Method and device for optical in ovo sex determination of fertilized and incubated birds' eggs

Patent metadata

Field	Value
Publication number	US-10852284-B2
Application number	US-201716090947-A
Country	US
Kind code	B2
Filing date	Mar 20, 2017
Priority date	Apr 4, 2016
Publication date	Dec 1, 2020
Grant date	Dec 1, 2020

Abstract

Method and a device for the optical in ovo sex determination of fertilized and incubated birds' eggs. The method includes monitoring formation of at least one identifiable blood vessel, creating a hole in the shell of the egg, finding and irradiating the blood vessel with at least one laser beam source emitting an excitation wavelength, recording backscatter radiation of the irradiated blood vessel and evaluating backscatter radiation from recorded spectral intensity of fluorescence radiation in a spectral range redshifted to an excitation wavelength. Sex-specific properties of male and female blood are contained in the intensity and spectral profile of the recorded fluorescence radiation. where intensity levels for male blood has an evaluable value distinct from that of female blood. Further, determining the sex of the bird egg from difference values of the fluorescence intensity and displaying the sex determined for the embryo in the egg.

First claim

The invention claimed is:

1. A method for optical in ovo sex determination of fertilized and incubated birds' eggs comprising:
monitoring a chronological progression of incubation until at least one identifiable blood vessel or a heart develops with flowing blood;
creating a hole in a shell of the bird egg by a hole-creating unit;
finding blood vessels developing in the bird egg, or finding the heart, using a vision system and a coaxial or lateral illumination with light in a visible wavelength range;
positioning at least one blood vessel or the heart in a laser focus of at least one laser beam source, either by moving the egg or by moving an objective lens that produces the laser focus;
irradiating the at least one blood vessel or the heart with at least one laser beam source emitting an excitation wavelength;
recording backscatter radiation of the irradiated at least one blood vessel or heart using at least one detector that is connected to at least one evaluation unit connected to a downstream amplification and detector unit, wherein during the recording a movement of the at least one blood vessel or the heart out of the laser beam takes place by a tracking of the at least one blood vessel or the heart or the objective lens;
wherein:
evaluation of the backscatter radiation, including fluorescence radiation, in the evaluation unit from a recorded spectral intensity of the fluorescence radiation in a spectral range redshifted to an excitation wavelength, wherein the sex-specific properties of male blood and of female blood are contained in the recorded spectral intensity and in a spectral profile of the recorded fluorescence radiation, and wherein at least one intensity level determined from measured spectral intensities of the fluorescence radiation, or values assigned thereto, for the male blood has an evaluable value distinct from at least one determined intensity level, or the values assigned thereto, for the female blood in the blood vessels or in the heart;
determination of the sex of the bird egg from a difference of at least one of values of the fluorescence intensity levels, or the values assigned thereto, in the evaluation unit; and
subsequent display of the sex determined in the evaluation unit for an embryo in the bird egg.

2. The method according to claim 1 ,
wherein
green light is used as light in the visible wavelength range, with which light a radiation device of the vision system is provided in order to irradiate the at least one blood vessel ( 22 , 24 ) or the heart.

3. The method according to claim 2 ,
wherein
if one laser beam source is used, the fluorescent intensity levels determined for a predefined spectral range from an established fluorescence intensity / wavenumber curve are respectively defined for the male blood and for the female blood such that they are combined by a logical operation for evaluation.

4. The method according to claim 1 ,
wherein
fluorescent intensities are evaluated in the evaluation unit using one of the following methods or a combination of the following methods, in order to determine the sex:
fluorescent intensities are compared with threshold values;
fluorescent intensities ratios are compared with threshold values;
fluorescent intensity sums or differences are compared with threshold values;
supervised classification methods, such as discriminant analyses, support vector machines, or neural networks, for example, are applied to the values of fluorescent intensity.

5. The method according to claim 2 ,
wherein,
if at least two laser beam sources are used, the fluorescent intensity levels determined for a predefined spectral range from multiple established fluorescence intensity / wavenumber curves are respectively defined in the evaluation unit for the male blood and for the female blood such that they are combined by a logical operation for evaluation.

6. The method according to claim 1 ,
wherein
fluorescence intensities are normalized, with vector normalization or area normalization, and are then mathematically evaluated in the evaluation unit with one of the following methods or a combination of the following methods, in order to obtain the sex:
normalized fluorescent intensities are compared with threshold values;
ratios between normalized fluorescent intensities are compared with threshold values;
sums or differences of normalized fluorescent intensities are compared with threshold values;
supervised classification methods, such as discriminant analyses, support vector machines, or neural networks, for example, are applied to values of the normalized fluorescent intensities.

7. The method according to claim 1 ,
wherein
a respectively evaluable distinct value for the fluorescent intensity levels is based at least on a measured threshold value or a predefined threshold value that is stored in the evaluation unit and assigned to the fluorescent intensity level.

8. The method according to claim 1 ,
wherein
the fluorescent intensity levels determined for a predefined spectral range from an established fluorescence intensity / wavenumber curves are defined in the evaluation unit as an integral intensity of the fluorescence radiation of the male blood and as an integral intensity of the fluorescence radiation of the female blood.

9. The method according to claim 1 ,
wherein
the fluorescent intensity levels determined for a predefined spectral range from an established fluorescence intensity / wavenumber curves are defined in the evaluation unit as an intensity maximum of the fluorescence radiation of the male blood and as an intensity maximum of the fluorescence radiation of the female blood.

10. The method according to claim 1 ,
wherein
the fluorescent intensity levels determined for multiple predefined and adjacently arranged spectral ranges from an established fluorescent intensity / number curves are defined in the evaluation unit as an integral intensity of the fluorescence radiation of the male blood and as an integral intensity of the fluorescence radiation of the female blood within the predefined spectral ranges.

11. The method according to claim 1 ,
wherein
if at least two laser beam sources are used, the fluorescent intensity levels determined for a predefined spectral range from multiple established fluorescence intensity / wavenumber curves are defined in the evaluation unit as integral intensities of the fluorescence radiation of the male blood and as integral intensities of the fluorescence radiation of the female blood.

12. The method according to claim 1 ,
wherein
the fluorescent intensity levels determined for a predefined spectral range from an established fluorescence intensity / wavenumber curves are defined in the evaluation unit as an integral intensity of the fluorescence radiation of the male blood and as an integral intensity of the fluorescence radiation of the female blood and Raman scattered radiation respectively overlapping the fluorescence radiation is defined for evaluation alone or in combination with other defined fluorescent intensity levels and assigned values, wherein if the fluorescent intensity levels and assigned values are combined, the Raman scatter radiation and the fluorescence radiation are evaluated together by a logical operation.

13. The method according to claim 1 ,
wherein,
if a pulsed laser beam from the laser beam source is used, the fluorescence intensity generated is measured in a time-resolved manner and, from a time constant T of a decay curve of the time-resolved fluorescence intensity, a sex determination is carried out, wherein a different time constant T male , T female , with T male

Assignees

• Univ Dresden Tech

Inventors

• Steiner Gerald
• Preusse Grit
• Galli Roberta
• Koch Edmund

Classifications

• G01N21/6408

  with measurement of decay time, time resolved fluorescence · CPC title

• A01K45/00

  Other aviculture appliances, e.g. devices for determining whether a bird is about to lay · CPC title

• G01N2333/465

  from birds · CPC title

• G01N33/08 Primary

  Eggs, e.g. by candling · CPC title

• G01N21/6486

  Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence) · CPC title