Grant Information
| Knowledge Area | Subject of Investigation | Field of Science | Percent |
|---|---|---|---|
| 301 - Reproductive Performance of Animals | 3010 - Honey bees | 1020 - Physiology | 60% |
| 314 - Toxic Chemicals, Poisonous Plants, Naturally Occurring Toxins, and Other Hazards Affecting Animals | 3010 - Honey bees | 1150 - Toxicology | 20% |
| 315 - Animal Welfare/Well-Being and Protection | 3010 - Honey bees | 1100 - Bacteriology | 20% |
Tetracyclines are the most commonly used antibiotic for treatment and prevention of disease inhumans and livestock, including honey bees. Like other antibiotics, tetracyclines have been classified as a chemical that negatively affects hormones because of its impact on reproductive cells. In mammals, oxytetracycline has been shown to decrease the movement of sperm and the amount of live sperm, it causes damage to testicles, and reduces the production of sperm and reproductive fertility overall. However, the impact of tetracyclines on honey bee reproduction has not been investigated. Historically, a honey bee queen's lifespan was 2-7 years, but now 50% of colonies replace their queens within six months. A colony will replace their queen for several reasons, including lack of stored sperm within the queen. Colonies that are failing have queens with low sperm counts and sperm counts are highly variable in drones (the male honey bee), but it is unclear why. Given the importance of queen reproductive success to honey bee colony survival, it is imperative to determine the factors contributing to diminished reproductive quality in both queens and drones. Additionally, the gut microbiome (community of bacteria that live in the gut) has been shown to play a role in growth and development of honey bee workers but its importance in queen and drone health and reproductive development are unknown. Here we aim to determine how antibiotics impact the reproductive health of honey bee 1) queens, and 2) drones, and 3) investigate the link between the microbiome and reproduction in honey bees.
The major goals of this projectare to 1) identify the cause for reproductive decline in honey bee populations, 2) update antibiotic treatment standards and regulations in beekeeping, 3) improve the standard of reproductive toxicology research methods, 4) close the research gap between in-hive treatments and fecundity in honey bees and other pollinators, and 5) highlight the need for future research on chemically induced reproductive toxicity in other agricultural livestock, declining pollinators, and organisms. I intend to accomplish these goals with a combination of research, outreach, teaching, learning, science communication, and collaboration with researchers, veterinarians, and beekeepers.To this aim, we will address three supporting objectives:
Objective 1:These results will enable us to determine if ingested oxytetracycline 1) can reach the reproductive tissue and at what concentration, 2) causes reproductive tissue damage, 3) affects internal organ pH, and 4) affects the native reproductive microbiome of queens.Mass spec methods:Afterreaching reproductive maturity the queens will be separated into cup cages accompanied by 20 NEWs and the cages split into control and treatment groups.The control group will be fed 0.5M filtered sugar syrup and the treatment group antibiotic suspended in 0.5M filtered sugar syrup for five days; 200 μg/mL oxytetracycline. After five days, queens will be anesthetized with CO2and sacrificed. The ovaries and spermatheca will be dissected under sterile conditions and stored at -80°C until analyzed.The samples will be weighed then homogenized in extraction buffer, after which, the samples will be purified for the chemical of interest by use of an Oasis HLB purification cartridge. The samples will then be evaporated by nitrogen stream and resuspended in LC/MS grade 10% methanol before analysis. Q-Exactive mass spectrometry will be used to identify and quantify oxytetracycline and its metabolite EOTCusing a predetermined 10-point calibration curve.Levels found in treated and control queens will be compared to account for potential preexisting environmental exposure and possible contamination.Histology methods:The spermatheca and ovaries will be fixed with 10% neutral formalin buffer and 70% molecular grade ethanol. The spermatheca will be orientated from spermatheca duct to the lateral end of the spermatheca and set in a block for paraffin' tissue sectioning. Ovaries will be orientated from the median oviduct to the cranial end of the ovary and set in a block for paraffin' tissue sectioning. A few sections from each of the reproductive organs will be set on a microscope slide and stained withHaemotoxylin and Eosin (H&E) for differentiation between nuclei, extracellular matrix, and cytoplasm. After imaging, pathology analysis will determine changes in the treatment tissue compared to control (i.e., necrosis and apoptosis). For further analysis, a few more sections of each organ will be placed on a microscope slide and stained using a TUNEL assay to determine if the antibioticcauses DNA fragmentation in the tissue.pH methods:Spermatheca and ovaries from control and treatment queens will be analyzed immediately after dissection with a micro pH electrode. The electrode will be used to puncture the septa of the spermatheca and ovaries to determine the internal organ's pH. When possible, the organ's pH will be measured in multiple positions. Antibiotictreated pH levels will be compared to control pH levels.Sperm viability methods:Spermatheca from control and treatment queens will be ruptured in buffer (releasing the housed sperm) and a sample will be taken for Live/Dead analysisto determine percent viability of each queen. Additionally, I will stain the semen samples with a mitochondrion-selective reagent (Rhodamine 123) to determine mitochondrial activity by the stains ability to bind to electronegative membrane potentials.Change in mitochondrial potential can be an early sign of apoptosis that would not be present in the Live/Dead staining and may give us further incite to how oxytetracycline is causing sperm death.Sequencing methods:The virgin and inseminated queens will be kept in cup cages accompanied by 20 NEWs. The cup cages will be split into control and treatment groups for both virgin and inseminated queens with 15 queens per group, replicated three times.The treatment group will be fed 200mg/mL oxytetracycline suspended in 0.5M filtered sugar syrup; controls only sugar syrup. Once the queens have been exposed to the antibiotic for five days, they will be anesthetized with CO2and sacrificed. The ovaries and spermatheca will be dissected under sterile conditions, homogenized, DNA will be extracted, and the microbiome composition will be determined by sequencing the V4 region of the 16S rDNA using Illumina iSeq 100. Sequencing data will be analyzed using the bioinformatic pipeline Qiime. The microbiome composition of the spermatheca and ovaries of antibiotic treated virgin and inseminated queens will be compared to that of control queens within the same group by community composition, relative and absolute abundance of species and alpha and beta diversity.Objective 2:The rational and experimental methods for investigating drone microbiome and fecundity are the same as Objective 1, except for the following differences.Sampling male reproductive system while the testis is viable will enable us to tease apart whetheroxytetracyclinereduces mature sperm viability, if it disrupts spermatogenesis, or both.Mas specmethods:Immature or mature drones will be fed200mg/mL oxytetracycline suspended sugar syrup for five days;controls only sugar syrup.Q-Exactive massspectrometry will be usedto quantify the amount of oxytetracycline and its metabolite 4-EOTC that is absorbed and targets the testis/seminal vesicle/mucus gland tissue. Levels found in treated and control drones will be compared to account for potential preexisting environmental exposure and possible contamination.Histology methods:The seminal vesicles will be fixed with 10% neutral formalin buffer and 70% molecular grade ethanol. The seminal vesicles will be orientated from the cranial to caudal end and set in a block for paraffin' tissue sectioning. The samples will be imaged and processed as explained in histology methods of Objective 1.pH methods:Testis from control and treatment drones will be analyzed immediately after dissection with a micro pH electrode. The electrode will be used to puncture the septa of the testis to determine the internal organ's pH.Antibiotic treated pH levels will be compared to control pH levels.Sperm viability methods:Only mature drones will be assessed for sperm viability after the treatment period.Seminal vesicles from control and treatment drones will be ruptured in buffer (releasing the housed sperm) and a sample will be taken for Live/Dead analysis.Iwill stain the semen samples with a mitochondrion-selective reagent (Rhodamine 123) to determine mitochondrial activity by the stains ability to bind to electronegative membrane potentials.Sequencing methods:I have already confirmed that drones possess a reproductive microbiome. The testis, seminal vesicles, and mucus glands of dissected drones will be treated and analyzed as stated in the sequencing methods of Objective 1.Objective 3:The role the gut microbiome plays in reproductive development and fitness is not well understood, and to our knowledge, it has never been studied in honey bees.Methods:During the above studies (Objective 1 and 2) the gut will be removed from queens and drones treated with antibiotics, control sugar syrup, germ free conditions and conventional conditions as well as all combinations of the groups (e.g. antibiotic and control treated germ-free and conventional bees). These organs will be kept at -80°C until processing. The gutswill be homogenized, DNA will be extracted, and the microbiome composition of each will be determined by sequencing the V4 region of the 16S rDNA using Illumina iSeq 100. Sequencing data will be analyzed using the bioinformatic pipeline Qiimeto characterize bacterial species. The microbial analysis of the gut will be compared to the reproductive findings from the same individual bees explored in Objective 1 and 2.?
Target Audience
Most of my reached target audience within this awardperiod was beekeepers. NC has the highest population of hobbyist beekeepers out of all 50 states. Beekeeping chapters across the state have been inviting me to travel and present my research. During this awardperiod I presented this research 17 times, including four county beekeeper presentations and one at the NC State Beekeeper meeting in 2021.In addition to beekeepers, I also got the opportunity to present my research to the local Audubon Society Chapter in Guilford County, NC, and to thecommunity at theGreensboro Science Pub event, Joymongers Brewing Greensboro, NC.I have now given 12invited oral presentations ranging from 45 minutes to 90 minutes. My secondary target audience is other scientists. I gave threeoral presentations at conferences during this awardperiod. The first at the Association of Southeastern Biologists (ASB) Annual Conference in Winston-Salem, NC, where I won first place in the microbiology section. The second presentation was at Carolina's Society of Environmental Toxicology and Chemistry Annual Meeting in Charleston, SC, where I had an accepted competition presentation. Lastly, I presented my dissertation defense in April of 2024.In addition to my oral presentations, I gave fourposter presentations within the awardperiod. I had an accepted poster at theNC Microbiome Symposium, RTP, NC, where I won a poster award. Secondly, I had anaccepted abstract and poster atthe American Society of Microbiology (ASM) Microbe conference in Washington D.C.. Thirdly,an accepted abstract and poster for competition at the Entomological Society of America (ESA) Annual Joint Meeting in Vancouver, CA, where I won second place. Lastly, an accepted abstract and poster at the Graduate Research and Creativity Expo hosted by UNCG in Greensboro, NC. Some minor target audience interactions during this awardperiod have been keeping up with my veterinarian contacts that help regulate antibiotic use in beekeeping, I continue correspondence to keep them up to date on my findings. Lastly, I mentor undergraduate students in our lab, but I also sat on a graduate panel about insect research for an insect apocalypse course.
Changes / Problems
While pursuing aim two, I have realized that there is an inhibitor in the honey bee queen reproductive organs (ovaries and spermathecae) that is inhibiting the amplification of bacteria via PCR. I have spent months trouble shooting with queen organs and have decided to move on to focus my dissertation and this grant on drones. Drones are the least studied honey bee and there is much to be done to understand how antibiotic hive treatments are affecting their development and overall reproductive capacity in honey bee populations. However, a successful drone rearing protocol did not exist. Creating these new methods took me the better part of two summers to complete. Additionally, drones are the hardest honey bee caste to work with. Workers only keep them Spring-Fall, they do not overwinter, and it is up to the workers if they want extra mouths to feed. This results in workers killing experimental drones often. To this issue, I wanted to determine if ingested oxytetracycline reduced sperm viability in mature drones, however, this experiment was never successful as it never resulted in an appropriate sample size of drones. Overall, I am satisfied with the discoveries I have made, even though it was a challenging PhD and study subject.?
Training & Professional Development
I have trained in the lab with Drs. David Tarpy and Brad Metz at NC State. I have also had training for multiped microscopes and cell readers at UNCG including at the Joint School of Nanoscience and Nanoengineering. I have sat on 11 committees and 2 panels. I taught general biology laboratory for science majors for three years. I have mentored 13 undergraduates and 6 graduate students throughout my PhD. I mentored new graduate instructors and created an introductory curriculum to help them establish productive teaching methods. I trained many new beekeepers. I was elected senator for biology then vice president of the Graduate Student Association at UNCG, of which I worked in for three years andconceptualized, organized, and hosted over 35 events for students. I have also volunteered at 23 events, including five science events for kids.?
Dissemination Streams
During this awardperiod I presented this research 17 times, including four county beekeeper presentations and one at the NC State Beekeeper meeting in 2021.In addition to beekeepers, I also got the opportunity to present my research to the local Audubon Society Chapter in Guilford County, NC, and to thecommunity at theGreensboro Science Pub event, Joymongers Brewing Greensboro, NC.I have now given 12invited oral presentations ranging from 45 minutes to 90 minutes. I gave threeoral presentations at conferences during this awardperiod. The first at the Association of Southeastern Biologists (ASB) Annual Conference in Winston-Salem, NC, where I won first place in the microbiology section. The second presentation was at Carolina's Society of Environmental Toxicology and Chemistry Annual Meeting in Charleston, SC, where I had an accepted competition presentation. Lastly, I presented my dissertation defense in April of 2024.In addition to my oral presentations, I gave fourposter presentations within the awardperiod. I had an accepted poster at theNC Microbiome Symposium, RTP, NC, where I won a poster award. Secondly, I had anaccepted abstract and poster atthe American Society of Microbiology (ASM) Microbe conference in Washington D.C.. Thirdly,an accepted abstract and poster for competition at the Entomological Society of America (ESA) Annual Joint Meeting in Vancouver, CA, where I won second place. Lastly, an accepted abstract and poster at the Graduate Research and Creativity Expo hosted by UNCG in Greensboro, NC.
Next Reporting Steps
Nothing Reported
Target Audience
Most of my reached target audience within this awardperiod was beekeepers. NC has the highest population of hobbyist beekeepers out of all 50 states. Beekeeping chapters across the state have been inviting me to travel and present my research. During this awardperiod I presented this research to beekeepers and scientists via invited oral 3times, contributed oral presentation 3 times, and 1 poster presentation.
Changes / Problems
While pursuing aim two, I have realized that there is an inhibitor in the honey bee queen reproductive organs (ovaries and spermathecae) that is inhibiting the amplification of bacteria via PCR. I have spent months trouble shooting with queen organs and have decided to move on to focus my dissertation and this grant on drones. Drones are the least studied honey bee and there is much to be done to understand how antibiotic hive treatments are affecting their development and overall reproductive capacity in honey bee populations. However, a successful drone rearing protocol did not exist. Creating these new methods took me the better part of two summers to complete. Additionally, drones are the hardest honey bee caste to work with. Workers only keep them Spring-Fall, they do not overwinter, and it is up to the workers if they want extra mouths to feed. This results in workers killing experimental drones often. To this issue, I wanted to determine if ingested oxytetracycline reduced sperm viability in mature drones, however, this experiment was never successful as it never resulted in an appropriate sample size of drones.
Training & Professional Development
During this reporting period, I trained two undergraduates and taught new students how to beekeep.
Dissemination Streams
I have presented at all the previously listed scientific conferences and to beekeeping communities in NC.
Next Reporting Steps
This award period was extended from 05/24 to 08/24, during this time I will be working on publishing my dissertation and three peer reviewed articiles. No further research will be done, only writing of results. <br><br>
<br>What was accomplished under these goals? I accomplished the following discoveries and am currently working on publishing the findings: 1) Characterization of the male honey bee (drone) microbiomes found in the gut, mucus glands, and seminal vesicles 2) Drones need social contact with workers in order toacquire thier core microbiome *1 and 2 accepted in mSphere as of 11/27/24 3) A common antibiotic (oxytetracycline) is highly toxic to mature drone sperm in vitro 4) After injesting less than field relative doses of oxytetracycline, it can be quanitified in queen and drone reproductive organs and guts 5) Determined the most successful in vitro rearing drone protocol 6) Oxytetracycline reduced the abundance of core microbiota in mature drones 7) Drone larvae are sensitive to oxytetracyline treatment and die if exposed to field relative doses <br><br><b>Publications</b><br>
Target Audience
Most of my reached target audience within this reporting period was beekeepers. NC has the highest population of hobbyist beekeepers out of all 50 states. Beekeeping chapters across the state have been inviting me to travel and present my research. During this reporting period I presented at the Rowan County Beekeepers Association and Henderson County Beekeepers Association. I also attended the North Caroline State Beekeepers Association annual meeting in July of 2022 where I networked with beekeepers and associated companies. In addition to beekeepers, I also got the opportunity to present my research to the local Audubon Society Chapter in Guilford County, NC. I have now given 10 invited oral presentations ranging from 45 minutes to 90 minutes. My secondary target audience is other scientists. I gave two oral presentations at conferences during this reporting period. The first at theAssociation of Southeastern Biologists (ASB) Annual Conference in Winston-Salem, NC, where I won first place in the microbiology section. The second presentation was at Carolina's Society of Environmental Toxicology and Chemistry Annual Meeting in Charleston, SC, where I had an accepted competition presentation. In addition to my oral presentations, I gave three poster presentations within the reporting period. I had an accepted abstract and poster at theAmerican Society of Microbiology (ASM) Microbe conference in Washington D.C., an accepted abstract and poster for competition at theEntomological Society of America (ESA) Annual Joint Meeting in Vancouver, CA, where I won second place. Lastly, an accepted abstract and poster at theGraduate Research and Creativity Expo hosted by UNCG in Greensboro, NC. Some minor target audience interactions during this reporting period have been keeping up with my veterinarian contacts that help regulate antibiotic use in beekeeping, I continue correspondence to keep them up to date on my findings. Lastly, I mentor undergraduate students in our lab, but I also sat on agraduate panel about insect research for an insect apocalypse course.
Changes / Problems
While pursuing aim two, I have realized that there is an inhibitor in the honey bee queen reproductive organs (ovaries and spermathecae) that is inhibiting the amplification of bacteria via PCR. I have spent months trouble shooting with queen organs and have decided to move on to focus my dissertation and this grant on drones. Drones are the least studied honey bee and there is much to be done to understand how antibiotic hive treatments are affecting their development and overall reproductive capacity in honey bee populations. Here are the new aims: 1) Identify how antibiotics impact the reproductive health ofdevelopingdrone honey bees in a controlled environment, 2) Identify how antibiotics impact the reproductive health ofmaturedrone honey bees in a controlled environment, and 3) Determine howcolony levelantibiotic treatments impact queen and drone reproduction.?
Training & Professional Development
I have received training from my PI, Kasie Raymann, on statistical analysis of my sequencing results to determine community dynamics such as but not limited to relative and absolute abundance. I have also been trained by Dr. Brad Metz from NC State on honey bee sperm buffer and staining protocols that included the use of his Cellometer for an already established sperm viability protocol, which I plan to use to determine sperm viability after antibiotic treatment of mature drones. Summer of 2022 I had the opportunity to learn and beekeep with expert beekeepers. This summer I have toured multiple apiaries and government labs (ex. EPA) to learn about their facilities and goals. Professional development has been the conferences and talks already mentioned.?
Dissemination Streams
? Beekeeping community - I continue to attend and participate in the North Carolina State Beekeepers Association (NCSBA) which is the largest state beekeeper's association in the country because of the high population of hobbyist beekeepers in North Carolina. I have given research talks/updates at two county (Henderson & Rowan) beekeeper chapters and will continue to travel NC to share my results. NC beekeepers are getting very interested in my results and have been sharing the information in their networks. Public community - Within this reporting period I was invited to the localT. Gilbert Pearson Auduban Society to talk to the public and since then have been invited to a local gardening group to give a talk. Science community - I have presented my results atAssociation of Southeastern Biologists, Carolina's Society of Environmental Toxicology And Chemistry, American Society of Microbiology (ASM) Microbe, Entomological Society of America (ESA) Annual Joint Meeting, and the Graduate Research and Creativity Expo at UNCG. Furthermore, I have meetings and keep up with multiple other scientists that are working on similar research. I also stay in touch with honey bee veterinarians to keep them up to date on my results and get updates from them on antibiotic regulations. Lastly, my research was featured in the UNCG research magazine:https://researchmagazine.uncg.edu/spring-2023/part-of-the-hive/beyond-the-gut/
Next Reporting Steps
I will continue to do interviews and other avenues to get my research information disseminated as much as possible, which includes the continuation of beekeeper and other community presentations. I will be finalizing the drone in-vitro diet soon and will publish a methods paper for in-vitro rearing of honey bee drones and two other papers on drone reproductive microbiomes. Lastly, I will continue my professional development through UNCG and traveling to work with collaborators. I am currently networking in the Research Triangle Park with BioTech companies, I am planning to stay in agriculture once I graduate in May.? <br><br>
<br>What was accomplished under these goals? I have determined that 1) drone reproductive microbiomes change and stabilize with maturity, 2) drones have essentially no reproductive microbiome when they emerge as adults and accumulate it over time, 3) a single antibiotic treatment reduces overall diversity and abundance in the reproductive organs of mature drones, 4) some of the same bacteria taxa are found in the drone's gut as their reproductive organs, 5) the concentration (200μg/mL) of a single antibiotic treatment kills drone larvae within four days, half of the recommended dose (100μg/mL) killed the drone larvae in seven days, only a quarter of the recommended dose (50μg/mL) survived in comparison with control, 6) drone's exposed to low amounts of oxytetracycline (50μg/mL) are 1-3 days developmentally delayed and eclose as smaller drones with smaller testes, 7) I have established drone in-vitro rearing methods that I will soon publish which will be the first of its kind in decades, this could lead to the ability to in-vitro rear mites which is of #1 importance to bee research right now.? <br><br><b>Publications</b><br>
Target Audience
My main target audience is beekeepers, mostly commercial beekeepers who use oxytetracycline prophylactically to treat their honey bee colonies in the Spring and Fall each year. The North Carolina State Beekeepers Association (NCSBA) is the largest state beekeeper's association in the country because of the high population of hobbiest beekeepers in North Carolina. I was awarded the John T. Ambrose Apiculture Student Researcher Award by the NCSBA on November 20, 2021. In addition to attending the conference for the award ceremony I was also invited to give a research talk to over 400beekeepers and other bee researchers that were in attendance. Since that talk and within this reporting period, I have given research talks/updates at two county (Gaston & Guilford)beekeeperchapters and have recently been invited to two more (Rowan & Wake). NC beekeepers are getting very interested in my results and have been sharing the information in their networks. Although beekeepers are my primary target audience, my research trying to understandantibiotic effects on reproduction can be extrapolated to other insects, pollinators, and animals, including humans. Due to the interest in declining reproductive health across most species, I was invited to give a talk titled, "Antibiotics Induce Infertility from Honey Bees to Humans", by the Greensboro Science Pub (GSP). GSP is a donation funding organization in Greensboro, NC that aims to promotescience literacy and conversation within our community. All of their events are free and located in breweries down town Greensboro that makes science accessible to most. Relaying science to the public usingaccessible language and materials is important to bridge the gap between our communities and scientists, additionally, it is great science communication practice for the scientists. My last target audience is veterinarians.Of all the existing honey bee medications, antibiotics are currently the only regulated substance. These regulations were introduced in 2017 with the FDAs implementation of the Veterinary Feed Directive (VFD) which included bees as livestock.My PI, Kasie Raymann, and myself have kept in close contact with a couple Honey Bee Veterinarians within this reporting period since our assistance in writting a textbook for veterinary schools that was published before this reporting period in January of 2021. Kasie (with my assistance) wrote achapter in, "Honey Bee Microbiota and the Physiology of Antimicrobial Resistance",to providean overview of the honey bee gut microbiome and its importance for bee health and the potential negative consequences of the use of antibiotics in beekeeping, which included some of my results supported by this fellowship.
Changes / Problems
Changes - No major changes have been made to the project narrative at this time. Problems - While pursuing aim two, I have realized that there is some inhibitor in the honey bee queen reproductive organs (ovaries and spermathecae) that is inhibiting the amplification of bacteria via PCR. I have investigated the problem for a month with no resolve, we have currently moved on to focus on finishing aim one and will return to this issue once aim one is complete. If bacterial amplification cannot be conducted in honey bee queen reproductive organs, we will still be able to complete the other questions under aim two such as how much of the antibiotic can reach the reproductive organs after oral ingestion (via mass spectrometry) and sperm viability in treated vs control queens (via cellular staining and fluorescent microscopy).
Training & Professional Development
I have received training from my PI, Kasie Raymann, on statistical analysis of my sequencing resultsto determine community dynamics such as but not limited to relative and absolute abundance. Within the reporting period, I have been trained on a new microscope (Cytation 5) that I will utilize for sperm viability imaging. I have also been trained by Dr. Brad Metz from NC State on honey bee sperm buffer and staining protocols that included the use of his Cellometer for an already established sperm viability protocol. Professional development has been the conferences and talks already mentioned. However, at the NCSBA conference in November 2021, I was introduced to two other bee researchers that developed my knowledge of the USDA in terms of a future career and government options compared to academic positions.
Dissemination Streams
Beekeeping community - The North Carolina State Beekeepers Association (NCSBA) is the largest state beekeeper's association in the country because of the high population of hobbiest beekeepers in North Carolina. I was awarded the John T. Ambrose Apiculture Student Researcher Award by the NCSBA on November 20, 2021. In addition to attending the conference for the award ceremony I was also invited to give a research talk to over 400beekeepers and other bee researchers that were in attendance. Since that talk and within this reporting period, I have given research talks/updates at two county (Gaston & Guilford)beekeeperchapters and have recently been invited to two more (Rowan & Wake). NC beekeepers are getting very interested in my results and have been sharing the information in their networks. Public community - My research trying to understandantibiotic effects on reproduction can be extrapolated to other insects, pollinators, and animals, including humans. Due to the interest in declining reproductive health across most species, I was invited to give a talk titled, "Antibiotics Induce Infertility from Honey Bees to Humans", by the Greensboro Science Pub (GSP). GSP is a donation funding organization in Greensboro, NC that aims to promotescience literacy and conversation within our community. All of their events are free and located in breweries down town Greensboro that makes science accessible to most. Relaying science to the public usingaccessible language and materials is important to bridge the gap between our communities and scientists, additionally, it is great science communication practice for the scientists. The first publication from this funding will be about honey bee drone's reproductive microbiome and is coming soon!
Next Reporting Steps
I was recently (in the current reporting period) interviewed for the UNCG Research Magazine where my research findings and funding source information will be reviewed. The magazine is interested in covering information on my predoctoral fellowship since I am the first student at UNCG to get a USDA predoctoral fellowship, in addition to my unique situation of creating my own dissertation plan instead of being handed a project as a PhD student. I will continue to do interviews and other avenues to get my research information disseminated as much as possible, which includes the continuation of beekeeper presentations. I will be finalizing the drone in-vitro diet soon and will publish a methods paper for in-vitro rearing of honey bee drones. Within the next reporting period, my aim one should be complete, and two to three papers will be in preparation for publication. Lastly, I will continue my professional development through UNCG and traveling to work with collaborators. I recently attended ASM Microbe in June 2022 and presented my research via a poster presentation and am awaiting an acceptance letter from the Entomology Society of America where I hope to present a competition poster in November 2022. <br><br>
<br>What was accomplished under these goals? I have determined that 1) drone reproductive microbiomes change and stabilize with maturity, 2) a single antibiotic treatment reduces overall diversity in the drone's reproductive microbiome and most likely reduces overall bacteria abundance in the reproductive organs, 3) some of the same bacteria taxa are found in the drone's gut as their reproductive organs - we are now working on classifying down to the species level and plan to look at functionality differences even at strain level, and 4) the concentration (200µg/mL) of a single antibiotic treatment kills drone larvae within four days, half of the recommended dose (100µg/mL) killed the drone larvae in seven days, only a quarter of the recommended dose (50µg/mL) survived in comparison with control. <br><br><b>Publications</b><br>