Method for detecting sex of hatching eggs at early hatching stage

Abstract

The invention discloses a method for detecting the sex of hatching eggs at the early hatching stage. The method comprises the following steps: extracting allantoic fluid for hatching early hatching eggs through an extraction device; carrying out metabonomics analysis on allantoic fluid to obtain allantoic fluid metabolite characteristics, and carrying out bioanalysis in combination with in-vitro cell culture to confirm gender difference metabolites; carrying out concentration grading treatment on the allantoic fluid to obtain low-abundance allantoic fluid containing gender difference metabolites; performing spectral comparative analysis on the to-be-detected allantoic fluid so as to construct a multi-machine learning allantoic fluid gender classification model; the sex of the hatching eggs in the early hatching period is detected through the model, and rapid sex detection of the hatching eggs is achieved. According to the full-chain process of early-stage minimally invasive sampling, differential metabolite screening, low-abundance component enrichment and multispectral machine learning, breakthrough of egg gender detection in multiple dimensions of time window, accuracy, cost controllability and the like is achieved.

Description

Technical Field

[0001] The invention relates to an egg sex detection method, relates to the technical field of agricultural and livestock product detection, and in particular to an early hatching breeding egg sex detection method. Background Art

[0002] Eggs are an important agricultural product, a daily staple food and a major source of protein. Egg production has always accounted for a major proportion of poultry egg production. However, a key problem that plagues the laying hen industry is that a number of day-old roosters are prematurely eliminated every year because they cannot lay eggs and have poor market applicability. Faced with resource waste and animal welfare issues, the core of the research is to conduct early, accurate and rapid sex identification of chicken embryos. As an exogenous sample of the embryo, allantoic fluid has the characteristics of relatively easy sampling, low invasiveness and high hatching rate. At present, some scholars extract allantoic fluid from the eggs on the 7th to 10th day of incubation, determine the concentration of estrone sulfate in the allantoic fluid, and use enzyme immunoassay ELISA (Enzyme-Linked ImmunoSorbent Assay) to determine the concentration of estrone sulfate, which is then used to determine the sex of the chicken embryo in the egg. Other scholars have designed a method for egg sex identification, which includes the collection, transportation, pretreatment and detection of estrogen concentration in the allantoic fluid of chicken embryos. This provides valuable information for understanding the sex determination of chicken embryos. However, the composition of allantoic fluid is complex and dynamic, and the existing research foundation of allantoic fluid is relatively weak, especially in balancing the needs of cost, rapid detection and low invasiveness, which faces major challenges. Therefore, it is of great significance to mine the information of gender differences in allantoic fluid and build an efficient and accurate rapid detection model based on it. Summary of the invention

[0003] In order to solve the problems existing in the background technology, the present invention provides a method for detecting the sex of hatching eggs in the early stage of incubation. The method of the present invention obtains the metabolite characteristics of the allantoic fluid after performing metabolomics analysis on the allantoic fluid, and confirms the sex-differentiated metabolites by combining bioinformatics analysis with in vitro cell culture; the allantoic fluid is further concentrated and graded to obtain low-abundance allantoic fluid containing sex-differentiated metabolites; the allantoic fluid to be detected with multiple components is subjected to spectral comparative analysis, thereby constructing a multi-component-multi-machine learning allantoic fluid sex classification model, and realizing the rapid classification of the sex of chicken embryos.

[0004] The technical solution adopted by the present invention is: The method for detecting the sex of early hatching eggs of the present invention comprises: Step 1: Build an extraction device, and use the extraction device to extract the allantoic fluid from several early-stage hatching eggs of known gender.

[0005] Step 2: Metabolomics analysis and bioinformatics analysis were performed on the allantoic fluid of each egg to confirm the sex-differentiated metabolites.

[0006] Step 3: The allantoic fluid of each breeding egg is concentrated and fractionated to obtain the allantoic fluid containing sex-differentiated metabolites.

[0007] Step 4: Obtain the fingerprint spectral feature data of the allantoic fluid containing sex-differentiated metabolites of each breeding egg and add the labels of their respective known sexes, thereby constructing a training set to input into a multi-machine learning model for training, testing, and verification, and obtaining a trained multi-machine learning allantoic fluid sex classification model.

[0008] Step 5: Extract the allantoic fluid of the early-stage hatching eggs to be tested through the extraction device, and then perform the same processing as steps 3 and 4 to obtain the fingerprint spectral feature data of the allantoic fluid to be tested containing gender-differentiated metabolites, and input it into the multi-machine learning allantoic fluid gender classification model. After processing, the gender of the early-stage hatching eggs to be tested is obtained, thereby realizing rapid gender detection of early-stage hatching eggs.

[0009] In the step 1, the extraction device comprises a light source, an extractor, a laser hole opener and an egg tray. The early incubation breeding eggs are cleaned and placed vertically on the egg tray. The light source is placed on the eggshell of the top surface of the early incubation breeding eggs and vertically downward toward the inside of the early incubation breeding eggs, so as to see through the positions of the allantoic fluid, chicken embryo, blood vessel, yolk and air chamber inside the early incubation breeding eggs, so as to avoid other parts and facilitate the extraction of the allantoic fluid. When the allantoic fluid is extracted, the early incubation breeding eggs are in a dark environment, the position of the allantoic fluid of the early incubation breeding eggs is seen through the light source, and then the laser hole opener is used to perform laser hole opening on the eggshell of the early incubation breeding eggs to connect the position of the allantoic fluid, and finally the allantoic fluid is extracted out of the breeding eggs by the extractor, and finally liquid wax is used to seal the micropores to ensure the normal hatching of the chicken embryo.

[0010] In step 1, the early incubation period specifically refers to the 5-10 days of incubation of the eggs.

[0011] In the step 2, the allantoic fluid of each breeding egg is subjected to a metabolomics analysis. First, each metabolite in the allantoic fluid of each breeding egg of known sex is obtained by liquid chromatography and tandem mass spectrometry method LC-MS / MS (Liquid Chromatography-Tandem Mass Spectrometry) in metabolomics analysis. Then, based on each metabolite in the allantoic fluid of each known sex, a bioinformatics analysis method is used to screen out gender-differentiated metabolites from each metabolite in the allantoic fluid.

[0012] In the step 2, the sex-differentiated metabolites include L-ornithine, proline and cytidine.

[0013] In the step 3, the allantoic fluid of the breeding eggs to be tested is first concentrated, and then graded according to the abundance to obtain high-abundance allantoic fluid, medium-abundance allantoic fluid and low-abundance allantoic fluid. The low-abundance allantoic fluid is the allantoic fluid containing gender-differentiated metabolites, so as to enrich the metabolite information of the low-abundance allantoic fluid.

[0014] In the step 4, for the allantoic fluid containing sex-differentiated metabolites of each breeding egg, the allantoic fluid is enriched on the surface of a crystal or a glass slide and then spectral scanning and collection is performed to obtain fingerprint spectral characteristic data of the allantoic fluid.

[0015] The minimally invasive allantoic fluid extraction device designed by the method of the present invention can ensure the hatching rate of chicken fetuses and achieve rapid collection. In the early stage of hatching eggs, the method also confirmed the sexual dimorphism of chicken embryo allantoic fluid metabolites and sex-differentiated metabolites through liquid chromatography and tandem mass spectrometry technology LC-MS / MS combined with bioinformatics analysis. The concentration and classification strategy designed by the method effectively enriched the low-abundance substances in the allantoic fluid, and constructed a multi-component-multi-machine learning spectral sex classification model. The results proved the contribution of the fingerprint spectral characteristics of low-abundance molecules in the allantoic fluid in sex classification, and achieved rapid classification of the sex of chicken embryos.

[0016] The early-stage hatching egg sex detection system of the present invention comprises: The data acquisition unit is used to confirm the allantoic fluid containing gender-differentiated metabolites, obtain the fingerprint spectral feature data of the allantoic fluid containing gender-differentiated metabolites and add gender labels, so as to construct a training set.

[0017] The model training unit is used to construct a multi-machine learning model and train it through a training set to obtain a trained multi-machine learning allantoic fluid gender classification model.

[0018] The gender detection unit is used to detect the fingerprint spectral feature data of the allantoic fluid containing gender-differentiated metabolites to be detected through a multi-machine learning allantoic fluid gender classification model, obtain the gender of the early-incubation breeding eggs to be detected and display it on a display.

[0019] The electronic device of the present invention comprises: a memory and a processor coupled to each other, wherein the memory stores program data, and the processor calls the program data to execute the method as described above.

[0020] The computer-readable storage medium of the present invention stores program data thereon, and when the program data is executed by a processor, the method described above is implemented.

[0021] The beneficial effects of the present invention are: The present invention identifies L-ornithine, proline and cytidine as sex dimorphism markers in allantoic fluid for the first time (the area under the curve (AUC) reaches 0.87), and verifies their function of regulating sex gene expression through in vitro gonadal cell culture, providing a molecular basis for sex identification.

[0022] The present invention systematically reveals the gender-specific metabolite characteristics by integrating allantoic fluid metabolomics with infrared fingerprint spectroscopy analysis. The construction of gender-specific spectral fingerprints makes up for the shortcomings of metabolomics in analyzing the global chemical composition.

[0023] The present invention integrates the concentration and classification method, spectral technology and machine learning to construct a sex classification model for multi-component allantoic fluid, which proves the effectiveness of the fingerprint spectral characteristics of low-abundance substances in allantoic fluid in sex classification, and is helpful for the rapid detection of sex in the early stage of chicken embryo incubation.

[0024] The full-chain process of "early minimally invasive sampling-differential metabolite screening-low-abundance component enrichment-multispectral machine learning" designed in the present invention has achieved breakthroughs in multiple dimensions of poultry egg sex detection in terms of time window, accuracy, cost controllability, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is a flow chart of the method for detecting the sex of breeding eggs of the present invention; Figure 2 is a schematic diagram of the minimally invasive extraction device of the present invention; Figure 3 is a graph showing the classification results of the allantoic fluid metabolomics of the present invention; Figure 4 It is a graph of the gender difference metabolite module and the receiver operating characteristic curve ROC (Receiver Operating Characteristic Curve) of the present invention; Figure 5 This is a graph showing the results of the multi-machine learning gender classification model of the present invention; In the figure: 1. light source, 2. breeding eggs, 201. allantoic fluid, 202. chicken embryo, 203. eggshell, 204. blood vessels, 205. yolk, 206. air chamber, 3. extractor, 4. laser hole opener, 5. egg tray. DETAILED DESCRIPTION

[0026] The present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0027] like Figure 1 As shown, the method for detecting the sex of early hatching eggs of the present invention is as follows: First, build the extraction device, such as Figure 2As shown, the extraction device includes a light source 1, an extractor 3, a laser hole opener 4 and an egg tray 5. The early incubation breeding eggs 2 are cleaned and placed vertically on the egg tray 5. The light source 1 is placed on the eggshell 203 on the top surface of the early incubation breeding eggs 2 and vertically downward toward the inside of the early incubation breeding eggs 2, so as to see through the positions of the allantoic fluid 201, the chicken embryo 202, the blood vessels 204, the yolk 205 and the air chamber 206 inside the early incubation breeding eggs 2, so as to avoid other parts and facilitate the extraction of the allantoic fluid 201; when the allantoic fluid 201 is extracted, the extraction device 1 includes a light source 1, an extractor 3, a laser hole opener 4 and an egg tray 5. When extracting 201, the early incubation breeding eggs 2 are in a dark environment, the position of the allantoic fluid 201 of the early incubation breeding eggs 2 is viewed through the light source 1, and then the laser hole opener 4 is used to perform laser hole opening on the eggshell 203 of the early incubation breeding eggs 2 to connect the position of the allantoic fluid 201, the angle range of the extractor 3 is controlled within the range of 30-120° of the micropore section, and finally the allantoic fluid 201 is extracted from the breeding eggs 2 by the extractor 3, and finally liquid wax is used to seal the micropores to ensure the normal hatching of the chicken embryo 202. The allantoic fluid 201 in several early incubation breeding eggs 2 of known sex is extracted respectively by the extraction device, and the early incubation period is specifically 5-10 days of the breeding egg hatching.

[0028] Then, the allantoic fluid 201 of each egg 2 is subjected to metabolomics analysis and bioinformatics analysis in turn to confirm the metabolites of gender differences. The allantoic fluid 201 of each egg 2 is subjected to metabolomics analysis. First, each metabolite in the allantoic fluid 201 of each egg 2 of known gender is obtained by liquid chromatography and tandem mass spectrometry method LC-MS / MS in metabolomics analysis. Then, based on each metabolite in the allantoic fluid 201 of each known gender, the bioinformatics analysis method is used to screen out gender-difference metabolites from each metabolite in the allantoic fluid 201. Metabolomics analysis is used to compare the metabolites of the allantoic fluid 201 of male and female chicken embryos 202. Then, machine learning and weighted gene co-expression network analysis methods are used to screen gender-difference metabolite modules. The gender-difference metabolite modules are L-ornithine, proline, and cytidine, etc. The module is used for the gender classification result of chicken embryos 202. The area under the curve value AUC is 0.87.

[0029] Then, the reliability of sex-differential metabolites was verified by in vitro culture of gonadal cells. In vitro culture of gonadal cells verification refers to setting different concentrations of L-ornithine, proline and cytidine for interference treatment of gonadal and mesonephric cells of chicken embryo 202, culturing in vitro for 24-48 hours, and detecting the expression of sex-specific genes based on real-time quantitative polymerase chain reaction PCR (Polymerase Chain Reaction) to verify the validity of sex-differential metabolites. Sex-differential metabolites include L-ornithine, proline and cytidine.

[0030] The allantoic fluid 201 of each egg 2 is concentrated and graded, and the allantoic fluid 201 of the egg 2 to be tested is first concentrated, and the concentration includes evaporation concentration, ultrafiltration concentration and dialysis concentration but is not limited to this range; and then the allantoic fluid 201 with high abundance, medium abundance and low abundance is obtained after the graded treatment according to the abundance, and the allantoic fluid 201 containing sex difference metabolites is obtained, wherein the low abundance allantoic fluid is the allantoic fluid 201 containing sex difference metabolites, so as to enrich the metabolite information of the low abundance allantoic fluid. The high abundance allantoic fluid is the allantoic fluid 201 with a molecular weight greater than 30 KDa, the medium abundance allantoic fluid is the allantoic fluid 201 with a molecular weight between 30 KDa and 10 KDa, and the low abundance allantoic fluid is the allantoic fluid 201 with a molecular weight less than 10 KDa. The concentration and classification method of the allantoic fluid 201 is to perform ultrafiltration concentration and classification strategy on the sample, and to pre-concentrate the allantoic fluid 201 of different components onto the surface of a crystal or a glass slide, and determine the optimal spectral parameters to obtain the fingerprint spectrum. In the specific implementation, the infrared spectral fingerprint characteristic peak of the allantoic fluid 201 is 1650 cm - ¹ is attributed to the amide I antiparallel β-helix, indicating protein CO stretching vibration. Lipid-specific bands include 1454 cm - The band at ¹ corresponds to the CH 2 Bend, and 1390 cm - ¹, COO of fatty acids and amino acids - Symmetrical stretching. At 1210 cm - The band at ¹ is consistent with PO in phospholipids and nucleic acids. 2 - The antisymmetric stretching is related to the - The band at ¹ is derived from PO in the diionic phosphomonoester found in phosphorylated proteins and nucleic acids. 2 - Symmetrical stretch. Span 675-900 cm - The spectral region of ¹ is where aromatic compounds are most prominent. At about 918 cm - ¹、845cm - ¹、676 cm - ¹ and 486 cm - The spectral changes observed at ¹ can be attributed to the out-of-plane bending vibrations of the CH groups in the substituted benzene rings, indicating aromatic compounds. In summary, the classification accuracy achieved can be attributed to changes in protein conformation and changes in the intensity of lipid, amino acid and nucleic acid features.

[0031] Then, the allantoic fluid 201 containing sex-differentiated metabolites of each egg 2 is obtained, enriched on a crystal or a glass slide surface, and then spectral scanning and collection are performed. The spectrum includes infrared spectroscopy, Raman spectroscopy and other spectral techniques, so as to obtain the fingerprint spectral characteristic data of the allantoic fluid 201; infrared spectroscopy can provide an in-depth understanding of molecular vibration, rotation and extension, and the allantoic fluid concentration and fractionation strategy enriches the low-abundance substances in the allantoic fluid on the crystal surface, which helps to gain a deeper understanding of the absorption peaks unique to chicken embryo AF metabolites. In specific implementation, the infrared spectral differentiation region shows correlation with the classification of the identified metabolites, such as protein-related metabolites: in the analysis of hatching days D8 and D9, amino acids such as proline, L-ornithine and L-phenylalanine were significantly increased in males. The protein structural characteristics of these amino acids can be seen by the 1650 cm - The amide peak at 1454 cm indicates a change in protein conformation. Lipid-related metabolites: In the analysis of incubation day D9, 3-hydroxybutyrate and D-(+)-malate were significantly increased in males. These lipid metabolites can be characterized by the 1454 cm - ¹ and 1390 cm - Nucleic acid-related metabolites: In the analysis of D10, uridine and cytidine were significantly increased in females. The characteristics of these nucleic acid metabolites can be seen by the 1210 cm - ¹ and 1040 cm - PO at ¹ 2 - The stretching vibration peaks reflect gender differences in nucleic acid metabolism.

[0032] The fingerprint spectral feature data are then added with respective known gender labels to construct a training set, and the training set is input into the multi-machine learning model for training, testing and verification to obtain a trained multi-machine learning allantoic fluid gender classification model.

[0033] In the specific implementation of the present invention, the differential metabolites of the allantoic fluid 201 of male and female chicken embryos 202 are studied based on non-targeted metabolomics. The allantoic fluid 201 extracted from Jinghong No. 1 breeding eggs 2 in actual breeding scenarios was selected as the test object of the embodiment. First, liquid chromatography and tandem mass spectrometry technology LC-MS / MS were used to analyze the metabolites in the allantoic fluid 201 of chicken embryos 202 at different incubation days. The classification results of orthogonal partial least squares discriminant analysis OPLS-DA (Orthogonal Partial Least Squares Discriminant Analysis) are as follows Figure 3 As shown in the figure, the orthogonal component scores and principal component scores of males and females can be obtained. On the 9th day of incubation, the explanatory parameter R 2 =0.997, prediction parameter Q 2=0.779, indicating that the metabolites of the allantoic fluid 201 of the chicken embryo 202 have sexual dimorphism. Then, after 200 permutation tests, the permutation retention, explanatory parameters and prediction parameters were obtained. The explanatory parameter R 2 = 0.99, prediction parameter Q 2 =-0.0207, which verifies the reliability of the model and ensures that there is no overfitting. The metabolites screened by weighted gene co-expression network analysis and orthogonal partial least squares discriminant analysis OPLS-DA were combined to construct a machine learning data set to obtain the characteristic importance evaluation results of metabolites, such as Figure 4 As shown, metabolites with higher contribution were selected as gender difference metabolites, including L-ornithine, proline and cytidine. The results of gender difference metabolites used for gender classification showed that the area under the receiver operating characteristic curve (ROC) was 0.84.

[0034] Then, in vitro cell culture was used to verify the effectiveness of sex-differential metabolites. First, the gonadal and mesonephric tissues of chicken embryo 202 were extracted and cultured in a sterile environment. The gonadal cells were treated with different concentration gradients of sex-differential candidate metabolites. The cell counting kit CCK8 (Cell Counting Kit-8) was used to determine that the concentration of metabolites most suitable for cell proliferation was 200 μM.

[0035] The difference in the expression of sex marker genes between gonadal cells treated with 200 μM L-ornithine, proline and cytidine and untreated control cells was analyzed based on real-time quantitative polymerase chain reaction (PCR). The results showed that cytidine (test result parameter P<0.001) and L-ornithine (test result parameter P<0.001) treatment significantly reduced the expression of anti-Müllerian hormone (AMH) in gonadal cells of male and female chicken embryos 202, further expanding the difference in the expression of anti-Müllerian hormone (AMH) between the two sexes. Cytidine (test result parameter P<0.01) treatment significantly downregulated the expression of DMRT1 (Double sex and mab-3 relatedtranscription factor 1) in the gonads of male chicken embryos 202, while proline (test result parameter P<0.05) induced upregulation, further increasing the difference in DMRT1 expression between males and females. In the gonadal cells of female chicken embryo 202, cytosine (test result parameter P<0.01) and L-ornithine (test result parameter P<0.01) both significantly down-regulated the expression of CYP19A1 (cytochrome P450, family 19, subfamily A, polypeptide 1), while proline up-regulated its expression in male cells. Cytidine and proline treatments attenuated the difference in CYP19A1 expression between the two sexes. Cytidine (test result parameter P<0.01) and L-ornithine (test result parameter P<0.01) also significantly down-regulated the expression of FOXL2 (forkhead box L2) in female gonads, while proline up-regulated its expression in males. These treatments also attenuated the difference in FOXL2 expression between the two sexes.

[0036] like Figure 5As shown, the present invention uses a variety of machine learning to construct a spectral classification model based on the fingerprint spectral data of multi-component allantoic fluid: random forest, support vector machine and extreme gradient boosting are deeply trained, and five parameters are used, including the area under the receiver operating characteristic curve ROC, classification accuracy, precision, recall rate, and receiver operating characteristic curve to comprehensively evaluate the classification performance of the spectrum. In specific implementation, the allantoic fluid 201 of male chicken embryos 202 is used as the experimental group and the female is used as the control group. The gender support vector machine SVM (Support Vector Machine) classification results of the chicken embryos 202 under different concentration and classification parameters are as follows: 1) low abundance: precision: 88%, recall rate: 92%, accuracy: 88%, area under the ROC curve AUC: 0.96; 2) medium abundance: precision: 78%, recall rate: 83%, accuracy: 80%, area under the ROC curve AUC: 0.92; 3) high abundance: precision: 70%, recall rate: 73%, accuracy: 68%, area under the ROC curve AUC: 0.79. The results show that: the high-abundance substances in the allantoic fluid 201 will mask the spectral signals of the low-abundance substances, thereby limiting their contribution to the gender classification spectrum. When there are only spectral signals of low-molecular-weight substances in the allantoic fluid 201, the spectral classification results are significantly improved. The research results of the present invention emphasize that the spectral classification results of the low-abundance allantoic fluid 201 after ultrafiltration concentration are better than the spectral classification results of the untreated and high-abundance allantoic fluid 201.

[0037] Finally, the allantoic fluid 201 of the early-incubation breeding eggs 2 to be tested is extracted by the extraction device, and its fingerprint spectral feature data is obtained, which is then input into the multi-machine learning allantoic fluid gender classification model for processing to obtain the gender of the early-incubation breeding eggs 2 to be tested, thereby realizing rapid gender detection of the early-incubation breeding eggs 2.

[0038] The use of non-targeted metabolomics methods combined with bioinformatics analysis proved the sex dimorphism of allantoic fluid 201 metabolites in chicken embryo 202, providing a reference for the feasibility of using low-abundance substances for sex classification. The significant effect of sex-differentiated metabolites on sex marker genes was verified by in vitro cell culture experiments. In view of the complexity of allantoic fluid 201 samples, designing a concentration and classification strategy during the early incubation process can effectively simplify complex mixtures such as allantoic fluid 201, enrich low-abundance substances, and ensure the characteristic spectral contribution of low-abundance substances in allantoic fluid 201. The present invention realizes high-precision and non-destructive detection of early sex of breeding eggs through innovative minimally invasive sampling, sex-differentiated metabolite screening and multimodal data analysis technology, which has both scientific value and industrial application prospects, and provides technical solutions for improving the efficiency of poultry farming and ethical disputes (such as eliminating male chicks).

[0039] The present invention also establishes a sex detection system for early-stage hatching eggs, including a data acquisition unit, a model training unit and a sex detection unit. The data acquisition unit is used to confirm the allantoic fluid 201 containing sex difference metabolites, obtain the fingerprint spectral feature data of the allantoic fluid 201 containing sex difference metabolites and add a sex label, thereby constructing a training set; the model training unit is used to construct a multi-machine learning model and train it through the training set to obtain a trained multi-machine learning allantoic fluid sex classification model; the sex detection unit is used to detect the fingerprint spectral feature data of the allantoic fluid 201 containing sex difference metabolites to be detected through the multi-machine learning allantoic fluid sex classification model, obtain the sex of the early-stage hatching eggs 2 to be detected and display it on a display.

[0040] The method of the present invention reveals the sexual dimorphism of allantoic fluid metabolites. The concentration and classification method designed based on this method combined with multi-machine learning proves the contribution of the spectral characteristics of low-abundance substances in allantoic fluid to sex classification. This research method can also provide new insights for the development of sex identification technology for poultry eggs.

[0041] It should be understood by those skilled in the art that the embodiments of the present application may be provided as methods, systems or computer program products. Therefore, the present application may take the form of a complete hardware embodiment, a complete software embodiment or an embodiment combining software and hardware. Moreover, the present application may take the form of a computer program product implemented on one or more computer-usable storage media (including but not limited to disk storage, optical storage, etc.) containing computer-usable program codes. The schemes in the embodiments of the present application may be implemented in various computer languages. The present application is described according to the flowcharts of the methods, systems and computer program products of the embodiments of the present application.

[0042] Although the preferred embodiments of the present application have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the present invention is intended to be interpreted as including the preferred embodiments and all changes and modifications falling within the scope of the present application.

[0043] Obviously, those skilled in the art can make various changes and modifications to the present application without departing from the spirit and scope of the present application. Thus, if these modifications and variations of the present application belong to the scope of equivalent technologies of the present invention, the present application is also intended to include these modifications and variations.

Claims

1. A method for detecting the sex of early hatching eggs, characterized in that: include: Step 1: constructing an extraction device, and using the extraction device to extract the allantoic fluid (201) from a number of early hatching eggs (2) of known gender; Step 2: performing metabolomics analysis and bioinformatics analysis on the allantoic fluid (201) of each egg (2) to confirm the metabolites with gender differences; Step 3: Concentrating and fractionating the allantoic fluid (201) of each breeding egg (2) to obtain the allantoic fluid (201) containing sex-differentiated metabolites; Step 4: Obtain the fingerprint spectral feature data of the allantoic fluid (201) containing sex-differentiated metabolites of each breeding egg (2) and add the respective known sex labels, thereby constructing a training set to be input into a multi-machine learning model for training, thereby obtaining a trained multi-machine learning allantoic fluid sex classification model; Step 5: The allantoic fluid (201) of the early-stage hatching eggs (2) to be tested is extracted by an extraction device, and then the fingerprint spectral feature data of the allantoic fluid (201) to be tested containing gender-differentiated metabolites is obtained by the same processing as in steps 3 and 4, and the data is input into a multi-machine learning allantoic fluid gender classification model. After processing, the gender of the early-stage hatching eggs (2) to be tested is obtained, thereby realizing rapid gender detection of the early-stage hatching eggs (2).

2. The method for detecting sex of early hatching eggs according to claim 1, wherein: In the step 1, the extraction device comprises a light source (1), an extractor (3), a laser hole opener (4) and an egg tray (5). The early incubation breeding egg (2) is vertically placed on the egg tray (5), and the light source (1) is placed on the eggshell (203) on the top surface of the early incubation breeding egg (2) and vertically downward toward the interior of the early incubation breeding egg (2). When extracting the allantoic fluid (201), the early incubation breeding egg (2) is in a dark environment, and the position of the allantoic fluid (201) of the early incubation breeding egg (2) is seen through the light source (1). Then, the laser hole opener (4) is used to perform a laser hole opening on the eggshell (203) of the early incubation breeding egg (2) to connect the position of the allantoic fluid (201). Finally, the allantoic fluid (201) is extracted from the breeding egg (2) by the extractor (3).

3. The method for detecting sex of early hatching eggs according to claim 1, wherein: In step 1, the early incubation period specifically refers to the 5-10 days of incubation of the eggs.

4. The method for detecting sex of early hatching eggs according to claim 1, wherein: In the step 2, a metabolomics analysis is performed on the allantoic fluid (201) of each breeding egg (2). First, each metabolite in the allantoic fluid (201) of each breeding egg (2) of known sex is obtained by liquid chromatography and tandem mass spectrometry (LC-MS / MS) in metabolomics analysis. Then, based on each metabolite in the allantoic fluid (201) of each known sex, a bioinformatics analysis method is used to screen out sex-differentiated metabolites in the allantoic fluid (201).

5. The method for detecting sex of early hatching eggs according to claim 1, wherein: In the step 2, the sex-differentiated metabolites include L-ornithine, proline and cytidine.

6. The method for detecting sex of early hatching eggs according to claim 1, wherein: In the step 3, the allantoic fluid (201) of the breeder eggs (2) to be tested is first concentrated, and then graded according to the abundance to obtain high-abundance allantoic fluid, medium-abundance allantoic fluid and low-abundance allantoic fluid, wherein the low-abundance allantoic fluid is the allantoic fluid (201) containing sex-differentiated metabolites.

7. The method for detecting sex of early hatching eggs according to claim 1, wherein: In the step 4, for the allantoic fluid (201) containing sex-differentiated metabolites of each breeding egg (2), the allantoic fluid (201) is enriched on a crystal or a glass slide surface and then spectral scanning and collection is performed, thereby obtaining fingerprint spectral characteristic data of the allantoic fluid (201).

8. A system for detecting sex of early hatching eggs according to any one of claims 1 to 7, characterized in that: include: A data acquisition unit is used to confirm the allantoic fluid (201) containing gender-differentiated metabolites, obtain fingerprint spectral feature data of the allantoic fluid (201) containing gender-differentiated metabolites and add gender labels, thereby constructing a training set; A model training unit, used for constructing a multi-machine learning model and training it through a training set to obtain a trained multi-machine learning allantoic fluid gender classification model; The sex detection unit is used to detect the fingerprint spectral feature data of the allantoic fluid (201) containing sex-differentiated metabolites to be detected by using a multi-machine learning allantoic fluid sex classification model, obtain the sex of the early hatching eggs (2) to be detected and display it on a display.

9. An electronic device, characterized in that: include: A memory and a processor coupled to each other, wherein the memory stores program data, and the processor calls the program data to execute the method according to any one of claims 1 to 7.

10. A computer-readable storage medium having program data stored thereon, characterized in that: When the program data is executed by a processor, the method according to any one of claims 1 to 7 is implemented.

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