RAW MATERIAL FOR ANIMAL NUTRITION COMPRISING AN ORGANO-MINERAL COMPLEX CONTAINING DIETARY PHOSPHATE AND A HUMIC SUBSTANCE

Abstract

The invention relates to a dietary raw material for animal nutrition, comprising an organo-mineral complex containing a dietary phosphate and a humic substance. The raw material improves the digestibility of the ration, absorbs mycotoxins and increases zootechnical performance.

Claims

1. A dietary raw material for animal nutrition, comprising an organo-mineral complex containing a dietary phosphate and a humic substance, said humic substance forming physicochemical bonds with phosphorus atoms.

2. The raw material according to claim 1, characterised in that the dietary phosphate is chosen from calcium phosphate, magnesium phosphate, monosodium phosphate, calcium-sodium phosphate and mixtures thereof.

3. The raw material according to claim 1, characterised in that the dietary phosphate is chosen from monocalcium phosphate, dicalcium phosphate, monodicalcium phosphate, tricalcium phosphate, and mixtures thereof.

4. The raw material according to claim 1, characterised in that the dietary phosphate is in the form of tri-magnesium phosphate.

5. The raw material according to claim 1, characterised in that the humic substance is a potassium humate or a sodium humate.

6. A method for manufacturing the dietary raw material according to claim 1, comprising a step of preparing a dispersion or aqueous solution containing the humic substance, followed by mixing this dispersion or this solution with a source of phosphorus and with a source of calcium, magnesium or sodium.

7. The manufacturing method according to claim 6, characterised in that the source of phosphorus is phosphoric acid.

8. The manufacturing method according to claim 6, characterised in that the source of calcium is calcium carbonate, quicklime or slaked lime.

9. The manufacturing method according to claim 6, characterised in that the humic substance is extracted from leonardite.

10. The manufacturing method according to claim 6, characterised in that the mixing of the aqueous dispersion or aqueous solution containing the humic substance, with the source of phosphorus and with the source of calcium, magnesium or sodium, is carried out at a temperature ranging from 50 C. to 80 C.

11. A method for improving the bioavailability of mineral phosphorus in a monogastric livestock animal, comprising utilizing the dietary raw material according to claim 1.

12. A method for the nutrition of a livestock animal comprising a step of incorporating the dietary raw material according to claim 1 in the ration of the animal.

13. The raw material intended for animal feed obtainable by preparing an aqueous solution comprising a water-soluble humic substance, a water-soluble source of mineral phosphorus, and optionally a water-soluble source of mineral calcium.

Description

DESCRIPTION OF THE FIGURES

[0076] FIG. 1 shows the x-ray diffraction diagram for a reference product containing monocalcium phosphate, devoid of humic substance (MCPH0).

[0077] FIG. 2 shows the x-ray diffraction diagram for a product of the invention containing 1.5% humates (MCPH15).

[0078] FIG. 3 compares the two diffraction diagrams of preceding FIGS. 1 and 2.

[0079] FIG. 4 shows the .sup.31P NMR spectra of a reference product containing monocalcium phosphate devoid of humic substance (MCPH0), and that of the product of the invention containing 1.5% humates (MCPH15).

[0080] FIG. 5 shows the .sup.31P NMR spectrum of a reference product containing monocalcium phosphate devoid of humic substance (MCPH0).

[0081] FIG. 6 shows the .sup.31P NMR spectrum of the product of the invention containing 1.5% humates (MCPH15).

[0082] FIG. 7 shows the effect of the invention on the change in the pH of the ruminal medium from 2.0 to 6.5 hours incubation under in vitro fermentation conditions.

[0083] FIG. 8 shows the effect of the invention on the concentrations of VFA of the ruminal medium from 2.0 to 6.5 hours incubation under in vitro fermentation conditions.

[0084] FIG. 9 shows the effect of different doses of the invention on the rate of absorption of various mycotoxins in the stomach of monogastric animals under in vitro conditions.

[0085] FIG. 10 shows the effect of different doses of the invention on the rate of absorption of various mycotoxins in the intestine of monogastric animals under in vitro conditions.

[0086] FIG. 11 shows the average consumption index (CI) at the end of finishing (CIO-35 between D0 and D35) for batches of chickens for which the feed has been enriched with different MCP containing or not containing different doses of humates.

[0087] FIG. 12 shows the ash content and the quantity of calcium and phosphorus of the tibias of chickens taken at D35 as a function of the treatments.

[0088] FIG. 13 shows the breaking force of the tibias of chickens taken at D35 as a function of the treatments.

[0089] FIG. 14 shows, for each treatment, the deviation (%) in pH of chicken meat between the measured value and the expected value after 15 minutes maturation (pH=6.4) or 7 days storage (pH=5.6).

[0090] FIG. 15 shows the mean consumption index (CI) at the end of finishing (CIO-35 between D0 and D35) for batches of chickens for which the feed has been enriched with a standard calcium phosphate or the product of the invention MCPH15.

[0091] FIG. 16 shows the quantity of P.sub.2O.sub.5 in the litter on D42 of chickens having received, in their feed, a standard calcium phosphate or the product of the invention MCPH15.

Example 1: Preparation of a Complex of Humates and Calcium Phosphate and Then Characterisation

Preparation of the Complex

[0092] Several aqueous solutions of sodium humates with variable concentration of humates ranging from 100 g/L to 250 g/L are prepared by dissolving sodium humates in water, preferably water heated to 50 C.
Sodium humate (CAS Number 68131-04-4) is commercially available under the reference Nut Mordan manufactured by Humatex. The pH of this product diluted to 10% in water is between 10.0 and 12.0 and its content of humic acid by dry weight is greater than 60%. The humic acid content by dr y weight is equal to the complement to 100% of the ash content measured after calcination of the sample in an electric furnace at 500 C. for 60 minutes and at 805 C.-825 C. for 60 minutes (according to the manufacturer).
A solution of Israeli green phosphoric acid (P.sub.2O.sub.5 total=54.2%) diluted to 50% P.sub.2O.sub.5 with water heated to 50 C. is prepared.
Each aqueous solution of humates is then poured, under stirring, into the solution of phosphoric acid, the stirring being maintained in order to disperse the solid formed and the temperature maintained at 50 C. Calcium carbonate (having a total CaO content greater than or equal to 53%) is then poured into the dispersion, then the mixture is stirred vigorously, crumbled, granulated and then dried.
Alternatively, the phosphoric acid is not diluted in water and it is poured simultaneously with the solution of sodium humates onto the calcium carbonate.
The weight ratio between the undiluted phosphoric acid at 54% P.sub.2O.sub.5 in water and the calcium carbonate was 2.5 for producing the MCP.
By varying the concentration of the solution of humates, monocalcium phosphates containing 1.0% (denoted MCPH10), 1.5% (denoted MCPH15), 2.0% (denoted MCPH20), 2.5% (denoted MCPH25), 5.0% (denoted MCP50) and 10.0% (denoted MCP100) of humates could be prepared.
Dicalcium phosphates containing 0% (denoted DCP), 1.0% (denoted DCP10), 5.0% (denoted DCP50) and 10.0% (denoted DCP100) of humates by applying a weight ratio [phosphoric acid at 54%P.sub.2O.sub.5/calcium carbonate]=1.3 could also be prepared. In the majority of tests, and in order to limit the number of invention samples studied, the choice was made for doses between 1.0% and 2.5%, preferably 1.5%.

Characterisation by XRD and NMR

Experimental Conditions of the XRD Analysis

[0093] The diffraction diagrams were recorded on a diffractometer (X'Pert Pro from PANalytical) in reflection mode and equipped with a copper tube (K=0.1542 nm, 45 kV and 40 mA) and a PIXcel linear detector (active length=3.3472). On the x-ray beam path, 0.04 rad Soller slits, a 1/16 divergence slit, a 10 mm mask and a anti-scatter slit were mounted as primary optics, and a nickel filter and 0.04 rad Soller slits as secondary optics. After fine grinding, the samples were compacted in a rear-loading sample holder.

Results of the XRD Analysis

[0094] The peaks of the diffraction diagram showed that the MCP without humate (MCPH0) consisted of two crystalline phases: in the majority monohydrated monocalcium phosphate (Ca(H.sub.2PO.sub.4).sub.2,H.sub.2O) and as a minority calcite (calcium carbonate; CaCO.sub.3). However, three very low intensity peaks could be identified after 65, the reflections of the reference card only having been recorded up to 64 (FIG. 1). This indicates that an additional crystalline phase is present in the MCPH0 without being identifiable by XRD.
The peaks of the diffraction diagram for MCPH15 show that this sample contains mainly monohydrated monocalcium phosphate (Ca(H.sub.2PO.sub.4).sub.2,H.sub.2O), probably anhydrous dicalcium phosphate (monetite Ca(HPO.sub.4)) and possibly calcite (calcium carbonate; CaCO.sub.3) and anhydrous monocalcium phosphate (Ca(H.sub.2PO.sub.4).sub.2 (FIG. 2). However, the diffraction diagram of MCPH15 shows diffraction peaks that are less intense than that of MCPH0, which indicates that this sample is less well crystallised. In order to confirm this, when the two diffraction diagrams of the two samples are superimposed, it can be observed that the baseline of the MCPH15 sample, between 15 and 40, is higher than that of the MCPH0 sample (FIG. 3). This thus indicates the amorphous presence in the MCPH15 sample. In addition, the organic matrices cannot be identified by x-ray diffraction and thus appeared as an amorphous phase. This drop in intensity of the peaks and this higher baseline of MCPH15 are explained by the presence of humates which correspond to a non-crystalline organic product and an amorphous phase that is not identified by XRD. In general, the organo-mineral complexes characterised by XRD show a broadening of the curves which indicates the presence of an amorphous product in the sample.

Experimental Conditions for the NMR Analysis

[0095] After fine grinding, the samples (powders) were introduced (packed) into a cylindrical sample holder (rotor) made of zirconium oxide, 4 mm in diameter and closed by a Kelefl plug. The analyses were carried out at room temperature using a Bruker Avance II 400 MHz spectrometer (Larmor frequency (.sup.31P)=103.85 MHz), equipped with a Bruker 4 mm double channel measurement head. The .sup.31P MAS (Magic Angle Spinning) spectra were recorded at a rotation speed of 12 kHz with a pulse angle equal to a duration of 3 microseconds and a recycling time of 200 seconds. The chemical shifts are expressed relative to an aqueous solution at 85% H.sub.3PO.sub.4. After analysis, the samples were fully recovered.

Results of the NMR Analysis

[0096] FIG. 4 shows the superposition of the .sup.31P NMR spectra for the two samples analysed. More precisely, FIG. 5 shows that after deconvolution, the spectrum for MCPH0 has four signals at 0.75, 0.15, 1.45 and 4.65 ppm. The signals at 0.15 and 4.65 correspond to the presence of monohydrated monocalcium phosphate (Ca(H.sub.2PO.sub.4).sub.2,H.sub.2O) as majority compound (80%), as demonstrated by the XRD analysis. In addition, these two signals correspond to two H.sub.2PO.sub.4.sup.groups which are differentiated, with the group at 4.65 being an acceptor of hydrogen bonds with the protons of the water molecule, whereas that at 0.15 would be a hydrogen bond donor. The two largest signals at 1.45 and 0.75 ppm characterise the crystallographic sites of the dicalcium phosphate (CaHPO.sub.4) not revealed by XRD and being the minority quantity in this sample (20%).
FIG. 6 shows that, after deconvolution, the spectrum for MCPH15 has five signals at 0.1, 0.2, 1.75, 4.75 and 10.0 ppm. As in the MCPH0 sample, the characteristic signals of monohydrated monocalcium phosphate (Ca(H.sub.2PO.sub.4).sub.2,H.sub.2O) are observed at 0.2 and 4.75 ppm, but in lower proportion (40% of the sample), their corresponding peaks being lower than for MCPH0. The peak at 1.75 ppm corresponds to the peak at 1.45 ppm of MCPH0, it therefore involves the presence of dicalcium phosphate (CaHPO.sub.4), at a level of 10%. However, for MCPH15, it is impossible to distinguish the equivalent of the peak at 0.75 ppm for MCPH0, which also reveals the presence of this dicalcium phosphate. Thus, the particularly broad signal at 0.1 ppm would correspond to the superposition of several chemical elements, namely; the missing portion of monohydrated monocalcium phosphate

[0097] (Ca(H2PO4)2,H2O; i.e. 20%), the invisible resonance of this dicalcium phosphate (CaHPO.sub.4; i.e. 10%) and a hidden portion of anhydrous monocalcium phosphate (Ca(H.sub.2PO.sub.4); at a height of 5% to 10%) the only phosphate component, for which the chemical shifts are in the region around 0.1 ppm. Although in a very low quantity, this last compound can only be present in amorphous form, explaining the characteristic shape of the very broadened signal. Finally, the very broad signal between 6.0 and 12.0 ppm corresponds to amorphous -Ca.sub.2P.sub.2O.sub.7 pyrophosphates, present at around 15% in the sample. This amorphous characteristic is determined by the width of the signal which indicates that the environment of the phosphorus is very disordered and therefore modified. This peak reflects the new environment of the phosphate bonded by the Ca-humic acid bond sites, highlighting the complexing of humic acids with minerals (Ca or P). The broadening of these two peaks, which reveal amorphous phases, determines a chemical reaction between the sources of phosphorus and the humates indicating the presence of complexes.

Thus, the results of the analyses by XRD and .sup.31P NMR of the two samples are consistent and complementary. With regard to the MCPH0 sample, the two methods confirm the presence of crystallised monohydrated monocalcium phosphate

[0098] (Ca(H.sub.2PO.sub.4).sub.2,H.sub.2O). Contrary to the XRD, the .sup.31P NMR determines the presence of dicalcium phosphate, otherwise referred to as monetite (CaHPO.sub.4), in the MCPH0 and presumably in amorphous form (broad NMR signals at 1.45 and 0.75 ppm) because it was not detected by XRD. This means that the 3 unidentified peaks beyond 64 do not correspond to this monetite. On the other hand, XRD reveals that crystallised calcite (CaCO.sub.3) remains in MCPH0. Finally, .sup.31P NMR makes it possible to estimate that the relative proportions of phosphorus are distributed in the following manner: as majority phase 80% as Ca(H.sub.2PO.sub.4).sub.2,H.sub.2O and approximately 20% as CaHPO.sub.4.

With regard to the MCPH15 sample, the .sup.31P NMR can confirm the presence of crystallised phases observed by XRD, namely: monohydrated monocalcium phosphate (Ca(H.sub.2PO.sub.4).sub.2,H.sub.2O) which is in the majority, followed by monetite (CaHPO.sub.4). Amorphous or very slightly crystallised (because visible by XRD) anhydrous monocalcium phosphate (Ca(H.sub.2PO.sub.4).sub.2) is also present. The .sup.31P NMR also determines the presence of a new species, (-Ca.sub.2P.sub.2O.sub.7) calcium pyrophosphates, which are in amorphous form since they are not identified by XRD. According to the XRD, residual traces of calcite also appear to be present. Other types of amorphous phases have been identified. Some amorphous phases, which are not perfectly identified by NMR, are linked to the presence of complexes in the sample, which determines that a chemical reaction has taken place between the sources of phosphorus and the humates. Physicochemical bonds between the humate type organic matrix and the minerals are present in the system (calcium and phosphorus). Finally, .sup.31P NMR can evaluate, in a very relative manner, the distribution of phosphorus in the following way: majority phase, more than 50% Ca(H.sub.2PO.sub.4).sub.2,H.sub.2O, several minority phases (less than 50%) have been identified for CaHPO.sub.4, amorphous pyrophosphates and partially crystallised Ca(H.sub.2PO.sub.4).sub.2, also an amorphous phase represented by the broadening of the spectra (curves) corresponding to the applicant's organo-mineral complex.
Thus, the two analyses, XRD and NMR, reveal the presence of amorphous phases equivalent to an organo-mineral complex originating from chemical reactions in the medium between humic acids and inorganic compounds (calcium phosphates).

Example 2: In Vitro Test on an Artificial Fermenter: Impact of the Product of the Invention on the Fermentation Parameters

[0099] A test has been carried out in an artificial fermenter with four products of the invention MCPH0, MCPH10, MCPH5 and MCPH100 prepared according to Example 1.

Experimental Protocol

[0100] The principle of artificial fermentation relies on the incubation of rumen fluid under anaerobic conditions at 39 C. (Menke and Steingass, 1988). The bacterial flora in the presence of the appropriate substrate reproduces the ruminal fermentations, which enables the gas production to be monitored as well as the end products such as volatile fatty acids (VFA), NNH.sub.3 etc. according to the objectives of the study.

[0101] The device consists of 21250 ml flasks containing 150 ml of an inoculum based on rumen fluid and buffered artificial saliva (1:2). This inoculum is incubated under anaerobic conditions at 39 C. without substrate (blank) or with 1.25 g of substrate (corn silage, hay, concentrate: 50/30/20, provides 2.2 g of P/kg) alone (CT control) or with the substrate supplemented with the appropriate dose of each product tested, in order to have an identical phosphorus intake (MCPH0 to MCPH100). The flask sampling system makes it possible to carry out kinetics at 2 hours, 4 hours and 6 hours 30 minutes. Table 1 reports the experimental design of the test.

TABLE-US-00001 TABLE 1 Experimental design of the in vitro fermentation test in the presence of products of the invention Concentration Product dose Number Product Substrate P(%) (g/D/VL) of flasks CT EM/F/C 4 MCPH0 50/30/20 22.4 100 4 MCPH10 22.5 99.6 4 MCPH5 21.9 102.3 4 MCPH100 21.7 103.2 4 Blank 1 Total 21

Results

[0102] FIG. 7 shows the change in pH at the start and end of fermentation. The treatments and the time have a significant effect (p.val<0.001). The high dose (10%) of humic acid in MCP increases the pH relative to the control and the other treatments. Moreover, the treatment*time interaction is significant (p.val=0.06). It therefore appears that the impact of the treatments varies according to the kinetic points.

[0103] FIG. 8 shows that the concentrations of VFA increase over time (p.val<0.001). Some treatments in particular tend to increase the total production of VFA relative to the CT control (p.val<0.10). The statistical study of this parameter shows a significant interaction between the treatments and the times (p.val=0.02). The results of the statistical study at each time show that after 2 hours fermentation, the high doses of humic acids tend to reduce production of VFA (p.val=0.07). After 4 hours, the treatments have no effect (p.val>0.10). Finally, after 6 hours, the addition of MCP increases the production of VFA and the formulas with 1% and 10% humic acids promote this effect (p.val<0.01).

CONCLUSION

[0104] This in vitro study has evaluated the effects of MCP-humate complex on fermentations of the rumen. The addition of MCP-humates complex limits the drop in pH during fermentation under these closed batch conditions. Moreover, it improves the production of VFA and therefore the energy intake for the ruminant.

Example 3: Monogastric Digestion Test In Vitro: Impact of the Product of the Invention on the Digestibility of Minerals Ca, P and Mg

[0105] The objective of this in vitro digestion test is to evaluate the digestibility of the minerals Ca, P and Mg for the products of the invention from Example 1. In order to confirm the results, the test was repeated once. The results of the two in vitro test performed are thus presented: test 1 and test 2 below.

Experimental Protocol

[0106] The device used is a bain-marie consisting of 15 places that can each receive a 50-ml Eppendorf tube each containing a feed sample of 1.0 g. The samples are incubated at 40 C. A test takes place in 3 steps in order to mimic the digestion of growing chickens, in the crop (amylase, pH: 5.8, duration: 35 min), then the succenturiate ventricle (pepsin, pH: 2.7, duration: 42 min) and finally in the intestine up to the ileal phase (pancreatin, pH: 6.5, duration: 170 min).

[0107] The bain-marie can accommodate 15 samples, a test is carried out over 1 day in order to have a sufficient number of repetitions per feed (n=3). From the 15 tubes, one tube is used to check the absence of contamination by minerals (blank, no sample) and one tube is used to receive wheat bran (laboratory standard) in order to check the proper proceeding of the test. Nine tubes are used to receive the samples of complete feed for growing chickens, which contain the products of the invention with the following doses of humates: 0% and 1.5%.

[0108] The chemical composition of the feeds to be digested is given in Table 2.

TABLE-US-00002 TABLE 2 Nutritional composition of two experimental feeds for growing chickens. % MCPH0 feed MCPH15 feed Humidity 17.5 11.0 MS 82.5 89.0 MM 6.0 5.7 MO 94.0 94.3 Proteins 18.8 18.8 Ca 0.84 0.78 P 0.49 0.48 Mg 0.17 0.16 Na 0.17 0.16 Ca/P 1.71 1.63
The digestibility in vitro is expressed by calculating the solubility of the minerals. In other words, the greater the quantity of minerals in the food bolus fluid (ileal fluid) the greater the absorption potential of these minerals. The solubility is calculated as: Solubility (%)=quantity of mineral in the ileal fluid/quantity of mineral in 1 g of feed to be digested.

Results

[0109] An in vitro digestion test has been carried out, in which MCPH0 and MCPH15 were compared. The results were as follows.

[0110] The solubility of the chicken feeds provided with MCPH0 or MCPH15 produced according to Example 1 is given in Table 3.

TABLE-US-00003 TABLE 3 Solubility of the minerals from the monogastric in vitro digestion test as a function of the treatments based on not based on the product of the invention (Avr S.D.). Ca solubility P solubility Mg solubility Feed n (%) (%) (%) MCPH0 2 59.6 (0.74) 68.2 (3.15) 57.8 (2.99) MCPH15 3 62.9 (0.91) 72.2 (0.92) 62.2 (0.69) p.val 0.02 0.11 0.08

[0111] The solubility of calcium is significantly affected by the presence of humates (p.val=0.02). In addition, the solubility of calcium is significantly higher by 3 points in the presence of 1.5% humates than in its absence. Similarly, the solubility of phosphorus is not-significantly higher (p.val=0.11) by 4 points in the presence of humates than in its absence. Finally, the solubility of magnesium is also not-significantly higher (p.val=0.08) by 4 points in the presence of humates than in its absence.

CONCLUSION

[0112] The results have shown that the solubilities Ca and P are affected in the feed of the test by the presence of MCP provided with humates. These results show that the solubilities of Ca and P are significantly better in the presence of humates (+3 and 4 points respectively). The solubility of Mg is systematically significantly better (from 1 to 4 points) in the presence of humates, whatever the digestion test.

Example 4: In Vitro Study of the Mycotoxin Absorption Capacity of the Product of the Invention in the Digestive Tract of Monogastric Animals

[0113] The goal of this in vitro study is to evaluate the capacity of the product of the invention to absorb mycotoxins under the digestion conditions of monogastric animals. The principle consists in placing the product in contact with a given dose of mycotoxins at a given temperature and pH.

Experimental Protocol

[0114] Four mycotoxins were chosen according to the recurrence of their presence in the feeds or plant raw materials intended for monogastric animals, but also according to the sensitivity of poultry and pigs (young and adults) towards these. The dose of each of the four mycotoxins was determined according to the European regulations (aflatoxin B1) and the sensitivity of the two species (zearalenone, ochratoxin A and deoxynivalenol). Thus, three doses (0.5%, 0.8% and 1.0%) of the product of the invention MCPH15 were placed in contact with 100 to 900 ng/ml of mixed mycotoxins (Table 4).

TABLE-US-00004 TABLE 4 Doses of mycotoxins and product of the invention introduced into the monogastric digestion digestion system in vitro Dose of mycotoxin Dose of product of the Mycotoxin (ng/ml) invention (g/100 g) aflatoxin B1 20 0.5 zearalenone 250 0.8 ochratoxin A 100 1.0 deoxynivalenol 900

[0115] The concentration of mycotoxins was measured after each incubation time in order to determine the quantity absorbed by the product.

[0116] The concentrations of mycotoxins were measured by UHPLC-MS/MS. More precisely, a 0.1 M buffered phosphate solution was prepared at pH 2.500.05 (average pH.sub.stomach between the pigs and poultry) and enriched with mycotoxins to the concentrations given above. The sample was added to the solutions enriched with mycotoxins and the suspension was then incubated at 40 C. for 1 hour. The suspension was then centrifuged and the supernatant was sampled. The remaining supernatant was rejected, and the test material was then placed in suspension in a buffered 0.1 M solution of phosphate at pH 6.800.05 (average PH.sub.intestine between pigs and poultry) before being incubated at 40 C. for an additional 3 hours. The suspension was centrifuged again and the supernatant was sampled. The sample solutions were enriched with isotopically labelled internal standards before being analysed by UHPLC-MS/MS. The quantification was performed using external calibration solutions enriched with isotopically labelled internal standards. All the samples were prepared in quintuplicate.

[0117] The absorption rate is calculated as follows:

Absorption rate stomach (%)=([mycotoxin][mycotoxin].sub.stomach supernatant)/[mycotoxin].sub.initial;

Final absorption rate (%)=1(([mycotoxin].sub.stomach supernatant+[mycotoxin].sub.intestine supernatant)/[mycotoxin].sub.initial).

Results

[0118] The results show that the higher the product dose in the system, the higher the rate of absorption. The absorption rates at low pH (stomach level) are likewise higher than at high pH (intestine level). Passing into the intestine, the rise in pH results in a certain desorption of mycotoxins. More precisely, the absorption capacity is, in decreasing order: aflatoxin>zearalenone>ochratoxin>deoxynivalenol. Once in the intestine, the desorption rate (quantity of mycotoxins released into the system because not retained by the binder) is respectively 20, 37, 50 and 100% for deoxynivalenol, aflatoxin, zearalenone and ochratoxin. The product does not have any absorption capacity in the intestine for the latter (FIGS. 9 and 10).

Example 5Test 1 In Vivo on Meat Poultry: Effect of the Product of the Invention on Performance, Metabolism and Meat Quality

Study Conditions

[0119] Zootechnical test on 48 cages of 40 male chickens inside a conventional rearing building having a standard consumption index (CI D35=1.50).

Feed

[0120] Distribution of six feeds (i.e. 8 repetitions per feed) to meat chickens of the strain Ross 308 during 35 days of rearing.
The feed was manufactured so as to incorporate the tested products during mixing of the raw materials.
The feed does not contain microbial phytase and the raw materials chosen are low in endogenous phytases.
Products tested: monocalcium phosphates provided or not provided with humates. The doses of humates in the product of the invention vary from 1.0 to 2.5%.

Measurements

[0121] The following measurements were performed during rearing: [0122] Collective weight at D0, D7, D13, D21 and D28 (total weight of the animals/number of individuals) and individual weight at D35; [0123] ADG (average daily gain) at D7, D13, D21, D28 and D35 (average weight/number of days in the period); [0124] Feed consumption at D7, D13, D21, D28 and D35 (weight of leftover feed at each age, each bag of feed was weighed and identified upstream); [0125] CI (consumption index) at D7, D13, D21, D28 and D35 (ADG/weight of feed consumed for each period); [0126] Dose of blood minerals at D35 [0127] Dose of minerals in the tibia at D35 [0128] Quality of tibias: [0129] Latency-to-lie test at D27 [0130] Post-mortem bone force and strength [0131] Post-mortem meat quality
The conditions of the study are summarised in Table 5 below.

TABLE-US-00005 TABLE 5 Test conditions on meat chickens in vivo Rearing characteristics Standard consumption index (CI) Feed phases 3 phases: Start-up (D0-D13), Growth (D13-D28) and Finishing (D28-D35) Feed Recommendations Ross: 95% metabolisable energy, characteristics proteins and lipids Intake of P in the form of monocalcium phosphate (MCP) Start-up: 100% of digestible P in the six feeds Growth and finishing: 100% of digestible P with MCP in feed positive control 80% of digestible P with MCP in feed negative control Replacement of MCP by the product of the invention at a level of 80% digestible P in the last four feeds. Products tested: products of the invention containing 1.0 to 2.5% humates. Zootechnical Weight, Average Daily Gain, Feed consumption, CI measurements Litter quality Individual Weight at D35 measurements Metabolism Blood doses D35 measurements Bone measurements Doses of minerals in the tibias at D35 Latency-to-lie test at D27 Post-mortem bone strength measurements Meat quality Weight of carcass and meat pieces Evaluation of wooden breast and white stripping pH of the meat Number of animals 8 repetitions of 40 males

Results of the Study In Vivo

[0132] The results have shown the effects on the consumption index [FIG. 11], quality [FIG. 12] and bone strength [FIG. 13] and pH of the meat [FIG. 14].

[0133] At a lower dose of digestible P in the feed, the CI is better by 2 points (1%) with MCPH10 and MCPH15 compared with the positive control (p.val=0.05) [FIG. 11]. Similarly, the pH of the meat is closest to the objective at the end of 15 minutes maturation (pH=6.4) with the intake of MCPH20 (p.val=0.02) whereas at 7 days storage (pH=5.6) this is the case with the intake of MCPH10 (p.val=0.13) [FIG. 14].

At an equal dose of digestible P in the feed, the breaking force of the tibias is 10% higher and 12% higher respectively with MCPH10 and MCPH20 compared with the negative control (p.val=0.004) [FIG. 13]. In addition, the ash content and the quantities of Ca and P in these same tibias are higher by 0.4 (p.val=0.001), 0.8 (p.val=0.004) and 1.7% (p.val=0.04) respectively with the intake of MCPH10 compared to the negative control [FIG. 14]. Thus, at equal quantity of nutrients, 80% of digestible P in the ration taken in with the help of the product of the invention makes it possible to obtain a better CI than with 100% digestible P intake with a standard monocalcium phosphate. Similarly, at an equal quantity of nutrients including digestible P, the product of the invention makes it possible to obtain better results for bone quality and meat preservation.

Example 6Meat Poultry Test In Vivo: Effect of the Product of the Invention on Performance, Leg Quality and Litter

Study Conditions

[0134] Zootechnical test on 16 cages of 30 male chickens within a conventional rearing building having a prevalence of at least 70% pododermatitis and a standard consumption index (CI=1.50 to D35 and 1.65 to D42).

Feed

[0135] Distribution of two feeds (i.e 8 repetitions per feed) to meat chickens of the strain Ross 308 during 42 days of rearing. The feed was manufactured so as to incorporate the tested products during mixing of the raw materials. At the end of production, a phytase is added on-top in the feeds at an equivalent dose of 500 FTU/kg. Products tested: monocalcium phosphates provided or not provided with humates (Control feed: MCPH0; Test feed: MCPH15).

Measurements

[0136] The following measurements were performed during rearing: [0137] Collective weight at D0, D7, D13, D21, D28 and D35 (total weight of the animals/number of individuals) and individual weight at D42; [0138] ADG (average daily gain) at D7, D13, D21, D28, D35 and D42 (average weight/number of days in the period); [0139] Feed consumption at D7, D13, D21, D28, D35 and D42 (weight of leftover feed at each age, each bag of feed was weighed and identified upstream); [0140] CI (consumption index) at D7, D13, D21, D28, D35 and D42 (ADG/weight of feed consumed for each period); [0141] Litter quality at D7, D13, D21, D28, D35 and D42 (litter score according to the grid of the ITAVI (French poultry technical institute)); [0142] Pododermatitis at D21 and D42 (score according to the ITAVI score grid); [0143] Residual potassium in the litter at D21 and D42 (2 samples per cage, one sample at mid-distance between the edge of the cage on the sleeping side and the pipette line and at one third of the way between the feeder and the edge of the cage; one sample at mid-distance between the edge of the cage on the sleeping side and the pipette line and two thirds of the way between the feeder and the edge of the cage) by assay of potassium, using an inductively coupled plasma atomic emission spectrometer (ICP-AES), after mineralisation in hot aqua regia.

[0144] The conditions of the study are summarised in Table 6 below.

TABLE-US-00006 TABLE 6 Test conditions on meat chickens in vivo Rearing 70% prevalence of pododermatitis characteristics Standard consumption index (CI) Feed phases 3 phases: Start-up (D0-D13), Growth (D13-D28) and Finishing (D28-D42) Feed Recommendations Ross: 6% metabolisable characteristics energy, 10% raw proteins, 10% digestible lysine, 6% calcium, 10% phosphorus, 11% Ca/P ratio relative to needs Intake of P in the form of monocalcium phosphate (MCP) Standard dose (500 FTU/kg) of phytase on-top. Start-up: 100% digestible P, in the two feeds Growth and finishing: Replacement of the MCP by the product of the invention at a level of 80% digestible P (22.7% P, 17.0% Ca and 0.0% Na) in one of the two feeds. Products tested: MCPH0 and MCPH15 Zootechnical Weight, Average Daily Gain, Feed consumption, CI measurements Litter quality Individual Weight at D42 and pododermatitis measurements measurements Litter measurements Humidity Phosphorus Number of animals 8 repetitions of 30 males

Results of the Study In Vivo

[0145] Among the results obtained, significant effects were observed on the consumption index (CI) over the period 0-35 days, as illustrated in FIG. 15.
This shows that the CI is significantly lower by 2 points (1%; p.val=0.056) over the period 0-35 days with the product of the invention (MCPH15), compared with the control MCPH0. This result confirms that which was already observed in the previously presented test. Moreover, at an equal dose of digestible phosphorous in the feed, the product of the invention can significantly reduce (p.val=0.001) by 14% the quantity of P.sub.2O.sub.5 in the litter at D42 relative to the intake of a standard monocalcium phosphate (FIG. 16).