FARDILHA'S
LAB

SpermXYSort aims to develop and validate the first cost-effective, immunological-based sperm sexing kit specifically designed for cuniculture (rabbit farming), with potential applications in other livestock sectors.

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Current sexing technologies, such as flow cytometry, are prohibitively expensive and complex for many animal industries. Our project proposes a disruptive solution: a user-friendly kit that uses magnetic bead technology to separate X-chromosome-bearing sperm (female) from Y-chromosome-bearing sperm (male) prior to artificial insemination.

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By enabling farmers to pre-select the sex of the offspring, SpermXYSort addresses critical economic inefficiencies and ethical concerns regarding "surplus animals," promoting a more sustainable and humane animal production system.

 

The Challenge & Motivation

In animal production, one sex is often preferred over the other depending on the production goal (e.g., females for reproduction/milk, males for meat).

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• The Problem: In the context of breeding doe production, about 50% of the progeny are males. These surplus animals have low commercial value, leading to economic losses and raising significant ethical issues regarding their culling or disposal.

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• The Gap: There is currently no commercially profitable sexing technology for rabbits. Existing methods used in cattle are too expensive, invasive, or damaging to sperm quality.

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• The Opportunity: Rabbits are the 3rd most slaughtered species globally. A low-cost sexing tool would revolutionize the industry by optimizing resources and aligning production with market demands.

 

Scientific Innovation

Our approach is based on the identification of a specific biomarker. Research by our team has identified a possible transmembrane protein specific to the X-chromosome-bearing spermatozoa in rabbits.

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How it works:

1. Targeting: We utilize high-affinity antibodies designed to recognize specific epitopes of the specific protein on the sperm surface.

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2. Magnetic Separation: These antibodies are coupled to magnetic beads. When mixed with semen, the beads bind specifically to the X-sperm.

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3. Sorting: Under a magnetic field, the X-sperm are separated from the Y-sperm.

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4. Result: A semen sample highly enriched in the desired sex, ready for artificial insemination.

This method is estimated to be 4 times cheaper than flow cytometry and is non-invasive to the sperm cell.

 

Objectives

1. Protein Characterization: Determine the 3D structure of the sex-specific protein using Cryo-Electron Microscopy (CryoEM) to identify conserved epitopes.

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2. Antibody Development: Create high-specificity antibodies to target these surface proteins.

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3. Prototype Development: optimize the magnetic-activated cell sorting protocol to ensure high purity (>90%) and sperm viability.

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4. Validation:

    ◦ In Vitro: Assess fertilization rates through In Vitro Embryo Production (IVP).

    ◦ In Vivo: Conduct a Proof-of-Concept with live animals to validate fertility rates, sex ratios, and offspring health (absence of malformations).

 

Impact & Sustainability

SpermXYSort aligns directly with the UN Sustainable Development Goals (SDGs 8, 9, and 12):

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• Environmental: Reduces the environmental footprint by ensuring resources (feed, water, energy) are invested only in productive animals.

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• Ethical (Animal Welfare): drastically reduces the birth of unwanted animals, mitigating the need for culling surplus males.

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• Economic: Increases profitability for farmers by maximizing the output of high-value animals.

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• Scalability: While developed for rabbits, the technology targets conserved proteins, meaning it has the potential to be adapted for bovine, porcine, and small ruminant industries.

 

The Consortium

This project brings together multidisciplinary expertise in veterinary sciences, biomedicine, and proteomics.

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Leading Institution:

• University of Trás-os-Montes and Alto Douro (UTAD) - CECAV: Led by Prof. Bruno Colaço, responsible for animal experimentation, in vivo trials, and project management.

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Partner Institution:

• University of Aveiro (UA) - iBiMED: Led by Prof. Margarida Fardilha, responsible for sperm biochemistry, proteomics, and in vitro technology development.

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Key Team Members:

• Patrícia Pinho: Lead Researcher on sperm surface markers and proteomics.

• Rosário Pinto Leite: Consultant (Genetics).

• Francis Impens: Consultant (Proteomics).

• Magda Henriques: Junior Researcher. 

• Renato M. Rodrigues: Research Fellow. ​

• Cândida Cruz: PhD Student.

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Industry Partner:

• Cortagri: A national leader in cuniculture, providing biological samples and support for future industrial validation.

 

Publications & Outputs

• European Patent Application EP24160720.9 (Pending) - Sex-specific sperm membrane proteins for sperm sexing.

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• The surface proteome of rabbit spermatozoa (Animal Reproduction Science, 2025).

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•Comparative Bioinformatic Analysis of the Proteomes of Rabbit and Human Sex Chromosomes (Animals, 2024).

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SpermXYSort, formerly known as RabbitXYSort, is the winner of the INOV@UTAD 2023 Business Idea Competition. This project is a direct outcome of the doctoral thesis developed by Dr. Patrícia Pinto Pinho.