Progress 05/01/17 to 12/31/21
Outputs
Target Audience:Despite the pandemic, during 2021 we succeeded in reaching all of our target audiences via Zoom and other virtual platforms. In terms of colleagues in the scientific community, we published papers derived directly or indirectly from USDA support in high-impact journals, including Annual Review of Entomology and PNAS and disseminated our findings in meetings, conferences, and symposia (including the Entomological Society of America). As an outgrowth of the ESA meeting, Dr. Berenbaum is now collaborating with an international group of scientists in preparing an op-ed recommending revisions to the EPA pesticide approval process to take into account the unique pesticide sensitivities of eusocial insect pollinators. Other professional meetings on Zoom included the USDA ARS Grand Challenge Workshop: Creating Pollinator Landscapes and Beekeeping Practies for a Changing Climtae, May 17, 2021; USDA/EPA Pollinator Workshop, SM OEC Strategic Pollinator Report, September 13-14, 2021; Pollinators and Pesticides: Chemical Roulette, Entomological Society of America, October 13, 2021. Our outreach activities were limited during 2021 due to pandemic social-distancing requirements, but we were able to connect virtually with many of our audiences, including beekeepers . Outreach efforts in 2021 relevant to this project include: Scientists Decry Death by 1,000 Cuts for World's Insects, January 11, 2021 (NBC News); Daniel Cross, The Media Will Need to Heed the Plight of the Pollinators, Sustainability Times January 21, 2021; Dave Dahl from WTAX on bee decline in February 2021; 2021: Interview with Travis Andrews, Washington Post, TikTok Viral Beekeeper Erika Thompson is Getting a Lot of Buzz, March 20, 2021; and the CU Illini After-5 Rotary, Are Pollinators Feeling the Sting of Climate Change? November 11, 2021. We have recently resumed in-person activities on a scaled-down basis at the University of Illinois with programming at the University of Illinois Pollinatarium, including a hands-on socially distanced 4-H class on beekeeping. Changes/Problems:The departure of co-PI Bernarda Calla for a position at the USDA ARS laboratory in Corvallis, Oregonslowed down progress in preparing and submitting manuscripts, but we have made considerable progress via long-distance electronic communication. What opportunities for training and professional development has the project provided?During 2021, this project provided training and professional development for Bernarda Calla, a former postdoctoral student and now research scientist, who expanded her genomic expertise beyond Lepidoptera to acquire familiarity with bees. Postdoctoral student and now research scientist Ling-Hsiu Liao expanded her experience with honey bee behavior to encompass honey bee physiology, biochemistry, toxicology, and pathology. Will Montag completed his master's thesis on small hive beetles. gaining experience not only with cultivating and identifying fungal species in beehives but also with using molecular techniques for fungus identification. Daniel Bush expanded his knowledge of honey bee research techniques as part of his doctoral work examining adaptations of a distinct strain of the fungus Aspergillus flavus to bee bread in beehives. In addition, Wen-Yen Wu began his second year in the Entomology doctoral program at the University of Illinois to work on this project in August 2021, focusing on chemical characterization of honey and effects of phytochemicals and pesticides on longevity. How have the results been disseminated to communities of interest?Results of our work have been disseminated to the academic community in scholarly journals and book chapters (see publications) and in meeting presentations (e.g., Entomological Society of America, poster and platform presentations every year of the project to date). From May 2020 through April 2021, our communications with academic colleagues were limited by the COVID-19 pandemic and most of our activities shifted to virtual formats. In 2021, Dr. Berenbaum delivered a seminar on bee health for the Department of Entomology at the University of Maryland in March 2021. A presentation on "Environmental Effects on Plants and Insect Pollinators" May 17, 2021, was delivered during the virtual USDA ARS Grand Challenge Workshop: Creating Pollinator Landscapes and Beekeeping Practices for a Changing Climate." Dr. Berenbaum also participated in a USDA/EPA Pollinator workshop, SM OEC Strategic Pollinator Report, September 13-14, 2021, and, on October 13, 2021, Dr. Berenbaum gave a talk on "Pollinators and Pesticides: Chemical Roulette" in a program symposium at the Entomological Society of America annual meeting and a talk, "Honey -- A New Look at the World's Processed Food," for the Horizons in Bee Research webinar series, Solatina (Latin American Society for Bee Research), November 3, 2021. In terms of outreach, Dr. Berenbaum also answered bee-related queries from multiple media outlets, including the Associated Press, "Scientists Decry Death by 1,000 Cuts for World's Insects," January 11, 2021 (NBC News), Daniel Cross, "The Media Will Need to Heed the Plight of the Pollinators," Sustainability Times January 21, 2021, Dave Dahl from WTAX on bee decline in February 2021 and Travis Andrews of the Washington Post on a "TikTok Viral Beekeeper" in March 2021 and CU-Illini After 5 Rotary, "Are Pollinators Feeling the Sting of Climate Change?", November 11, 2021. Our Annual Review of Entomology (Berenbaum and Calla 2021) was covered by bbc.com, inverse.com, and Knowable Magazine (https://knowablemagazine.org/article/living-world/2021/bee-gold-honey-superfood ) and Smithsonianmag.com. A note in Nature (2021) coauthored by Dr. Berenbaum and Dr. Liao was featured by Am. Bee J. https://geneticliteracyproject.org/2022/03/23/viewpoint-how-do-pesticides-impact-bees-influential-university-of-illinois-entomologist-mae-berenbaum-callsfor-pesticide-approvals-to-include-pre-release-real-world-monitoring/ . Dr. Berenbaum was also invited to contribute a paper to American Bee Journal, titled, "Bee Suits and Law Suits: A Sad History of Bees and Pesticides in U.S. Agriculture" to appear in March of 2022. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
With respect to all five of our objectives, to simulate variable honey composition, we conducted assays in which adult workers consumed monofloral honeys -- Nyssa ogeche (tupelo), Robinia pseudoacacia (black locust), and Fagpyrum esculentum (buckwheat) -- individually and in combination and evaluated phytochemical content and antioxidant capacity in relation to adult worker gene expression, survival, and longevity with and without topically applied bifenthrin. Enhanced insecticide tolerance was associated with honey phytochemical diversity; buckwheat honey possessed the highest phytochemical diversity and antioxidant capacity, supported the highest bifenthrin LD50 , and produced the greatest number of differentially expressed transcripts compared with bees on a phytochemical-free sugar diet. RNA-Seq analysis revealed five types of events in guts of workers consuming tupelo or buckwheat honey compared to a sugar diet: Increased expression of oxidoreductase molecular function pathways (including CYP6AS and CYP9Q genes that metabolize natural and synthetic toxins) and reduced expression of pathways of carbohydrate metabolism, protein degradation, ion and protein transport, and neurotransmission, suggestive of a trade-off between metabolizing toxins to reduce mortality and metabolizing sugars for acquiring energy. That phytochemical mixtures in honey collectively induce detoxification genes and pathways may be a critical component of bee survival and resilience in environments contaminated with synthetic insecticides.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Althaus, S., M.R. Berenbaum, J. Jordan and D.A. Shalmon. 2021. No buzz for bees: Media coverage for bees. Proc. Natl. Acad. Sci. USA, 118 (2) e2002552117; https://doi.org/10.1073/pnas.2002552117.
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Berenbaum, M.R., Bush, D.S. and Liao, L.H. 2021. Cytochrome P450-mediated mycotoxin metabolism by plant-feeding insects. Current Opinion in Insect Science, 43: 85-91.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2021
Citation:
Montag W., L.-H. Liao and M.R. Berenbaum. 2021. Sexually dimorphic responses to monofloral honeys in the small hive beetle Aethina tumida. J Apicul. Review (Under Revision).
|
Progress 05/01/20 to 04/30/21
Outputs
Target Audience:Despite the pandemic, during 2020-2021 we succeeded in reaching all of our target audiences via Zoom and other virtual platforms. In terms of colleagues in the scientific community, we published papers derived directly or indirectly from USDA support in high-impact journals, including Annual Review of Entomology (journal impact factor 13.8) and PNAS (JIF 9.4), and disseminated our findings in meetings, conferences, and symposia (including the Entomological Society of America). As an outgrowth of the ESA meeting, Berenbaum is now collaborating with an international group of scientists in preparing an op-ed recommending revisions to the EPA pesticide approval process to take into account the unique pesticide sensitivities of eusocial insect pollinators. Our outreach activities were limited during 2020-2021 due to pandemic social-distancing requirements, but we were able to connect virtually with many of our audiences, including beekeepers (via, e.g., the virtual Garfield Park Beekeeping Forum) and interested members of the general public (e.g. Illinois Master Gardeners). We have recently resumed in-person activities on a scaled-down basis at UIUC with programming at the UI Pollinatarium, including an upcoming hands-on socially distanced 4-H class on beekeeping. Moreover, the COVID-19 pandemic provided an opportunity for research scientist Liao to expand our outreach to beekeepers and interested members of the public in Taiwan, where she was able to present three invited talks: Invited Talkto beekeepers and public, 2020, Miaoli District Agricultural Research and Extension Station, Council of Agriculture, Taiwan. (29June) Talk Title: "Mechanisms of Colony Resistance to Environmental Toxicants in the Honey Bee"(In Mandarin) Invited Talkto students and professors, 2020, Entomology Department,National Chung Hsing University, Taiwan. (20 May) Talk Title: "From Me to Us: Mechanisms of Colony Resistance to Environmental Xenobiotics in the Honey Bee, Apis mellifera"(In Mandarin) InvitedTalkto students and professors, 2020, Biology Department, Tunghai University, Taiwan (15 April) Talk Title: "Mechanisms of Colony Resistance to Environmental Xenobiotics in the Honey Bee,Apis mellifera" Changes/Problems:We did not anticipate the extent to which seasonal variation would affect honey bee physiological and toxicological responses to natural and synthetic chemicals and our ability to replicate this work was complicated by a project scientist's continuing visa issues and exacerbated by the COVID-19 lockdown, which effectively prevented us from carrying out any experiments during the 2020 field season. We made progress in winter 2020-2021 with the use of caged "winter bees" housed in the UI Bee Research Facility and have recently succeeded in ramping up field activities in Spring-Summer 2021 What opportunities for training and professional development has the project provided?During 2020-2021, this project provided training and professional development for Bernarda Calla, a former postdoctoral student and now research scientist, who expanded her genomic expertise beyond Lepidoptera to acquire familiarity with bees. Postdoctoral student and now research scientist Ling-Hsiu Liao expanded her experience with honey bee behavior to encompass honey bee physiology, biochemistry, toxicology, and pathology. Graduate student Daniel Pearlstein gained expertise in honey bee physiology, anatomy, and behavior, along with beekeeping techniques, culminating in completion of his master's thesis. Will Montag completed his master's thesis on small hive beetles. gaining in the process gaining experience not only with cultivating and identifying fungal species in beehives but also with using molecular techniques for fungus identification. Daniel Bush expanded his knowledge of honey bee research techniques as part of his doctoral work examining adaptations of a distinct strain of the fungus Aspergillus flavus to beebread in beehives.In addition, Wen-Yen Wu began his doctoral studies at UIUC to work on this project in August 2020, focusing on chemical characterization of honey and effects of phytochemicals and pesticides on longevity. How have the results been disseminated to communities of interest?Results of our work have been disseminated to the academic community in scholarly journals and book chapters (see publications) and in meeting presentations (e.g., Entomological Society of America, poster and platform presentations every year of the project to date). From May 2020 through April 2021, our communications with academic colleagues were limited by the COVID-19 pandemic and most of our activities shifted to virtual formats. Berenbaum gave a plenary address for the Virtual Annual Meeting of the North Carolina Entomological Society and the Sociedad Colombiana de Entomologica in October 2020 as well as a seminar on our bee research for the Texas A&M Aggie Women in Entomology the same month. Liao, Wu, and Berenbaum presented a talk on the effects of fungicides on honey bee flight performance at the November 2020 Entomological Society of America annual (virtual) meetingand Berenbaum delivered a seminar on bee health for the Department of Entomology at the University of Maryland in March 2021. Also worthy of mention is that our USDA-AFRI funded research was featured in Journal of Experimental Biology in February 2020 in an article by G. Manteca titled, "Honeybees on treadmills uncover natural remedy for fungicide effects" (J Exp Bio 223: jeb211474). In terms of outreach to the public (similarly limited by COVID-19 to online events), Berenbaum presented a talk on applied bee science at the Garfield Park Conservatory Bee Forum in May 2020, on the pollinator-pocalypse for Illinois Extension in June 2020, and on bee health as the featured speaker at the Illinois Master Gardener Virtual Awards Ceremony and Education in October 2020, Berenbaum also answered bee-related queries from multiple media outlets, including the Associated Press, "Scientists decry death by 1,000 cuts for world's insects," January 11, 2021 (NBC News), Daniel Cross, "The media will need to heed the plight of the pollinators," Sustainability Times January 21, 2021, Dave Dahl from WTAX on bee decline in February 2021 and Travis Andrews of the Washington Post on a "TikTok viral beekeeper" in March 2021. What do you plan to do during the next reporting period to accomplish the goals?To achieve all of our objectives in full, we have several experiments (e.g., RNA-Seq) that require additional replication (due to seasonal variation in honey bee physiology and behavior) as well as experiments to expand on preliminary work. Work on Objectives 4 and 5 remain to be consolidated and incorporated into a manuscript.
Impacts
What was accomplished under these goals?
During 2020-2021, we encountered multiple unexpected obstacles to conducting several experiments the results of which were necessary for finishing the project. We did not anticipate the extent to which seasonal variation would affect honey bee physiological and toxicological responses to natural and synthetic chemicals and our ability to replicate this work was complicated by a project scientist's continuing visa issues and exacerbated by the COVID-19 lockdown, which effectively prevented us from carrying out any experiments during the 2020 field season. We made progress in winter 2020-2021 with the use of caged "winter bees" housed in the UI Bee Research Facility and have recently succeeded in ramping up field activities in Spring-Summer 2021.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Berenbaum, M.R., D.S. Bush and L-H Liao, 2021. Cytochrome P450-mediated mycotoxin metabolism by plant-feeding insects. Current Opinion in Insect Science 43: 85-91.
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Berenbaum, M.R. and B. Calla, 2020. Honey as a functional food for honey bees. Annual Review of Entomology 66: https://doi.org/10.1146/annurev-ento-040320-074933.
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Liao, L.-H., D.J. Pearlstein, W.-Y. Wu, A. Kelley, W. Montag, E.M. Hsieh and M.R. Berenbaum. 2020. Increase in longevity and amelioration of pesticide toxicity by natural levels of dietary phytochemicals in the honey bee, Apis mellifera. PLoS ONE 15(12): e0243364. https://doi.org/10.1371/journal.pone.0243364.
|
Progress 05/01/19 to 04/30/20
Outputs
Target Audience:To share information with scientific peers, we have published our findings in peer-reviewed scientific journals and disseminated our findings in meetings, conferences, and symposia (including the Entomological Society of America and the American Bee Research Conference). Berenbaum also engaged in a range of outreach activites that, in 2019-2020, included "Networks of pollinators", Phil Theta Kappa International Honor Society, Parkland College, June 1, 2019; Domesticated insects, Human-Animal Institute, Levis Center, UIUC Urbana, IL, July 16, 2019; "Honey bee health and female empowerment", Women's Business Council, Illinois Terminal, Champaign, IL, August 22, 2019; "Fearing the 'Insect Apocalypse'? Renowned entomologist says 'Get rid of your lawn'", interview with Alex Ruppenthal, WTTW News, October 2, 2019; "Bye Bye Bugs", interview with Seth Borenstein, PCT Magazine, October 4, 2019; "Asian giant hornet invasion threatens honey bees in Pacific Northwest" interview with Neil Vigdor, New York Times, December 24, 2019; "HoneyLove", Los Angeles CA, January 18, 2020; "Straight talk on bees, silkworms, and human survival", Interview with David Templeton, Pittsburgh Post-Gazettte, February 4, 2020; "Bumblebees are dying across North America and Europe as the climate warms", Interview with Chris Mooney, Washington Post, February 6, 2020; "What is a Murder Hornet?", Interview with Aila Slisco, Newsweek.com, May 4, 2020; "Murder hornets", Stevie Jay ESPN 93.5 AM, May 5, 2020; "Can CRISPR/AI/IoT/IPM or any other acronym save the honey bees, or Why applied bee science needs basic bee science", Beekeeping Forum, Garfield Park Conservatory, Chicago, IL, May 24, 2020 (Zoom). At UIUC, we continued programming at the UI Pollinatarium, hosting ca. 2,000-3,000 visitors ranging from schoolchildren to seniors; Illinois Master Naturalists Annual meeting (Zoom), June 17. Changes/Problems:We did not anticipate the extent to which seasonal variation would affect honey bee physiological and toxicological responses to natural and synthetic dietary chemicals and our ability to replicate this work was complicated in 2019-2020 by visa issues experienced by a project scientist and exacerbated by the COVID-19 lockdown, which effectively prevented us from carrying out any experiments during the current field season. What opportunities for training and professional development has the project provided?This project provided training and professional development for Bernarda Calla, a former postdoctoral student and now research scientist, who expanded her genomic expertise beyond Lepidoptera to acquire familiarity with bees; as well postdoctoral student and now research scientist Ling-Hsiu Liao expanded her experience with honey bee behavior to encompass honey bee physiology, biochemistry, toxicology and pathology. Two graduate students learned new skills as well, with Michael Wong becoming familiar with bioassay and toxicology and Will Montag learning rearing techniques for small hive beetle and gaining experience with cultivating and identifying fungal species found in beehives. Liao and Calla also learned and put into practice public engagement skills by developing a board game about bees and pollination for Urbana Middle School's Students Playing and Learning After School Hours program. How have the results been disseminated to communities of interest?Results of our work have been disseminated to the academic community in scholarly journals and book chapters (see publications) and in meeting presentations (e.g., Entomological Society of America, poster and platform presentations every year of the project to date, Society of Toxicologic Pathology, Sigma Xi, Duquesne University, HoneyLove (Los Angeles CA)).Outreach to the beekeeping community was carried out through presentations in person (American Bee Research Conference, UIUC Beekeeping Short Course) and online (Garfield Park Conservatory Beekeeping Forum). We also including summaries of our research in a number of public outreach events (e.g., Peggy Notebaert Museum, North American Pollinator Protection Campaign, Women's Business Council of Champaign, Osher Lifelong Learners, Phi Theta Kappa, National Academy of Sciences World of Genomics public event, Pollen Power campus event). What do you plan to do during the next reporting period to accomplish the goals?To achieve all of our objectives in full, we have several experiments that require additional replication (due to seasonal variation in honey bee physiology and behavior) as well as experiments to expand on preliminary work. Work on Objectives 4 and 5 remain to be consolidated and incorporated into a manuscript: with respect to Objectives 1-3: Obj. 1: Determine whether there is differential transcription of genes involved in detoxification, immunity, and longevity according to honey chemistry; this RNA-Seq analysis would be strengthened by additional replication, due to seasonal variation in the initial work. Obj. 2: Determine whether detoxification of pesticides (and hence toxicity of pesticides) varies with honey chemistry; although experimentshave been carried out with phytochemical components, bioassays with different monoflorals remain to be completed. Obj. 3: Assess whether defense against fungal pathogens varies with honey chemistry; we would like to expand this work beyond our initial work with A. flavus. Obj. 4: Determine whether efficacy of defense against oxidative stress varies with honey chemistry: and Obj. 5. Determine whether worker longevity varies with honey chemistry (particularly in accordance with antioxidant activity).
Impacts
What was accomplished under these goals?
Obj. 1: To quantify differences in gene expression according to honey chemistry, day-old adult bees were assigned to five diets; each of three diets contained one of three monofloral honeys: tupelo. buckwheat, black locust, with a mixed treatment and sugar control. Guts of bees consuming locust honey had the fewest transcripts changing in abundance (threegenes down-regulated vs control). Buckwheat honey changed the transcription of the highest number of genes when compared to sugar. The highest statistically represented term was 'xenobiotic metabolism' biological process in the genes upregulated by buckwheat, tupelo, and the mix of three honeys compared to the sugar control. Several other oxido-reduction, detoxification- and secondary metabolism-related terms were present in these comparisons. The honey mix also showed enrichment for ATPase activity and transmembrane transporting activity. For Obj. 2: to determine whether pesticide toxicity varies with honey chemistry, we conducted longevity assays with adult bees on diets with bifenthrin, chloranthraniliprole, or imidacloprid in the presence and absence of p-coumaric acid and quercetin and demonstrated amelioration of toxicity of bifenthrin and chloranthraniliprole. For Obj. 3, to assess whether defense against fungal pathogens varies with honey chemistry, we focused on Aspergillus flavus (stonebrood), measuring tolerance of a strain found in beehives and a freeliving strain to survive under physicochemical hive conditions and tmetabolize hive xenobiotics (e.g., propolis, low pH, high osmotic pressure). In all cases. A. flavus from hives displayed superior tolerance to hive biotic and abiotic stress factors. We will continue this work, examining fungal detoxification of specific honey and propolis phytochemicals. For Obj. 4 and 5, to determine if antioxidant defense varies with chemistry, we characterized honey phenolic acids and flavonoids from buckwheat, tupelo and black locust honey by HPL-MS, identifying dramatic differences in the phytochemical content and composition of the three monoflorals and measured antioxidant capacity of seven monofloral honeys (buckwheat, tupelo, locust, almond, carrot, goldenrod and soy honeys) using relative DPPH radical scavenging capacity and oxygen radical absorbance capacity assays and found that antioxidant capacity of buckwheat honey is highest, followed by antioxidant capacity of almond honey and carrot honey, soy and locust honey (both of which are legumes) have the lowest antioxidant capacity.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Liao, L.-H., W.-Y. Wu, A. Dad and M.R. Berenbaum. 2019. Fungicide suppression of flight performance in the honeybee (Apis mellifera) and its amelioration by quercetin. Proc. Roy. Soc. B 286: https://doi.org/10.1098/rspb.2019.2041.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Berenbaum, M.R. and Liao, L.H. 2019. Honey bees and environmental stresstoxicologic pathology of a superorganism. Toxicologic Pathology 47(8): 1076-1081.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2020
Citation:
Berenbaum, M.R. and B. Calla, 2020. Honey as a functional food for honey bees. Annual Review of Entomology 66: (In Press).
- Type:
Journal Articles
Status:
Under Review
Year Published:
2020
Citation:
Liao, L.-H., D.J. Pearlstein, W.-Y. Wu, A. Kelley, W. Montag, E.M. Hsieh and M.R. Berenbaum. 2020. Optimal range of concentration for amelioration of xenobiotic toxicity by honey phytochemicals in Apis mellifera L. PLoS ONE.
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Berenbaum, M.R. 2019. Catching ZZZZs. American Entomologist 65: 220-223.
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Progress 05/01/18 to 04/30/19
Outputs
Target Audience:Target audiences include the scientific community (entomologists, toxicologists, geneticists, phytochemists) and the apiculture community. Changes/Problems:We are also examining the growth of yeast cultures from the small hive beetle, Aethina tumida (Murray), on various culture media (phytonic, cornmeal, oatmeal/salt, starch, and corneal-glucose-peptone) in preparation for isolating cultures of the fungus Kodamaea ohmeri for comparing effects of monofloral honeys on growth of this fungus. These studies are ongoing. In a complementary series of assays, we have examined the feeding responses of Ae. tumida adults to 50% (w/w) water dilutions of white tupelo, locust, buckwheat honey, and a phytochemical-free sugar-based control diet over 24 or 72-hour intervals. Our preliminary data indicate that three factors - gender (Fisher exact test, p=0.04<0.05), hive of origin (p=0.02<0.05), and duration of assay (p=0.008<0.01) - affected orientation to the diet. Small sample size prevented us from detecting significant differences among treatments within the 72-hour interval assay, but trends suggest that males and females may differ in diet preferences. Whereas half of males tested displayed no responses to diet, the remainder selected either tupelo (33%) or locust honey (17%). In contrast, 43% of females exhibited a preference for buckwheat honey (43%), with only 23% displaying a preference for either tupelo or locust. Strikingly, no beetles displayed a preference for the phytochemical-free sugar control diet, suggesting that substances other than sugars may play a role in food-finding and diet selection in this species. These assays will be repeated to increase statistical power in summer 2019, as more adult beetles become available. What opportunities for training and professional development has the project provided?Three graduate students (William Montag, Daniel Bush, and Daniel Pearlstein) have gained experience designing and conducting behavioral and physiological experiments with honey bees. Postdoctoral associate Bernarda Calla and Ling-Hsiu Liao were involved with the RNA-Seq component of the project. How have the results been disseminated to communities of interest?In 2017-2018, the following activities involved dissemination to the scientific community: Symposium in honor of Rachel Galun, Israeli Entomological Society, Volcani Center, Rishon LeTsiyon, Israel, October 18, 2017; "Genomic footprint of eusociality in bees: Floral foods and CYPome blooms", PBT Section Symposium: "Protecting bee pollinators with comparative toxicology and functional genomics", Entomological Society of America annual meeting, Denver, CO, November 5, 2017; Annual meeting, Chicago American Chemical Society, College of DuPage, Glen Ellyn, IL, November 2017; "Bees and pesticides" Heartland Apicultural Society, St. Louis, MO (also two panels: Pesticides in Agriculture and Future Technologies for Beekeeping), Washington University, July 12-14, 2018. Presentations for the general public include: Interview on neonicotinoid metabolism Catherine Offord, New Scientist, March 20, 2018; "Bees and pesticides - Old and new", Beekeeping Short Course, Urbana, IL, April 7, 2018; Interview with Neha Jain, GotScience blog, February 12, 2018; Interview, state of federal funding for agricultural research, Madelyn Beck, Tri-States Public Radio, Galesburg, IL, March 29, 2017; "Pets, not pests - Arthropods as companion animals", Second Annual Human-Animal Studies Summer Institute, University of Illinois, Urbana, IL, July 10, 2018; "Fungicide effects on bees," Heartland Apicultural Society, Washington University, St. Louis, MO, July 12, 2018; "That time I met President Obama," Pollen Power Camp, July 26, 2018; Interview with Sean, Apocalypse Now This, for documentary on the beepocalypse, July 25, 2018; "Bees and bourbon," Morton Arboretum, Lisle, IL, August 7, 2018; "Honey, I'm spirited: Honey and alcoholic beverages", Arcadia Café; Lumen Events, February 23, 2019; "The mystery of the disappearing bees - A case for the genome detectives",World of Genomics, National Academy of Sciences, April 13, 2019. What do you plan to do during the next reporting period to accomplish the goals?We plan to completeObjective One [RNASeq analysis], continue progress in evaluating monofloral honey suitability for hive fungi and for small hive beetle preference and performance, anddisseminate our results.
Impacts
What was accomplished under these goals?
Objective 1. Determine if there is differential transcription of genes involved in detoxification, immunity, and longevity with honey chemistry. One-day-old adult bees from a single healthy hive were assigned to five different diet treatments. All diets contained casein as a phytochemical-free protein source and were variously amended. Each of three diets contained one of the monofloral honeys being compared: tupelo (TUP), buckwheat (BUC), and black locust (LOC). In the fourth treatment, bees were allowed to choose freely among the three honeys (TLB). The final treatment comprised a diet representing the phytochemical-free "sugar control" (SUC), which was made up of 40% fructose:29% glucose:1% sucrose in water. This mixture approximates the average proportions of sugars found in the three types of honey compared. (White and Doner 1980; Pasini et al. 2013; Gardiner 2015). Buckwheat honey alone caused a reduction in lifespan compared to the sugar control. Bees consuming diet with buckwheat honey alone had six times the risk of death (hazard ratio, HR=6.16, 95% CI=1.8-21.2, P<0.01, Cox proportional hazards regression) as those consuming the sugar control diet. Bees from the TLB diet experienced the highest average survival time among all groups (mean= 13.81 ± 0.20 days). Total RNA was extracted from midguts dissected from workers collected after two weeks of feeding on the different diets and then sent for sequencing as 15 samples: 3 samples from each of the 5 treatments, where each sample was a pool of RNA from 3 individual midguts within a treatment. Each RNA sample was processed into a barcoded sequencing library and the 15 libraries were sequenced in one lane of Illumina HiSeq 4000 with 100 base pairs single-end reads. The sequencing resulted in 20 million reads per library, with most libraries having more than 25 million reads. Adapters were removed at the sequencing center, resulting in reads with average Phred-score quality > 30 along the whole read. The files containing FASTQ-formatted files were retrieved and transferred to our local server. Additional trimming was done to remove reads shorter than 80 bases and for which Phred scores were below 35 at the beginning or end of each read using Trimmomatic (Bolger et al. 2014). The reads were then mapped to the most recent version of the Apis mellifera genome assembly (v. 4.5) using STAR aligner (Dobin et al. 2013). Reads were quantitated using RSEM (Li and Dewey 2011) and differential expression was determined with the EdgeR Bioconductor package (Robinson et al. 2010). In total, 339 genes were differentially expressed at a fold-change >2 between two treatments (p < 0.001).Bees on locust honey had the fewest transcript changes (3 genes down-regulated vs sugar treatment and no up-regulated transcripts). Buckwheat honey changed transcription of the greatest number of genes vs sugar. Differentially expressed genes were annotated using the NCBI nr (non-redundant), Swissprot-uniprot and PFAM public databases. Gene ontology (GO) annotations were derived from matching Swissprot records and Pfam domain content. A GO term enrichment test was carried out with GOseq (Young et al. 2010). The highest statistically represented term was 'xenobiotic metabolism' biological (GO:0006805) process in the genes upregulated by buckwheat, tupelo, and the three-honey mix vs the sugar control (FDR-corrected p-value <0.05). Several other oxido-reduction, detoxification- and secondary metabolism-related terms were consistently present in these comparisons. The honey mix also was enriched for ATPase activity and transmembrane transporting activity. No GO terms were enriched in genes downregulated by buckwheat honey, locust honey, or the mix at our set cut-off FDR p-value for Fisher's exact test. Tupelo honey down-regulated genes were enriched for gene categories in hydrolysis. Objective 2. Measure effects of honey phytochemicals on pesticide toxicity. We performed bifenthrin LD50 assays for honey bees fed diets comprising the five treatments described earlier. One-day-old bees from three hives consumed diets in the five treatments for three days and then underwent topical treatments with 1 µl acetone containing bifenthrin at 0 ppb, 120 ppb, 150 ppb, 240 ppb, 300 ppb, 600 ppb, 1200 ppb, 1500 ppb, 2400 ppb, and 3000 ppb. LD50 values in bees on each of honey diets were higher than those for bees on the phytochemical-free diet at 24 and 48 hours. The LD50 value for bifenthrin was highest on buckwheat honey diet (1590.0 ppb) and was greater than the LD50 value for bees on the phytochemical-free sugar diet (SUC vs BUC, parallelism test p=0.01<0.05; relative median potency=0.75 (0.60-0.93, 95% CL). Hive 1 bees were most sensitive to bifenthrin, but diet had no effect on bifenthrin LD50 . In contrast, diet affected bifenthrin LD50 for Hive 2 bees (SUC vs BUC), and Hive 3 bees (SUC vs BUC and SUC vs TLB). Overall, buckwheat honey appears to confer some protection against bifenthrin, as the sole dietary honey or as one of a choice of honeys. Objective 3. Determine differences in fungistatic/fungitoxic properties of different honeys. To determine if As. flavus displays adaptation to hive conditions, growth of two strains - one from beebread (AFBB) and one from field environments (AF36) - was compared in the presence of propolis, low pH, and high osmotic pressure. Propolis retarded A. flavus growth, but AFBB grew ~35% faster over five days than AF36. AFBB also displayed faster growth at low pH. Finally, AFBB exhibited greater growth at low matric potential than did AF36, which did not produce hyphae at -60 MPa. Objective 4. Assess antioxidant capacity of different honeys. In addition to buckwheat, tupelo, and locust honey, we measured antioxidant capacity of almond, carrot, goldenrod and soy honeys using the DPPH radical scavenging capacity assay (Cheng et al. 2006) and found that antioxidant capacity of buckwheat honey is highest, followed by almond honey and carrot honey. Antioxidant capacity of goldenrod honey was comparable to that of tupelo honey and soy and locust honey have the lowest antioxidant capacity. Objective 5. Determine impacts of honey on adult bee lifespan. Because carbohydrate composition of the diet can elicit differential gene expression (Wheeler and Robinson 2014) and thus affect longevity, we controlled for carbohydrate variation in summer 2018 by extracting phytochemicals (Gheldof et. al. 2002) from the three monofloral honeys tested and adding each phytochemical extract into a phytochemical-free sugar control diet. An additional sucrose diet (70% sucrose water (w/v)) was included to simulate supplementary feeding practices and we also included the TLB choice diet. All experimental diets contained casein at 1:12 protein:carbohydrate. Three replicates of each treatment from each of three colonies from apiaries maintained by UIUC were used for this experiment, for a total of 9 replicated cages of 6 treatments. Each cage contained 30 one-day-old bees, as well as artificial queen mandibular gland pheromone strip to simulate the pheromonal attributes of a queenright colony. Effects of treatment factors on bee survival were analyzed using the Cox proportional hazards model and survival curves for each treatment group were obtained through the Kaplan-Meier estimator, with differences between the curves compared using the Tarone-Ware test. Bees consuming a diet self-selected among the three diets containing honey extracts exhibited the highest survivorship, with a 23% reduced risk. Surprisingly, bees on diets containing only sucrose exhibited higher survivorship than bees consuming a phytochemical-free sugar "basic" diet (mainly monosaccharides), suggesting that both phytochemical content and carbohydrate composition influence longevity.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2019
Citation:
Wong, M.J., L.H. Liao and M.R. Berenbaum. 2018. Biphasic concentration-dependent interaction between imidacloprid and dietary phytochemicals in honey bees (Apis mellifera). PLoS ONE. 13(11): e0206625.
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Progress 05/01/17 to 04/30/18
Outputs
Target Audience:Target audiences include the scientific community (entomologists, toxicologists, geneticists, phytochemists) and the apiculture community. Changes/Problems:Due to concerns about risk to other ongoing bee research projects posed by culturing chalkbrood, Ascophaera apis, we are working to secure space in a separate Biosafety 2 room on campus. Once space is secured, we will begin culturing chalkbrood in order to conduct fungal growth assays for this project in the upcoming year. As a side-project to the fungal growth assays, we have taken advantage of the monofloral honey resources already in place to test the survival response of the small hive beetle, Aethina tumida (Murray), to monofloral honey diets. A preliminary assay using white tupelo, Christmasberry, and multifloral honey has just been completed, with replicate assays to take place soon. All diets supported production of viable offspring and completion of a full lifecycle. We also intend to test the preferences of small hive beetle among monofloral honeys using in-hive trapping, with captured live specimenswhich will then be used for continuing diet assays. What opportunities for training and professional development has the project provided?Five graduate students (Ling-Hsiu Liao, William Montag, Michael Wong, Edward Hsieh, and Daniel Pearlstein) have gained experience designing and conducting behavioral and physiological experiments with honey bees. Liao completed a doctoral dissertation and has published two of six chapters; the remaining four will be submitted for publication soon. Wong completed his MS thesis and is preparing his thesis on effects of honey phytochemicals on imidacloprid toxicity for a submission to PLoS ONE (June 2018). Postdoctoral associate Bernarda Calla has been introduced to working with honey bees and is involved with the RNA-Seq component in the project; academic hourly Wen Yen Wu developed new approaches to separating and identifying phytochemicals in mixtures and assaying antioxidant activity. How have the results been disseminated to communities of interest?In 2017-2018, the following activities involved dissemination to the scientific community: Annual Research Day, Environmental and Molecular Toxicology Program Oregon State University, "Buzz-kill-can the honey bee survive the 21st century?" January 19, 2017; "Plenary Address, "Can Science Save the Honey Bees?" Annual meeting, American Association for the Advancement of Science, Boston, MA. February 18, 2017; American Chemical Society AGRO webinar "How to eat a plant--bees vs butterflies", March 16, 2017; "Can the honey bee survive the 21st century?" Dept. Biology, California State University-Northridge, May 5, 2017; Symposium in honor of Rachel Galun, Israeli Entomological Society, Volcani Center, Rishon LeTsiyon, Israel, October 18, 2017; Genomic footprint of eusociality in bees: floral foods and CYPome "blooms", PBT Section Symposium: Protecting bee pollinators with comparative toxicology and functional genomics", Entomological Society of America annual meeting, Denver, CO, November 5, 2017; Keynote, "Honey bees as chemists," Annual meeting, Chicago American Chemical Society, College of DuPage, Glen Ellyn, IL; "Genomics of dinner (if you're a vegetarian", Dinner keynote, Arthropod Genomics Symposium, Urbana, IL, June 8, 2018 (scheduled) In 2017-2018, the following activities involved dissemination to the public: Ask Me Anything Reddit--honeybees, February 14, 2017 (with Gene Robinson); "Bee-reft--why pollinator decline matters", St. Louis Zoo Lecture Series, St. Louis, MO, March 22, 2017; Honey bee health update-2017", Beekeeping Workshop, McLean County Extension, April 15, 2017; "Human impacts on insect affairs--turnabout is not fair play," Jane and Whitney Harris Lecture, Missouri Botanical Garden, St. Louis, MO How I became an entomologist", Pollen Power camp, Institute for Genomic Biology, Urbana, IL, July 11, 2017; "Colony collapse disorder follow-up", Interview with Alex Bjerga, Bloomberg News (Time Magazine), August 1, 2017; Jessica Kunz WCIA-TV, "Researchers helping bee populations" August 9, 2017; "Authors and appetites," Luncheon speaker, PEO, Urbana, IL September 23, 2017; "Pollinators in danger", Champaign Rotary West, Round Barn Center, Champaign, IL, September 27, 2017; Seth Borenstein, AP News, bumble bees and climate change," October 1, 2017; Interview, Cici Zhang, Scholastic Magazine, October 25, 2017; Science Fiction vs Science Fact panel Science Entertainment Exchange, Austin Film Festival, Austin, TX, October 27, 2017; Monofloral honeys, Power of the Alumni annual conference research presentation, Champaign, IL, February 11, 2018; Interview on neonicotinoid metabolism Catherine Offord, New Scientist, March 20, 2018; NPR interview on agricultural science funding, March 30, 2018; Interview, state of federal funding for agricultural research, Madelyn Beck, Tri-States Public Radio, Galesburg, IL, March 29, 2017; "Bees and pesticides, old and new," Beekeeping Short Course, Institute for Genomic Biology, April 7, 2018; "Bees on stamps," CU Stamp Club, Urbana Free Library, Urbana, IL, May 7, 2018 What do you plan to do during the next reporting period to accomplish the goals?Goals for the next reporting period include: 1. Completion of Objective 1: RNASeq analysis; 2.Progress toward completion of Objective 5: Continue longevity assays to ascertain phytochemical effects on monofloral honey viscosity, osmotic potential, and thixotropic properties; 3.Progress toward completion of Objective 3 in evaluating monofloral honey suitability for hive fungi and for small hive beetle preference and performance; and 4.Dissemination plans include Summer 2018 presentation for the scientific community at the Arthropod Genomics Symposium, Urbana, IL; Summer 2018presentations for the public at Heartland Apicultural Society July 8, 2018 and the UI Pollinatarium; Fall 2018presentation for the scientific community at the annual meeting of the Entomological Society of America, Vancouver, BC, Canada.
Impacts
What was accomplished under these goals?
We evaluated impacts of three monofloral honeys, white tupelo (Nyssa ogeche), black locust (Robinia pseudoacacia), and buckwheat (Fagopyrum esculentum), on honeybee health. To confirm phytochemical differences in composition, these honeys were analyzed with LC-MS and were confirmed to vary in total phenolic content, phenolic acid diversity, and flavonoid diversity. Whereas buckwheat honey is rich in phenolic acids, tupelo honey has comparatively high content of abscisic acid and flavonols and black locust honey has high concentrations of hyperoside (3-O-galactoside of quercetin) and two propolis-associated flavanones. Obj. 1--To test for differential transcription of genes involved in detoxification, immunity, and longevity with honey chemistry we conducted an experiment with honeybees in an apiary at the University of Illinois Bee Research Facility. A capped brood frame was collected from a single naturally mated queen colony and then incubated in a dark room at 34 degrees C to obtain newly emerged adult workers. These bees were randomly and evenly introduced into five cages in groups of 3-20 individuals. One-tenth of a strip of commercialized artificial queen mandibular pheromone was also introduced into each cage at the same time. The caged bees were provided with water and one of fivedifferent diets: sugar water (control), buckwheat honey, tupelo honey, locust honey, and a combination of the three honeys. Three hives were used, and each diet treatment was repeated three times within a hive. After 14 days, bees were sacrificed and midguts dissected, individually collected, and flash-frozen in liquid nitrogen. Total RNA was extracted from the dissected guts (total n =156). Extractions were carried out with the NucleoSpin RNA kit. Guts were lysed by adding 300 ul of RA1 buffer with 5 ul of 1M DTT to the frozen sample and disrupted with Biomashers pestles. From that point, extractions were carried out following kit manufacturer protocols. RNA was eluted in a final volume of 30 ul of nuclease-free water. DNAse treatment was performed as per kit protocol before washing and elution. The RNA was quantified in a Nanodrop spectrophotometer. Concentrations were uniform across extractions and the ratio of 280/260 nm absorbance was >2.1 for all the samples, indicating that extractions are DNA-free. To confirm quality for sequencing, the RNA will be assessed with a Bioanalyzer using a nano RNA chip. The RNA will be used to prepare 15 barcoded libraries, which will be sequenced in one lane of an Illumina HiSeq 4000 with 100 base pair single-end reads. We expect to generate approximately 20 million reads per sample. Obj. 2. To measure effects of honey phytochemicals on pesticide toxicity, we compared worker longevity on three monofloral honeys alone and a sugar control. Six capped brood frames were collected, two frames each of three naturally mated queen colonies, and then incubated in a dark room at 34 degrees C to collect adults within 24 hoursof emergence . These adults were randomly introduced into eight cages in groups of 30 along with 0.1 strip of pheromone as described. Caged bees received water and one of the four diets. After 24-36 hoursof feeding, bees from each diet treatment were divided into two groups (by cages) and topically treated with 0.00146 ug/bee bifenthrin ( 0.1 LD50) or a solvent control (acetone). On the tupelo honey and sugar diets, survival was not affected by exposure to sublethal bifenthrin application. However, buckwheat and locust honey interacted with bifenthrin such that sublethal bifenthrin exposure enhanced survival. This result was not anticipated and may result from enhanced upregulation of detoxification genes by simultaneous exposure to pesticide and phytochemicals. Effects of quercetin and p-coumaric acid on toxicity of two additional pesticides (the acaricide tau-fluvalinate and the neonicotinoid imidacloprid) were also tested. We performed 24- and 48-hour LD50 assays on tau-fluvalinate in the presence and absence of quercetin and p-coumaric acid (replicated three times). One-day-old bees were fed a range of concentrations of phytochemicals for three days and then underwent topical treatments using tau-fluvalinate at a range of doses. Individually, each phytochemical had no effect on the LD50 of tau-fluvalinate. However, 1.50 mM p-coumaric acid plus 0.25 mM quercetin increased the tau-fluvalinate LD50 compared to 1.50 mM p-coumaric acid alone (from 1.76 to 2.78 ug/bee at 24 hours, and 1.40 to 2.34 ug/bee at 48 hours). Thus, depending on concentration, phytochemicals can interact to mitigate pesticide toxicity. With respect to imidacloprid, 0.25 mM quercetin and 0.5 mM p-coumaric acid, both individually and in combination, were tested in with imidacloprid at a range of concentrations; phytochemicals had no effect on the median-lethal concentration (LC50) of imidacloprid after 24 or 48 hours. In chronic toxicity bioassays, in the absence of imidacloprid, both phytochemicals individually, but not in combination, enhanced longevity. Consuming two phytochemicals with casein had an antagonistic effect on toxicity of p-coumaric acid or quercetin alone in the absence of pesticides. With low imidacloprid concentrations (15, and 45 ppb), both phytochemicals, individually and in combination, increased longevity but with 75 ppb imidacloprid, the two phytochemicals did not affect lifespan. Exposure to high imidacloprid concentrations after consuming p-coumaric acid alone and quercetin alone reduced bee longevity, suggesting these phytochemicals are protective only at low imidacloprid levels. All tested experimental factors affected bee longevity.p-Coumaric acidand quercetin had positive effects on longevity (with hazard ratios <1) at low imidacloprid concentrations. Imidacloprid had overall slightly negative effects on longevity (with hazard ratios 1.001>1.) but the effects were also concentration-dependent. Obj. 3--To determine differences in fungistatic/fungitoxic properties, we are currently setting up cultures of Kodamaea ohmeri in preparation for the monofloral honey assay on fungal growth. Due to concerns about risk to other bee research projects posed by chalkbrood, Ascophaera apis, we are securing space in a Biosafety 2 room on campusafter which we will culture chalkbrood to conduct fungal growth assays. Obj 4--To assess antioxidant capacity of different honeyswe measured antioxidant capacity in two ways, the relative DPPH radical scavenging capacity (RDSC) assay (Cheng et al., 2006) and the oxygen radical absorbance capacity (ORAC) assay (Gillespie et al., 2007). By both measures, locust honey has the lowest antioxidant capacity, whereas white tupelo levels are intermediate and buckwheat antioxidant capacity is highest by a factor of four-fold to ten-fold. Obj. 5--To determine impacts of honey on adult bee lifespan, we conducted longevity assays as described for Obj.1. The three monofloral honeys did not enhance longevity of adult bees in our trials. Without casein, both buckwheat and tupelo honey reduced lifespan (Tarone-Ware test; Sug vs Buc, χ2=45.40, p<0.001; Sug vs Tup, χ2=11.622, p<0.001). Casein in the diet extends lifespan overall (Cox model, HR: 0.594, p<0.001), as well as within the control treatment and the three honey treatments. Outcomes of all treatment groups differed significantly. In a second trial, tupelo and buckwheat honey alone reduced lifespan compared to the control (Sug vs Tup, log-rank test, χ2=17.14, p<0.001; Sug vs Buc, χ2=6.82, p<0.01) and the three honeys in combination (TLB; TLB vs Tup, χ2=16.02, p<0.001; TLB vs Buc, χ2=5.87, p<0.05). However, during the first 8 days, bees consuming buckwheat honey experienced increased longevity (hazard ratio: 0.311, p<0.05). Bees consuming the three-honey combination also had increased survival compared to bees consuming tupelo honey alone (log-rank test, χ2=4.16, p<0.05).
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
Calla, B., M. McLean, L.-H. Liao, I. Dhanjal, C. Tittiger, G.M. BLomquist and M.R. Berenbaum. 2018. Functional characterization of CYP4G11 - A highly conserved enzyme in the western honey bee Apis mellifera. Insect Molecular Biology (In Press).
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Liao, L.-H., W.-Y. Wu and M.R. Berenbaum, 2017. Behavioural responses of honey bees (Apis mellifera) to natural and synthetic xenobiotics in food. Scientific Reports 7: 15924.
- Type:
Journal Articles
Status:
Awaiting Publication
Year Published:
2018
Citation:
Johnson, R.M., A. Zayed, B. Harpur, K. Dogantzis and M.R. Berenbaum, 2018. Genomic footprint of evolution of eusociality in bees: Floral food use and CYPome blooms . Insectes Sociaux (Accepted Pending Revision).
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