Catherine Gulliver. Collaborators: Dr Edward Clayton, Dr Michael Friend, Dr John Wilkins, Dr Belinda King, Dr Susan Robertson
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1 Catherine Gulliver Collaborators: Dr Edward Clayton, Dr Michael Friend, Dr John Wilkins, Dr Belinda King, Dr Susan Robertson
2 Outline Background Why alter sex ratio? Factors affecting sex ratio? Maternal nutrition Polyunsaturated fatty acids (PUFAs) Omega-3, omega-6 and sex ratio Pen trial Take home messages
3 Why Alter Sex Ratio? Skew offspring sex ratio to favour preferred gender Prime lambs Maternal flocks Dairy cattle Male offspring Female offspring Female offspring
4 Factors Affecting Sex Ratio? X or Y sperm Genetic factors Environmental factors: Maternal nutrition Timing of insemination Time of season Litter size
5 Factors Affecting Sex Ratio? Adapted from Cameron (2004)
6 Factors Affecting Sex Ratio? Food Condition Weight Adapted from Cameron (2004)
7 Maternal Condition Strongest correlation with sex ratio when condition measured at/near conception (Cameron, 2004) Females in poorer condition at conception less male offspring (Review: Rosenfeld & Roberts, 2004) Deer, pigs, sheep
8 Maternal Diet Increased glucose in vitro favours male embryos (Cattle: Gutierrez-Adan et al. 2001) Increased glucose in vivo male-biased litters (Mice: Cameron et al. 2008) Mice fed high-fat / low carbohydrate diet male-biased litters (Rosenfeld et al. 2003) Mechanism?
9 Potential Mechanisms Pre-conception mechanisms: A) Differential motility/transport B) Selective fertilisation Post-conception mechanisms: C) Faster embryo development pre-implantation D) Developmental arrest of embryo pre-implantation (Rosenfeld & Roberts, 2004)
10 Polyunsaturated Fatty Acids (PUFAs) Long-Chain Omega-3 Long-Chain Omega-6
11 Polyunsaturated Fatty Acids (PUFAs) Omega-3 Fresh forage Omega-6 Grain α-linolenic acid (C18:3n-3) Short-chain fatty acids Linoleic acid (C18:2n-6) Eicosapentaenoic acid (EPA) Arachidonic Acid (AA) Long-chain fatty acids
12 PUFAs and Reproduction Improve fertility Reduce post-partum interval to ovulation (De Fries et al. 2008) Increase follicular growth (Lucy et al. 1991) Improve oocyte quality & embryo survival (Santos et al. 2008) Sperm transport (Rooke et al. 2001) Sex ratio?
13 PUFAs and Sex Ratio Omega-3 α-linolenic acid Omega-6 Linoleic acid Eicosapentaenoic acid (EPA) Arachidonic Acid (AA) Prostaglandin Series-3 (PG-3) ANTI - INFLAMMATORY Prostaglandin Series-2 (PG-2) PRO - INFLAMMATORY Improve survival of males? (Marsupials: Rosenfeld & Roberts, 2004) (Heifers: Wamsley et al. 2005) Loss of male embryos? (Mice; Fountain et al. 2008)
14 Research Questions 1. Can we alter sex ratio of offspring by altering ratio of omega-6:omega-3 in the maternal ewe diet? 2. Physiological mechanisms? a) Fatty acid concentrations in plasma and RBC b) Prostaglandins & inflammation Experiments Animal house trial prostaglandin response Pen trial sex ratio
15 Pen Trial - Aims Determine effect of high omega-3 diet on: 1. Blood fatty acid profile 2. Time to oestrus 3. Ovulation rate 4. Pregnancy rate 5. Sex ratio of offspring
16 Dietary Treatments High omega-3 88% Silage High omega-6 70% Oats Omega-6 : Omega : 1 Omega-6 : Omega : 1
17 Pens and Feeding Pen layout randomised block design 3 pens per treatment Total N = 148 per treatment Fed diets 6 weeks pre- and 3 weeks post mating
18 Measurements Live weight and condition score Blood samples for fatty acid analysis (every 2 weeks)
19 Mating Synchronised with CIDRS mated on 1 st natural oestrus
20 Mating Synchronised with CIDRS mated on 1 st natural oestrus Mating with crayon-harnessed rams (2 per pen)
21 Pregnancy and Lambing Transrectal-ultrasound ovulation rate & pregnancy Corpus luteum September lambing record sex of lambs
22 Plasma Omega-6:Omega-3 Ratio
23 Plasma Omega-6:Omega-3 Ratio Oats High Omega-6 p < 0.05 Silage High Omega-3
24 Prostaglandin (PGFM) Response
25 Prostaglandin (PGFM) Response Oats High Omega-6 Silage High Omega-3 p < 0.05
26 Time to Oestrus
27 Time to Oestrus Oats High Omega-6 Silage High Omega-3 p < 0.05 Day of Mating Cumulative Proportion Showing Oestrus (%)
28 Time to Lambing
29 Time to Lambing Oats High Omega-6 Silage High Omega-3 p < 0.05
30 Reproduction Results Dietary Treatment Reproduction Measure High Omega-3 High Omega-6 P-value Proportion of ewes ovulating (%)
31 Reproduction Results Dietary Treatment Reproduction Measure High Omega-3 High Omega-6 P-value Proportion of ewes ovulating (%) Mean ovulation rate: For ewes that ovulated 1.03 (± 0.02) 1.04 (± 0.02) For all ewes 0.77 (± 0.05) 0.83 (± 0.06) 0.190
32 Reproduction Results Dietary Treatment Reproduction Measure High Omega-3 High Omega-6 P-value Proportion of ewes ovulating (%) Mean ovulation rate: For ewes that ovulated 1.03 (± 0.02) 1.04 (± 0.02) For all ewes 0.77 (± 0.05) 0.83 (± 0.06) Proportion of ewes pregnant (%)
33 Reproduction Results Dietary Treatment Reproduction Measure High Omega-3 High Omega-6 P-value Proportion of ewes ovulating (%) Mean ovulation rate: For ewes that ovulated 1.03 (± 0.02) 1.04 (± 0.02) For all ewes 0.77 (± 0.05) 0.83 (± 0.06) Proportion of ewes pregnant (%) Mean foetal number: For ewes pregnant 1.19 (± 0.03) 1.15 (± 0.03) For all ewes 1.07 (± 0.04) 1.05 (± 0.04) 0.764
34 Reproduction Results Continued... Dietary Treatment Reproduction Measure High Omega-3 High Omega-6 P-value Pregnancy loss (%)
35 Reproduction Results Continued... Dietary Treatment Reproduction Measure High Omega-3 High Omega-6 P-value Pregnancy loss (%) Proportion of ewes lambing (%)
36 Reproduction Results Continued... Dietary Treatment Reproduction Measure High Omega-3 High Omega-6 P-value Pregnancy loss (%) Proportion of ewes lambing (%) Mean number of lambs born: For ewes lambing 1.26 (± 0.04) 1.26 (± 0.04) For all ewes 1.04 (± 0.05) 1.14 (± 0.05) 0.148
37 Sex Ratio Males Females Males Females Silage High Omega-3 Oats High Omega-6
38 Sex Ratio Males Females Males Females Silage High Omega-3 Oats High Omega-6
39 Sex Ratio Males Females Males Females Silage High Omega-3 Oats High Omega-6
40 Sex Ratio p = p = Males Females Males Females Silage High Omega-3 Oats High Omega-6
41 Sex Ratio p = p = p = Males Females Males Females Silage High Omega-3 Oats High Omega-6
42 Sex Ratio High Omega-6 Oats diet
43 Conclusions Ewes fed high omega-6 oats diet had: Higher omega-6 levels in blood Increased prostaglandin response Shorter time to oestrus and lambing Higher proportion of female lambs Pre-conception mechanism? Timing of oestrus in relation to ovulation and fertilisation?
44 Take Home Messages Omega-6:omega-3 ratio in feed important Further research: Mechanisms Pre- or post-conception nutrition? Prostaglandin and inflammation? Useful, practical and cost effective feed types
45 Acknowledgements WWAI Farm Staff: Greg Scott, Greg Clark, Steven Huckell, Michael Loiterton, Rex Edis John Wilkins ultrasounding John Piltz, Craig Rodham, Penny Philips, Geoff Casburn, Richard Meyer, Peter Hawkins, Jamie Ayton Brian Alston, Tony Hobbs, Brad Kelk, John Broster, Steph Knott CSU Students Jess & Alex Ausfarm Nutrition My family and anyone else that I ve forgotten!
46 Questions?
47 Contact Details Catherine Gulliver Mobile: Location: Orange, NSW
48 References Cameron, E. Z. (2004). "Facultative Adjustment of Mammalian Sex Ratios in Support of the Trivers-Willard Hypothesis: Evidence for a Mechanism." Proceedings: Biological Sciences 271(1549): Cameron, E. Z., P. R. Lemons, et al. (2008). "Experimental alteration of litter sex ratios in a mammal." Proceedings of the Royal Society of Biological Sciences 275: De Fries, C. A., D. A. Neuendorff, et al. (1998). "Fat supplementation influences postpartum reproductive performance in Brahman cows." Journal of Animal Science 76: Gutierrez-Adan, A., J. Granados, et al. (2001). "Influence of glucose on the sex ratio of bovine IM/IVF embryos cultured in vitro." Reproduction, Fertility and Development 13: Rooke, J. A., C. C. Shao, et al. (2001). "Effects of feeding tuna oil on the lipid composition of pig spermatozoa and in vitro characteristics of semen." Reproduction 121: Rosenfeld, C. S., K. M. Grimm, et al. (2003). "Striking variation in the sex ratio of pups born to mice according to whether maternal diet is high in fat or carbohydrate." Proceedings of the National Academy of Science (check - Proc. Natl Acad. Sci. USA) 100: Rosenfeld, C. S. and R. M. Roberts (2004). "Maternal diet and other factors affecting offspring sex ratio: A review." Biology of Reproduction 71(4): Santos, J. E., T. R. Bilby, et al. (2008). "Long chain fatty acids of diet as factors influencing reproduction in cattle." Reproduction in Domestic Animals 43 (Supplement 2): Trivers, R. L. and D. E. Willard (1973). "Natural Selection of Parental Ability to Vary the Sex Ratio of Offspring." Science 179(4068):
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