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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Biosciences & Biotechnology Laboratory » Research » Research Project #436982

Research Project: Cryoconservation of Ovarian Tissue and Primordial Germ Cells in the Turkey

Location: Animal Biosciences & Biotechnology Laboratory

Project Number: 8042-31000-110-002-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Aug 1, 2019
End Date: Jul 31, 2021

Objective:
The University of Guelph and the Agricultural Research Service (ARS) desire to enter into this Agreement for the purpose of supporting research to be carried out at ARS and Cooperator facilities. ARS desires the Cooperator to provide goods and services necessary to carry out research of mutual interest within the Beltsville Agricultural Research Center and the University of Guelph. Fertility rates from frozen/thawed semen are very low for the turkey, and avian oocytes/embryos cannot be cryopreserved because of the yolk material. An alternative strategy is to freeze immature ovarian tissue that can be thawed and transplanted into young recipient female turkeys. This approach has proven effective for the chicken but similar methods have not been developed for the turkey and many important details are unknown. Specific objectives that address the gap in knowledge include: 1) developing a vitrification protocol; 2) determining the appropriate donor age for tissue/cell recovery; and 3) determining the appropriate recipient age for tissue/cell transplantation.

Approach:
The Location is engaged in research including poultry germplasm conservation. Under the authority of 7 USC 3319a, ARS desires to acquire goods and personnel services from the Cooperator to further agricultural research supporting the independent interests of both parties. This Agreement serves as an order for services to be funded on an annual basis. Turkey lines housed at the University of Guelph will be used for this study. The first experiment will use an in ovo chicken assay for evaluating cell death and cell proliferation of frozen/thawed turkey ovarian tissue. Ovarian tissue isolated post-mortem from day-old poults will be cryopreserved using a protocol that has been successful in chickens and Japanese quail. After the warming procedure, turkey ovarian tissue will be cultured in Day 3 chicken embryos for 10 days. Tissue viability will be assessed using either the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. For the second experiment, ovarian tissue isolated post-mortem from poults will be cryopreserved. To test a possible donor age difference in turkeys, 4 age groups (1 day-, 3 day-, 5 day- and 7 day-old poults) will be evaluated. For each age group, ovaries will be cryopreserved, thawed, cultured in ovo and evaluated by the TUNEL assay to determine viability. For the third experiment, ovarian tissue isolated post-mortem from the appropriate donor age (determined in the second experiment), with or without cryopreservation, will be transplanted into recipient poults. To test a possible recipient age effect in turkeys, 3 age groups (3 day-, 5 day- and 7 day-old poults) of recipients will be evaluated. Recipient birds will be grown to sexual maturity to evaluate the functionality of the transplanted tissue.