Progress 10/01/19 to 09/30/20
Outputs
Target Audience:Pork is the number one consumed meat worldwide. The United States is a significant contributor to pork production. Nationally, North Carolina ranks second in pork production, employing more than 46,000 people and directly contributing over $2.5 billion (with a total economic impact estimated at $9 billion) to the state economy. Also noteworthy is that in North Carolina's Sampson and Duplin counties were the top two pork producing counties in the US in 2012. Maintaining healthy hogs and increasing production is important. Pre-weaning and post-weaning mortality and morbidity are major factors that negatively impact swine production. The purpose of this study is to investigate the impact that Moringa oleifera, given as a supplement to sows during gestation and lactation, has on quantity and quality of milk; and determine what influence it makes to intestinal health and growth of piglets before and during weaning. Milk is critically important for supporting neonatal growth, development, and health during both the pre- and post-weaning periods and a mother's nutrition during gestation and lactation periods significantly impact the quantity and quality of milk and thus the vitality and vigor of neonates. Moringa oleifera Lam also known as "Mother's best friend", is a tree with many reported medicinal properties. Nutritionally, it is packed with many vitamins, minerals, phytochemicals, and antioxidant compounds that promote health in humans and several livestock species (chickens, pigs, cows, and goats). In humans and ruminants, it has been shown to increase milk production; however, information relating to whether or how it influences milk production of pigs is lacking. Furthermore, what impact providing Moringa to sows during gestation and lactation has on Milk, health and growth of piglets is not known. With this in mind, the primary target audience for this project are persons with interests and steaks in the pork industry, i.e., researchers and producers. A second audience of relevance are researchers (biomedical and animal), consumers, and health care providers that are interested in maternal-fetal health and understanding the connections between nutrition, milk, and neonatal health. Finally, those interested understanding the health impacts of Moringa in animals and humans would also be interested in the activities of this study. This could also help to increase the consumption of Moringa as well as its production. To complete the aims of this study, on farm and in Lab experiments will be conducted as well as completion and submission of research and review articles research publications, presentations, and seminars. Changes/Problems:Due to the restrictions stemming from the pandemic progress on the project goals were hampered. The project timeline was significantly impacted and our collaborator at Purdue had to request a no cost extension of the funds to complete analysis of milk samples. A graduate student was registered for a conference to present work, but that travel was cancelled because of the SARS-CoV2 pandemic. Because of these challenges, safety concerns, and limited access to the lab stemming from the COVID -19 pandemic, we have made some alterations/additions to the scope of this project. Including, adopting an in vivo mouse model, an in vitro pig cell culture model system, and in vitro nutrition studies. We proposed these actions, because experimentation with swine required many people to work together in proximity and was not possible with the Pandemic related safety precautions and restrictions. What opportunities for training and professional development has the project provided?During the reporting year, this project provided training and laboratory experience to a total of 4 students (3 undergraduates and 4 MS student). The undergraduate students were trained in feed formulation and analysis, blood and milk collection, proper animal handling and restraint, and isolation of white blood cells from milk and blood, performing total and differential cell counting and measuring immunoglobulin and other proteins levels in samples using enzyme linked immunosorbent assays (ELISA). In addition to this, one graduate student was trained in proteomic analysis, another performed batch culture studies with Moringa as a supplement to improve feed efficiency and reduce greenhouse gas emission. Three MS students have learned first-hand how to prepare literature review articles for publication. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?In the most immediate future, the project will include alterations/additions to the scope of this project to include an in vitro model system, whereby pig white blood cells will be treated in a cell culture dish in the presence of Moringa leaf extracts and evaluated for changes in functions. To evaluate the impact of Moringa on milk production quantity and quality (changes in milk components) we will use an in vivo mouse model system. For this, mice will be fed Moringa and the amount and quality of the milk investigated. We are proposing this action, because experimentation with swine requires many people to work together, this is not possible with the current safety precautions and restrictions. In contrast neither mouse nor in vitro require the same number or personnel and thus can be completed safely. We will continue to work on drafting of review and research articles for submission and performing glycoproteome analysis. We hope that some of these restrictions will be lifted soon and are hopeful that we will be able to begin conducting the trial in swine in year three of the project. We will need to train a new set of students and will require an extension to be able to complete all the analysis of the samples collected.
Impacts
What was accomplished under these goals?
Because of restrictions from the COVD-19 pandemic, we were not able to conduct the experiment as planned and therefore did not make progress on the aim of evaluating the impact of Moringa on sow and piglet performance. Although we were not able to conduct the Moringa feeding trial, because of social distancing and safety requirements and protocols, we have moved forward as we could. (1) We completed growing, drying, and processing Moringa oleifera leaves that will be used for feeding sows as soon as the restrictions are lifted. We used the time to draft literature reviews on the: (a) Impact of Moringa on immune cell function, (b) Use of Moringa as a galactagogue-to influence milk production and (c) use of Moringa in livestock production. All these papers are still in process and were very essential in uncovering many unanswered questions that have informed additional and related projects that are to be addressed in year three of the project. These review articles are also being led by MS students and are crucial for their successful completion of degree. From samples that we were able to collect as part of the activities in the 2018-2019 period, we generated and continue to generate interesting data on swine milk (non-moringa- supplemented). One of the most significant findings we have made to date is that there are many unique proteins expressed in milk at different stages. Milk is divided into three major phases- colostrum (day 1- early milk), transitional milk (day 2- d14) and then mature milk (day 14 on). The changes that occur during these phases help support the changing needs of the neonate. It is known that milk is a dynamic and rich fluid, but there is still a lot not known about it, what is in it and how it influences young. We have data showing that there are 121 uniquely expressed proteins in day 3 swine milk that were not expressed in colostrum or mature milk. We will investigate what these proteins are, and then work to determine their functions so we may predict what role they may have on the neonate/piglet. This is impactful, because it may help identify proteins that could help support health, growth and development of neonates and piglets.
Publications
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Progress 10/01/18 to 09/30/19
Outputs
Target Audience:Pork production is vital to the U.S. and global economies. Maintaining healthy hogs and increasing production is important. The primary target audience for this project are persons with interests and stake in the pork industry. While the activities will generate data that is specifically targeted toward research scientists (swine) as well as swine producers, the data generated may also be of benefit to biomedical research scientists as well as persons interested in health of other animals, livestock or companion. To complete the aims of this study, internships and in lab and on farm research training has been provided to undergraduate and graduate students. Changes/Problems:1) While a supplier for Moringa was identified we have only received 1/10 of the total Moringa ordered. If we do not receive all that we need, we will either order from an additional source (pending budget), postpose the study, or decrease the inclusion rate of the Moringa that was proposed in the study. 2) While we have identified and begun training students for this project, two of the students may not be able to participate in the project as planned because of summer classes. 3) Furthermore, we have not yet been successful in optimizing the protocol for isolating mononuclear cells from milk. Therefore, if we are not successful, we may not be able to complete the analysis of T-cells, B-cells and monocytes as proposed, but may instead focus on neutrophils. 4) Finally, from the training and dry runs at the Swine Unit we determined that conducting this study in the outdoor facility as proposed was not feasible and therefore have adjusted the study to be conducted in the indoor unit. What opportunities for training and professional development has the project provided?During the reporting year, this project provided training and laboratory experience to a total of 10 students (8 undergraduates and 2 MS student). The undergraduate students were trained in feed formulation and analysis, blood and milk collection, proper animal handling and restraint, protein isolation and western blotting, the technique for isolating white blood cells from milk and blood, performing total and differential cell counting and measuring immunoglobulin and other proteins levels in samples using enzyme linked immunosorbent assays (ELISA). In addition to this, one graduate student was able to complete two one-week summer internship training on how to collect milk from sows as well as how to processes the samples for proteomic analysis. Currently she is being mentored on how to analyze and perform bioinformatics and statistics on the proteomic data. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
As proposed, all experimental activities are scheduled to begin in year two of the project. As such there is no progress on the experimental aims to be reported. Year one was been dedicated to purchasing necessary equipment, procuring Moringa and training/optimization of animal handling and sample collection. We therefore report the following accomplishments. 1) Hammer mill, pelleter and vacuum sealer were purchased for processing and storing the Moringa. 2) A supplier for Moringa was identified and we have received 1/10 of the total Moringa order. 3) A supplier for the sows that will be used for this study has been identified. 4) Students have been trained in animal handling, sample collection, and processing. 5) Training for laboratory assays such as ELISA (enzyme-linked immunosorbentassay)and tissue culture was started. 6) From the training and dry runs at the Swine Unit we determined that conducting this study in the outdoor facility was not feasible and therefore have adjusted the study to be conducted in the indoor unit.
Publications
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