Allantoic Fluid-Based qPCR for Early Onset In Ovo Sexing.

The day-old male chick culling remains a welfare issue in the poultry industry. Several governments have prohibited this practice, pushing hatcheries to seek alternatives. Although different solutions exist for solving this problem, sex determination during the embryo's incubation (in ovo sexing) is considered the most suitable one among the consumers and industry. However, to be industrialized, in ovo sexing technologies must meet several requirements: compatibility with all egg colors and early developmental stages while maintaining a high hatchability rate and accuracy at low cost and high throughput. To meet these requirements, we studied the use of the sexual genes HINTW (female-specific) and DMRT-1 (both sexes) at incubation days 6-9. By utilizing the quantitative polymerase chain reaction in allantoic fluid (AF) samples, our study confirmed female-specific HINTW detection on all days without any significant detrimental effects on embryo development. We achieved 95% sexing accuracy using the HINTW cycle threshold (Ct) alone and 100% accuracy rate when using Δλ values (difference between the HINTW and DMRT-1 Ct). In conclusion, the developed assay can provide information about AF as a sample for in ovo sexing and open new industrial possibilities for faster and cheaper assays.

A review of the recent advances for the in ovo sexing of chicken embryos using optical sensing techniques.

The culling of day-old male chicks has caused ethical and economic concerns. Traditional approaches for detecting the in ovo sex of chicken embryos involve opening the eggshell and inner membrane, which are destructive, time-consuming, and inefficient. Therefore, noncontact optical sensing techniques have been examined for the in ovo sexing of chicken embryos. Compared with traditional methods, optical sensing can increase determination throughput and frequency for the rapid sexing of chicken embryos. This paper presented a comprehensive review of the different optical sensing techniques used for the in ovo sexing of chicken embryos, including visible and near-infrared (Vis-NIR) spectroscopy, hyperspectral imaging, Raman spectroscopy, fluorescence spectroscopy, and machine vision, discussing their advantages and disadvantages. In addition, the latest research regarding different detection algorithms and models for the in ovo sexing of chicken embryos was summarized. Therefore, this paper provides updated information regarding the optical sensing techniques that can be used in the poultry industry and related research.

Trends in in ovo sexing technologies: insights and interpretation from papers and patents.

Numerous researchers and institutions have been developing in ovo sexing technologies to improve animal welfare by identifying male embryos in an early embryonic stage and disposing of them before pain perception. This review gives a complete overview of the technological approaches reported in papers and patents by performing a thorough search using Web of Science and Patstat/Espacenet databases for papers and patents, respectively. Based on a total of 49 papers and 115 patent families reported until May 2023 worldwide, 11 technology categories were defined: 6 non-optical and 5 optical techniques. Every category was described for its characteristics while assessing its potential for application. Next, the dynamics of the publications of in ovo sexing techniques in both paper and patent fields were described through growth curves, and the interest or actual status was visualized using the number of paper citations and the actual legal status of the patents. When comparing the reported technologies in papers to those in patents, scientific gaps were observed, as some of the patented technologies were not reported in the scientific literature, e.g., ion mobility and mass spectrometry approaches. Generally, more diverse approaches in all categories were found in patents, although they do require more scientific evidence through papers or industrial adoption to prove their robustness. Moreover, although there is a recent trend for non-invasive techniques, invasive methods like analyzing DNA through PCR or hormones through immunosensing are still being reported (and might continue to be) in papers and patents. It was also observed that none of the technologies complies with all the industry requirements, although 5 companies already entered the market. On the one hand, more research and harmony between consumers, industry, and governments is necessary. On the other hand, close monitoring of the market performance of the currently available techniques will offer valuable insights into the potential and expectations of in ovo sexing techniques in the poultry industry.

Highly sensitive and quick in ovo sexing of domestic chicken eggs by two-wavelength fluorescence spectroscopy.

The in ovo sexing of chicken eggs is a current task and a prerequisite to overcome the mass killing of male day-old chicks from laying lines. Although various methods have been developed and tested in recent years, practicable methods for sex determination are still missing which can be applicated in poultry hatcheries before the chicken embryo is capable of nociception and pain sensation. Optical spectroscopic methods enable an early determination of the sex. In this study, a novel method based on two-wavelength in ovo fluorescence excitation is described. More than 1600 eggs were examined. In ovo fluorescence was sequentially excited at 532 nm and 785 nm. The fluorescence intensities of the spectral regions behave inversely with respect to sex. It is shown that the observed sex-related differences in the fluorescence intensities are based on the embryonic hemoglobin synthesis. The accuracy of sex determination is 96% for both sexes. The hatching rate is not reduced compared to an equivalent reference group.

In ovo sexing of eggs from brown breeds with a gender-specific color using visible-near-infrared spectroscopy: effect of incubation day and measurement configuration.

The culling of day-old male chicks is an animal welfare issue within the laying hen industry that raises substantial ethical concern. Alternative methods are sought to pre-select males during embryonic development. This method is called in ovo sexing and allows more humane male culling. A robust and non-invasive in ovo color sexing technique was developed and validated in this research. To this end, visible-near-infrared (vis-NIR) point spectroscopy was used, which has advantages over state-of-the-art hyperspectral imaging in terms of accuracy and cost. Two independent experiments were each conducted on a batch of 600 Isa Brown eggs. These eggs were individually illuminated on d 8 to 14, and d 18 of incubation by a halogen lamp and the signal was measured in the vis-NIR range from 300 to 1,145 nm. Next, optimal preprocessing strategies were applied and partial least squares discriminant analysis (PLS-DA) models were built and further optimized after performing a forward interval partial least squares (FiPLS) for variable selection. Results demonstrated that d 12 is too early for vis-NIR in ovo sexing, resulting in a prediction accuracy of 86.49%. However, after 13 d of incubation, an accuracy of 97.78% was obtained, increasing to 99.52% on d 14. Furthermore, these accuracies were higher than earlier reported percentages from hyperspectral imaging and successful sexing was expedited from d 14 to d 13. Moreover, prediction improvement up to 99.05% was obtained on d 13 by correcting for the variability in eggshell properties using d 0 eggshell corrections. Applying the method on d 18 resulted in a lower accuracy of 94.62% due to stronger light attenuation by the growing embryos. Finally, a reduced spectral range of 749 to 861 nm was found to be sufficient for correct classification of 98.46% of the eggs. This paves the way for high-throughput and cost-efficient usage of smaller and cheaper spectrophotometers in commercial hatcheries.

The Need for an Alternative to Culling Day-Old Male Layer Chicks: A Survey on Awareness, Alternatives, and the Willingness to Pay for Alternatives in a Selected Population of Dutch Citizens.

A concerning by-product of producing laying hen chicks are the hatched male layer chicks. As a consequence of their inability to lay eggs, these male chicks are culled as day-old chicks in the hatchery. To find an alternative for this ethical dilemma (generally), three alternatives are under study, namely, in ovo sex determination, using dual-purpose breeds, and the rearing of layer cockerels. In order to assess the awareness of this practice and preference for one of the alternatives, we conducted an online survey of the Dutch public. Most of the 259 respondents completing the survey were highly educated woman (HEW, n = 143) versus others (REST, n = 86). The questionnaire was divided into six topics: (1) general knowledge of the poultry industry, (2) awareness of culling male layer chicks (CMC), and (3) its acceptability, (4) alternatives to CMC, (5) willingness to pay (WTP) for eggs without CMC, and (6) WTP for cockerel meat. Awareness about CMC was 52%, and its acceptability was rejected by 78% (HEW) and 67% (REST). The level of acceptability increased when more salient facts were given, and almost all respondents agreed that an alternative was needed (90% HEW, 84% REST). For both groups of respondents, more than 50% preferred in ovo sex determination over keeping the current practice or using dual-purpose breeds or male layers. Furthermore, the majority of respondents were willing to pay more than double the price for eggs without CMC being involved. Roughly 40% would not buy processed cockerel meat burgers, most likely due to their vegan or vegetarian diet. Of the remaining respondents, half were willing to pay the current price or 1 euro more for processed cockerel meat burgers. The most important factors when buying poultry meat or eggs without CMC were food safety, animal friendliness (welfare), and the environment; price was the least important factor. Despite the skewed respondents' background, the results of our survey show that consumers are willing to pay more for poultry products that do not require culling day-old male chicks.

Review: What are the challenges facing the table egg industry in the next decades and what can be done to address them?

There has been a strong consumer demand to take welfare into account in animal production, including table eggs. This is particularly true in Europe and North America but increasingly around the world. We review the main demands that are facing the egg industry driven by economic, societal and sustainability goals. We describe solutions already delivered by research and those that will be needed for the future. Already table egg consumption patterns have seen a major shift from cage to non-cage production systems because of societal pressures. These often feature free-range and organic production. These changes likely signal the future direction for the layer sector with the acceleration of the conversion of cage to barn and aviary systems with outdoor access. This can come with unintended consequences from bone fracture to increased disease exposure, all requiring solutions. In the near future, the laying period of hens will be routinely extended to improve the economics and environmental footprint of production. Many flocks already produce close to 500 eggs per hens in a lifetime, reducing the number of replacement layers and improving the economics and sustainability. It will be a challenge for scientists to optimize the genetics and the production systems to maintain the health of these hens. A major ethical issue for the egg industry is the culling of male day-old chicks of layer breeds as the meat of the males cannot be easily marketed. Much research has and will be devoted to alternatives. Another solution is elimination of male embryos prior to hatching by in ovo sexing approaches. The race to find a sustainable solution to early stage sex determination is on. Methods based on sex chromosomes, sexually dimorphic compounds and spectral properties of eggs containing male or female embryos, are being researched and are reviewed in this article. Other proposed solutions include the use of dual-purpose strains, where the males are bred to produce meat and the females to produce eggs. The dual-purpose strains are less efficient and do not compete economically in the meat or egg market; however, as consumer awareness increases viable markets are emerging. These priorities are the response to economic, environmental, ethical and consumer pressures that are already having a strong impact on the egg industry. They will continue to evolve in the next decade and if supported by a strong research and development effort, a more efficient and ethical egg-laying industry should emerge.

In-ovo sexing of 14-day-old chicken embryos by pattern analysis in hyperspectral images (VIS/NIR spectra): A non-destructive method for layer lines with gender-specific down feather color.

Up to now there is no economically maintainable modality for chicken sexing in early embryonic stages (first 3 d) that is suitable for large-scale application in the commercial hatcheries. Hence, the culling of male day-old chicks of layer lines is still the normal procedure.In this paper we present a non-destructive optical technique for gender determination in layer lines with gender-specific down feather color. This particular chicken strain presents a sexual dimorphism in feather color, where the female day-old chicks have brown down feathers and the males have yellow down feathers.The eggs are candled with halogen lamps and a hyperspectral camera collects the transmitted light within the spectral range from 400 nm to 1,000 nm. For data analysis and classification, common methods like principal component analysis and linear discriminant analysis are used. The accuracy of gender determination was determined for 11- to 14-day-old embryos. At 14 d of incubation (7 d before hatch) the sex can be determined with an overall accuracy of approximately 97%.