show highly significant reductions in the numbers of bacteria, yeasts, fungi and oocytes in the batches having received the preparation of the invention in their feed at doses of 1 kilogram per metric ton and 2 kilograms per metric ton as compared with control batches having received a neutral feed. The batches having received the preparation of the invention in their feed at doses of 100 grams and 500 grams per metric ton of feed did not have any significant difference as compared with the control. The batches having received the antibiotic and the antiparasitic agent in their feed have a visible but non-significant reduction as compared with the controls. However, the numbers of bacteria, yeasts, fungi and oocytes in batches having received the preparation of the invention at the doses of 1 kilogram and 2 kilograms per metric ton of feed is significantly less than the number of bacteria, yeasts, fungi and oocytes recorded in the batches having received the antibiotic and the antiparasitic agent in the feed. These results show that the preparation according to the invention used as an additive in chicken and turkey feed has multiple activities like promoting growth (reduction in the intestinal flora), as an antiparasitic (anticoccidian) agent and as an antifungal agent significantly superior to the antibiotics and antiparasitic agents used in this test. The results of the autopsies also show that the intestinal lesion, kidney, lung, spleen and liver congestion scores are significantly lower in the batches having received the feed containing the preparation of the invention as compared with the batches having received the neutral feed or the feed containing the antibiotic and the antiparasitic agent. These results again show that the preparation of the invention used as an additive in chicken and turkey feed has activities protecting intestinal integrity, hepatobiliary protective activities, and activities purifying the lungs and kidneys and protecting the spleen. These results confirm that the preparation of the invention represents a solution with multiple activities for combating pathologies of chickens and turkeys which are multifactorial diseases.

(14) Batches of 25 chicks of one day of age were fed for 5 weeks on average with respectively: batch 1: a white broiler feed which contains neither antibiotics nor antiparasitic agents; batch 2: a feed containing a growth factor antibiotic, Flavomycin, in an amount of 200 milligrams per kilogram of feed and an antiparasitic agent, Salynomycin, in an amount of 70 milligrams per kilogram; batch 3: a feed containing the preparation of the invention in the feed at a proportion of 100 grams per metric ton of feed; batch 4: a feed containing the preparation of the invention in the feed at a proportion of 500 grams per metric ton of feed; batch 5: a feed containing the preparation of the invention in the feed at a proportion of 1 kg per metric ton of feed; and batch 6: a feed containing the preparation of the invention in the feed at a proportion of 2 kg per metric ton of feed.

(15) TABLE-US-00002 TABLE 2 Zootechnical and microbiological results of the broiler in vivo test Con- Mor- sump- Body Bacterial load Fungal load tality tion weight (CFU/g of (CFU/g of rate index (g) faeces) faeces Batch 1 0% 2.3 105 8.6 .sup.1 .Math. 10.sup.10 0.3 8.6 .Math. 10.sup.9 3.7 Batch 2 0% 2.2 110 6.4 4.9 .Math. 10.sup.9 2.1 1.6 .Math. 10.sup.9 0.3 Batch 3 0% 2.2 113 3.4 4.5 .Math. 10.sup.9 3.2 5.7 .Math. 10.sup.8 1.2 Batch 4 0% 2.1 121 4.5 3.2 .Math. 10.sup.8 4.1 1.3 .Math. 10.sup.8 0.3 Batch 5 0% 1.9 138 6.6 7.3 .Math. 10.sup.7 5.2 8.6 .Math. 10.sup.7 1.5 Batch 6 0% 1.8 157 2.2 2 .Math. 10.sup.7 1.70 .sup.5 .Math. 10.sup.7 1.1

(16) The in vivo test for turkeys gave similar results to those obtained in the broiler chicken in vivo test.

EXAMPLE 3: FIELD TESTS

(17) Field tests were carried out on a farm raising chicken broilers and a farm raising turkey broilers. In these tests, two batches of 13,500 chicks each received the preparation of the invention at the dose of two kilograms per metric ton of feed during the whole duration of the tape (40 days for chickens and 13 weeks for turkeys). Two other batches of 13,500 chicks each received the preparation of the invention at the dose of 1 kilogram per metric ton of feed and two other batches of 13,500 chicks each received a feed containing the antibiotic Flavomycin at 200 grams per metric ton of feed and the antiparasitic agent Salynomycin at 70 grams per metric ton of feed for chickens and monensin for turkeys. At the end of the strip, the zootechnical performances and the lesion scores were measured for each batch.

(18) The results show:

(19) Mortality: The percentage of dead chickens and turkeys during the raising cycle was 2.9% for chickens and 2.5% for turkeys for the batches having received 1 kilogram or 2 kilograms of preparation of the invention per metric ton of feed. For the batches having received the antibiotic and the antiparasitic agent in the feed, the mortality was 5.2% for turkeys and 5.9% for chickens.

(20) Average weight: The average weight of both batches having received 1 kilogram or 2 kilograms per metric ton of feed of the preparation of the invention is 2.3 kg per subject in chickens and 12.8 kg in turkeys; that of the two batches having received the antibiotic and the antiparasitic agent was 2.135 kg in chickens and 11.9 kg in turkeys.

(21) Consumption index: The consumption index for the batches having received two kilograms of the preparation of the invention per metric ton of feed was 1.68, that of the batches having received one kilogram of the preparation per metric ton of feed was 1.82, and that of the two batches having received the antibiotic and the antiparasitic agent was on the order of 1.98.

(22) The titers of the antibodies related to the vaccines used: The inhibition titer of hemagglutination as compared with the antibodies of the viruses of Newcastle's disease the batches having received the preparation in an amount of two kilograms and one kilogram per metric ton of feed was 64, 64, 64, 32 respectively and, for the two batches having received the antibiotic and the anti-coccidian agent, were 16 and 16.

(23) The lesional score of the intestine: The lesional scores of the intestine were lowest (1.22) for the batches having received the preparation of the invention in an amount of 2 kg and 1 kg while the scores for the batches having received the antibiotic and the anti-coccidian agent were on the order of 2.74 and 3.13.

(24) The score of the integrity of the organs: The aspect of the organs (liver, spleen, lungs, muscles and bones) were clearly more healthy (not showing any sign of congestion) for the batches having received the preparation of the invention in an amount of 2 kg and 1 kg per metric ton of feed, while congestion signs visible to the naked eye were ascertained in the majority of the autopsies in both batches having received the antibiotic and the antiparasitic agent.

(25) These results surprisingly show that the preparation of the invention exerts the multiple activities for which it was designed even under intensive raising conditions in the field.

EXAMPLE 4: INHIBITION AND ADSORPTION OF MYCOTOXINS IN FEED AND CORN

(26) Samples of corn and composed feed were treated with variable amounts of the preparation of the invention in the proportions of 2 kg, 4 kg and 6 kg per metric ton of feed. These samples were hermetically packaged and incubated at a temperature of 27.5 C. 50 gram aliquots were taken after four weeks for dosage of the mycotoxins; the results show that the corn samples having been treated by the preparation of the invention had significantly lower levels of DON (deoxynivalenol), ZON (Zearalenone) and ochratoxin than the control sample. This inhibition and adsorption effect on mycotoxins is increased according to the dose used of the preparation of the invention. A similar result was obtained for the composed feed sample treated by the preparation of the invention with respect to the untreated sample for mycotoxins, ochratoxin (ZON) and aflatoxin.

(27) These results show that the preparation of the invention also has the capability of protecting feed and corn from mycotoxins which represent a significant risk factor promoting the outbreak of diseases and causing, in acute cases, mortal intoxication of animals.

(28) TABLE-US-00003 TABLE 3 Percentage of reduction of the mycotoxins in corn treated with the preparation of the invention as compared with the untreated control. Preparation 0.2 g/100 0.4 g/100 0.6 g/100 Mycotoxins g of corn g of corn g of corn Zearalenone 56% 59% 61% DON 63% 73% 75% ochratoxin 13% 22% 27%

(29) TABLE-US-00004 TABLE 4 Reduction percentage of the mycotoxins in the treated composed feed with respect to the untreated control. Preparation 0.2 g/100 0.4 g/100 0.6 g/100 Mycotoxins g of feed g of feed g of feed Zearalenone 35% 63% 94% Aflatoxin 74.35% 86% 89%