Progress 05/01/23 to 04/30/24
Outputs
Target Audience:In the first year of this grant, we are ramping up research so the reporting to the target audience will increase in future years. Auburn's mission as a land-grant university encompasses research, teaching, and extension. Our target audience is poultry growers and private industry partners. Our work involves the development of novel inventions. During the first year, some initial steps were taken to promote the research we are doing and get feedback from industry stakeholders. In particular, meetings were held with the Director of Dairy, Pork, and Poultry of the Alabama Farmer Federation. These meetings entailed describing the research project and goals and setting up a network of communication to facilitate research on the project. In addition, we are in contact and in collaboration with Auburn's technology transfer offices. These professionals bridge the gap between faculty and industry sponsors. Furthermore, Dr. Macklin is frequently involved with outreach and Extension activities and directly interacts with poultry stakeholders on a day-to-day basis. As research scales up and results need to be communicated, we will present at the meeting of the American Association of Avian Pathologists and the Poultry Science Association, the largest poultry stakeholder meeting in the US. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has provided substantial opportunities for training and professional development, significantly contributing to the growth and capability of all team members involved. Central to our team's strategy is the training of three undergraduate students who have been meticulously taught to perform microinjections into litter beetle eggs--a procedure that demands precision and patience. This hands-on experience in advanced biotechnological techniques not only enhances their practical skills but also deepens their understanding of entomological research and its applications in biocontrol. Further enriching the project's educational impact, a PhD student from our lab has taken a leadership role, overseeing the training and daily operations of the undergraduate team. This responsibility has been instrumental in his professional development, enhancing his skills in leadership, mentorship, and project management. His ability to guide and inspire his team has not only boosted the project's productivity but has also prepared him to be a valuable asset in the agricultural workforce, where such expertise is crucial for advancing innovative solutions in pest management. Additionally, the recent appointment of a graduate research assistant (GRA) starting in Fall 2024 underscores our commitment to integrating fresh academic insights and research methodologies into our project. This position will allow the GRA to explore the intersections of pathobiology and poultry science, further expanding the project's scope and impact. In order to assemble such a diverse and capable team, we have developed and disseminated comprehensive job announcements and posters through a variety of channels. These materials were carefully crafted to attract candidates who are academically qualified and passionate about the field of agricultural science and biocontrol. The process of creating these announcements involved input from multiple team members, ensuring that they accurately reflected the project's goals and the specific skills needed. The dissemination strategy targeted platforms frequented by potential candidates with relevant backgrounds, facilitating a recruitment process that was both efficient and effective. Through these initiatives, the project has not only advanced its scientific objectives but has also played a crucial role in fostering the next generation of scientists. These training and professional development opportunities ensure that project participants are well-prepared to contribute to the scientific community and the broader field of agriculture, emphasizing the project's dual focus on achieving research excellence and enhancing educational outcomes. 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 next phase of our project, we are dedicated to continuing our rigorous schedule of microinjections and screenings to successfully establish a Wolbachia-infected line of litter beetles. This foundational work is critical as we advance towards our goal of biological control. Concurrently, we will initiate a series of environmental experiments and surveys on poultry farms across Alabama. These studies will include extensive travel and distance monitoring of beetles, utilizing mark-recapture techniques and trapping studies to develop and refine methodologies crucial for the potential implementation of the sterile insect technique directly on farms. Our research aims to determine whether litter beetles predominantly exist as isolated populations within barns or if they migrate between barns and surrounding wild areas. Understanding this behavior is pivotal; if beetles are found to primarily colonize barns as discrete populations, this could significantly enhance our ability to target and eradicate them effectively. Additionally, we plan to conduct RNAseq experiments to compare male and female beetles, aiming to identify sex-specific transcripts. These findings could lead to the development of RNAi-based strategies for sex-specific eradication, enhancing our capacity for sex sorting within beetle populations. As we gather and analyze data from these experiments, we will prepare to disseminate our findings through scientific manuscripts and detailed reports, ensuring that the knowledge gained from this research contributes broadly to the field and provides a robust foundation for future biocontrol strategies.
Impacts
What was accomplished under these goals?
Over the course of this grant, significant strides have been made toward realizing our ambitious goal to develop a reproductive biocontrol strategy for litter beetles, leveraging the cytoplasmic incompatibility (CI) effects of Wolbachia bacteria. This innovative approach, which centers on the hypothesis that Wolbachia CI can suppress litter beetle populations, has seen considerable advancements despite the inherent challenges and complexities of biocontrol research. To date, our team has successfully expanded our litter beetle colonies and refined our microinjection techniques, which are critical for introducing Wolbachia into beetle eggs. We have effectively increased both the rate of these injections and the size of our colonies, thereby enhancing our capacity for broader experimental endeavors. This expansion is crucial as it directly supports the first objective of our project: to optimize microinjections for generating Wolbachia infections in litter beetles. In our pursuit of this goal, we have tested two distinct Wolbachia strains, wMel and wHa, using innovative buffers like sucrose phosphate glutamate (SPG) to improve the viability and efficiency of the microinjections. Additionally, we have explored alternative methods, such as cytoplasmic transfers, providing a comparative basis to assess the most effective techniques for inducing CI. Moreover, we have advanced our molecular biology tools by developing specific PCR primers to screen for Wolbachia infections in beetles. Establishing robust positive and negative controls has solidified our foundation for accurately assessing infection statuses post-microinjection. In addressing our objective to explore sex sorting techniques, preliminary tests have been conducted on RNAi yeast plasmid constructs designed to target litter beetle females specifically. This innovative approach could revolutionize the way we manage beetle populations in the poultry industry. While the microinjections have not yet resulted in successful Wolbachia infections, our team remains undeterred. The recent acquisition of a Wolbachia-infected beetle line, Tribolium confusum, closely related to litter beetles, offers a promising new avenue for achieving our goals. The development of egg collection methodologies for this new beetle line is underway, and it is aimed at harnessing its Wolbachia for effective colonization of litter beetle ovaries. Looking ahead, we are poised to expand our hypothesis testing and further refine our techniques in the second year of the project. The accomplishments to date underscore our commitment and adaptability in navigating the complexities of biocontrol research. Despite the challenges, our team is optimistic and driven, determined to continue pushing the boundaries of what is possible in the biocontrol of litter beetles.
Publications
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