Transcript A tool to produce disease free embryos in wood bison (Bison bison

41st Annual Conference of the IETS, Versailles, France
January 10-13, 2015
Superovulation:
A tool to produce disease free
embryos in wood bison
(Bison bison athabascae)
Jesus Manuel Palomino DVM, MSc
PhD Candidate
University of Saskatchewan
Recipient of:
Peter W. Farin Trainee Travel Scholarship Award
The Problem
Genetic
diversity
Brucellosis and
tuberculosis
Disease-free
areas
Population
decline
Loss of genetic
diversity
Hartop et al., 2009
Reproductive
Technologies
Germ plasm
bio-bank
Disease-free
offspring
Reproductive technologies to produce
disease-free wood bison embryos

In vitro embryo production


Superstimulation and oocyte collection
In-vivo-derived embryos

Superovulation and embryo collection
Progression of thesis research
 Study 1. Inducing ovulation in Wood Bison

 Study 2. Effect of LH vs hCG and progesterone on

superovulation
 Study 3. Does eCG improve the superovulatory

response and embryo quality in wood bison?

Study 4. …

Study 5. …
What did we learn from Studies 1 to 3?
 Ovarian superstimulation:
- 8 vs 2 vs 1 administrations of FSH
 Induction of ovulation:
- hCG more effective than LH
 Adjunct treatments
- eCG: Did not improve superovulation or embryo quality
- Exogenous progesterone not needed: anovulatory season
Problem
 Number of embryos collected: 4 embryos/cow
 Number of Grade 1 & 2 (Freezable) embryos: 2 embryos/cow
Can we improve these numbers?
Study 4
Effect of lengthening of FSH treatment on
superovulation and embryo quality in wood
bison
Rationale:

In cattle, lengthening the period of FSH treatment induced
a greater number of ovulations and CL compared with
conventional treatment (Garcia Guerra et al., 2012)

Lengthening FSH treatment will rescue more subordinate
follicles in the wave and they will grow to a sufficient stage
of maturity to respond to LH and ovulate
Hypothesis

Lengthening FSH treatment will improve the ovulation
rate and embryo quality in superovulated wood bison.
Objective:

Determine the effect of lengthening of FSH treatment on
superovulatory response and embryo quality in wood
bison during the ovulatory and anovulatory season
Experiment 1: Anovulatory season
Experimental design:
Experiment 2: Ovulatory season
Experimental design:
Animals and facilities


Alternative Livestock Center, University of Saskatchewan
50 adult female and 4 male wood bison
Hydraulic chute
to handle bison
Synchronization: Follicular ablation
Day -1
Day 0
Superovulation
FSH 400 mg (Folltropin,
Bioniche)
Hyaluronic acid
(Map 5, Bioniche)
hCG 3000 IU
(Chorulon, Merck)
Artificial Insemination: Chilled semen



Semen collection: electroejaculation, 3 bulls, pooled
Extender: Tryladil (MOFA) + 20% egg yolk
Chilled semen 5º C (1-3 days)
Embryo collection: Non-surgical
Embryo classification (IETS, 2010)

cvzd
Table 1: Anovulatory season
FSH Short
FSH Long
16
16
10.8 ± 2.0
10.8 ± 1.1
52%
54%
(67/130)
(70/130)
Number of CL
4.5 ± 0.9
5.7 ± 1.0
Total ova-embryos
2.6 ± 0.5
3.2 ± 0.7
Grade 1 & 2
0.5 ± 0.3
0.3 ± 0.2
Grade 3
0.3 ± 0.2
0.1 ± 0.1
Grade 4
1.2 ± 0.4
1.8 ± 0.6
Recovery rate
48 %
51%
(# ova-embryos/# CL)
(26/54)
(35/68)
Number of bison
Number of follicles ≥9 mm on Day 5
Ovulation rate
(# ovulations / # follicles ≥9 mm)
Results are presented as Mean±SEM, otherwise stated.
Table 2: Ovulatory season
FSH Short
FSH Long
14
14
8.1 ± 1.1
9.8 ± 1.4
0.40
60%
74%
0.07
(67/130)
(73/98)
Number of CL
4.6 ± 0.6
6.5 ± 0.9
0.13
Total ova-embryos
2.2 ± 0.5
4.4 ± 0.7
0.04
Grade 1 & 2
1.1 ± 0.4
2.3 ± 0.6
0.09
Grade 3
0.2 ± 0.1
0.8 ± 0.4
0.03
Grade 4
0.8 ± 0.2
1.7 ± 0.6
0.05
Recovery rate
48 %
68%
0.06
(# ova-embryos/# CL)
(22/46)
(44/65)
Number of bison
Number of follicles ≥9 mm on Day 5
Ovulation rate
(# ovulations / # follicles ≥9 mm)
P - value
Results are presented as Mean±SEM, otherwise stated.
Garcia Guerra et al., 2012
Dias et al., 2013
Conclusions

Extended superstimulatory protocol improved
embryo production during the ovulatory season

Embryo quality was not affected by lengthening
the superstimulatory protocol

Repeat experiment with greater number of bison
Study 5:
Production of disease-free embryos from
superovulated wood bison
Objective:

Determine the effectiveness of washing procedures for
removing Brucella abortus from bison embryos exposed in
vitro
Abstract 140
Level 3 laboratory – Intervac/VIDO
Acknowledgments

Dr. Adams
Dr. Mapletoft
Dr. Woodbury
Dr. McCorkell
Dr. Muir/ Dr. Machin
Dr. Anzar
Dr. Mastromonaco

Miriam

Rodrigo, Hayder, Alisha
Nick, Brad
Kosala, Lyle




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Department of Veterinary
Biomedical Sciences