Source: UNIVERSAL PIG GENES INC submitted to NRP
OPTIMIZING BOAR SPERM FERTILIZATION COMPETENCY
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
SMALL BUSINESS GRANT
Reporting Frequency
Annual
Accession No.
1024101
Grant No.
2020-51103-32563
Cumulative Award Amt.
$649,866.00
Proposal No.
2020-06323
Multistate No.
(N/A)
Project Start Date
Sep 15, 2020
Project End Date
Sep 14, 2023
Grant Year
2020
Program Code
[8.3]- Animal Production & Protection
Recipient Organization
UNIVERSAL PIG GENES INC
3354 140TH ST
CLEARFIELD,IA 508408063
Performing Department
(N/A)
Non Technical Summary
Problem: U.S. swine herd reproductive performance is limited more by sire sub fertility than dam. Today's boar semen processing is largely unchanged since the 1980s. The newly identified, but longtime existing technical problem is current techniques doom a considerable portion of spermatozoa sub fertile and for premature death, reducing the overall pounds/sow/marketed due to inefficient use of superior genetics.Opportunity: Managing the identified problem can 1) increase sow litter sizes (opportunity cost of $1 billion per 5% increase in piglets per litter marketed) and 2) lead to better use of swine genetics (opportunity cost of $400 million).Project objectives: 1) Validate MU Vanguard Buffers in sow fertility trials. 2) Characterize field fertility of MU Vanguard buffers versus a commonly used industry extender for commercialization. 3) Characterize 30-day extended sperm lifespan of our product. 4) Validate industry extenders compatible with MU Vanguard Supplement.Description of effort: This Phase II project seeks to validate our Phase I findings and commericalize USDA-NIFA research from the University of Missouri (two patented/provisionally patented IPs). We will validate the technology in fertility trials and conduct lab experiments to characterize unprecedented 30-day shelf life and adapt our technology to existing products. Anticipated Results: Validation of our product with technical expected outcomes outlined in the proposal. Commercial application of the proposed research is high because semen extenders are used to produce 95% of the 121 million pigs produced annually. We have a historical track record in adopting swine artificial insemination into industry, we enthusiastically look forward to commercialization.
Animal Health Component
90%
Research Effort Categories
Basic
10%
Applied
90%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
30135301081100%
Knowledge Area
301 - Reproductive Performance of Animals;

Subject Of Investigation
3530 - Other swine products;

Field Of Science
1081 - Breeding;
Goals / Objectives
The overarching goal of this project is to validate Phase I in vitro optimized buffers with in vivo testing by field artificial insemination (AI), as well as characterize and build on the unprecedented long sperm life span attained in Phase I. Most of Phase II is field trial focused. Goals and objectives are split into in vivo and in vitro trials as follows:In Vivo:Goal #1: Validate MU Vanguard Storage and Activator Buffer in sow fertility trials.Objective 1: Do MU Vanguard Extenders promote reproductive success?Objective 2: Is the activator buffer necessary and if so which activator buffer is preferred?Goal #2: Characterize field fertility of MU Vanguard buffers versus commonly used industry extenders.Objective 3: How does the fertility of MU Vanguard Extender(s) compare to currently used extenders?In Vitro:Goal #3: Characterize 30-day extended sperm lifespan of MU Vanguard Storage Buffer.Objective 4: How does longer term (up to 30 days) incubation of MU Vanguard stored semen perform from a biomarker defined, sperm quality perspective?Goal #4: Validate industry extenders compatible with MU Vanguard Supplement.Objective 5: Which commonly used industry extenders are compatible with MU Vanguard in supplement form?
Project Methods
Objective #1All buffers will be mixed at Iowa State University and/or the University of Missouri and shipped by parcel to IBS with a final buffer osmolality of 290-300 mOsm/L. Semen will be collected following IBS standard operation procedure, maintained at 35°C to prevent temperature shock, and immediately diluted in the lab with our MU Vanguard Storage Buffer. Only raw semen with acceptable motility (>80%) and morphology (<15%) will be used. A final count of 2.5 billion spermatozoa per 45 mL storage buffer will be used per dose. Semen will be shipped by overnight parcel to the sow farm, utilizing proper packaging in relationship to route temperatures, as routinely done. Sows will be bred following our established and common protocol. Briefly, only sows displaying estrus post-weaning will be bred. Total inseminations/sow will be 2-3/estrus (24 hours apart until estrus no longer displayed). A total of 25 sows will be inseminated per treatment group. Pregnancy rates, farrowing rates, and litter sizes will be evaluated. Animals will be observed daily by farm staff and tended to by a veterinarian as needed.Objective #2This is a higher-powered sow fertility study of Task 1. This should not be viewed as overlap or repeating work, as this was designed to provide preliminary validation, work out logistics, ensure sow health, and inform any revisions that need made to study design of Task 2, such as decreasing sperm per dose or the number of inseminations. In pig extender fertility trials such as this, a high-powered study is typically required and results will support trademarking, commercialization, and marketing. Methodology remains the same as proposed in Task 1; however, this will be performed at two time points. We will include 100 sows/treatment (300/group). Methodology from Task 1 applies, including location where buffers are made, semen collected, and fertility trial being done.Objective #3This fertility trial will examine commonly used extenders with the best treatment from Task 2. We will inseminate 100 sows/treatment (300/group) in two replicates (600 total sows). Fresh extended boar semen reduces its fertility after three days of incubation, likely because of the storage-induced capacitation that lead to the development of MU Vanguard extenders. This task will be designed with that in mind, utilizing semen that is beyond 3 days of shelf life yet still maintains motility. Further, reduced fertility is realized with 1.0 billion sperm/insemination. To stress the extenders tested here and the commercial relevance of reduced sperm/dose, we will perform this trial with 1.0 billion sperm/insemination.Objective #4This task is solely in vitro. It will be initiated upon the completion of of Task 1. In this task, we desire to better characterize 30 day incubation of semen stored in our buffer/extender, using a biomarker strategy. Buffer will be prepared at Iowa State University and/or the University of Missouri, shipped overnight to IBS for semen extension, then shipped back to Iowa State University and/or University of Missouri for lab analysis. We will use the same sperm concentration as used in task 1 and 2. Traits will be analyzed on Day 1, 7, 14, 21, and 30. We will analyze: motility; pH stability; and biomarker analysis by flow cytometry with appropriate controls. A total of 6 boars per treatment will be analyzed in 3 replicates each. Spermatozoa will by analyzed flow cytometry or similar technologies high-throughput technologies allowing flouresecent biomarker analysis of: 1) Zn signature, 2) viability, 3) acrosome health and status, 4) live nuclear stain Hoechst 33342, 5) Ca2+ levels, 6) intracellular pH; 7) LCA revealed altered membranes during storage, 8) mitochondrial membrane potential, and 9) DNA fragmentation/integrity.Objective #5This task is being explored because a decreased barrier to entry of the boar semen extender market is selling a semen extender supplement, rather than an entirely new semen extender. We will validate whether our combination of supplements/ingredients can benefit currently used semen extenders. The same lab methods will be used as outlined in Objective #4, with days 1 and 7 analyzed.All objectives are subject to changes as suggested by reviewers.

Progress 09/15/22 to 09/14/23

Outputs
Target Audience:The main target audience for this project long term after phase II are boar stud managers that help make decisions with farmers, as well as sow farm decision makers in breeding management, to purchase the final product. Other target audiences include veterinarians. The efforts to deliver science-based knowledge will be through presentations, fact sheets, and/or working with industry labs and businesses to utilize and implement such findings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training and professional development of staff scientists was provided by this project. How have the results been disseminated to communities of interest?Results were shared at the 2021 Boar Stud Manager Conference in August of 2021 (the conference was originally scheduled for 2020, but delayed due to COVID-19) in an invited presentation. Additional results were shared one on one with communities of interest. What do you plan to do during the next reporting period to accomplish the goals?This is the final year, the project is done;therefore, no further reporting periods after this.

Impacts
What was accomplished under these goals? All goals and objectives were completed during this project.For objective #1, we confirmed that MU Vanguard Extenders promote reproductive success and no decreased fertility was observed in this validation trial.For objective #2, we observed the effects of activator buffers and which activator buffer method was preferred if one was to be used depending on customer potential preference. Objective #3 was complete during year 2, and additional trials were performed during year 3 to confirm and add increased statistical power. Wecharacterizedlonger-term incubation (Objective #4) and made refinements with new discoveries to increase this longer period extension. For objective #5, we observed which industry extenders are compatible with MU Vanguard in supplement form, observing that some extenders are more compatible than others. Details are included in the technical report

Publications

  • Type: Other Status: Other Year Published: 2023 Citation: Beings this is an SBIR Phase II grant, we were focused on the development and characterization of product we started developing in SBIR Phase I round, thus external publications are not being made.


Progress 09/15/20 to 09/14/23

Outputs
Target Audience:The main target audience for this project long term after phase II are boar stud managers that help make decisions with farmers, as well as sow farm decision makers in breeding management, to purchase the final product. Other target audiences include veterinarians. The efforts to deliver science-based knowledge will be through presentations, fact sheets, and/or working with industry labs and businesses to utilize and implement such findings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training and professional development of staff scientists was provided by this project. How have the results been disseminated to communities of interest?Results were shared at the 2021 Boar Stud Manager Conference in August of 2021 (the conference was originally scheduled for 2020, but delayed due to COVID-19) in an invited presentation. Additional results were shared one on one with communities of interest. What do you plan to do during the next reporting period to accomplish the goals?This is the final year, the project is done; therefore, no further reporting periods after this.

Impacts
What was accomplished under these goals? All goals and objectives were completed during this project. For objective #1, we confirmed that MU Vanguard Extenders promote reproductive success and no decreased fertility was observed in this validation trial. For objective #2, we observed the effects of activator buffers and which activator buffer method was preferred if one was to be used depending on customer potential preference. For objective #3 trials were performed to characterize field fertility of MU Vangaurd buffers versus commonly used industry extenders for which we saw increase in average reproductive traits.We characterized longer-term incubation (Objective #4) and made refinements with new discoveries to increase this longer period extension. For objective #5, we observed which industry extenders are compatible with MU Vanguard in supplement form, observing that some extenders are more compatible than others. Details are included in the technical report

Publications

  • Type: Other Status: Other Year Published: 2023 Citation: Beings this is an SBIR Phase II grant, we were focused on the development and characterization of product we started developing in SBIR Phase I round, thus external publications are not being made. We will work with collaborators and universities to make future publications, either in fact sheet form or peer reviewed publications.


Progress 09/15/21 to 09/14/22

Outputs
Target Audience:The main target audience for this project long term after phase II are boar stud managers that help make decisions with farmers, as well as sow farm decision makers in breeding management, to purchase the final product. Other target audiences include veterinarians. The efforts to deliver science-based knowledge from phase III will be through presentations, research submitted to leading journals in the field, and/or working with industry labs and businesses to utilize and implement such findings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Further training and professional development of staff scientists was provided by this project. How have the results been disseminated to communities of interest?Results were shared at the 2021 Boar Stud Manager Conference in August of 2021 (the conference was originally scheduled for 2020, but delayed due to COVID-19) in an invited presentation. Additional results are being shared one on one with communities of interest. What do you plan to do during the next reporting period to accomplish the goals?We plan to perform additional fertility trials.

Impacts
What was accomplished under these goals? Objective #1 was completed during the first (previous) reporting period. Objective #2 was completed during this period. We observed the effects of activator buffers and which activator buffer method was preferred if one was to be used depending on customer potential preference. Objective #4 was completedduring the first (previous) reporting period. Objective #5 was completed during this period. We performed experiments to observe which industry extenders are compatible with MU Vanguard in supplement form, observing that some extenders are more compatible than others. Details are included in the technical report.

Publications

  • Type: Other Status: Other Year Published: 2022 Citation: Beings this is an SBIR Phase II grant, we are focusing on the development and characterization of product we started developing in SBIR Phase I round, thus external publications are not being made.


Progress 09/15/20 to 09/14/21

Outputs
Target Audience: The main target audience for this project long term after phase II are boar stud managers that help make decisions with farmers, as well as sow farm decision makers in breeding management, to purchase the final product. Other target audiences include veterinarians. The efforts to deliver science-based knowledge from phase III will be through presentations, research submitted to leading journals in the field, and/or working with industry labs and businesses to utilize and implement such findings. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Further training and professional development of staff scientists was provided by this project. How have the results been disseminated to communities of interest?Results were shared at the 2021 Boar Stud Manager Conference in August of 2021 (the conference was originally scheduled for 2020, but delayed due to COVID-19) in an invited presentation. What do you plan to do during the next reporting period to accomplish the goals?We will finish Objective #2 as originally planned to span year one and two along with start Objective #3. Given new discoveries that affected longer-term incubation, we will finish characterizingthis longer period shelf life.Tasks for Objective #5areplanned for this next reporting period.

Impacts
What was accomplished under these goals? For objective #1, we confirmed that MU Vanguard Extenders promote reproductive success and no decreased fertility was observed in this validation trial. This prompted a higher power study, as originally proposed, and is currently underway. Elucidating the activating buffer affect and a higher-powered study under Objective #2 was initialized,spaning years oneand twoas originally planned. Tasks for Objective #3 areplanned for year two. We have been characterizing longer-term incubation (Objective #4) and making refinements with new discoveries to increase this longer period extension.Tasks for Objective #5areplanned for year two.

Publications

  • Type: Other Status: Other Year Published: 2021 Citation: Beings this is an SBIR Phase II grant, we are focusing on the development and characterization of product we started developing in SBIR Phase I round, thus external publications are not being made while we are finalizing product development.