Non-invasive methods, systems, and their use for determining the properties of chicken eggs and / or chicken embryos inside eggs using near-IR spectroscopy.

Abstract

To provide a convenient, rapid, and reliable method for determining one or more properties of a bird egg, especially a chicken egg, and / or one or more properties of a bird embryo inside the egg, especially a chicken embryo. [Solution] A non-invasive method for determining one or more properties of a bird egg, particularly a chicken egg, and / or one or more properties of a bird embryo inside the egg, particularly a chicken embryo, using near-IR spectroscopy is described herein. More specifically, a non-invasive method for determining the sex of a bird embryo inside a bird embryo, particularly a chicken embryo, is described herein. Furthermore, a system for non-invasively determining one or more properties of a bird egg, particularly a chicken egg, and / or one or more properties of a bird embryo inside the egg, particularly a chicken embryo, is described herein, as is the use of a spectrometer selected from a multi-channel spectrometer, a miniature diffraction grating spectrometer, and a monolithic miniature spectrometer, and in particular a computer program therefor, in the system and / or method described herein.

Description

[Technical Field]

[0001] The present invention relates to a non-invasive method for determining one or more properties of a bird egg, particularly a chicken egg, and / or one or more properties of a bird embryo inside the egg, particularly a chicken embryo, using near-IR spectroscopy. More specifically, the present invention relates to a non-invasive method for determining the sex of a bird embryo inside an egg, particularly a chicken embryo. The present invention further relates to a system for non-invasively determining one or more properties of a bird egg, particularly a chicken egg, and / or one or more properties of a bird embryo inside an egg, particularly a chicken embryo, and to the use of a spectrometer selected from a multi-channel spectrometer, a miniature diffraction grating spectrometer, and a monolithic miniature spectrometer in the system and / or method of the present invention. [Background technology]

[0002] The production of poultry eggs, particularly chicken eggs (used herein synonymously with the term "chicken eggs"), for human food consumption or chicken breeding is now often carried out on an industrial basis. Due to the large volume of eggs produced, there is a need for rapid and reliable methods to control the quality and properties of the eggs and the embryos inside them. Another industrial sector that requires controlled quality poultry eggs, particularly chicken eggs, is the production of vaccines, especially influenza vaccines. So-called "egg-based vaccine manufacturing processes" are currently used to produce inactivated vaccines as well as attenuated live vaccines.

[0003] Therefore, in egg production, chicken breeding, and egg-based vaccine manufacturing processes, there is a very important interest in obtaining rapid and reliable information regarding the properties of the egg, such as the fertilization status (fertilized or unfertilized) or the (internal) embryo load of the egg. Primarily in chicken egg production and chicken breeding, there is also a more specific interest in the properties of the chicken embryo inside the egg, such as the developmental stage of the chicken embryo, the vitality of the chicken embryo, and the sex of the chicken embryo. With the properties of the chicken egg or the chicken embryo inside determined, eggs containing chicken embryos with desirable properties can be identified and further used.

[0004] For example, breeding egg-laying hens is generally not well-suited for meat production due to economic and product quality considerations. Consequently, the male chicks from these egg-laying hens are rarely raised and are currently slaughtered after hatching. This existing practice is not only economically inadequate but also highly undesirable from an ethical standpoint, and efforts have been made for some time to avoid culling male chicks after hatching.

[0005] Several studies have already dealt with methods for hatching bird eggs that include a step of determining one or more properties of the bird egg and / or one or more properties of the bird embryo inside the egg, including the following: Reference US 5,575,237 discloses a method for hatching bird eggs. Reference WO 2010 / 150265 A2 concerns hyperspectral identification of egg fertilization and sex. Reference WO 2014 / 033544 A2 deals with spectrophotometric analysis of chicken embryo feather color. Reference WO 2019 / 174661 A1 describes a device for testing fertilized eggs. D. Gohler et al. report on the sex determination of chicken embryos at 14 days of age using a non-destructive method of pattern analysis with hyperspectral imaging (VIS / NIR spectrum) for laying hens that have sex-specific downy coloration. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] US 5,575,237 [Patent Document 2] WO 2010 / 150265 A2 [Patent Document 3] WO 2014 / 033544 A2 [Patent Document 4] WO 2019 / 174661 A1

Non-Patent Literature

[0007]

Non-Patent Literature 1

Summary of the Invention

Problems to be Solved by the Invention

[0008] In view of existing prior art, there remains a need for a convenient, rapid, and reliable method for determining one or more properties of bird eggs, particularly chicken eggs, and / or one or more properties of the avian embryo inside the egg, particularly the chicken embryo.

[0009] Correspondingly, a main object of the present invention is to provide a convenient, rapid, and reliable method for determining one or more properties of bird eggs, particularly chicken eggs, and / or one or more properties of the avian embryo inside the egg, particularly the chicken embryo. A more specific object of the present invention relates to a non-invasive method for determining the sex of an avian embryo inside an egg, particularly a chicken embryo.

[0010] Another object of the present invention is to provide a simple and inexpensive system for non-invasively determining one or more properties of bird eggs, particularly chicken eggs, and / or one or more properties of the avian embryo inside the egg, particularly the chicken embryo.

[0011] Yet another object of the present invention relates to expanding the application field of a certain type of spectrometer.

Means for Solving the Problems

[0012] The main object and other objects of the present invention are achieved by at least the following steps: M1) Obtaining an egg from a bird egg, preferably a chicken (egg-laying hen); M2) Irradiating the egg obtained in step M1) with light having a spectrum ranging from at least ≥700 nm to ≤900 nm from a light source for egg inspection. Step M3): a step of capturing light transmitted through the egg, which is a part of the light used for inspecting and irradiating the egg in step M2); Step M4): a step of obtaining, for each case, the transmission spectrum of the transmitted light captured in step M3) based on one or more specific wavelength ranges that are predetermined wavelength partial ranges within the wavelength range of ≧700 nm to ≦900 nm determined in step M2); and Step M6): a step of determining one or more properties of the bird egg, preferably a chicken egg, and / or one or more properties of the bird embryo inside the egg, preferably a chicken embryo, based on the transmission spectrum obtained in step M4), preferably based only on one or more specific wavelength ranges of this transmission spectrum; It has now been found that it can be achieved by a non-invasive method for determining one or more properties of a bird egg, preferably a chicken egg, and / or one or more properties of the bird embryo inside the egg, preferably a chicken embryo, comprising the steps above.

[0013] The present invention and its preferred variations and preferred combinations of parameters, properties, and elements are defined within the scope of the claims. Further, the preferred embodiments, details, and modifications of the present invention are defined and described in the following description and the examples shown below.

[0014] It has been experimentally found that the non-invasive method according to the present invention is very suitable for detecting or determining the fertilization status (fertilized or unfertilized) of an unhatched bird (preferably chicken) egg, the (internal) germ load of an unhatched bird (preferably chicken) egg, the viability of an unhatched bird (preferably chicken) embryo inside the egg, the developmental state of an unhatched bird (preferably chicken) embryo (inside the egg), and the gender of an unhatched bird (preferably chicken) embryo.

[0015] In step M1) of the non-invasive method according to the present invention, a bird egg is obtained. A preferred bird egg is an egg from a chicken (i.e., an egg of a laying hen). In a preferred variation of the method of the present invention, as will be described in more detail below, the egg is obtained from a breed of chicken that produces brown or brownish feathers on one gender and white or yellowish feathers on the opposite gender.

[0016] In step M2) of the non-invasive method according to the present invention, the eggs are candled with light from a light source having a spectrum in the wavelength range of at least ≥700 nm to ≤900 nm. Preferably, the eggs are candled from below. It is preferable to transmit the light from the light source through the egg along its long axis or short axis.

[0017] In step M3) of the non-invasive method according to the present invention, the light transmitted through the egg is preferably captured by a light capture means as described below. The captured transmitted light is a portion of the light used to candling the egg in step M2), and preferably the residual light (not captured) or at least a large portion of it is absorbed by the egg or its contents. In this case, the egg can be candled so that light from the light source used for candling directly enters the egg. However, in other cases, the light source used for candling can be positioned at a non-zero distance from the egg so that light from it does not directly enter the egg. More generally, in particular when the light is not directly captured on the surface of the egg, and / or when the light used to candling the egg is not a focused and / or directed beam of light that is taken into place, some of the light may enter the egg and then exit the egg again without being captured. Therefore, it is preferable that the residual light mentioned above, that is, the light that is not absorbed, comprises light absorbed by the egg or its contents, and light that did not enter the egg or entered the egg but was emitted from the egg without being absorbed.

[0018] In step M4) of the non-invasive method according to the present invention, the transmission spectrum of the transmitted light captured in step M3) is acquired. Preferably, the transmission spectrum is acquired by a suitable spectrometer in which the transmitted light is preferably guided by a suitable light guide, as will be described in more detail below. Acquiring the transmission spectrum of the transmitted light captured in step M3) based on one or more singular wavelength ranges may, in one example, mean that the acquired spectrum comprises only the singular wavelength ranges. More generally, the transmission spectrum may be acquired with higher precision in the singular wavelength ranges than anywhere else in the acquired transmission spectrum, and precision may mean, for example, the absence of loss determined with respect to the transmitted light or the fidelity of the acquired spectrum to the actual or true spectrum of the transmitted light. In fact, acquiring the transmission spectrum of the transmitted light captured in step M3) based on one or more singular wavelength ranges may also mean that the acquired transmission spectrum preferably comprises at least the singular wavelength ranges with sufficient precision and / or fidelity. In either case, how the transmission spectrum is acquired, and therefore the acquired transmission spectrum, will generally depend on the technical means used to acquire the transmission spectrum, preferably the specifications of the spectrometer, but also on, for example, the light guide means. Therefore, these technical means, i.e., the spectrometer and / or, for example, the light guide means, can be adapted to enable the acquisition of transmission spectra based on a specific wavelength range.

[0019] In step M6) of the non-invasive method according to the present invention, one or more properties of a bird egg, preferably a chicken egg, and / or one or more properties of a bird embryo inside the egg, preferably a chicken embryo, are determined. Preferably, a determination unit (described in more detail below) is used to determine these properties in step M6) (including preferred variations of step M6), as discussed herein. Preferably, the determination unit comprises a data processing unit adapted for this purpose. Preferably, the data processing unit further comprises software suitable for the purposes of the present invention.

[0020] Preferred items are, One or more properties of a bird egg, preferably a chicken egg, are selected from the group consisting of the fertilization state of the egg and the (internal) germ load of the egg, and / or One or more properties of the bird embryo inside the egg, preferably a chicken embryo, are selected from the group consisting of the developmental stage of the bird embryo, preferably a chicken embryo, the vitality of the bird embryo, preferably a chicken embryo, and the sex of the bird embryo, preferably a chicken embryo, and / or The following additional stage M5): M5) A step in which the transmission spectrum of the transmitted light obtained in step M4) or the absorbance spectrum based thereon is compared with the corresponding transmission spectrum or absorbance spectrum in the same 1 or 2 or more specific wavelength ranges (preferably only 1 or 2 or more defined specific wavelength ranges) from a predetermined database, where known values ​​of 1 or 2 or more properties of a bird egg, preferably a chicken egg, and / or 1 or 2 or more properties of the bird embryo inside the egg, preferably a chicken embryo, are particularly defined in the 1 or 2 or more specific wavelength ranges and the corresponding transmission spectrum or the corresponding absorbance spectrum. It includes and / or The following stage M6): M6) Determining one or more properties of a bird egg, preferably a chicken egg, and / or one or more properties of the bird embryo inside the egg, preferably a chicken embryo, based on the transmission spectrum obtained in step M4), and / or determining based on the results of comparing the transmission spectrum of the transmitted light obtained in step M4) or the absorbance spectrum based thereon with the corresponding transmission spectrum or corresponding absorbance spectrum determined in step M5) in each of the same one or two or more specific wavelength ranges from a predetermined database, in one or more specific wavelength ranges (preferably limited to one or more specific wavelength ranges). The present invention is a method comprising (or a preferred method according to the present invention as described herein) that includes

[0021] In step M5) of the non-invasive method according to the present invention, the transmission spectrum of the transmitted light obtained in step M4) or the absorbance spectrum therefrom is compared with the corresponding transmission spectrum or absorbance spectrum in each of the same 1 or 2 or more singular wavelength ranges from a predetermined database, in 1 or more singular wavelength ranges, which determine a known value of 1 or more properties (described above or below) to be determined. Preferably, in step M5) (comprising a preferred variation of step M5) as described herein), a determination unit (described in more detail below) is used to compare the spectra (as described above or below). Preferably, the determination unit comprises a data processing unit adapted for this purpose. Preferably, the data processing unit further comprises software suitable for the purposes of the present invention.

[0022] The comparison in step M5) compares only a portion of the spectrum instead of the entire spectrum, and therefore requires processing less data, thus enabling the efficient determination of one or more properties in question. Preferably, the singular wavelength ranges are predetermined based on the type of property to be determined, i.e., also based on the type of egg and / or bird, and based on the acquired transmission spectra from a predetermined database that can be considered to further form training data. When the property to be determined is known (generally by using means other than spectral analysis, such as conventional sex determination of chicks after hatching, when the property to be determined is the sex of the embryo inside the egg), and this knowledge is part of the training data, analysis of training data comprising spectra acquired with respect to eggs can reveal that certain wavelength ranges in the spectrum specifically indicate the property to be determined, and these wavelength ranges can then be selected as singular wavelength ranges. This analysis can be performed by a human or artificial intelligence. For example, a machine learning architecture can be trained on a defined spectral database to accept acquired spectra (described above or below) defined in steps M1) to M4) as input and provide as output one or more properties determined for each egg and / or embryo inside it. However, the comparison step in step M5) described above can be brought about by many other methods. Generally, the comparison step can be understood as determining the similarity between spectra acquired (as defined above or below) in steps M1) to M4) and not in the database, and corresponding spectra from the database that are in the same one or more singular wavelength ranges. If the similarity exceeds a given threshold, it can be assumed that the values ​​of one or more properties determined by each spectrum from the database also apply to the acquired spectra. Basically any similarity measure between two (individual) functions can be used to measure similarity.

[0023] The non-invasive method according to the present invention is particularly preferred for determining one or more properties of a chicken egg and / or one or more properties of a chicken embryo inside the egg.

[0024] In the method of the present invention, in step M5) defined above, it is preferable to compare the absorbance spectrum based on the transmission spectrum of the transmitted light obtained in step M4) with the corresponding absorbance spectrum in one or more specific wavelength ranges from a predetermined database, each in the same one or more specific wavelength ranges (preferably only one or more predetermined specific wavelength ranges), which determines known values ​​for one or more properties of a bird egg, preferably a chicken egg, and / or one or more properties of a bird embryo inside the egg, preferably a chicken embryo.

[0025] In the method of the present invention, in step M6) defined above, it is preferable to determine one or more properties of a bird egg, preferably a chicken egg, and / or one or more properties of the bird embryo inside the egg, preferably a chicken embryo, based on the results of comparing the absorbance spectrum based on the transmission spectrum of the transmitted light obtained in step M4) in one or more unique wavelength ranges (preferably limited to one or more unique wavelength ranges) with the corresponding absorbance spectra defined in step M5) in each of the same one or more unique wavelength ranges from a predetermined database.

[0026] In the method of the present invention, the “absorption spectrum” as defined herein is preferably determined based on the transmission spectrum obtained in step M4) and the calibration spectrum, which is the measured spectrum of the light used to illuminate the eggs in step M2), and preferably the transmission spectrum on which the absorption spectrum is determined is corrected based on the dark current spectrum corresponding to the spectrum obtained under conditions that are equivalent to the transmission spectrum of step M4) except that the light does not pass through the eggs. In one example, I0 refers to the calibration spectrum, and I tr This refers to the acquired transmission spectrum, I dark When it points to the dark current spectrum, the absorption spectrum I ab to I ab =I0-(Itr -I dark ) can be considered to be calculated as. However, the absorption spectrum can be considered to be defined or calculated differently. When the transmission spectrum is obtained in a state where no ambient light exists, it may be considered that there is no need to measure the dark current spectrum and use it to calculate the absorption spectrum. The term "absorption spectrum" used in this specification can be considered to actually mean absorbance, that is, the absorbance spectrum. In one example, I0 refers to the calibration spectrum, and I tr refers to the acquired transmission spectrum, and I dark refers to the dark current spectrum, the absorbance spectrum can be calculated from the transmission spectrum as I ab = log 10 [I0 / (I tr - I dark )].

[0027] The non-invasive method of the present invention, particularly in its specific or more specific variations defined herein, and more specifically in this method, is very suitable for determining the gender of the avian embryo inside the egg, particularly the chicken embryo, when eggs from birds that produce feathers of one color on one gender and feathers of another color on the opposite gender are used. It has been found by self-experiment that such birds are preferably examples of chickens of breeds that produce feathers of one color on one gender and feathers of another color on the opposite gender, particularly chickens of breeds that produce brown or brownish feathers on one gender and white or yellowish feathers on the opposite gender. Such breeds of chickens are known in the art, for example, chickens of the brown egg-laying breeds known as "Hy-Line Brown", "Roman Brown" (Lohmann Tierzucht GmbH, Germany), or "ISA Brown". "ISA Brown" is a crossbred chicken with a color scheme related to gender. "ISA" is the initial of "Institut de Selection Animale", the company that developed this crossbred in 1978.

[0028] Therefore, in specific modifications, preferably a non-invasive method for determining the sex of a chicken embryo, As stage M1) M1) The stage of obtaining eggs from chicken breeds that produce feather color differentiation based on sex. It includes and / or As an additional stage (M5) M5) A step in which the transmission spectrum or absorption spectrum based thereon obtained in step M4) is compared with a corresponding transmission spectrum or corresponding absorption spectrum that determines the known sex of the chicken embryo, which is in the same 1 or 2 or more specific wavelength ranges from a predetermined database, preferably only in the 1 or 2 or more specific wavelength ranges, in one or more specific wavelength ranges. It includes and / or Stage M6) M6) Determine the sex of the chicken embryo based on the transmission spectrum obtained in step M4), and / or determine the sex based on the results of comparing the transmission spectrum of the transmitted light obtained in step M4) or the absorption spectrum based thereon with the corresponding transmission spectrum or corresponding absorption spectrum defined in step M5) in each of the same one or two or more specific wavelength ranges from a predetermined database, preferably only in one or two or more defined specific wavelength ranges. Equipped with, The above method according to the present invention (or the method according to the present invention described herein as preferred) is preferred.

[0029] In the method of the present invention, in step M5) defined above, it is preferable to compare the absorbance spectrum based on the transmission spectrum of the transmitted light obtained in step M4) with a corresponding absorbance spectrum that determines the known sex of the chicken embryo, which is in the same 1 or 2 or more peculiar wavelength ranges from a predetermined database, preferably only in the 1 or 2 or more predetermined peculiar wavelength ranges, in one or more peculiar wavelength ranges.

[0030] In the method of the present invention, in step M6) defined above, it is preferable to determine the sex of the chicken embryo based on the results of comparing the absorbance spectrum, which is based on the transmission spectrum of the transmitted light obtained in step M4), with the corresponding absorbance spectrum defined in step M5) in each of the same one or more specific wavelength ranges from a predetermined database, in one or more specific wavelength ranges, preferably only in one or more predetermined specific wavelength ranges.

[0031] A more specific variation is a non-invasive method for determining the sex of a chicken embryo, M1) The stage of obtaining eggs from chicken breeds that produce feather color differentiation based on sex. M2) A step in which the eggs obtained in step M1) are irradiated with light having a spectrum that extends at least in the wavelength range from ≥700nm to ≤900nm. M3) This stage involves capturing the light that has passed through the egg, which is a portion of the light used to illuminate the egg in stage M2. M4) A step in which the transmission spectrum of the transmitted light captured in step M3) is acquired in each case based on one or more specific wavelength ranges which are predetermined wavelength subranges within the wavelength range of ≥700 nm to ≤900 nm defined in step M2), M5) A step in which the absorbance spectrum based on the transmission spectrum of the transmitted light obtained in step M4) is compared in one or more specific wavelength ranges with corresponding absorbance spectra from a predetermined database that are only present in each of the same one or more specific wavelength ranges, preferably one or more predetermined specific wavelength ranges, which determine the known sex of the chicken embryo (i.e., the corresponding absorbance spectrum acts as a reference spectrum), and M6) A step in which the sex of a chicken embryo is determined based on the results of comparing the absorbance spectrum obtained in step M4) from a predetermined database with the corresponding absorbance spectrum determined in step M5) in each of the same one or two or more specific wavelength ranges, preferably only in one or two or more predetermined specific wavelength ranges, in one or more specific wavelength ranges. The above method according to the present invention (or the method according to the present invention described herein as preferred) is preferred.

[0032] Another preferred method is a non-invasive method for determining the sex of a chicken embryo inside an egg, wherein the egg obtained in stage M1) Obtained from chicken breeds that produce brown or brownish feathers, preferably brown or brownish down, on one sex and white or yellowish feathers, preferably white or yellowish down, on the other sex, and / or Preferably, obtained from brown egg-laying chickens selected from the group consisting of Highline Brown chickens, Roman Brown chickens, and ISA Brown chickens, and / or These are fertilized eggs. The above method according to the present invention (or the method according to the present invention described herein as preferred).

[0033] It has been found that using the method according to the present invention yields particularly good results in determining the sex of the bird embryo inside the egg, especially the chicken embryo, when the feather color of the bird embryo, especially the chicken embryo, is expressed to a degree that allows for sufficiently accurate detection of feather color by the method at the time of application. In many cases, the feather color of the bird embryo, especially the chicken embryo, more specifically the chicken embryo of the chicken breed preferred by the method of the present invention (as described above), is sufficiently expressed when incubated for a period of ≥9 to ≤15 days after laying, preferably ≥12 to ≤14 days, and more preferably ≥13 to ≤14 days. More generally, the incubation period of the bird egg before applying the method of the present invention (as described above or below) can be selected depending on the time required for the bird embryo inside the egg to express characteristics that exhibit the properties to be determined, thereby affecting the optical properties of the embryo and / or egg, particularly the transmission properties, in one or more specific wavelength ranges. Bird embryos, particularly chicken embryos, can be cited as examples of such characteristics because the color of their downy feathers can depend on the sex of the embryo.

[0034] Furthermore, in the variant of the present invention relating to determining the sex of a bird embryo inside an egg, particularly a chicken embryo, when eggs are used from birds, particularly chicken breeds, that produce one color of feathers on one sex and another color of feathers on the other sex, it has been found in our experience that the most significant spectral information is obtained from the wavelength range ≥700 nm to ≤900 nm or a preferred sub-range thereof, particularly from the defined specific wavelength (sub-range) range described herein. These findings are particularly applicable when the eggs used in the method of the present invention are obtained from chicken breeds that produce brown or brownish down on one sex and white or yellowish down on the other sex.

[0035] the result, The eggs (preferably chicken eggs) obtained in step M1) have been kept warm for a period of time ranging from ≥9 to ≤15 days after laying, preferably from ≥12 to ≤14 days, more preferably from ≥13 to ≤14 days, and / or The light used to irradiate the eggs in step M2) is light having a spectrum that extends at least from ≥720nm to ≤870nm, preferably >750nm or from ≥750nm to ≤870nm. The method according to the present invention (or the method according to the present invention as described herein as preferred) is preferred.

[0036] Our experience has shown that, regarding the capture of light transmitted through the egg in step M3) of the method of the present invention, the diameter of the measurement spot on the surface of the egg must be selected to be sufficiently large so that occasional inaccurate placement of the egg within the carrier or carrier rack compartment (described below) does not adversely affect the measurement results to a degree that is considered significant in practice. The measurement spot can be defined as the region on the surface of the egg from which the light transmitted through the egg is captured.

[0037] Therefore, equally preferred methods (with respect to all variations of the methods of the present invention described herein) are, At stage M3), on the surface of the egg, Having a diameter ranging from ≥0.5cm to ≤2.5cm, preferably in the range of ≥1cm to ≤2.3cm, and / or ≥0.2cm on the surface of the egg 2 from ≤5cm 2 Preferably, up to ≥0.8cm 2 from ≤4cm 2 The area extends to the range of The present invention involves capturing light that has passed through an egg within a designated measurement spot (or the preferred method according to the present invention as described herein).

[0038] In particular, during our experiments, we have found that when processing large quantities of eggs, or to reduce the amount of scattered light that may otherwise interfere with the measurement, it is preferable in the method of the present invention to place the eggs used in a carrier, especially in a separate compartment of a carrier rack. Preferably, to further reduce the amount of scattered light, the partition walls, i.e., the partition walls of the compartments, are colored black. Preferably, the carrier or carrier rack is configured to allow illumination of the eggs using a light source preferably from below in step M2) of the non-invasive processing according to the present invention, and further in step M3) to allow light transmitted through the eggs to be taken in preferably from the opposite side (preferably from above) from the side on which the light source is located.

[0039] A preferred method is that (with respect to all variations of the method of the present invention described herein) step M1) The step of providing a carrier, preferably a carrier rack, having a plurality of compartments, each configured to be suitable for receiving bird eggs, preferably chicken eggs, and separated from each other by partition walls to reduce the amount of scattered light, Preferably, the carrier, preferably the carrier rack, is configured to enable the illumination of bird eggs, preferably chicken eggs, placed in a compartment with a light source, and to further enable the capture of light transmitted through the eggs (preferably comprising a step of enabling light shielding coupling of the light source to the eggs). The above steps and, A carrier, preferably a carrier rack, is prepared by placing bird eggs, preferably chicken eggs, in one compartment of the carrier. The present invention further comprises (or the present invention as described herein as preferred).

[0040] In a more specific variation of the method according to the present invention, a carrier, preferably a carrier rack, can be used, which is disclosed and described in more detail in the published document WO 2019 / 174661 A1 (as "Trager(3)").

[0041] A light source suitable for irradiating eggs obtained in step M1) of the non-invasive method according to the present invention in step M2) comprises light having a spectrum in its emitted electromagnetic radiation spectrum that extends in the wavelength range of at least ≥700 nm to ≤900 nm, preferably a continuous spectrum that extends in the wavelength range of at least ≥700 nm to ≤900 nm. In this sense, suitable light sources include incandescent bulbs and halogen lamps, particularly tungsten halogen lamps. Typically, a light source having a power of 35 W and / or a luminous intensity of 1100 dc is suitable for the purposes of the method of the present invention. In particular, in variations of the method of the present invention for determining the sex of a bird embryo inside an egg, preferably a chicken embryo, a light source having higher power and / or higher luminous intensity can be advantageous. In particular, higher power lamps with higher luminous intensity of ≥35 W, preferably ≥40 W, more preferably ≥50 W, preferably ≤75 W, allow for shorter integration times and, as a result, higher processing speeds.

[0042] Therefore, a preferred method is that the light source for irradiating the eggs in step M2) Power of ≥35W, preferably ≥40W, more preferably ≥50W, preferably ≤75W, and / or Luminous intensity ≥1000cd, preferably ≥1100cd, more preferably ≥1200cd, even more preferably ≥1300cd, The method according to the present invention (or the method according to the present invention as described herein as preferred) is a halogen lamp having a halogen lamp, preferably a tungsten halogen lamp.

[0043] In a preferred variation of the present invention, eggs from birds, preferably chicken breeds, that produce feathers of a certain color on one sex and feathers of a different color on the other sex are used, and based on the transmission spectrum of the transmitted light taken in step M3) and acquired in step M4), wherein the one or more singular wavelength ranges of step M4) are the wavelength ranges of ≥720nm to ≤760nm, ≥730nm to ≤830nm, ≥750nm to ≤870nm, and / or ≥800nm ​​to ≤870nm, as will be specified in more detail below, we have found in our experience that particularly good results are achieved in determining the sex of the bird embryo inside the egg, preferably a chicken embryo.

[0044] Furthermore, particularly good results in these preferred variations of the method of the present invention relating to determining the sex of a bird embryo inside an egg, preferably a chicken embryo, are obtained in a more preferred variation of the invention, comprising step M5), wherein step M5) compares the transmission spectrum of the transmitted light obtained in step M4) with the corresponding transmission spectra that determine the known sex of the chicken embryo in each of the same one or more singular wavelength ranges selected from the wavelength ranges of ≥720 nm to ≤760 nm, ≥730 nm to ≤830 nm, ≥750 nm to ≤870 nm, and ≥800 nm to ≤870 nm. These findings are particularly applicable when the eggs used in the method of the present invention are from chicken breeds that yield brown or brownish down on one sex and white or yellowish down on the other sex.

[0045] Therefore, a preferred method is preferably a non-invasive method for determining the sex of a chicken embryo. One or more singular wavelength ranges of stage M4) Wavelength range from ≥720nm to ≤760nm, Wavelength range from ≥730nm to ≤830nm, Wavelength ranges from ≥750nm to ≤870nm, preferably >750nm or ≥750nm to ≤830nm, and Wavelength range from ≥800nm ​​to ≤870nm, Selected from a group consisting of, Preferably, one or more singular wavelength ranges of step M4) Wavelength ranges from ≥750nm to ≤870nm, preferably >750nm or ≥750nm to ≤830nm, and Wavelength range from ≥800nm ​​to ≤870nm, Selected from the group consisting of, and / or Stage M5) The transmission spectrum of the transmitted light obtained in step M4) or the absorbance spectrum based thereon, Wavelength range from ≥720nm to ≤760nm, Wavelength range from ≥730nm to ≤830nm, Wavelength ranges from ≥750nm to ≤870nm, preferably >750nm or ≥750nm to ≤830nm, and Wavelength range from ≥800nm ​​to ≤870nm, Selected from the group consisting of, preferably Wavelength ranges from ≥750nm to ≤870nm, preferably >750nm or ≥750nm to ≤830nm, and Wavelength range from ≥800nm ​​to ≤870nm, In one or more singular wavelength ranges of step M4) selected from the group consisting of, preferably only one or more singular wavelength ranges, A step of comparing a corresponding transmission spectrum or corresponding absorbance spectrum in the same one or more specific wavelength ranges from a predetermined database with a corresponding transmission spectrum or corresponding absorbance spectrum that determines the known sex of a chicken embryo. It includes and / or Stage M6) M6) The sex of the bird embryo inside the egg, preferably a chicken embryo, The transmission spectrum of the transmitted light obtained in step M4) or the absorbance spectrum based thereon is used in one or more specified singular wavelength ranges defined in step M5), The known sex of the chicken embryo is determined in each of the same one or more specific wavelength ranges, using the corresponding transmission spectrum or corresponding absorption spectrum determined in step M5), A step in which a determination is made based on the results of comparing, preferably, only these spectra. Equipped with The above method according to the present invention (or the method according to the present invention described herein as preferred).

[0046] In the method of the present invention, in step M5) defined above, it is preferable to compare the absorbance spectrum based on the transmission spectrum of the transmitted light obtained in step M4) with the corresponding absorbance spectrum from a predetermined database that is in the same one or two or more specific wavelength ranges (defined above), preferably only within one or two specific wavelength ranges, and which determines the known sex of the chicken embryo.

[0047] In the method of the present invention, in step M6) defined above, the absorbance spectrum based on the transmission spectrum of the transmitted light obtained in step M4) is compared with the corresponding absorbance spectrum defined in step M5) for the known sex of the chicken embryo in the same one or two or more defined specific wavelength ranges defined in step M5), preferably, the sex of the chicken embryo is determined based on the results of comparing only these spectra.

[0048] For example, in a preferred variation of the method of the present invention as defined above, in step M6) to determine the sex of the bird embryo inside the egg, preferably a chicken embryo, the transmission spectrum of the transmitted light obtained in step M4) is compared with the corresponding transmission spectrum that determines the known sex of the chicken embryo in the same wavelength range of ≥800nm ​​to ≤870nm in the wavelength range of ≥800nm ​​to ≤870nm.

[0049] To further improve the results of the method according to the present invention, it is preferable to obtain a calibration spectrum and / or a dark current spectrum.

[0050] A preferred method comprises step M5), in which step M6) the sex of the bird embryo, preferably the sex of a chicken embryo, is determined based on the absorbance spectrum determined based on the transmission spectrum obtained in step M4) and the calibration spectrum, which is the measured spectrum of the light used to candle the egg in step M2). Preferably, the transmission spectrum, which is used to determine the absorbance spectrum, is corrected based on the dark current spectrum corresponding to the spectrum obtained under conditions where the transmission spectrum is equal to that of step M4) except that light does not pass through the egg. The present invention relates to a method (or a method described herein as preferred, preferably a method for determining the sex of a chicken embryo).

[0051] Another preferred method is the method according to the present invention (or the method according to the present invention as described herein as preferred, preferably the method for determining the sex of a chicken embryo), comprising step M6), wherein step M6) is the step of determining the sex of a chicken embryo, and the step is to process the absorbance spectrum based on the absorbance spectrum (determined based on the transmission spectrum obtained in step M4) and the calibration spectrum, which is the measured spectrum of the light used to illuminate the egg in step M2) as defined above, preferably based on the spectral absorption function determined by taking the derivative of the absorbance spectrum and / or smoothing the absorbance spectrum.

[0052] In the non-invasive method according to the present invention, excellent results were obtained when principal component analysis was performed on the spectral absorption function, as outlined in more detail below, using eggs from a breed of bird, preferably a chicken (as defined herein or as preferred herein), that produces feathers of a certain color for one sex and feathers of a different color for the opposite sex, in order to determine the sex of the bird embryo inside the egg, preferably a chicken embryo.

[0053] Therefore, a preferred method comprises step M6), in which the sex of a bird embryo, preferably a chicken embryo, is determined by determining whether the combination value of the combination of spectral absorption function values ​​at different wavelengths is greater than or less than a predetermined threshold. Preferably, the combination value is a linear combination of spectral absorption function values ​​at different wavelengths, and the coefficients of the linear combination are determined from the coefficients of one or more principal components obtained from principal component analysis performed on a statistical population of chicken embryos, and preferably, a predetermined threshold is zero. The present invention relates to a method (or a method described herein as preferred, preferably a method for determining the sex of a chicken embryo).

[0054] Analyzing the spectral transmittance and / or absorbance spectra measured for eggs using the principal components of these spectra derived from a training dataset has been found to enable a particularly efficient, yet still reliable, determination of egg properties. In this way, using principal component analysis allows a given measured spectrum or any (individual) function derived therefrom (such as the spectral absorption function referred to herein) to be converted into a set or list of undivided random variables {I} corresponding to the measured (light) intensity at different wavelengths (or each derived value of the function derived therefrom). λ This corresponds to viewing it as {I}. A set or list of random variables can be equivalently viewed as a single-multivalent, i.e., vector-like random variable. The underlying statistical population is a given set of eggs having a determining property, and by measuring the spectrum of each egg, a certain spectrum can be associated with each egg, and therefore with the corresponding values ​​of multiple single-valent random variables (or equivalently, the single "multivalent" of a multi-valent random variable). The property that distinguishes some eggs from others will result in differences in the spectra of the statistical population, insofar as it has an effect on the optical properties of the eggs, especially their transmission properties, and therefore in terms of differences in the values ​​of each (single-valent) random variable corresponding to the spectrum. These differences are the random variable {I}. λThe covariance matrix of {I} can be used to incorporate the properties. The overall properties will usually be reflected by the differences between different parts of the spectrum, each of which is relatively small and therefore may not be suitable as an indicator of the properties to be determined. It is desirable to find an indicator that will allow us to reliably estimate the properties to be determined from a given spectrum. Preferably, such an indicator will be a quantity that differs as much as possible between different spectra for each property. Principal component analysis is performed on the random variable {I}. λ This allows for rearranging linear combinations of} according to changes in random variables within a statistical population, thus enabling the deriving of an efficient selection of indicators from the spectrum to determine the properties of each egg based on the indicators.

[0055] Preferably, the "different wavelengths" in the "combination of spectral absorption function values ​​at different wavelengths" defined above (or below) are: Wavelength range from ≥720nm to ≤760nm, Wavelength range from ≥730nm to ≤830nm, Wavelength ranges from ≥750nm to ≤870nm, preferably >750nm or ≥750nm to ≤830nm, and Wavelength range from ≥800nm ​​to ≤870nm, It is selected from a group of wavelength ranges consisting of the following.

[0056] Furthermore, a preferred method comprises step M6), in which the sex of a bird embryo, preferably a chicken embryo, is determined based on a combination of values ​​relating to combinations of values ​​of the spectral absorption function at different wavelengths. Preferably, the combined value is determined from one or more principal components of the spectral absorption function, which is determined from a principal component analysis performed on a statistical population of bird embryos inside an egg, preferably chicken embryos. The present invention relates to a method (or a method described herein as preferred, preferably a method for determining the sex of a chicken embryo).

[0057] Similarly, a preferred method is one in which one or more principal components obtained from principal component analysis are included in the step of determining the combined value, and at least one of the one or more principal components is selected from the group consisting of a first principal component and a third principal component, preferably, One or more principal components are the first principal component and the third principal component, and / or One or more main components do not include a second main component. The present invention relates to a method (or a method described herein as preferred, preferably a method for determining the sex of a chicken embryo).

[0058] Similarly, the present invention relates to a system for non-invasively determining one or more characteristics of a bird egg (preferably a chicken egg) and / or one or more characteristics of a bird embryo (preferably a chicken embryo) inside the egg, preferably a system for non-invasively determining the sex of a chicken embryo, at least Element S1) A light source for irradiating eggs with light having a spectrum extending at least in the wavelength range from ≥700nm to ≤900nm. A light-up means (preferably one or more light-up means) for capturing transmitted light, which is the portion of light that has passed through the egg, from the light used to irradiate the egg for candling, having a spectrum defined by element S2) and element S1), Element S3) A spectrometer for acquiring the transmission spectrum of the transmitted light taken up by element S2), based on one or more singular wavelength ranges which are predetermined wavelength subranges within the wavelength range from ≥700nm to ≤900nm, and Element S4) is a decision unit, Bird eggs, preferably chicken eggs, possessing one or more properties and / or bird embryos inside the eggs, preferably chicken embryos, The above determination unit for more precisely determining the sex of a chicken embryo based on a transmission spectrum, The above-mentioned system is equipped with the above-mentioned features.

[0059] In general, in the context of non-invasive methods for determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg, all aspects of the present invention discussed herein apply with necessary modifications to the system for non-invasively determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg as defined above.

[0060] Preferably, the determination unit (element S4)) of the system according to the present invention is connected to a spectrometer (element S3)) and preferably to a data processing unit connected to a light source (element S1) or comprising such a unit. Preferably, the data processing unit is adapted to carry out all variations and preferred variations of steps M5) and M6) of the non-invasive method of the present invention disclosed herein, including step M5) for comparing the transmission spectra of transmitted light acquired in step M4) of the non-invasive method of the present invention, and step M6) for determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg based on the transmission spectra acquired in step M4). Preferably, the data processing unit comprises software suitable for the purposes of the present invention.

[0061] The preferred system is, The light-taking means S2) is preferably on the surface of the egg. Having a diameter ranging from ≥0.5cm to ≤2.5cm, preferably in the range of ≥1cm to ≤2.3cm, and / or ≥0.2cm on the surface of the egg 2 from ≤5cm 2 Preferably, up to ≥0.8cm 2 from ≤4cm 2 The area extends to the range of Adapted to capture transmitted light within a defined measurement spot (preferably only within this range), and / or Spectrometer S3) Transmitted light is adapted to detect regardless of its source, and / or Adapted to detect transmitted light without spatial resolution, and / or Selected from the group consisting of charge-coupled device detectors and photodiode array detectors, Equipped with a detector, This is a system according to the present invention (or a system according to the present invention described herein as preferred).

[0062] If the spectrometer (S3)) of the system according to the present invention is equipped with a detector adapted to detect transmitted light regardless of its source, the transmitted light is preferably the portion of the light used to illuminate the egg that has passed through the egg (element S2) as defined above, and has the spectrum (element S1) as defined with respect to the light source.

[0063] Preferably, the light acquisition means (see element S2) of the system according to the present invention comprises a lens, preferably an optical lens, for focusing or directing the acquired light to a light guide means (described below) or a spectrometer.

[0064] A similarly preferred system is one or more further elements S5) Light guiding means (preferably one or more light guiding means) for guiding the captured transmitted light from the light acquisition means S2) to the spectrometer S3), Preferably comprising one or more optical fibers adapted or optimized to transmit light with wavelengths in the range of ≥700nm to ≤900nm, preferably ≥720nm to ≤870nm, more preferably ≥750nm to ≤870nm, and even more preferably 750nm or ≥750nm to ≤870nm. The above-mentioned light guiding means, and / or S6) A carrier having a plurality of compartments, each configured to receive bird eggs, preferably chicken eggs, and separated from each other by partition walls to reduce the amount of scattered light, preferably a carrier rack, Preferably, it is configured to be suitable for irradiating eggs (preferably chicken eggs) placed in a compartment, and for capturing the light that has passed through the eggs using a light capture means. More preferably, it is configured to enable light-shielding coupling of a light source or light guide means to an egg (preferably a chicken egg), The above carrier (or the above carrier rack), The present invention comprises a system (or a system described herein as preferred) that includes the above.

[0065] Furthermore, a preferred system is one in which the light source (element S1) is Power of ≥35W, preferably ≥40W, more preferably ≥50W, preferably ≤75W, and / or Luminous intensity ≥1000cd, preferably ≥1100cd, more preferably ≥1200cd, even more preferably ≥1300cd, A halogen lamp having a tungsten halogen lamp, preferably a tungsten halogen lamp or the same. This is a system according to the present invention (or a system according to the present invention described herein as preferred).

[0066] Another preferred system is the system according to the present invention (or the system according to the present invention described herein as preferred), characterized in that the spectrometer S3) is selected from the group consisting of a multi-channel spectrometer, a miniature diffraction grating spectrometer, and a monolithic miniature spectrometer. More generally, it may be preferred to use a spectrometer that resolves incident light only spectrally and not spatially as the spectrometer S3). In other words, the spectrometer S3) does not necessarily need to be suitable for supplying an image, particularly a spectral image, based on the incident light. Preferably, the light acquisition means, light guide means, and / or spectrometer are adapted to the specific wavelength range referred to herein.

[0067] Those skilled in the art were particularly surprised to learn that such a relatively simple spectrometer as used in the present invention (i.e., a multi-channel spectrometer, a monolithic miniature spectrometer, or a miniature diffraction grating spectrometer) could be used to obtain very high accuracy in predicting the sex of a chicken embryo in a chicken egg (in this case, a chicken egg from a breed of chicken that produces feather color differentiation based on sex). Similar methods known in the prior art usually teach that a relatively complex hyperspectral camera is required for their respective purposes.

[0068] For industrial applications aiming for high throughput, a spectrometer with sufficient light sensitivity is advantageous. Our own experiments have found that multi-channel spectrometers (MCS) or compact diffraction grating spectrometers (CGS) very well satisfy these requirements. To further enhance industrial applicability, the inventors have found it advantageous to use a spectrometer or spectrometer system equipped with a charge-coupled device sensor and / or having ≥2 channels, preferably ≥5 channels, more preferably ≥5 and ≤12 channels, and even more preferably 10 channels.

[0069] Therefore, the preferred ones are, The spectrometer (element S3) A spectrometer selected from the group consisting of multi-channel spectrometers and miniature diffraction grating spectrometers, or A spectrometer system comprising one or more spectrometers selected from the group consisting of multi-channel spectrometers and miniature diffraction grating spectrometers, and or equipped with Preferably, A spectrometer and / or spectrometer system comprises ≥2 channels, preferably ≥5 channels, more preferably ≥5 and ≤12 channels, and even more preferably 10 channels. A spectrometer or spectrometer system comprising one or more spectrometers, or at least one or more spectrometers, includes one or more (preferably one) charge-coupled device sensors. This is a system according to the present invention (or a system according to the present invention described herein as preferred).

[0070] For example, in one preferred variation of the present invention, the system according to the present invention comprises a multichannel spectrometer having 1 or more than 10 channels as a spectrometer (element S3)) and equipped with 1 or more charge-coupled device sensors. For example, in a further preferred variation of the present invention, the system according to the present invention comprises a spectrometer system having 10 channels as a spectrometer (element S3)) and comprising several miniature diffraction grating spectrometers, all of which are equipped with charge-coupled device sensors.

[0071] When the system according to the present invention includes a spectrometer or spectrometer system having two or more channels as a spectrometer, preferably, electronic multiplexing is applied to process these two or more channels.

[0072] In one preferred variation of the system of the present invention, the system of the present invention (or the system of the present invention as described herein as preferred) is combined with or comprises an element thereof of a device for inspecting fertilized eggs disclosed in document WO 2019 / 174661 A1. For example, in such a variation, the carrier of the system of the present invention may be designed as the carrier (3) disclosed in document WO 2019 / 174661 A1, and the system of the present invention may comprise an egg transport unit designed as the egg transport unit (12) disclosed in document WO 2019 / 174661 A1. Such an egg transport unit may be connected to a determination unit (element S4) of the system of the present invention to form a device or system for determining one or more properties of a bird egg (preferably a chicken egg) and / or one or more properties of a bird embryo (preferably a chicken embryo) inside the egg, and for sorting eggs according to the results of determining these one or more properties.

[0073] The present invention also relates to the use of a spectrometer selected from the group consisting of a multichannel spectrometer, a miniature diffraction grating spectrometer, a monolithic miniature spectrometer, and a combination thereof, in a system and / or method for non-invasively determining one or more characteristics of a bird egg, preferably a chicken egg, and / or one or more characteristics of a bird embryo inside the egg, preferably a chicken embryo, preferably for non-invasively determining the sex of a bird embryo inside the egg, preferably a chicken embryo. Preferably, the eggs are chicken eggs obtained from chicken breeds that produce feather color differentiation based on sex. Regarding the use described above.

[0074] In general, all aspects of the present invention discussed herein in the context of non-invasive methods for determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg, and in the context of a system according to the present invention, are applicable with modifications necessary for the use of the spectrometer defined above.

[0075] The present invention also relates to a computer program for determining one or more characteristics of a bird egg and / or one or more characteristics of a bird embryo inside the egg based on a transmission spectrum obtained according to steps M1) to M4) of the method defined above, particularly in any of its specific implementations, wherein the computer program comprises instructions to cause the computer to perform steps M5) and / or M6) of the method defined above, particularly in any of its specific implementations. The “computer” that runs the computer program may also correspond to one or more data processing units. For example, the computer program may execute the determination unit S4) of the system defined above, in which case the determination unit may comprise one or more data processing units.

[0076] Computer programs can sometimes be stored / distributed on suitable media such as optical storage media or solid-state media supplied together with or as part of other hardware, but they can also be distributed in other forms, such as through the internet or other wired or wireless telecommunication systems.

[0077] In this specification, parameter ranges, particularly the lower and upper limits of the wavelength range, are specified, and the symbols ≧ and ≦ are used to indicate that the respective limit values ​​are included in the specified range, while the symbols > and < indicate that the respective limit values ​​are not included.

[0078] The present invention will be further explained and illustrated with the accompanying drawings, which briefly describe it below. [Brief explanation of the drawing]

[0079] [Figure 1] This figure shows a part of the measuring instrument for carrying out a non-invasive method for determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg according to the present invention. An element of the system for non-invasively determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg according to the present invention is shown, namely a carrier rack (element S6)) having multiple compartments, each configured to be suitable for receiving a chicken egg and separated from each other by partition walls to reduce the amount of scattered light. Furthermore, a light acquisition means (two detector heads above the carrier rack, element S2)) is shown for guiding the acquired transmitted light from the light acquisition means to a spectrometer (element S3) not shown in Figure 1. [Figure 2]This figure shows a detail of Figure 1, in which two chicken eggs from a brown-egg laying hen (Roman Brown) are placed in two different compartments of a carrier rack (element S6). The carrier rack is configured to allow the chicken eggs placed in the compartments to be illuminated (see the opening at the bottom of the compartment to allow for the mounting of the light source). Above the eggs in the compartments are two detector heads (light capture means, element S2) for capturing the light that has passed through the eggs. [Figure 3] This figure shows the details of the measuring equipment shown in Figure 1. To allow us to see the three halogen lamps (35W) (light source, element S1)) located below the carrier rack and connected to a determination unit (element S4) not shown in Figure 3, the carrier rack (element S2)) is absent in Figure 3. The halogen lamps can be blocked by a rotating shutter. Above the halogen lamps, two detector heads (light acquisition means, element S2)) can be seen. [Figure 4] Figures 1 to 3 show two exemplary transmission spectra obtained using the measuring instruments shown. Transmission spectrum 401 was obtained for a chicken egg containing a male embryo, and transmission spectrum 402 was obtained for a chicken egg containing a female embryo. [Figure 5] Figure 4 shows an example absorbance spectrum calculated from the transmission spectrum, where absorbance spectrum 501 is calculated from transmission spectrum 401, and absorbance spectrum 502 is calculated from transmission spectrum 402. [Figure 6] This figure shows exemplary absorbance spectra calculated from transmission spectra obtained for the entire (training) statistical population of chicken eggs. Two spectral bands, 601 and 602, can be roughly identified, but they are not clearly separated. Spectral band 601 consists of absorbance spectra calculated for eggs containing male chicken embryos, and spectral band 602 consists of absorbance spectra calculated for eggs containing female chicken embryos. [Figure 7]This figure shows the spectral absorption function corresponding to the absorbance spectrum in Figure 6 after processing including smoothing and the formation of the first derivative. Spectral band 701 was formed by processing the absorbance spectrum within band 601, and spectral band 702 was formed by processing the absorbance spectrum within band 602. By comparing Figure 7 with Figure 6, it can be seen that the separability of these spectral bands has been improved by processing over at least a large portion of the illustrated wavelength range. [Figure 8] This figure exemplifies the loading of the first principal component ("PC-1") determined from the processed absorbance spectrum shown in Figure 7, which is the spectral absorption function. [Figure 9] This figure exemplifies the loading of the third principal component ("PC-3") determined from the processed absorbance spectrum shown in Figure 7, which is the spectral absorption function. [Figure 10] This figure shows a point group where each point corresponds to the spectral absorption function expressed with respect to the first and third principal components, i.e., the processed absorbance spectrum. The circular ("f") dots and triangular ("m") dots belong to the absorbance spectral bands 702 and 701 shown in Figure 7, respectively. These absorbance spectra form the basis for training, i.e., for determining the principal component loadings by performing principal component analysis, while the square dots ("0") correspond to the spectral absorption function determined in the same way as for the absorbance spectra in bands 701 and 702 used for training, i.e., the processed absorbance spectrum, but the underlying transmission spectrum is obtained using eggs in which the sex of the chicken embryos contained within was unknown, thereby forming a control group or test group. The line 1001 shown in Figure 10 is selected to most clearly separate the circular ("f") dots from the triangular ("m") dots, and therefore, preferably, to equally clearly separate the square ("0") dots into "female dots" and "male dots". [Figure 11]Exemplary transmission spectra 1101, 1102, and 1103 obtained for chicken eggs incubated over various periods are shown, illustrating the significant decrease in overall light transmitted through the egg as the embryo develops within the egg. [Modes for carrying out the invention]

[0080] Examples The following embodiments further illustrate and illustrate the present invention without limiting its scope.

[0081] Brown chicken eggs were obtained from Lohmann Tierzucht GmbH and used in all experiments in these examples. All eggs came from a breed of chicken (brown egg-laying hen) that produced brown or brownish down in female chicks and white or yellowish down in male chicks. Therefore, the sex of the chicks could be determined or confirmed based on the feather color after hatching.

[0082] The following measurement system was used in the experiments for the examples in this section. Measurement system 1 : Spectrometer: A multi-channel spectrometer ("MCS") with a CCD ("charge-coupled device") sensor, e.g., Carl Zeiss MCS FLEX CCD. Wavelength range: 190~980nm 2 channels, integration time: Channel 1: 500ms, Channel 2: 1500ms Light guide means: Near-IR single-core 600μm optical fiber cable Measurement system 2 : Spectrometer: A monolithic miniature spectrometer (MMS, e.g., made by Carl Zeiss) with a PDA ("photodiode array") sensor. Wavelength range: 300~1100nm 2 channels, integration time: Channel 1: 10000 ms, 1 × Light guide means: Near-IR single-core 600μm optical fiber cable A standard 35W halogen lamp with a cold light reflector (Osram) was used in all experiments. [Examples]

[0083] Example 1: Optimization of experimental parameters (including "training execution") A. Experimental setup: The measuring equipment used in the experiments in these embodiments consisted of a carrier rack with multiple compartments for egg placement separated from each other by partition walls. Eggs were placed in these compartments and illuminated from below using a light source (specified above). A detector head was positioned above the eggs to capture the light transmitted through them. The diameter of the measurement spot on the egg surface allowed by the detector head was approximately 2 cm. This relatively large diameter of the measurement spot has been found to reduce the sensitivity of the measurement to inaccurate placement of eggs within the carrier rack compartments (which occasionally occurred). The detector head was connected to a spectrometer (measurement system 1 specified above) through a light guide (specified above) to obtain the transmission spectrum of the captured transmitted light. The spectrometer and light source were connected to a data processing unit.

[0084] To improve measurement quality, a calibration spectrum was obtained and dark current measurements were performed before the actual measurements. The calibration spectrum was obtained as a reference for the absorbance spectrum of the egg being measured. For this purpose, the light source spectrum was measured without the sample (egg), and the absorbance of the sample (egg) was determined by comparing it with the sample (egg) spectrum. In the measurement of the calibration spectrum, the light intensity was reduced to a level that could be processed by the spectrometer by using neutral glass filters (Schott NG4 and Schott NG9).

[0085] B. Measurement of the spectrum: 1191 chicken eggs, which had been kept warm for 13 to 14 days, were removed from the incubator and placed in compartments of a carrier rack (40 eggs per carrier rack). These eggs were illuminated from below with a light source, and the light transmitted through the eggs was captured within a designated measurement spot on the egg surface (a measurement spot with a diameter of approximately 2 cm). This light was then transmitted to a spectrometer (measurement system 1) via a light guide (optical fiber cable) and recorded. The received transmission spectra were further analyzed as described below.

[0086] C. Confirmation of the sex of chickens after hatching. After the measurements were taken (as described under item B above), the eggs were kept incubated further until the chicks hatched, and after hatching, the sex of the chickens was determined by control methods known in the art. The following results have been found. TIFF2026136151000002.tif32153 The information obtained in this way was used to further analyze the received spectrum.

[0087] D. Analysis of the spectrum (obtained according to the method described under item B above)

[0088] D.1 The measured transmission spectra were obtained as spectral files in ASCII format. For analysis, these files were imported into data analysis software (Unscrambler by Camo Analytics). Next, a reference dataset was added using known transmission spectra of male embryos inside male eggs and female embryos inside female eggs. In the raw spectra accepted as described above, the transmission range between 620 nm and approximately 980 nm was focused on, but the spectral range between 180 nm and 620 nm was not. It was found that the raw spectra obtained from eggs containing male embryos tended to show higher light transmittance than the spectra obtained from eggs containing female embryos.

[0089] D.2 Next, a training dataset was generated from the combined data of groups 1 and 2 (see above), and the absorbance spectra (raw transmission spectra and calibrated spectra, see above) were calculated for each. Regarding the absorbance spectra, there was no clear separation between the two groups, but a tendency for higher light absorption was observed in eggs with female embryos compared to eggs with male embryos. Two exemplary raw transmission spectra are shown in Figure 4, where spectrum 401 refers to an egg with a male chicken embryo, and spectrum 402 refers to an egg with a female chicken embryo. Figure 5 illustrates absorbance spectra calculated from the transmission spectra shown in Figure 4 within a narrow wavelength range where substantial transmission was observed. Absorbance spectrum 501 was calculated from transmission spectrum 401, i.e., corresponding to an egg with a male chicken embryo, and absorbance spectrum 502 was calculated from transmission spectrum 402, i.e., corresponding to an egg with a female chicken embryo.

[0090] D.2.1 Next, the absorbance spectra were smoothed according to the Savitsky-Gorey method, and the first derivatives were formed therefrom. In this exemplary analysis, more precisely, the absorbance spectra were first smoothed according to the Savitsky-Gorey method, and then the first derivatives of each smoothed spectrum were formed. This results in a considerably improved separation between the data points accepted for eggs with female embryos and the data points accepted for eggs with male embryos, as can be seen by comparing Figure 6, which shows the absorbance spectra corresponding to the training dataset, with Figure 7, which shows the processed absorbance spectra, i.e., the first derivatives of the smoothed absorbance spectra, which may commonly be called spectral absorption functions. In Figure 6, the spectral band 601 of the spectrum corresponding to the egg containing a male chicken embryo substantially overlaps with the spectral band 602 of the spectrum corresponding to the egg containing a female chicken embryo. However, in Figure 7, the spectral band 701 of the processed spectrum corresponding to the egg containing a male chicken embryo substantially overlaps with the spectral band 702 of the processed spectrum corresponding to the egg containing a female chicken embryo only in a relatively narrow intermediate wavelength range of approximately 775 nm.

[0091] Similar results were accepted in each experiment using measurement system 2 (as defined above).

[0092] D.2.2 For further analysis, spectral ranges were defined according to spectral characteristics (position of spectral bands), and these were used to test the quality of sex determination of chicken embryos inside eggs. The first analysis included spectral ranges within ≥620nm to ≤980nm, ≥810nm to ≤850nm, ≥800nm ​​to ≤870nm, and ≥820nm to ≤840nm. Subsequently, principal component analysis was performed for the ranges within ≥620nm to ≤980nm, ≥810nm to ≤850nm, ≥800nm ​​to ≤870nm, and ≥820nm to ≤840nm. In the first-order approximation, the clearest difference between egg groups containing female embryos and egg groups containing male embryos was observed in the spectral range ≥800nm ​​to ≤870nm, where the separation between spectral bands 701 and 702 was particularly clear, as already shown in Figure 7. In this case, the difference between sexes lies on the first principal component, illustrating that the greatest change in optical properties between eggs in groups 1 and 2, on which the training datasets were based, was attributable to the sex of the chicken embryo contained within the egg.

[0093] Therefore, in this case, the first principal component can be considered a suitable indicator of the sex of the chicken embryo inside each egg, but there may also be other principal components that can determine the sex of the chicken embryo inside each egg, or that are suitable for determining other properties of each egg. For example, the loading of the seventh principal component from each spectral intensity, i.e., the coefficients of the linear combinations that form the seventh principal component, cannot be ignored, and these coefficients form a peak, for example, around 815 nm. Therefore, it is possible to still use the seventh principal component as an indicator of any property that affects the transmission and / or absorption characteristics of the egg at wavelengths around 815 nm. In fact, the transmission spectra obtained for groups 1 and 2 have a major peak near this wavelength, as can be seen exemplarily in Figure 4, and therefore, the combination of the first and seventh principal components was also considered for determining the sex of the chicken embryo.

[0094] By using principal components that indicate the properties of the egg or the chicken embryo inside it, the corresponding (single) values ​​can be used as representative values ​​for each spectrum for the purpose of determining these properties. Therefore, once the loadings of the principal components indicating the properties are determined from the training dataset, it is possible to avoid analyzing the entire spectrum and focus on analyzing only one or more (monovalent) principal components that are relevant, thereby enabling high computational efficiency.

[0095] To determine the sex of a chicken embryo, one might consider focusing only on the first principal component of each spectrum, and determining the sex depending on whether this component is greater than or below a predetermined threshold. For the overall statistical population of eggs, it can be assumed that the sexes of chicken embryos inside the egg are distributed almost evenly, and therefore the predetermined threshold can be set to zero. More generally, to determine the sex of a chicken embryo, one might consider focusing on the first and seventh principal components of each spectrum, and determining the sex depending on whether the combination of the first and seventh principal components, for example, a linear combination, is greater than or below a straight line positioned based on training data in the plane spanned by the first and seventh principal components. This determination can often be seen as equivalent to determining whether the ratio of the first and seventh principal components, one of which is appropriately shifted at a predetermined time, is greater than or below a predetermined threshold.

[0096] During this analysis, it was found that the second principal component may not be a precise indicator of the sex of the chicken embryo. Instead, it is expected that the second principal component will show changes in the transmission spectrum due to various measurement channels related to different spectral sensors. In fact, it is generally considered preferable to determine one or more properties without relying on known wavelength subranges or principal components, regarding changes in the acquired transmission spectrum that do not show a determined property and sometimes show known characteristics of the applied measurement procedure.

[0097] D.2.3 Using the results from spectral analysis by the method under section D.2, a preliminary estimate of the high-probability predictive accuracy for the sex of chicken embryos inside the eggs was made for groups 1 and 2, based on information from the first principal component. The following accuracy for proper classification was predicted according to this preliminary estimate. Male embryos: Of the 428 embryos, 420 were correctly classified as male, and 8 were incorrectly classified as female. This number corresponds to approximately 98% being correctly classified as male. Female embryos: Of the 447 embryos, 422 were correctly classified as female, and 25 were incorrectly classified as male. This number corresponds to approximately 94% being correctly classified as female.

[0098] D.3 For further optimized spectral analysis, another training dataset was generated, computed, and smoothed for eggs from groups 1 and 2 as described above under item D.2, the absorbance spectrum of each was calculated, and its first derivative was formed according to Savitsky-Gorey.

[0099] D.3.1 Next, the spectral range was defined as described above under item D.2.2, and the quality of sex determination based thereon was tested. The spectral ranges included in this experiment were (i) ≥ 720 nm to ≤ 760 nm, and (ii) ≥ 800 nm to ≤ 870 nm. Thus, a significantly improved separation was found between the data points accepted for eggs containing female embryos and the data points accepted for eggs containing male embryos. The spectral ranges ≥ 730 nm to ≤ 830 nm and ≥ 750 nm to ≤ 870 nm, preferably > 750 nm or ≥ 750 nm to ≤ 830 nm, particularly the spectral region around the 750 nm wavelength (i.e., approximately ±15 nm, ±10 nm, or ±5 nm), provided particularly significant information regarding the sex of chicken embryos. This information corresponds to the total absorption near characteristic peaks in the transmission spectra illustrated in Figure 4, particularly the absorption that causes the dips between these peaks.

[0100] Principal component analysis was performed on the spectral range of the further optimized spectral analysis described in D.3.2 and D.3.2. The obtained loadings found for the first and third principal components are shown in Figures 8 and 9, respectively. The magnitudes of the respective loadings illustrate that the first principal component is between ≥730 nm and ≤830 nm, which can be substantially related to the sex-specific total absorption that appeared before and after two characteristic peaks in this region of the transmission spectrum, particularly as exemplified in Figure 4, and the third principal component is between the two characteristic peaks, which can be substantially related to the dip around the wavelength of 750 nm. This optimized analysis shows that sex-specific information can be particularly accurately represented using a combination of principal components 1 and 3, i.e., another improvement over the results from the experiments described above under item D.2. Principal component 1 was found to represent the nearly higher absorption of eggs with female embryos compared to eggs with male embryos in this analysis, while principal component 3 was found to represent yet another absorption characteristic that distinguishes eggs with female embryos from eggs with male embryos in this analysis.

[0101] When arriving at an index for a property determined by a combination of principal components, the property can be determined based on whether this combination of principal components is greater than or less than a predetermined threshold. Generally, principal components can be combined in any way that forms an appropriate index. For example, a property can be determined based on a linear combination of principal components.

[0102] As a result of optimization analysis, the first and third principal components of each spectrum can be considered to determine the sex of a chicken embryo, and it has been found that the sex can be determined depending on whether the combination of the first and seventh principal components, for example, a linear combination, is greater than or less than a straight line positioned based on the training data in the plane spanned by the first and third principal components. This determination can often be seen as equivalent to determining whether the ratio of the first and third principal components, one of which is potentially shifted appropriately in advance, is greater than or less than a predetermined threshold.

[0103] D.3.3 Using the results from spectral analysis by the method described in section D.3, an optimized estimation of the high-probability prediction accuracy for the sex of chicken embryos inside eggs was performed for groups 1 and 2, incorporating information from the first and third principal components (i.e., principal components 1 and 3 described above), as further detailed below with reference to Figure 10. This optimized estimation predicted the following accuracy for proper classification. Male embryos: Of the 428 embryos, 415 were correctly classified as male, and 13 were incorrectly classified as female. This number corresponds to approximately 97% being correctly classified as male. Female embryos: Of the 447 embryos, 425 were correctly classified as female, and 22 were incorrectly classified as male. This number corresponds to approximately 95% being correctly classified as female. Overall: Of the 875 eggs, 840 were correctly classified as either male or female, representing approximately 96% of the eggs correctly classified. [Examples]

[0104] Example 2: Prediction of the sex of chicken embryos inside control eggs using the method of the present invention ("Verification run") Referring to Figure 10 and further detailing below, the sex of the chicken embryos inside the eggs of group 5 (control group) was predicted according to the method described above under item D.3. Next, the predicted results were compared with the sex of the chicks confirmed after hatching. The following accuracy of chicken sex classification was found based on this prediction. Male embryos: Of the 103 embryos, 96 were correctly classified as male and 7 were incorrectly classified as female. This number corresponds to approximately 93% being correctly classified as male. Female embryos: Of the 105 embryos, 101 were correctly classified as female, and 4 were incorrectly classified as male. This number corresponds to approximately 96% being correctly classified as female. Overall: 197 out of 208 eggs were correctly classified as either male or female, which corresponds to approximately 99% of correctly classified eggs.

[0105] Figure 10 shows the corresponding spectral absorption functions, i.e., the first and second principal components obtained from principal component analysis performed on the training dataset for each processed absorbance spectrum, plotted in comparison. The triangular dots correspond to the spectra calculated for training data from group 1 (i.e., eggs containing male embryos), and the circular dots correspond to the spectra calculated for training data from group 2 (i.e., eggs containing female embryos). The plots in Figure 10 also include square dots corresponding to the spectral absorption functions calculated from data measured for group 5 (i.e., eggs whose internality is unknown, whether male or female), i.e., the processed absorbance spectra. The principal components for these processed absorbance spectra, which can be considered test spectra or control spectra, were calculated using loadings obtained from principal component analysis performed on the training data. The dividing line 1001 shown in Figure 10 represents the straight line that most clearly separates the triangular dots from the round dots, and the most clear separation can generally mean the absolute distance or the sum of the squares of the distances from each dot to the line. The slope of the line illustrates that both the first and third principal components are required for the optimal separation between the triangular dots and the round dots, and thus for the optimal distinction between eggs containing male embryos and eggs containing female embryos. The offset of the line from the origin mainly reflects the situation where the egg groups 1 and 2 are not of equal size. The sex of the embryos inside the eggs of group 5 (control group), represented by square dots in Figure 10, can be predicted based on the side of the line to which each square dot is located. Generally, determining the side of the line to which a point is located can be understood as determining whether it is greater than or less than a defined threshold given by this line in a rotated coordinate system. In this particular case shown in Figure 10, it can be expected that the square dot located on the lower right side of the line corresponds to an egg containing a male embryo, and the square dot located on the upper left side of the line corresponds to an egg containing a female embryo.Therefore, the offset from the origin and the slope of line 1001 represent a linear combination of the first and third principal components selected as indicators of the sex of the embryo inside the egg.

[0106] From the results of Examples 1 and 2, it can be seen that excellent results regarding the prediction of the sex of the chicken embryo inside the egg are available when the chicken eggs are obtained from a breed of chicken that produces brown or brownish feathers on one sex and white or yellowish feathers on the other sex, and when a multichannel spectrometer (preferably having a CCD sensor) or a monolithic miniature spectrometer (preferably having a PDA sensor) is used to acquire the transmission spectrum in step M3). Similar results can be expected according to preliminary experiments conducted by the inventors, where a miniature diffraction grating spectrometer (preferably having a CCD sensor) is used to acquire the transmission spectrum in step M3).

[0107] Those skilled in the art would not have assumed that the accuracy of predicting the sex of a chicken embryo inside an egg could be achieved with the relatively simple spectrometers used in these experiments (i.e., multi-channel spectrometers, monolithic miniature spectrometers, or miniature diffraction grating spectrometers) as demonstrated by the inventors in these experiments. Therefore, these results are surprising. Similar experiments conducted prior to these experiments used relatively complex hyperspectral cameras for their respective purposes.

[0108] The results of Experiments 1 and 2 show that the method optimized according to item D.3 of Example 1 allows for more accurate prediction of the sex of the chicken embryo inside the egg than, for example, the preliminary method according to item D.2 of Example 1. Therefore, it is considered preferable to use the combination of the first and third principal components of the acquired spectra to predict the sex of the chicken embryo inside the egg, and to determine the principal component loadings from the corresponding spectra acquired with respect to the training data. [Examples]

[0109] Example 3: Identification of immature chicken embryos inside an egg by the method of the present invention. Chicken eggs containing various types of immature chicken embryos were obtained as follows. The first group of eggs was incubated for less than one day so that the chicken embryos inside were almost undeveloped when the measurement method of the present invention was performed. The second group of eggs contained chicken embryos that were in a developmental state equivalent to about 9 to 10 days of incubation when the measurement method of the present invention was performed.

[0110] Next, the transmission spectra of eggs from both groups were recorded as described above in items A and B of Example 1.

[0111] In the first egg group (see above), the spectrometer became saturated due to extremely high light transmittance. This state can be identified by the horizontal line in the spectrum. Very similar results were observed when unfertilized eggs were used instead of eggs that had been incubated for less than a day.

[0112] In the second egg group, the light transmittance observed for eggs containing chicken embryos whose developmental stage was equivalent to that of eggs kept warm for approximately 9 to 10 days was considerably higher than that observed for eggs containing chicken embryos whose developmental stage was equivalent to that of eggs kept warm for approximately 13 to 14 days. On the other hand, the light transmittance observed for eggs containing chicken embryos whose developmental stage was equivalent to that of eggs kept warm for approximately 9 to 10 days was considerably lower than that observed for eggs kept warm for less than one day or for unfertilized eggs.

[0113] Therefore, it is thought that the first egg group, the second egg group, and eggs containing chicken embryos whose developmental stage is equivalent to that of eggs kept in a warm environment for approximately 13 to 14 days can be distinguished from each other by the light transmittance they allow. Light transmittance decreases as the embryo develops. Light transmittance can be measured in terms of defined characteristics of the transmission spectrum obtained for each egg, such as maximum transmittance, average transmittance, or total transmittance, and / or whether the transmission spectrum contains a horizontal line. For example, maximum transmittance can correspond to the peak height in each transmission spectrum. A horizontal line in the transmission spectrum, which is thought to exist instead of a peak in the spectrum, indicates that the spectrometer is saturated due to a large amount of light passing through the egg, and can indicate that the egg belongs to group 1 (or that the egg is an unfertilized egg). Figure 11 illustrates exemplary transmission spectra, where spectrum 1101 corresponds to eggs from group 1 and / or unfertilized eggs, spectrum 1102 corresponds to eggs from group 2, and spectrum 1103 corresponds to eggs containing chicken embryos at approximately 13 to 14 days of development.

[0114] Therefore, the results of the experiment in this Example 3 show that eggs containing immature chicken embryos can be easily and accurately identified by the method of the present invention. Accordingly, the method of the present invention is very suitable for detecting or determining the fertilization status of an egg (fertilized or unfertilized), the vitality of the chicken embryo inside the egg, and the developmental status of an unhatched chicken embryo (inside the egg).

[0115] Furthermore, the developmental stage of a bird embryo inside an egg, particularly a chicken embryo, can be determined using the method according to the present invention. For example, for this purpose, the transmission spectrum of a chicken embryo can be compared to a reference spectrum of similar chicken embryos in different developmental stages, for example, with respect to various incubation periods after laying.

Claims

1. A non-invasive method for determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg, At least the following steps: M1) The stage of obtaining bird eggs, M2) A step in which the eggs obtained in step M1) are irradiated with light from a light source having a spectrum extending over a wavelength range of at least ≥700 nm to ≤900 nm. M3) A step in which light transmitted through the egg is taken in, wherein the taken-in transmitted light is a part of the light used to irradiate the egg in step M2), M4) A step of obtaining the transmission spectrum of the transmitted light captured in step M3) based on one or more specific wavelength ranges which are predetermined wavelength subranges within the wavelength range of ≥700 nm to ≤900 nm as defined in step M2) in each case, and M6) A step in which the one or more properties of the bird egg and / or the one or more properties of the bird embryo inside the egg are determined based on the transmission spectrum obtained in step M4), A method characterized by comprising:

2. One of the aforementioned properties of the bird egg, or one of more than the aforementioned properties, is selected from the group consisting of the fertilization state of the egg and the embryo load of the egg, and / or One of the aforementioned properties or more of the aforementioned properties of the bird embryo inside the egg is selected from the group consisting of the developmental stage of the bird embryo, the vitality of the bird embryo, and the sex of the bird embryo, and / or The following stages are added as an additional stage (M5): M5) A step in which the transmission spectrum of the transmitted light within the one or more unique wavelength ranges obtained in step M4) or the absorption spectrum based on the transmission spectrum is compared with a corresponding transmission spectrum or corresponding absorption spectrum from a predetermined database within the same one or more unique wavelength ranges, wherein the corresponding transmission spectrum or corresponding absorption spectrum within the unique wavelength range determines known values ​​for the one or more properties of the bird egg, preferably a chicken egg, and / or the bird embryo inside the egg, preferably a chicken embryo. It includes and / or Stage M6) consists of the following stages: M6) A step of determining the 1 or 2 or more properties of the bird egg, preferably the chicken egg, and / or the 1 or 2 or more properties of the bird embryo, preferably the chicken embryo, inside the egg, based on the transmission spectrum obtained in step M4), and / or based on the result of comparing the transmission spectrum of the transmitted light within the 1 or 2 or more defined specific wavelength ranges obtained in step M4) or the absorption spectrum based on the transmission spectrum with the corresponding transmission spectrum or the corresponding absorption spectrum from a predetermined database within the same 1 or 2 or more specific wavelength ranges, as determined in step M5), Equipped with, The method according to claim 1, characterized by the features described above.

3. The method is a non-invasive method for determining the sex of the chicken embryo inside the egg. Preferably, Stage M1) consists of the following stages: M1) The step of obtaining eggs from a breed of chicken that produces differentiation of the chicken's feather color based on the chicken's sex, and / or The following stages are added as an additional stage (M5): M5) A step of comparing the transmission spectrum of the transmitted light within the one or more specific wavelength ranges obtained in step M4) or the absorption spectrum based on said transmission spectrum with the corresponding transmission spectrum or corresponding absorption spectrum from a predetermined database within the same one or more specific wavelength ranges that determines the known sex of the chicken embryo inside those eggs, and / or Stage M6) consists of the following stages: M6) A step of determining the sex of the chicken embryo inside the egg based on the transmission spectrum obtained in step M4), and / or based on the result of comparing the transmission spectrum of the transmitted light within the one or two or more defined specific wavelength ranges obtained in step M4) or the absorption spectrum based on the transmission spectrum with a corresponding transmission spectrum or corresponding absorption spectrum from a predetermined database within the same one or two or more specific wavelength ranges, as determined in step M5), Equipped with, The method according to claim 1 or 2, characterized by the features described herein.

4. The eggs obtained in step M1) are Obtained from chicken breeds that produce brown or brownish feathers for one sex and white or yellowish feathers for the opposite sex, and / or Preferably obtained from a breed of chicken that is a brown egg-laying breed selected from the group consisting of Highline Brown chickens, Roman Brown chickens, and ISA Brown chickens, and / or These are fertilized eggs. The method according to any one of claims 1 to 3, characterized by...

5. The eggs obtained in step M1) are kept warm for a period of time ranging from ≥9 to ≤15 days after laying, preferably from ≥12 to ≤14 days, more preferably from ≥13 to ≤14 days, and / or The light used to irradiate the eggs in step M2) is light having a spectrum that extends over a wavelength range of at least ≥720 nm to ≤870 nm, preferably >750 nm or ≥750 nm to ≤870 nm. The method according to any one of claims 1 to 4, characterized by...

6. The light passing through the egg is captured in step M3) within a predetermined measurement spot on the surface of the egg, The aforementioned measurement spot is Having a diameter in the range of ≥0.5 to ≤2.5 cm, preferably ≥1 to ≤2.3 cm, and / or The above egg surface is ≥0.2 to ≤5 cm 2 Preferably ≥0.8 to ≤4 cm 2 Extending over an area within the range, The method according to any one of claims 1 to 5, characterized by...

7. Stage M1) is A step of providing a carrier having multiple compartments, preferably a carrier rack, wherein each compartment is configured to receive a bird egg, preferably a chicken egg, and the compartments are separated from each other by partition walls to reduce the amount of scattered light, and preferably the carrier, preferably the carrier rack, is configured to allow the bird eggs, preferably chicken eggs placed in the compartments to be illuminated with a light source and to capture the light that has passed through the eggs, and The step of placing the bird eggs, preferably chicken eggs, in a compartment of the carrier, preferably the carrier rack, It also has, The method according to any one of claims 1 to 6, characterized by...

8. The light source for irradiating the eggs in step M2) is a halogen lamp, preferably a tungsten halogen lamp. The aforementioned halogen lamp is Power of ≥35W, preferably ≥40W, more preferably ≥50W, and preferably ≤75W, and / or Luminous intensity of ≥1000 cd, preferably ≥1100 cd, more preferably ≥1200 cd, even more preferably ≥1300 cd, Having, The method according to any one of claims 1 to 7, characterized by...

9. The one or more singular wavelength ranges of step M4) are, Wavelength range from ≥720 nm to ≤760 nm, Wavelength range from ≥730 nm to ≤830 nm, Wavelength ranges from ≥750 nm to ≤870 nm, preferably >750 nm or ≥750 nm to ≤830 nm, Wavelength range from ≥800 nm to ≤870 nm, Selected from the group consisting of, and / or The method is as follows, as stage M5: M5) Wavelength range from ≥ 720 nm to ≤ 760 nm, Wavelength range from ≥730 nm to ≤830 nm, Wavelength ranges from ≥750 nm to ≤870 nm, preferably >750 nm or ≥750 nm to ≤830 nm, Wavelength range from ≥800 nm to ≤870 nm, In the step of comparing the transmission spectrum of the transmitted light obtained in step M4) or the absorption spectrum based on the transmission spectrum within the one or more specific wavelength ranges selected from the group of wavelength ranges comprising the same one or more specific wavelength ranges, with the corresponding transmission spectrum or corresponding absorption spectrum that determines the known sex of the chicken embryo inside those eggs from a predetermined database within the same one or more specific wavelength ranges, It includes and / or The method proceeds as follows, in stage M6): A step in which the sex of the bird embryo inside the egg, preferably the chicken embryo, is determined based on the results of comparing the transmission spectrum of the transmitted light obtained in step M4) within one or more defined specific wavelength ranges as defined in step M5) or the absorption spectrum based on the transmission spectrum within one or more defined specific wavelength ranges with the corresponding transmission spectrum or corresponding absorption spectrum that determines the known sex of the chicken embryo inside the egg within the same one or more specific wavelength ranges as defined in step M5). Equipped with, The method according to any one of claims 1 to 8, characterized by...

10. The method comprises step M5), in which step M6) the sex of the bird embryo, preferably the sex of the chicken embryo, is determined based on the absorbance spectrum determined based on the transmission spectrum obtained in step M4) and the calibration spectrum which is the measured spectrum of the light used to illuminate the egg in step M2), Preferably, the absorbance spectrum and the transmission spectrum determined thereon are corrected based on the dark current spectrum, the dark current spectrum corresponding to the spectrum obtained under conditions equal to the transmission spectrum of step M4), except that light does not pass through the egg. A method according to any one of claims 2 to 9, preferably the method according to any one of claims 3 to 9.

11. In step M6), the sex of the bird embryo, preferably the chicken embryo, is determined by determining whether the combination value is above or below a predetermined threshold, the combination value means a combination of spectral absorption function values ​​at different wavelengths, the spectral absorption function is determined based on the absorbance spectrum by taking the derivative of the absorbance spectrum and / or smoothing the absorbance spectrum, Preferably, the combination value means a linear combination of the values ​​of the spectral absorption function at different wavelengths, the coefficients of the linear combination are determined from the coefficients of one or more principal components obtained from principal component analysis performed on the spectral absorption function determined with respect to a statistical population of chicken embryos inside those eggs, and preferably the predetermined threshold is zero, and / or In step M6), the sex of the bird embryo, preferably the chicken embryo, is determined based on a combination value, which means a combination of the values ​​of the spectral absorption function at different wavelengths. Preferably, the combination value is determined from one or more principal components of the spectral absorption function, where the one or more principal components represent principal component analysis performed on the spectral absorption function determined with respect to a statistical population of bird embryos inside those eggs, preferably chicken embryos. The method according to the present invention, characterized by the present invention.

12. One or more principal components obtained from principal component analysis are involved in the step of determining the combination value, and at least one of the one or more principal components is selected from the group consisting of a first principal component and a third principal component. Preferably, The one or more principal components are the first principal component and the third principal component, and / or The aforementioned one or more main components do not include a second main component. The method according to 11, characterized by the features described above.

13. A system for non-invasively determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg, preferably for non-invasively determining the sex of a chicken embryo inside the egg, At least the following elements: S1) A light source for irradiating the eggs with light having a spectrum extending over a wavelength range of at least ≥700 nm to ≤900 nm, S2) A light-taking means for taking in transmitted light, wherein the taken-in transmitted light is a portion of the light for irradiating the egg with an egg inspection, having a spectrum as defined in element S1), and the portion of the light-taking means passes through the egg. S3) A spectrometer for obtaining the transmission spectrum of the captured transmitted light as defined in element S2), wherein the transmission spectrum is based on one or more singular wavelength ranges, and the one or more singular wavelength ranges are predetermined wavelength subranges within the wavelength range from ≥700 nm to ≤900 nm in each case, and S4) A determination unit for determining the properties of the bird egg, preferably the chicken egg, and / or the bird embryo inside the egg, preferably the chicken embryo inside the egg, based on the transmission spectrum, more preferably for determining the sex of the chicken embryo inside the egg. Equipped with, Preferably, the spectrometer S3) is selected from the group consisting of a multi-channel spectrometer, a miniature diffraction grating spectrometer, and a monolithic miniature spectrometer. A system characterized by the following features.

14. The use of a spectrometer selected from the group consisting of a multichannel spectrometer, a miniature diffraction grating spectrometer, a monolithic miniature spectrometer, and a combination thereof, in a system and / or method for non-invasively determining one or more properties of a bird egg, preferably a chicken egg, and / or one or more properties of a bird embryo inside the egg, preferably a chicken embryo, and preferably for non-invasively determining the sex of a bird embryo inside the egg, Preferably, the egg is a chicken egg obtained from a chicken breed that produces a differentiation of the chicken's feather color based on the chicken's sex. A characteristic use.

15. A computer program for determining one or more properties of a bird egg and / or one or more properties of a bird embryo inside the egg based on a transmission spectrum obtained according to steps M1) to M4) of a method as defined in any one of claims 1 to 12, An instruction that causes a computer to perform steps M5) and / or M6) of the method as defined in any one of claims 1 to 12 when the program is executed by the computer, A computer program characterized by having the following features.

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