Composition comprising a mixture of bacillus subtilis and lactic acid bacterium intended for improving the health of young oviparous animals

Abstract

The invention relates to a composition comprising a mixture of at least one strain of a bacterium of the genus Bacillus and at least one strain of lactic acid bacterium for use in the prevention of infections in farmed oviparous animals.

Claims

1. A method for the prevention of infections or the improvement of zootechnical performance levels in oviparous animals, comprising a step of placing in direct contact with the eggs of farmed oviparous animals, a composition essentially consisting of a mixture of: a. the following three strains of Bacillus subtilis: NOL01, NOL02, NOL03, said strains being deposited at the CNCM under the respective numbers: CNCM I-4606, CNCM I-5043 and CNCM I-4607, and b. the Lactotoccus lactis spp lactis 1 strain of lactic acid bacterium, strain NOL11, said strain being deposited at the CNCM under number CNCM I-4609.

2. The method according to claim 1, wherein said composition comprising from 10.sup.4 to 10.sup.11 bacterial colonies of Bacillus and from 10.sup.4 to 10.sup.11 bacterial colonies of lactic acid bacterium, the bacterial colonies being per mL or g of composition.

3. The method according to claim 1, wherein the strains of Bacillus are in sporulated form and/or in vegetative form.

4. The method according to claim 1, wherein said composition is placed in contact with said eggs by spraying, incubation, soaking, spreading or powder dusting.

5. The method according to claim 1, wherein said composition further being in contact with the newborns having hatched from said eggs from said farmed oviparous animals and/or in contact with the food supplied to the newborns having hatched from said eggs from said farmed oviparous animals.

6. The method according to claim 1, wherein said farmed oviparous animals are birds and said composition is applied on the shell of the eggs of said birds.

7. The method according to claim 1, wherein said farmed oviparous animals are aquatic oviparous animals and said composition is dispersed in the water of the rearing tanks of said aquatic oviparous animals in order to be placed in contact with said eggs, or in order to be placed directly in contact with said eggs.

Description

LEGEND OF THE FIGURES

(1) The graph of FIG. 1 shows the cumulative mortality of fries after hatching in troughs in two tests (diamonds and squares) and in the control trough (triangles). This cumulative mortality is expressed in percentage of the number of eggs hatched as a function of time (in days).

(2) The graph of FIG. 2 shows the same graph as FIG. 1, in which the pathological events have been mentioned. A: Very severe costiasis, B: Very severe septicemic flavobacteriosis/costiasis, C: Severe costiasis, and ID: Very severe costiasis.

(3) FIG. 3 shows the average weight of the fries in the different lots. The x-axis shows the time in days, and the y-axis shows the average weight of the fries in grams. The diamond-shaped points show the average weights of the fries belonging to the control lot, the triangles show that of the lot which received soaking of the eggs followed by an application of the composition in the environment, and the circles show that of the lot having received soaking of the eggs followed by an application of the composition in the food.

(4) FIG. 4 shows the cumulative mortality of the fries over time in the different lots. The x-axis shows the time in days, and the y-axis shows the mortality percentage. The diamond-shaped points show the mortality in the control lot, the triangles show the mortality in the lot having received soaking of the eggs followed by an application of the composition in the environment, and the circles show the mortality of the lot having received soaking of the eggs followed by an application of the composition in the food. The curves show the statistical models associated with the actual mortalities with, in thin dotted lines, the model of the control lot, in solid lines, the model of the lot receiving application by soaking of the eggs and in the environment, and lastly, in thick dotted lines, the lot for application by soaking of the eggs and in the food.

EXAMPLES

Example 1—Efficacy of the Application of the Composition on Rainbow Trout Fries by Soaking of the Eggs and in the Environment

(5) The aim of this test is to determine the efficacy of the composition according to the invention on rainbow trout fries. The composition was applied on the eggs and in the environment of the animals.

(6) Protocol:

(7) The experiment is conducted on a farm on a production line.

(8) The line corresponds to 3 hatching troughs with 60,000 eggs per trough. One will serve as control and the other two will receive the applications of the composition.

(9) Both types of application are done:

(10) Soaking of the Eggs:

(11) The eggs are soaked for 4 minutes in 2 L of river water for the control lot and in 2 L of river water mixed with a half-dose of product for the test lot.

(12) A dose of composition according to the invention was prepared from two

(13) separate 20 mL vials: Vial 1 containing Bacilli NOL01, NOL02 and NOL03, at a concentration of about 10.sup.9 CFU/mL of the 3 Bacilli, Vial 2, containing the Lactococci NOL11, at a concentration of about 10.sup.9 CFU/mL.

(14) Spraying on the Environment:

(15) The troughs and trays of the test lots are treated with 2 L of river water mixed with a half-dose of product each before being placed in the water.

(16) This treatment was repeated just before hatching, at the end of hatching, at the end of vitelline resorption, then once per week until the transfer date of the fries into the grow-out area. For each of these treatments, the water of the 3 troughs is decreased as much as possible so as not to harm the health of the fries, then increased to its normal level after the treatment of the test troughs. The total length of the test is about 2 months, period between the arrival of the eggs on the rearing farm and the transfer of the fries. This makes for a total of about 10 treatments.

(17) The spraying operations were done using a manual sprayer (garden sprayer type).

(18) Measurements and Results

(19) During the test, the daily mortality in the three troughs is read, the health events are also noted.

(20) FIG. 1 shows that the cumulative mortality appears to be approximately equivalent between the various troughs until September 10, where a less significant increase in mortality is observed in the test troughs. The mortality gap between the test troughs and the control trough deepens even more around September 19. Ultimately, 3,000 fewer deaths are observed in the test troughs than in the control trough on August 21, or a drop in mortality of 20 to 25%.

(21) These two dates correspond to health episodes that are a septicemic flavobacteriosis and a costiasis, as shown by FIG. 2.

(22) The water diverted from the river arriving in the troughs being the same for all 3 troughs, the pathogenic pressure is also equivalent. These results therefore show that the application of the composition as mentioned above makes it possible to avoid the mortality of the fries during a health episode.

(23) Conclusion

(24) The application of the composition by soaking on the eggs followed by an application in the environment of the rainbow trout fries makes it possible to limit the impacts of a pathogenic pressure on the health of the fries.

Example 2—Efficacy of the Application of the Composition on the Rainbow Trout Fries by Soaking of the Eggs, and in the Environment or Via the Food

(25) The aim of this test is to determine the efficacy of the composition according to the invention on the rainbow trout fries. The application of the composition being done on the eggs, and in the environment of the animals or on the eggs and via the food.

(26) Protocol:

(27) The experiment is done on a rearing farm on a production line.

(28) The line corresponds to 3 hatching troughs with 60,000 eggs per trough. One serves as control (trough 5) and the other two (trough 2 and trough 3) receive the applications of the composition according to the invention.

(29) The performed applications are as follows:

(30) Soaking of the Eggs:

(31) The eggs were soaked for 5 minutes in 2 L of source water for the control lot and in 2 L of source water mixed with a half-dose of product for the test lots, as defined in example 1.

(32) Application in the Environment:

(33) Before filling the troughs with water:

(34) Test trough 3: the day before the eggs arrive, the trough is filled with water with a dose of flora, then recirculated for 1 day.

(35) After filling the troughs with water:

(36) Test troughs 2, 3 and control trough 5: before each application, the water of the troughs is decreased as much as possible so as not to harm the health of the fries, then increased to its normal level after application of the flora in test trough 3.

(37) Test trough 3: The walls of the trough that are not submerged in the water are sprayed with 1 L of spring water mixed with a half-dose of product.

(38) This treatment is repeated just before hatching, at the end of hatching, at the end of vitelline resorption, then once per week until the transfer date of the fries into the grow-out area. The spraying operations were done using a manual sprayer (garden sprayer type).

(39) Application in the Food:

(40) Test trough 2: Once per week, as of the first day of feeding, a half-dose of flora will be mixed by hand in the food before feeding the fries. The flora will be incorporated into the food about 1 hour before distribution. The half-dose of flora will be mixed with the food in a bucket before distribution.

(41) Measurements and Results

(42) During the test, the daily mortality and the average weekly weight of the fries were read in each trough, as well as the health events.

(43) TABLE-US-00001 Average weight of the fries by trough (g) D27 D34 D41 D48 D55 D62 D69 D75 Trough 2 0.156 0.25 0.35 0.515 0.687 0.826 1.22 1.45 Trough 3 0.161 0.25 0.34 0.495 0.644 0.704 1.16 1.4 Trough 5 0.172 0.217 0.32 0.526 0.635 0.752 1.09 1.35

(44) FIG. 3 shows that the application of said composition by soaking of the eggs and via the food or in the environment improves the average weight of the fries, with a gain at the end of hatching respectively of 7.4% and 3.7% additional weight compared to the fries in the control group.

(45) FIG. 4 shows that the two lots with the application of said composition by soaking of the eggs and via the food or in the environment were less affected by the health episode that occurred at about 33 days in all of the troughs. This is reflected by a significantly lower mortality of the fries (between 12 and 16% lower mortality relative to the control lot).

(46) Conclusion

(47) The application of the composition according to the invention by soaking on the eggs followed by an application in the environment or in the food of the rainbow trout fries improves the weight gain of the fries and reduces the mortality of the animals.

Example 3—Poultry Application

(48) Experiments on poultry (hen) eggs were done.

(49) The invention is not limited to the described embodiments, and other embodiments will appear clearly to one skilled in the art.