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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Biosciences & Biotechnology Laboratory » Research » Publications at this Location » Publication #314079
Title: Modulating sperm membrane lipids in domestic turkeys to improve semen cryopreservation for valuable genetic stocks

Author
item LIU, JIANAN - US Department Of Agriculture (USDA)
item VELLEMAN, SANDRA - The Ohio State University
item GRAHAM, JAMES - The Ohio State University
item Long, Julie

Submitted to: Society for the Study of Reproduction Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 3/7/2018
Publication Date: 6/18/2015
Citation: Liu, J., Velleman, S.G., Graham, J.K., Long, J.A. 2015. Modulating sperm membrane lipids in domestic turkeys to improve semen cryopreservation for valuable genetic stocks. Society for the Study of Reproduction Annual Meeting. 93:160.

Interpretive Summary:

Technical Abstract: There is a pressing need for preserving germplasm from research poultry lines, which are valuable genetic stocks for the research community and poultry industry. Semen cryopreservation can be a feasible strategy for maintaining threatened poultry lines, but the quality of frozen/thawed poultry semen with current methods is not reliable enough for germ-line retrieval, especially from turkeys. It has been suggested that poultry sperm cryosurvival is affected by the lipid composition of plasma membrane, which can be modified by inclusion of polyunsaturated fatty acids (PUFAs) in the diet. We tested the hypothesis that diets enriched with n-3 or n-6 PUFAs can modify the lipid composition of turkey sperm membranes and improve sperm cryosurvival. Using a standard formulation as the base diet, the following control and treatments were evaluated: no additives (Group 1, control); additional (120 mg/kg) vitamin E (Group 2); 5% Arasco® oil (n-6 PUFA) + vitamin E (Group 3); 5% Dhasco® oil (n-3 PUFA) + vitamin E (Group 4); 5% soybean oil (n-6 PUFA) + vitamin E (Group 5); and 2.5% soybean/2.5% Arasco® oil + vitamin E (Group 6). Four unique research turkey lines from Ohio State University (OSU) were evaluated. RBC1 and RBC2 are random-bred control lines; the E and F lines are sub-lines of RBC1 and RBC2, respectively, that have been selected for production traits. Sixty males from each line (n = 10/group) were fed the control or modified diets for 6 wks. Semen was frozen (8% dimethylacetamide; 0.5-ml straw; 2-min vapor freeze) after 3 and 6 wks of diet consumption and subsequently thawed (30 sec; 5°C) to assess membrane integrity, lipid peroxidation (LPO) and fertility. Sperm membrane integrity was determined using the SYBR/PI dual stain and assessed by flow cytometry. In RBC2 line, the percentage of sperm with intact plasma membranes was greater (P<0.05) for Groups 2-6 (range: 33.9-47.0%) than the control (31.0%). In RBC1 line, three diet treatments (Groups 4-6) improved (P<0.05) sperm membrane integrity (range: 32.5-36.8%) compared to the control (28.3%). For the E and F lines, not all diets improved sperm membrane integrity compared to control. Sperm that were subjected to LPO were detected by C11-BODIPY581/591/PI stain and flow cytometry. In RBC2 line, all diet treatments reduced (P<0.05) the percentage of peroxidized sperm (range: 10.1-33.0%) compared to the control (37.2%); a similar trend was seen in E line (control: 66.0%; Groups 2-6: 32.7-46.1%). For the RBC1 and F lines, only Groups 5 and 6 reduced (P<0.05) LPO in sperm compared to the control. For fertility trials, commercial hens (n=5/diet group; 30/OSU line) were inseminated with 300x106 sperm for two consecutive days. Eggs were collected for 9 wks, candled on Day7 of incubation and allowed to hatch. Line-specific differences in fertility rates were observed for the first 4 wks of egg production. For RBC2 line, Groups 5 and 6 supported higher fertility (P<0.05) than the control, Group 2 and Group 4. None of the diets improved (P>0.05) the fertility of frozen/thawed E line semen. For RBC1 line, more viable embryos (P<0.05) were observed for Group 5 than the control, Group 2 or Group 3; whereas no differences (P>0.05) occurred among diet treatments for the F line. Fertility persisted after wk 5, however, all embryos died within 48 hr of incubation. Taken together, the results suggest that PUFA-enriched diets may provide an alternative strategy to meet the immediate needs of poultry cryoconservation. Research supported by USDA Grant 2013-67015-2108. Reference ID: 0540-000500