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Allantoic Fluid-Based qPCR for Early Onset In Ovo Sexing
Simão Monteiro Belo Santos1, Matthias Corion1, Bart De Ketelaere1
1Department of Biosystems─Biosensors Group, KU Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium.
Journal of Agricultural and Food Chemistry
|April 13, 2024
Summary
Male chick culling is a welfare concern. This study developed an accurate in ovo sexing method using HINTW and DMRT-1 genes in allantoic fluid, offering a humane alternative for the poultry industry.
Area of Science:
- Poultry Science
- Animal Welfare
- Molecular Biology
Background:
- Male chick culling is a significant animal welfare issue in the poultry industry, leading to governmental bans and a search for alternatives.
- In ovo sexing (determining embryo sex during incubation) is the most viable solution for consumers and industry, but requires specific technological advancements.
Purpose of the Study:
- To develop and validate an industrializable in ovo sexing technology.
- To meet requirements for compatibility with all egg colors, early developmental stages, high hatchability, accuracy, low cost, and high throughput.
Main Methods:
- Investigated the use of female-specific HINTW and male-specific DMRT-1 genes for sex determination.
- Utilized quantitative polymerase chain reaction (qPCR) on allantoic fluid (AF) samples from incubation days 6-9.
- Assessed the impact of the assay on embryo development and hatchability.
Main Results:
- Confirmed female-specific HINTW gene detection in AF across all tested days (6-9) without adverse effects on embryo development.
- Achieved 95% sexing accuracy using HINTW cycle threshold (Ct) values alone.
- Reached 100% sexing accuracy when incorporating Δλ values (difference between HINTW and DMRT-1 Ct values).
Conclusions:
- The developed qPCR assay using AF is a reliable method for in ovo sexing.
- This technology offers a promising pathway for industrial applications, enabling faster and more cost-effective sex determination in poultry.
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