Gender, viability and/or developmental stage determination of avian embryos in ovo

Abstract

The present invention relates to a process for the non-destructive determination of gender, developmental stage and/or viability of an avian embryo in an egg, comprising (a) detecting at least a first developmental marker compound selected from sugars and/or amino acids, precursors and metabolites thereof in an egg at a time period of from the beginning of the incubation of the egg until the hatching; (b) measuring the amount of the at least first detected developmental marker compound, and (c) comparing the amount to a base line established for male and female, developmental stage of the embryo, and/or alive and deceased or non-developed embryo, to determine whether the embryo is viable, male and/or female, and/or the developmental stage of the embryo.

1 . A process for the non-destructive determination of gender, developmental stage and/or viability of an avian embryo in an egg, comprising
 (a) detecting at least a first developmental marker compound selected from sugars and/or amino acids, precursors and metabolites thereof in an egg at a time period of from the beginning of the incubation of the egg until the hatching;   (b) measuring the amount of the at least first detected developmental marker compound, and   (c) comparing the amount to a base line established for male and female, developmental stage of the embryo, and/or alive and deceased or non-developed embryo, to determine whether the embryo is viable, male and/or female, and/or the developmental stage of the embryo.   
     
     
         2 .- 34 . (canceled) 
     
     
         35 . The process according to  claim 1 , wherein the at least first developmental marker compound is detected in an embryonic fluid of an egg, 
     
     
         36 . The process according to  claim 1 , wherein the embryonic fluid is the allantoic fluid. 
     
     
         37 . The process according to  claim 1 , wherein at least a second marker is detected in step (a), and wherein the at least first and second markers are analysed and compared to the base line, and to each other to establish a developmental marker ratio. 
     
     
         38 . The process according to  claim 1 , wherein the developmental markers are selected from Glucose, Choline and/or Valine. 
     
     
         39 . The process according to  claim 38 , wherein the avian species is  Gallus gallus domesticus , and wherein a first or further developmental marker is selected from at least one of:
 Glucose in absolute amount for a female embryo in the range of from 30 μM/ml to 70 μM/ml, and for a male embryo of from 1 μM/ml to 30 μM/ml;   Choline in an absolute amount for a female embryo in the range of from 1 10 μM/ml to 130 μM/ml, and for a male embryo of from 90 μM/ml up to, but not including 1 10 μM/ml; or   Valine in an absolute amount for a female embryo in the range of from 1 10 μM/ml to 130 μM/ml, and for a male embryo of from 90 μM/ml up to, but not including 1 10 μM/ml.   
     
     
         40 . The process according to  claim 38 , wherein at least a first and a second developmental marker are detected and analysed, and wherein the absolute amounts and the ratio of the at least first to second markers is employed to determine the gender and/or viability. 
     
     
         41 . The process according to  claim 1 , comprising the step of analysing the embryonic fluid invasively or non-invasively. 
     
     
         42 . The process according to  claim 41 , wherein the analysis is performed by a magnetic resonance imaging method including nuclear resonance methods; by a spectral resonance method including infrared or Raman spectroscopy; and/or an analytical method such as GC or HPLC coupled with suitable detectors, fluorescence spectroscopy and/or an enzyme-linked immunosorbent assay. 
     
     
         43 . The process according to  claim 1 , further comprising at least one of;
 (i) determining whether an embryo in an egg is viable and male, or viable and female, and separating a multitude of viable male eggs from a multitude of viable female eggs, to form an predominantly male or predominantly female egg selection;   (ii) subjecting the viable female or male egg selections to an incubation and hatching process to form a predominantly female or male chick population;   (iii) determining the developmental stage of the embryo and the time until hatching is likely to occur, and separating eggs into multitudes of eggs with similar or equal developmental stage;   (iv) determining the viability of the embryo, and separating eggs into multitudes of viable eggs and non-viable eggs;   (v) subjecting the viable egg selections to an incubation and hatching process in line with the expected hatching date, to form a chick population of predominantly the same developmental stage; and   (vi) injecting a virus or virus-like material into each egg identified as containing a live embryo and male or female, and/or of a certain developmental stage.   
     
     
         44 . The process according to  claim 43 , further comprising isolating the obtained vaccine from the incubated eggs. 
     
     
         45 . A chick or a chick population obtainable by the process according to  claim 43 . 
     
     
         46 . The process according to  claim 37 , wherein the avian species is  Gallus gallus domesticus , and wherein the developmental markers are selected from at least one of:
 Trimethylglycine; Aspartate and/or Asparagine; Glutamate and/or Glutamine, and/or Proline.   
     
     
         47 . The process according to  claim 1 , further comprising at least one of:
 (i) selecting eggs of essentially the same development stage;   (ii) subjecting the egg selection to an incubation and hatching process in line with the developmental stage, to form a chick population of essentially the same age;   (iii) determining whether an embryo in an egg is viable, and separating non-viable eggs prior to hatching and/or vaccine injection steps;   (iv) determining whether an embryo is male, or female, and separating a multitude of male eggs from a multitude of female eggs, to form a predominantly male or predominantly female egg selection; and   (v) injecting a virus or virus-like material into each egg identified as containing a live embryo and male or female, in line with the developmental stage of the embryo.   
     
     
         48 . The process according to  claim 47 , further comprising isolating the obtained vaccine from the incubated eggs. 
     
     
         49 . A multitude of eggs obtainable from the process of  claim 48 . 
     
     
         50 . The multitude of eggs of  claim 49  for animal and/or human feed production, for the production and/or isolation of cosmetic, medical and/or nutritional compounds, for methane production through fermentation, and/or high quality fertilizer production.