A tool to produce disease free embryos in wood bison B ison bison athabascae Jesus Manuel Palomino DVM MSc PhD Candidate University of Saskatchewan 41st Annual Conference of the ID: 174255
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Superovulation: A tool to produce disease free embryos in wood bison (Bison bison athabascae)
Jesus Manuel Palomino DVM, MSc PhD CandidateUniversity of Saskatchewan
41st Annual Conference of the
IETS, Versailles, FranceJanuary 10-13, 2015
Recipient of:
Peter
W.
Farin
Trainee Travel Scholarship
AwardSlide2
The ProblemHartop et al., 2009Brucellosis and tuberculosis
Disease-free areasPopulation decline
Loss of genetic diversity
Reproductive Germ plasm Disease-free Technologies bio-bank offspring
Genetic diversitySlide3
Reproductive technologies to produce disease-free wood bison embryosIn vitro embryo productionSuperstimulation and oocyte collectionIn-vivo-derived embryosSuperovulation and embryo collectionSlide4
Progression of thesis researchStudy 1. Inducing ovulation in Wood BisonStudy 2. Effect of LH vs hCG
and progesterone on superovulationStudy 3. Does eCG improve the superovulatory response and embryo quality in wood bison?Study 4. …
Study 5. …
Slide5
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 treatmentseCG: 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?Slide6
Study 4 Effect of lengthening of FSH treatment on superovulation and embryo quality in wood bisonRationale: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 ovulateSlide7
HypothesisLengthening 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 seasonSlide8
Experiment 1: Anovulatory seasonExperimental design:Slide9
Experiment 2: Ovulatory seasonExperimental design:Slide10
Animals and facilitiesAlternative Livestock Center, University of Saskatchewan50 adult female and 4 male wood bison
Hydraulic chute to handle bisonSlide11
Synchronization: Follicular ablationDay -1Day 0Slide12
Superovulation
Hyaluronic acid (Map 5, Bioniche)
hCG 3000 IU (Chorulon, Merck)
FSH 400 mg (
Folltropin
,
Bioniche
)Slide13
Artificial Insemination: Chilled semenSemen collection: electroejaculation, 3 bulls, pooledExtender: Tryladil (MOFA) + 20% egg yolkChilled semen 5
º C (1-3 days)Slide14
Embryo collection: Non-surgicalSlide15
Embryo classification (IETS, 2010)cvzdSlide16
Table 1: Anovulatory seasonFSH ShortFSH LongNumber of bison1616Number of follicles ≥9 mm on Day 510.8 ± 2.010.8 ± 1.1Ovulation rate (# ovulations / # follicles ≥9 mm)52%
(67/130)54%(70/130)Number of CL4.5 ± 0.95.7 ± 1.0Total ova-embryos2.6 ± 0.53.2 ± 0.7 Grade 1 & 20.5 ± 0.3
0.3 ± 0.2 Grade 3
0.3 ± 0.20.1 ± 0.1 Grade 41.2 ± 0.41.8 ± 0.6Recovery rate (# ova-embryos/# CL)
48 %
(26/54)
51%
(35/68)
Results are presented as
Mean±SEM
, otherwise stated.Slide17
Table 2: Ovulatory seasonFSH ShortFSH LongP - valueNumber of bison1414Number of follicles ≥9 mm on Day 58.1 ± 1.1 9.8 ± 1.40.40Ovulation rate
(# ovulations / # follicles ≥9 mm) 60% (67/130) 74% (73/98)0.07Number of CL4.6 ± 0.6 6.5 ± 0.90.13Total ova-embryos
2.2 ± 0.5 4.4
± 0.70.04 Grade 1 & 21.1 ± 0.42.3 ± 0.60.09 Grade 30.2
± 0.1
0.8
± 0.4
0.03
Grade 4
0.8
± 0.2
1.7
± 0.6
0.05
Recovery rate
(# ova-embryos/# CL)
48 %
(22/46)
68%
(44/65)
0.06
Dias et al., 2013
Garcia Guerra et al., 2012
Results are presented as
Mean±SEM
, otherwise stated.Slide18
ConclusionsExtended 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 bisonSlide19
Study 5: Production of disease-free embryos from superovulated wood bisonObjective:Determine the effectiveness of washing procedures for removing Brucella abortus
from bison embryos exposed in vitroAbstract 140Slide20
Level 3 laboratory – Intervac/VIDOSlide21
Acknowledgments Dr. Adams Dr. MapletoftDr. WoodburyDr. McCorkellDr. Muir/ Dr.
MachinDr. AnzarDr. Mastromonaco
Miriam
Rodrigo, Hayder, AlishaNick, Brad Kosala, Lyle
Department of Veterinary Biomedical Sciences