Biological and environmental factors influencing the health of the honey bee, Apis mellifera

BIOLOGICAL AND ENVIRONMENTAL FACTORS INFLUENCING THE HEALTH OF THE HONEY BEE, APIS MELLIFERA

Sponsoring Institution

National Institute of Food and Agriculture

Project Status

NEW

Funding Source

HATCH

Reporting Frequency

Annual

Accession No.

1001975

Grant No.

(N/A)

Project No.

TEX09557

Proposal No.

(N/A)

Multistate No.

(N/A)

Program Code

(N/A)

Project Start Date

Nov 13, 2013

Project End Date

Nov 5, 2018

Grant Year

(N/A)

Project Director
Rangel-Posada, JU, .

Recipient Organization
TEXAS A&M UNIVERSITY
750 AGRONOMY RD STE 2701
COLLEGE STATION,TX 77843-0001

Performing Department
Entomology

Non Technical Summary
Problem Statement The honey bee, Apis mellifera L., is arguably the most important insect pollinator in North America, contributing an estimated $17 billion annually to the U.S. economy, primarily through crop pollination (Morse and Calderone 2000, Calderone 2012). Despite the economic importance of honey bees, the number of managed colonies available for pollination services has dropped dramatically during the last decade, threatening the production of many bee-dependent crops across the country (Rucker et al. 2012). The most influential culprit in such declines continues to be the ectoparasitic mite, Varroa destructor, a pest of honey bees that sucks the hemolymph of brood and adults, and vectors several bee viruses (Chen et al. 2004). Recent surveys across the country have indicated that not only Varroa-associated problems, but multiple ailments including poor nutrition, failing queens, pesticide contamination, and genetic inbreeding, among others, are the most commonly reported problems facing apiculture today (vanEngelsdorp et al. 2008, 2009, 2010, 2013). Thus, as the demand for healthy honey bee colonies is expected to increase along with food demands, the health and number of managed honey bee colonies available for pollination in the United States continues to decline (vanEngelsdorp et al. 2011, 2012), threatening the production of many crops dependent on honey bees for successful crop set. Thus, improving the health and productivity of managed and feral honey bees as important pollinators of major crops is of key importance for food security in both Texas, and the United States. Honey bees are also biologically significant because they serve as a paradigm of organization and cooperation, because they exhibit strict reproductive division of labor (Winston 1987, Seeley 1995). Non-reproductive tasks in the colony (e.g., nest construction, brood rearing, foraging, and defense) are carried out by tens of thousands of sterile female workers, while reproductive tasks are carried out by one female queen (i.e., the mother of all individuals in the colony) and a few thousand male drones that are produced seasonally. Given that such few individuals are responsible for the overall reproductive fitness of a colony, it is important to understand the factors that influence the reproductive health of honey bee queens and drones. Colony fitness is affected by the reproductive success of its queen (i.e., measured in terms of mating success, mating frequency, oviposition rate, and overall health and longevity of the queen), and the reproductive success of its drones (i.e., proportion of total number of males produced by a colony per year that successfully mate and inseminate queens). Fitness is also broadly affected at the colony level by the overall success of the colony as a whole (i.e., measured in terms of number of derivative colonies produced during colony fissioning, as well as colony growth and longevity), which is highly dependent on the worker population size. Thus, fitness is initially dependent on the reproductive quality of the queen and drones that sire a colony's worker force. Based on the demand for more basic research exploring the effects of reproductive quality, as well as the need for more applied research on the effects of crop quality on colony health, my (non exhaustive) 5-year research program plans to incorporate various basic and applied projects regarding the reproductive health of managed and feral honey bee colonies, as well as the overall health and productivity of honey bee colonies in cotton agroecosystems. To answer the questions we will conduct field and laboratory studies on the behavioral ecology, toxicology, microbiology, molecular biology, pollination biology, and physiology, of feral and managed honey bee populations. More information for each project will be outlined separately.

Animal Health Component

50%

Research Effort Categories

Basic

80%

Applied

20%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
211	3010	1130	100%

Knowledge Area
211 - Insects, Mites, and Other Arthropods Affecting Plants;

Subject Of Investigation
3010 - Honey bees;

Field Of Science
1130 - Entomology and acarology;

Keywords

Goals / Objectives
To explore the biological and environmental factors that influence the reproductive health of honey bee queens and drones in managed honey bee populations, and how such reproductive health affects colony growth, productivity and longevity To assess the health and Africanization levels of feral honey bee populations in South Texas To determine the reproductive quality of honey bee queens and drones in Texas apiaries To uncover the effect of pollination in cotton agroecosystems on the growth and overall health of managed honey bee colonies

Project Methods
Research Approach Biological and environmental factors that influence the reproductive health of honey bee queens and drones. This line of research is aimed at understanding what biological and environmental factors affect honey bee queen and drone reproductive health, and how reproductive quality ultimately influences colony-wide productivity and fitness. First, we will explore the biological factors that influence queen reproductive health by evaluating the effects of a queen's physiology (i.e., as measured by her mating frequency, genotype, among others) on her acceptance and attractiveness to workers in a colony. We will also explore how these factors might affect whether a queen is accepted by the workers, or is replaced with a new queen--a process known as queen supersedure--, a significant problem affecting honey bee colonies today. We will also look at the effects of queen quality on colony phenotype in terms of growth, productivity, and longevity. Second, we will explore the biological factors that influence drone reproductive health by examining a drone's genotype on his reproductive quality (i.e., as measured by spermatozoa morphology, motility, gene expression, among others). Consequently, we also want to determine if colonies headed by queens of varying reproductive quality produce drone populations with varying reproductive competitiveness. Lastly, we will explore the potential sub-lethal effects of commonly used in-hive chemicals used to control the ectoparasitic mite Varroa destructor, on queen and drone reproductive physiology. For instance, we want to find out if exposure to in-hive miticides cause physiological changes in the reproductive system of drones and queens that might lead to poor quality and viability of spermatozoa and sperm cell storage. We also want to explore whether these chemicals cause behavioral changes that decrease the flight performance of and sexual competitiveness of queens and drones during mating. Answering these questions will help us uncover the biological and environmental factors that influence the reproductive quality of managed honey bee queens and drones. Health and patterns of Africanization of the feral honey bee population in South Texas. This line of research will look at the feral honey bee population of South Texas. We will be locating feral honey bee colonies living in tree cavities at the Welder Wildlife Refuge near Sinton, TX, and build on previous work conducted at that site regarding the genetic lines of bees present in the feral population in that area of the state. First, we will determine the current hybridization pattern between the tropical-evolved African subspecies Apis mellifera scutellata brought from South Africa into South America in 1956, and the temperate Apis subspecies previously imported from Europe into North America in the 1800s (Pinto et al. 2004, 2005). We will conduct the genetic studies necessary to determine whether the African-European honey bee hybrid zone is still present at this site. Second, we will also collect honey bee samples to assay levels of common honey bee pathogens in the feral population. For instance, we measure the infestation levels of Varroa mites present in these colonies, as well as the infection rates of the gut microsporidia Nosema ceranae and Nosema apis, which cause one of the most prevalent diseases affecting adult honey bees (Matheson 1993, Fries 1993, 1996, Traver and Fell 2011 a, b). Results from these surveys might indicate that feral honey bees might have special adaptations to overcome biological and environmental as nuances compared to managed honey bee populations. Reproductive health of honey bee queens and drones in Texas apiaries. Collaborating with the Texas Apiary Inspection Service and Texas Beekeepers Association, this line of research will take advantage of information and bee samples provided by beekeepers across Texas to examine reproductive quality in managed queens and drones produced in our state. We will assess mating quality, sperm viability, and overall reproductive health of queens and drones obtained from commercial beekeepers that produce queens for sale in and out of state. Similar methodologies outlined above will be used to assess reproductive quality. d. Effects of honey bee pollination on colony heath and productivity in cotton agroecosystems. This line of research will explore the effects of pollen foraging on honey bee colony health. Cotton is one of the crops that benefits from honey bee pollination (McGregor 1959). About 20 percent of cotton seed production depends on insect pollination, of which, up to 80 percent is attributed to honey bees (Morse and Calderone 2000, Calderone 2012). Honey bees are known to visit cotton flowers, mainly to collect nectar (Loper and DeGrandi-Hoffman 1994), but also pollen, especially during certain times of the year (Moffett et al. 1975). But despite the evidence showing the direct benefits to cotton by honey bee pollination, there have not been any studies looking at the effects of cotton pollination by honey bees on the health and productivity of their colonies. Honey bees collect pollen as the colony's sole source of protein, which is mainly used for brood rearing and glandular development of nurse worker bees (Winston 1987, Seeley 1995). Thus, colony growth and health depends on the quantity, as well as the quality, of the resources provided by foragers. For instance, it has been shown that diet severely affects immunocompetence in honey bees (Alaux et al. 2010). Incidentally, choice experiments have demonstrated that honey bees have strong preferences for some pollen sources (Cook et al 2003). The preference is always for pollen that contains the most essential amino acids needed by honey bees (i.e., valine, leucine, and isoleucine; deGroot 1953). When offered cotton pollen in another choice experiment, honey bees showed reduced effectiveness in packing the pollen, perhaps because the grains are spiny and hard to manipulate (Vassière et al. 1994), or because they are not nutritious enough, or both (Roulston & Cane 2000). Besides these studies, little else is known about the effects of pollen quality on honey bee foraging biology on cotton. We thus propose to use our current knowledge regarding the foraging biology of honey bees in cotton agroecosystems to conduct a set of experiments that will help us better understand the relationship between honey bees, colony health, and cotton. Specifically, we propose to (1) test the effects of cotton pollen on the health and productivity of honey bee colonies, (2) examine the nutritional and physiological properties of cotton pollen, and (3) explore any differences in pollen between conventional and Bt cotton. These projects will help us better determine key avenues of research that should be explored in future studies regarding the role of honey bees in cotton agroecosystems.

Progress 10/01/16 to 09/30/17

Outputs
Target Audience:The target audience for this proposed work includes the following: • The scientific community interested in the fields of honey bee biology, apiculture, social insects, animal behavior, behavioral ecology, toxicology, entomology • The beekeeping community in the state of Texas and in the United States • The farming community that is directly or indirectly involved in using honey bees for crop pollination, or those who own or operate farms where pesticides might be used to control crop pests • The students at Texas A&M University who will be involved in designing, carrying out, analyzing, and presenting experiments as well as taking courses relavant/related to this grant • The general public, as an increase in honey bee health will help increase food security in the country Changes/Problems:A major change in the last couple of years is that we have not been able to secure funding to conduct experiments for Objective 4. To uncover the effect of pollination in cotton agroecosystems on the growth and overall health of managed honey bee colonies. Therefore I would like to remove this objective from the list of objectives for my Hatch Project. What opportunities for training and professional development has the project provided?Professional development activities for Juliana Rangel Posada since last progress report Participant, PESCA Grant Writer's workshop. TAMU, ILSB 3147. 7 September 2017. Participant, P&T Candidate Dossier Preparation Workshop. AGLS 129. 5 April 2017. Pedagogy Lunch Forum. Topic: Disciplines related to Graduate and Professional Programs. TAMU, Rudder 601. 22 November 2016. Negotiations Skills for Women in Academia. TAMU, MSC 2406B. 10 November 2016. Interactive Teaching Methods/Approaches to Engage Students. MSC 2405. 8 Nov. 2016. Pedagogy Lunch Forum #4 - Disciplines related to Biological and Life Sciences. TAMU, Rudder 601. 7 November 2016. ENTO 320 Course Observation by Center for Teaching Excellence. Done by Nate Poling. 102 Heep, TAMU. 29 October 2016. Membership in professional / scientific societies [9] Entomological Society of America (ESA) Member, 2006-Present. [8] American Association of Professional Apiculturists (AAPA). 2010-Now. President '13-14. [7] Texas Beekeepers Association Member, January 2013 - Present. [6] Honorary Member, Williamson County (TX) Beekeepers Association, Mar 2013 - Present. [5] Eastern Apicultural Society (EAS) Member, 2009 - 2012. [4] International Union for the Study of Social Insects (IUSSI) Member, 2005 - Present. [3] Animal Behavior Society (ABS) Associate Member, 2005 - Present. [2] International Society for Behavioral Ecology (ISBE) Member, 2005 - Present. [1] ADVANCE Scholars Program. Texas A&M University. 2013 - 2014. How have the results been disseminated to communities of interest?The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented oral talks or posters at at least one scientific conference per year, in one of the following conferences: • Annual National and Regional meetings of the Entomological Society of America (ESA), 2014, 2015, 2016, 2017 • American Bee Research Conference 2014, 2015, 2016, 2017, 2018 • Graduate Research Forum, Department of Entomology, Texas A&M University, 2014, 2015, 2016, 2017, 2018 • London Honey Show (2016) • American Beekeeping Federation Annual meeting, 2014, 2015, 2016, 2018 Texas Beekeepers Association annual convention, 2014, 2015, 2016, 2017, 2018 • Texas Beekeepers Association summer clinic, 2014, 2015, 2016, 2017, 2018 • Austin Area Beekeepers Association Bee School, 2014, 2015, 2016, 2017, 2018 • Central Texas Beekeepers Association Bee School, 2014, 2015, 2016, 2017, 2018 • Central Texas Beekeepers Association Bee School, 2014, 2015, 2016 The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented oral talks or posters at least one beekeeping meeting per year to share our results with the beekeeping community, at: • Texas Beekeepers Association Summer Clinic or Annual Convention • American Beekeeping Federation • Local, county beekeepers association in TX • Heartland Beekeepers Association or any State or Regional Beekeeping conference The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects will attend and present oral talks or posters at least one university-wide conference or meeting per year to share our results with the local academic/student community, which can be any combination of the following conferences: • Texas A&M University Department of Entomology's Annual Graduate Research Forum • Texas A&M University Department of Entomology's Seminar Series • Ecological Symposium, Texas A&M University • Student Research Week, Texas A&M University The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented in at least one community outreach event to share our results with the public and communicate our work through outreach activities to our local community, which can be any combination of the following conferences: • Brazos Valley Worldfest • Brazos Valley Museum of Natural History's Booneville Days • Brazos Valley Museum of Natural History's Butterfly Release Exhibit • Texas A&M University's College of Agricultural and Life Sciences Annual Tailgate What do you plan to do during the next reporting period to accomplish the goals?We will continue to train undergraduate and graduate students, research staff and postdoctoral research associates to accomplish the research, service and outreach goals set by this Hatch Project, as we have been doing for the last 4 years. 1. We will change knowledge of the students and postdocs involved in the projects through: • Development, execution, and delivery of research project • Completion of graduate level courses at Texas A&M University that will help increase the knowledge of students in the field of Entomology and Apiculture • Biweekly laboratory meetings with all members of the Rangel Lab to assess progress in all objectives • Instruction of Honey Bee Biology, and Honey Bee Management courses at the undergraduate level at Texas A&M University • Development and execution of several scientific techniques/tools to accomplish research goals 2. We will evaluate the outputs involved in the projects by developing and executing the following experimental procedures: • Insect source. We will use Italian honey bee colonies and queens (Apis mellifera Ligustica) for all experiments, to be purchased from nearby producers in the city of Navasota, TX. We will label all queens to ensure they do not get inadvertently replaced by Africanized bee stocks. All queens heading source larvae for grafting will be super-sisters to each other. To create super-sister queens, all queens will be daughters of the same mother who will be single-drone inseminated (SDI), following the technique described by Laidlaw (1977), which has been used in similar studies • Grafting. Experimental queens will be raised by transferring young larvae from a colony to cells in another queenless colony to initiate queen-rearing behavior, a common procedure known as "grafting" • Observation colonies. We will create medium-sized honey bee colonies from larger colonies that live in nearby bee yards, all headed by artificially inseminated super-sister Italian queens. • Genotyping. To calculate queen mating frequency we will collect emerging brood from a queen's colony and extract DNA from their thoraces using DNeasyTM kits. The DNA samples will be screened at 6 highly polymorphic microsatellite loci • Spermatozoa viability analysis. We will analyze spermatozoa viability following a protocol similar to that described by Collins and Donoghue (1999). This will be done in collaboration with the Theriogenology Laboratory directors at Texas A&M University. • Statistical analysis. To test the effects of the queen-rearing environment on the morphological characteristics of queens, we will conduct non-parametric Kruskal-Wallis and ANOVA tests to analyze data • Drone rearing. We will use plastic frames sold commercially with hexagonal cell templates to rear drones in laboratory conditions. 3. We will evaluate our progress in the following general ways. We hope to be successful in grafting queens for all experiments. We also hope that drone rearing will occur early in each year's spring so that they can be produced early in the year, not only for drone quality measurements, but also to enable successful mating frequency of queens. We also hope that RNA can be extracted in large numbers to create the microRNA and mRNA libraries for the fertility fingerprint. 4. We will produce the following data and deliverables: • The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects will produce at least two of the following tangible products per year to share our results with the public: • Scientific articles in peer-reviewed scientific journals • Articles in Trade magazines such as American Bee Journal, Bee Culture • Columns in the Texas Beekeepers Association Journal • Blogs and posts in social media sites such as the Facebook page of the TAMU honey bee lab, or its website • Oral presentations, posters, and outreach events outlined above

Impacts
What was accomplished under these goals? Discovered that fungicides, insect growth regulators, and acaricides used in almond orchards during bloom negatively affect honey bee worker longevity Received the 2016 "Dean's Outstanding Award in Early Career Research" from TAMU's College of Agricultural and Life Sciences Discovered that the prevalence of the levels of the microsporidian gut pathogens Nosema ceranae and N. apis have changed over the last 20 years in a feral honey bee population in South Texas, but have remained relatively at low numbers, and that N. apis disappeared from this population since 1998 Discovered that the combination of miticides fluvalinate, amitraz and coumaphos in wax significantly decreases sperm numbers and viability in queens Discovered that approximately 95% of the feral honey bees at Welder Wildlife Refuge is of Africanized maternal descent, and 60% of nuclear DNA contains alleles of African descent Helped collect data for the national survey of colony losses led by the Bee Informed Partnership at the University of Maryland Discovered that the chemical β-ocimene helps regulate foraging behavior of the honey bee Discovered that endopolyploidy changes with age-related polyethism in honey bees Discovered that exposure to in-hive miticides during queen development (used to control the ectoparasitic mite Varroa destructor) negatively affects worker behavior Showed that the miticides fluvalinate and coumaphos significantly decrease span of sexual maturity and lower sperm viability in drones We are mapping the location of feral honey bees at the Powdermill Nature Reserve in Pennsylvania and looking at feral bees in New Mexico, determining the mitochondrial DNA strains of these colonies, and their level of Nosema infections [in progress] We are finding out severe negative synergistic effects of fungicides used in almond orchards during bloom on worker bee longevity We discovered that managed honey bees in Dominica, W.I., are not Africanized, have low levels of Nosema spp., and some have high levels of Varroa mites [in progress]. Mentored three students (so far) enrolled in TAMU's Undergraduate Scholars Program

Publications

Type: Journal Articles Status: Published Year Published: 2016 Citation: Walsh E*, Rangel J (2016) Local honey bee queen production and quality. Bee World. 93(2): 30-32. DOI: 10.1080/0005772X.2016.1211501.
Type: Journal Articles Status: Published Year Published: 2017 Citation: Fisher II A*, Coleman JC, Hoffmann C, Fritz B, Rangel J (2017) The synergistic effects of almond protection fungicides on honey bee (Apis mellifera) forager survival. Journal of Economic Entomology. DOI: 10.1093/jee/tox031.
Type: Journal Articles Status: Published Year Published: 2017 Citation: Lau P*, Bryant V, Rangel J (2017) Determining the minimum number of pollen grains needed for accurate honey bee (Apis mellifera) colony pollen pellet analysis. Palynology. DOI: 10.1080/01916122.2017.1306810.
Type: Journal Articles Status: Published Year Published: 2017 Citation: Kulhanek K, Steinhauer N, Rennich K, Pettis J, Ellis JD, Sagili RR, Wilson ME, Tarpy DR, Rose R, Lee K, Rangel J, vanEngelsdorp D (2017) A national survey of managed honey bee 2015-2016 annual colony losses in the USA. Journal of Apicultural Research. DOI: 10.1080/00218839.2017.1344496.
Type: Journal Articles Status: Published Year Published: 2017 Citation: Gonzalez AN, Ing N, Rangel J (2017) Up-regulation of antioxidant genes in the spermathecae of honey bee (Apis mellifera) queens after mating. Apidologie. DOI: 10.1007/s13592-017-0546-y.
Type: Journal Articles Status: Published Year Published: 2017 Citation: Walsh E*, Rangel J (2018) Queen pheromones and mandibular gland dissection. Bee World. DOI: 10.1080/0005772X.2017.1373511.
Type: Journal Articles Status: Published Year Published: 2018 Citation: Fisher II A*, Coleman C, Hoffmann C, Fritz B, Rangel J (2018) The effects of the insect growth regulators methoxyfenozide and pyriproxyfen and the acaricide bifenazate on honey bee (Hymenoptera: Apidae) forager survival. Journal of Economic Entomology. DOI: 10.1093/jee/tox347.
Type: Journal Articles Status: Awaiting Publication Year Published: 2018 Citation: Ma R*, Villar G, Grozinger CM, Rangel J (2017) Larval pheromones act as colony-wide regulators of collective foraging behavior in honey bees. Behavioral Ecology. In revision.
Type: Journal Articles Status: Awaiting Publication Year Published: 2018 Citation: Rangel J, Ward L (2017) Evaluation of the predatory mite Stratiolaelaps scimitus for biological control of the honey bee ectoparasitic mite Varroa destructor. Journal of Apicultural Research. In review.
Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Fisher II A*, Harrison K*, Rangel J (2017) Spatio-temporal variation in honey bee (Apis mellifera) drone spermatozoa viability in central Texas apiaries. Southwestern Entomologist. In review.
Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Payne AN*, Rangel J (2017) The effect of queen insemination volume on honey bee (Apis mellifera) colony growth. Apidologie. In review.
Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Rangel J, Camp C**, Gonzalez A, Stoner** M, Hatter** A, Traver BE (2017) Genetic diversity and prevalence of Varroa destructor, Nosema apis and N. ceranae in managed honey bee (Apis mellifera) colonies in the Caribbean island of Dominica, West Indies. Journal of Apicultural Research. In review.

Progress 10/01/15 to 09/30/16

Outputs
Target Audience:The target audience for this proposed work includes the following: • The scientific community interested in the fields of honey bee biology, apiculture, social insects, animal behavior, behavioral ecology, toxicology, entomology • The beekeeping community in the state of Texas and in the United States • The farming community that is directly or indirectly involved in using honey bees for crop pollination, or those who own or operate farms where pesticides might be used to control crop pests • The students at Texas A&M University who will be involved in designing, carrying out, analyzing, and presenting experiments as well as taking courses relavant/related to this grant • The general public, as an increase in honey bee health will help increase food security in the country Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Professional development activities for Juliana Rangel Posada [30] Candidate dossier preparation workshop, COALS. TAMU. 30 March 2016. [29] Workshop on understanding the P&T process, COALS. TAMU. 23 March 2016. [28] Mentoring Graduate Students and Post-Doctoral Scholars Workshop. Faculty Teaching [27] Mentoring Graduate Students and Post-Doctoral Scholars Workshop. Faculty Teaching Academy. Rudder 701. College Station, TX. 29 January 2016. [26] Best Practices in Mentoring. Faculty Teaching Academy. MSC 2406. 16 Oct. 2015. [25] Annual Biomimicry Education Summit. Austin, TX. 4 October 2015. [24] EndNote Tutorial. Medical Sciences Library. 25 September 2015. [23] ADVANCE Women in Science Writing Club weekly meetings. Spring 2013 - Present. [22] Lecturing Well Workshop. Center for Teaching Excellence, Instructed by Carolyn Sandoval. 401 Rudder, TAMU. College Station, TX. 23 September 2014. [21] Course observation by Jean Layne during my ENTO 320 lecture to receive feedback on course and my teaching style. 102 HEEP, TAMU. 2 February 2014. [20] Consultation with Jean Layne to go over ENTO 320 syllabus and receive feedback. Center for [19] Teaching Excellence, 103 HEND, TAMU. College Station, TX. 7 January 2014. [18] Course Syllabus Course. Center for Teaching Excellence, Instructed by Jean Layne. 103 HEND, TAMU. College Station, TX. 11 December 2013. [17] Faculty Writing Workshop, Promoting Outstanding Writing for Excellence in Research (POWER) Program, TAMU. HLTH 632, College Station, TX. 6 December 2013. [16] Effective Student Mentoring Techniques workshop. Center for Teaching Excellence, TAMU. MSC 2406B, College Station, TX. 19 November 2013. [15] Teaching Statement, Teaching Portfolio, Course Portfolio workshop. Center for Teaching Excellence, Instructor: Jean Layne. 103 HEND, TAMU. 23 October 2013. [14] Universal Design for Learning workshop for course design. Center for Teaching Excellence, Instructed by Carolyn Sandoval. 103 HEND, TAMU17 October 2013. [13] Getting Started with eCampus Webinar. TTVN WebMeeting System. 4 September 2013. [12] New graduate student orientation. Entomology, TAMU. 20 August 2013. [11] Attended and networked with fellow honey bee researchers at the Meso-American Conference on Native Bees, National University, Heredia, Costa Rica. 24-31 Aug 2013. [10] Teaching Statement workshop. Center for Teaching Excellence, 103 HEND. 13 Jun 2013. [9] Selected as 2013 - 2014 ADVANCE Scholar, TAMU. Internal Mentor: Dr. Patricia Pietrantonio. External Mentor: Dr. Marla Spivak, Distinguished McKnight Professor, Department of Entomology, University of Minnesota. 4 June 2013. [8] ADVANCE Scholars Retreat, ADVANCE Center, TAMU. Bryan, TX. 23 May 2013. [7] NSF-REU's mentoring meeting. Instructed by Dr. Kevin Heinz and Ms. Rebecca Hapes. Department of Entomology, TAMU. College Station, TX. 8 May 2013. [6] Individual training on honey bee queen instrumental insemination. Instructor: Sue Cobey. College Station, TX 23-25 April 2013. [5] Roadmap to Success Workshop. ADVANCE Center. 8 - 9 April 2013. [4] TAMU's IPM Professional Development meeting. New Braunfels, TX. 28 March 2013. [3] Promotion & Tenure workshop, Dean of Faculties. Rudder Tower 501. 20 March 2013. [2] "Awesome" writing group. PI: Dr. Nancy Ing. 101 Kleberg. College Station, TX. February 2013 - Present. [1] Certificate in Teaching Training. Office of Postdoctoral Affairs, NCSU. Nov. 2011. Membership in professional / scientific societies [9] Entomological Society of America (ESA) Member, 2006-Present. [8] American Association of Professional Apiculturists (AAPA). 2010-Now. President '13-14. [7] Texas Beekeepers Association Member, January 2013 - Present. [6] Honorary Member, Williamson County (TX) Beekeepers Association, Mar 2013 - Present. [5] Eastern Apicultural Society (EAS) Member, 2009 - 2012. [4] International Union for the Study of Social Insects (IUSSI) Member, 2005 - Present. [3] Animal Behavior Society (ABS) Associate Member, 2005 - Present. [2] International Society for Behavioral Ecology (ISBE) Member, 2005 - Present. [1] ADVANCE Scholars Program. Texas A&M University. 2013 - 2014. How have the results been disseminated to communities of interest?The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented oral talks or posters at at least one scientific conference per year, in one of the following conferences: • Annual National and Regional meetings of the Entomological Society of America (ESA), 2014, 2015, 2016 • American Bee Research Conference 2014, 2015, 2016 • Graduate Research Forum, Department of Entomology, Texas A&M University, 2014, 2015, 2016 • London Honey Show (2016) • American Beekeeping Federation Annual meeting, 2014, 2015, 2016 • Texas Beekeepers Association annual convention, 2014, 2015, 2016 • Texas Beekeepers Association summer clinic, 2014, 2015, 2016 • Austin Area Beekeepers Association Bee School, 2014, 2015, 2016 • Central Texas Beekeepers Association Bee School, 2014, 2015, 2016 The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented oral talks or posters at least one beekeeping meeting per year to share our results with the beekeeping community, at: • Texas Beekeepers Association Summer Clinic or Annual Convention • American Beekeeping Federation • Local, county beekeepers association in TX • Heartland Beekeepers Association or any State or Regional Beekeeping conference The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects will attend and present oral talks or posters at least one university-wide conference or meeting per year to share our results with the local academic/student community, which can be any combination of the following conferences: • Texas A&M University Department of Entomology's Annual Graduate Research Forum • Texas A&M University Department of Entomology's Seminar Series • Ecological Symposium, Texas A&M University • Student Research Week, Texas A&M University The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented in at least one community outreach event to share our results with the public and communicate our work through outreach activities to our local community, which can be any combination of the following conferences: • Brazos Valley Worldfest • Brazos Valley Museum of Natural History's Boom Days • Texas A&M University's College of Agricultural and Life Sciences Annual Tailgate What do you plan to do during the next reporting period to accomplish the goals?We will continue to train undergraduate and graduate students, research staff and postdoctoral research associates to accomplish the research, service and outreach goals set by this Hatch Project, as we have been doing for the last 4 years. 1. We will change knowledge of the students and postdocs involved in the projects through: • Development, execution, and delivery of research project • Completion of graduate level courses at Texas A&M University that will help increase the knowledge of students in the field of Entomology and Apiculture • Biweekly laboratory meetings with all members of the Rangel Lab to assess progress in all objectives • Instruction of Honey Bee Biology, and Honey Bee Management courses at the undergraduate level at Texas A&M University • Development and execution of several scientific techniques/tools to accomplish research goals 2. We will evaluate the outputs involved in the projects by developing and executing the following experimental procedures: • Insect source. We will use Italian honey bee colonies and queens (Apis mellifera Ligustica) for all experiments, to be purchased from nearby producers in the city of Navasota, TX. We will label all queens to ensure they do not get inadvertently replaced by Africanized bee stocks. All queens heading source larvae for grafting will be super-sisters to each other. To create super-sister queens, all queens will be daughters of the same mother who will be single-drone inseminated (SDI), following the technique described by Laidlaw (1977), which has been used in similar studies • Grafting. Experimental queens will be raised by transferring young larvae from a colony to cells in another queenless colony to initiate queen-rearing behavior, a common procedure known as "grafting" • Observation colonies. We will create medium-sized honey bee colonies from larger colonies that live in nearby bee yards, all headed by artificially inseminated super-sister Italian queens. • Genotyping. To calculate queen mating frequency we will collect emerging brood from a queen's colony and extract DNA from their thoraces using DNeasyTM kits. The DNA samples will be screened at 6 highly polymorphic microsatellite loci • Spermatozoa viability analysis. We will analyze spermatozoa viability following a protocol similar to that described by Collins and Donoghue (1999). This will be done in collaboration with the Theriogenology Laboratory directors at Texas A&M University. • Statistical analysis. To test the effects of the queen-rearing environment on the morphological characteristics of queens, we will conduct non-parametric Kruskal-Wallis and ANOVA tests to analyze data • Drone rearing. We will use plastic frames sold commercially with hexagonal cell templates to rear drones in laboratory conditions. 3. We will evaluate our progress in the following general ways. We hope to be successful in grafting queens for all experiments. We also hope that drone rearing will occur early in each year's spring so that they can be produced early in the year, not only for drone quality measurements, but also to enable successful mating frequency of queens. We also hope that RNA can be extracted in large numbers to create the microRNA and mRNA libraries for the fertility fingerprint. 4. We will produce the following: • The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects will produce at least two of the following tangible products per year to share our results with the public: • Scientific articles in peer-reviewed scientific journals • Articles in Trade magazines such as American Bee Journal, Bee Culture • Columns in the Texas Beekeepers Association Journal • Blogs and posts in social media sites such as the Facebook page of the TAMU honey bee lab, or its website • Oral presentations, posters, and outreach events outlined above

Impacts
What was accomplished under these goals? Major accomplishments with respect to our research at TAMU Discovered that the prevalence of the levels of the microsporidian gut pathogens Nosema ceranae and N. apis have changed over the last 20 years in a feral honey bee population in South Texas, but have remained relatively at low numbers, and that N. apis disappeared from this population since 1998 Discovered that the combination of miticides fluvalinate, amitraz and coumaphos in wax significantly decreases sperm numbers and viability in queens Discovered that approximately 95% of the feral honey bees at Welder Wildlife Refuge is of Africanized maternal descent, and 60% of nuclear DNA contains alleles of African descent Helped collect data for the national survey of colony losses led by the Bee Informed Partnership at the University of Maryland Discovered that the chemical β-ocimene helps regulate foraging behavior of the honey bee Discovered that endopolyploidy changes with age-related polyethism in honey bees Discovered that exposure to in-hive miticides during queen development (used to control the ectoparasitic mite Varroa destructor) negatively affects queen retinue response [in progress] Showed that the miticides fluvalinate and coumaphos significantly decrease span of sexual maturity and lower sperm viability in drones [in progress] We are mapping the location of feral honey bees at the Powdermill Nature Reserve in Pennsylvania and looking at feral bees in New Mexico, determining the mitochondrial DNA strains of these colonies, and their level of Nosema infections [in progress] We are finding out severe negative synergistic effects of fungicides used in almond orchards during bloom on worker bee longevity [in progress] We discovered that the managed honey bee population in Dominica, W.I., is not Africanized, has low levels of Nosema spp., some colonies have high levels of Varroa mites [in progress]. Mentored one student that is in the 2015-2016 Undergraduate Scholars Program Received "Best Paper in 2013" award by the editors of the Journal Insectes Sociaux Mentored three NSF-REU students in Summer 2013, all of which produced very interesting results, and two of who received travel grants and presented at ESA's 2013 Annual Meeting Mentored one undergraduate student in Fall 2013 and Spring 2014, who presented his research at three different scientific conferences, including the SWB of ESA meeting in TX Initiated a collaborative effort with graduate students in the laboratories of Dr. Ulrich Muller and Dr. Nancy Moran, and Shalene Jha, University of Texas at Austin Created a collaborative interaction with the members of the Texas Beekeepers Association Trained 9 undergraduate research assistants whose help was essential in projects Served as Faculty Instructor in three Beekeeping Schools, where I showed a lot of my own research as part of the classes I instructed I have given or have been co-author in over 40 scientific talks, including 3 international and several invited speaker presentations at conference symposia

Publications

Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J, Baum K, Rubink WL, Coulson, RN, Johnston JS, Traver BE (2015) Prevalence of Nosema species in a feral honey bee population: A 20-year survey. Apidologie. 47(4): 561-571. DOI: 10.1007/s13592-015-0401-y.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J, Strauss K**, Hjelmen CE, Johnston JS (2015) Endopolyploidy changes with age-related polyethism in the honey bee, Apis mellifera. PLoS ONE. DOI: 10.1371/journal.pone.0122208.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Ma R*, Rangel J, Ulrich M (2015) The role of ?-ocimene in regulating foraging behavior of the honey bee, Apis mellifera. Apidologie. 47(1): 135-144. DOI: 10.1007/s13592-015-0382-x.
Type: Journal Articles Status: Published Year Published: 2016 Citation: Rangel J, Tarpy DR (2016) The effects of miticides on the mating health of honey bee (Apis mellifera L.) queens. Journal of Apicultural Research. 54: 275-283. DOI: 10.1080/00218839.2016.1147218.
Type: Journal Articles Status: Published Year Published: 2016 Citation: Rangel J, Giresi M*, Pinto MA, Baum KA, Rubink WL, Coulson RN, Johnston JS (2016) Africanization of a feral honey bee (Apis mellifera) population in South Texas: Does a decade make a difference? Ecology and Evolution. 6(7): 2158-2169. DOI: 10.1002/ece3.1974.
Type: Journal Articles Status: Published Year Published: 2016 Citation: Seitz N*, Traynor KS, vanEngelsdorp D, Steinhauer N, Rennich K, Wilson M, Ellis J, Rose R, Tarpy D, Sagili R, Caron D, Delaplane K, Rangel J, Lee K, Bayliss K, Wilkes J, Skinner J, Pettis J. (2016) A national survey of managed honey bee 2014 - 2015 annual colony losses in the USA. Apidologie. 46: 292. DOI:10.1007/s13592-015-0356-z.
Type: Journal Articles Status: Published Year Published: 2016 Citation: ] Rangel J, B�r�czky K, Schal C, Tarpy DR (2016) Honey bee (Apis mellifera) queen reproductive potential affects queen mandibular gland pheromone composition and worker retinue response. PLoS ONE. 11(6): e0156027. DOI:10.1371/journal.pone.0156027.
Type: Journal Articles Status: Published Year Published: 2016 Citation: Rangel J, Tarpy DR (2016) In-hive miticides and their effect on queen supersedure and colony growth in the honey bee (Apis mellifera). Journal of Environmental and Analytical Toxicology. 6: 377. doi:10.4172/2161-0525.1000377.
Type: Journal Articles Status: Published Year Published: 2016 Citation: Walsh E*, Rangel J (2016) Local honey bee queen production and quality. Bee World. 93(2): 30-32. DOI: 10.1080/0005772X.2016.1211501.
Type: Journal Articles Status: Awaiting Publication Year Published: 2017 Citation: Fisher II A*, Coleman JC, Hoffmann C, Fritz B, Rangel J (2016) The synergistic effects of almond protection fungicides on honey bee (Apis mellifera) forager survival. Journal of Economic Entomology. In Press.
Type: Journal Articles Status: Submitted Year Published: 2017 Citation: Rangel J, Ward L, Berg A**, Johnson M**, Johnston JS (2016) Evaluation of the predatory mite Stratiolaelaps scimitus for biological control of the honey bee ectoparasitic mite Varroa destructor. Biocontrol. Submitted.
Type: Journal Articles Status: Submitted Year Published: 2017 Citation: Kulhanek K, Steinhauer N, Rennich K, Pettis J, Ellis JD, Sagili RR, Wilson ME, Tarpy DR, Rose R, Lee K, Rangel J, vanEngelsdorp D (2016). A national survey of managed honey bee 2015-2016 annual colony losses in the USA. Journal of Apicultural Research. Submitted.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Newsletter. J Rangel. 2015. Texas Beekeepers Association Bimonthly Column. 6 columns per year.
Type: Journal Articles Status: Published Year Published: 2016 Citation: Newsletter. J Rangel. 2016. Texas Beekeepers Association Bimonthly Column. 6 columns per year.

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:The target audiences for this proposed work includes the following: • Scientific community interested in the fields of honey bee biology, apiculture, social insects, animal behavior, behavioral ecology, toxicology • The beekeeping community in the state of Texas and in the United States • The farming community that is directly or indirectly involved in using honey bees for crop pollination, or those who own or operate farms where pesticides might be used to control crop pests • The students at Texas A&M University who will be involved in designing, carrying out, analyzing, and presenting experiments as well as taking courses relavant/related to this grant • The general public, as an increase in honey bee health will help increase food security in the country Changes/Problems:We are not planning on working on Goal 4, "To uncover the effect of pollination in cotton agroecosystems on the growth and overall health of managed honey bee colonies." This is because we were unable to procure funding to explore this goal, so we decided not to pursue it in 2015. We will try to obtain funding in 2016 and subsequent years, which will allow us to pursue this research question. What opportunities for training and professional development has the project provided? We have been able to train 9 undergraduate students working on independent research projects on one or multiple of these research goals. We have recruited and continue to train three graduate students working on research projects on one or multiple of these research goals. We have recruited one postdoctoral research associate and one apiary manager/extension educator, both of whom are helping with a lot of the research goals We have given dozens of training beekeeping talks at various beekeeping and community events including: Texas Beekeepers Association Summer Clinic Texas Beekeepers Association Annual Convention Austin Area Beekeepers Association Bee Day Central Texas Beekeepers Association Spring Bee School Regional and county beekeepers association in TX Beekeepers Association Meetings in several states across the United States Heartland Beekeepers Association or any State or Regional Beekeeping conference American Beekeeping Federation American Honey Producers Association Brazos Valley Worldfest Brazos Valley Museum of Natural History's Boom Days Texas A&M University's College of Agricultural and Life Sciences Annual Tailgate How have the results been disseminated to communities of interest? We have been able to train 9 undergraduate students working on independent research projects on one or multiple of these research goals. We have recruited and continue to train three graduate students working on research projects on one or multiple of these research goals. We have recruited one postdoctoral research associate and one apiary manager/extension educator, both of whom are helping with a lot of the research goals We have given dozens of training beekeeping talks at various beekeeping and community events including: Texas Beekeepers Association Summer Clinic Texas Beekeepers Association Annual Convention Austin Area Beekeepers Association Bee Day Central Texas Beekeepers Association Spring Bee School Regional and county beekeepers association in TX Beekeepers Association Meetings in several states across the United States Heartland Beekeepers Association or any State or Regional Beekeeping conference American Beekeeping Federation American Honey Producers Association Brazos Valley Worldfest Brazos Valley Museum of Natural History's Boom Days Texas A&M University's College of Agricultural and Life Sciences Annual Tailgate What do you plan to do during the next reporting period to accomplish the goals?We will continue to train undergraduate and graduate students, as well as postdoctoral research associates to accomplish our reserach goals and to train and educate beekeepers and the public on topics related to honey bee health. We will continue to procure extramural funding from federal and private agencies as well. We will wrap up all finished projects and write up reports for trade magazines, as well as manuscripts to be submitted to peer-reviewed scientific journals.

Impacts
What was accomplished under these goals? To explore the biological and environmental factors that influence the reproductive health of honey bee queens and drones in managed honey bee populations, and how such reproductive health affects colony growth, productivity and longevity. ACCOMPLISHMENTS: We found that the combination of field-relevant concentrations of the miticides fluvalinate, coumaphos, and amitraz, severely decrease the number and viability of honey bee sperm both in mated queens and in sexually mature drones We found that the percentage of rearing success and longevity of queens and drones that are raised in a beeswax environment contaminated with combination of field-relevant concentrations of the miticides fluvalinate, coumaphos, and amitraz, is significantly lower than those raised in miticide-free beeswax. We have a couple of manuscripts in preparation to publish these results, and will continue to conduc To assess the health and Africanization levels of feral honey bee populations in South Texas ACCOMPLISHMENTS: The arrival to the United States of the Africanized honey bee (AHB), a hybrid between European honey bees and the African honey bee Apis mellifera scutellata, is a remarkable model for the study of biological invasions. This immigration has created an opportunity to study the dynamics of secondary contact of honey bee subspecies from African and European lineages in a feral population in South Texas. An 11-year survey (1991-2001) of this population showed that mitochondrial haplotype frequencies changed drastically over time from a resident population of eastern and western European maternal ancestry, to a population dominated by African haplotypes. A subsequent study of the nuclear genome showed that the Africanization process included bidirectional gene flow between European and Africanized honey bees, giving rise to a new panmictic mixture of A. m. scutellata- and European-derived genes. In this study, we examined gene flow patterns in the same population 23 years after the first hybridization event occurred. Of 89 tree cavities known to shelter colonies from earlier surveys, 28 were inhabited in 2013. Of these, 25 colonies were of A. m. scutellata maternal ancestry, and 3 colonies were of western European maternal ancestry. No colonies of eastern European maternal ancestry were detected, although they were present in the earlier samples. Nuclear DNA revealed an increase in the introgression of A. m. scutellata-derived genes into the population. Our results suggest this feral population is an admixed swarm with a now greater presence of African-derived nuclear and mitochondrial genetic composition. This manuscript was submitted in 2015 and by December 2015, a revision to the manuscript is being reviewed in the journal Ecology and Evolution. To determine the reproductive quality of honey bee queens and drones in Texas apiaries. ACCOMPLISHMENTS: We found that the viability and number of spermatozoa in drones from different apiaries varies significantly based on the geographic location of the apiary, which suggests that there may be environmental factors such as nutritional availability, that influence drone reproductive quality. We have not had as much luck in 2015 to raise a sufficiently large number of queens to determine if there are differences in queen quality based on the geographic location of the apiary around the Central Texas area. We will continue to perform experiments on these topics, and plan on getting a couple of manuscripts in preparation to publish these results in 2016. To uncover the effect of pollination in cotton agroecosystems on the growth and overall health of managed honey bee colonies. ACCOMPLISHMENTS: We were unable to procure funding to explore this goal, so we decided not to pursue it in 2015. We will try to obtain funding in 2016 and subsequent years, which will allow us to pursue this research question.

Publications

Type: Journal Articles Status: Published Year Published: 2015 Citation: Ma R, Mueller UG, Rangel J (2015) Assessing the role of B-ocimene in regulating foraging behavior of the honey bee, Apis mellifera. Apidologie. DOI: 10.1007/s13592-015-0382-x
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J, Strauss K, Hjelmen CE, Johnston JS (2015) Endopolyploidy changes with age-related polyethism in the honey bee, Apis mellifera. PLoS ONE. DOI: 10.1371/journal.pone.0122208
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J, Baum K, Rubink WL, Coulson RN, Johnston JS, Traver BE (2015) Prevalence of Nosema species in a feral honey bee population: a 20-year survey. Apidologie. DOI: 10.1007/s13592-015-0401-y
Type: Journal Articles Status: Awaiting Publication Year Published: 2016 Citation: Rangel J, Tarpy DR (2015) The effects of miticides on the mating health of honey bee (Apis mellifera L.) queens. Journal of Apicultural Research. In press.
Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Rangel J, B�r�czky K, Schal C, Tarpy DR (2016) Honey bee (Apis mellifera) queen reproductive potential affects queen mandibular gland pheromone composition and worker retinue response. PLoS ONE. Accepted with minor revisions.
Type: Journal Articles Status: Accepted Year Published: 2016 Citation: Rangel J, Giresi M, Pinto MA, Baum KA, Rubink WL, Coulson RN, Johnston JS (2015) Africanization of a feral honey bee (Apis mellifera) population in South Texas: Does a decade make a difference? Ecology and Evolution. Accepted with minor revisions.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J. 2015. Honey Bee Lab Update Column. Texas Beekeepers Association Journal. Mar-Apr issue.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J. 2015. Honey Bee Lab Update Column. Texas Beekeepers Association Journal. May-June issue.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J. 2015. Honey Bee Lab Update Column. Texas Beekeepers Association Journal. Jul-Aug issue.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J. 2015. Honey Bee Lab Update Column. Texas Beekeepers Association Journal. Sep-Oct issue.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J. 2015. Honey Bee Lab Update Column. Texas Beekeepers Association Journal. Nov-Dec issue.
Type: Journal Articles Status: Published Year Published: 2015 Citation: Rangel J. 2015. Honey Bee Lab Update Column. Texas Beekeepers Association Journal. January-February issue.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Rangel J (2015) Alternative IPM methods for control of Varroa destructor mites in Texas apiaries. Invited Symposium presentation. Electronic conference proceedings (abstract), annual meeting of the Entomological Society of America Southwestern Branch. Tulsa, OK.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Ward L, Rangel J (2015) Observations of the predatory mite Stratiolaelaps scimitus: a potential biocontrol agent of Varroa destructor. Electronic conference proceedings (abstract), annual meeting of the Entomological Society of America Southwestern Branch. Tulsa, OK.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Lau P, Rangel J, Bryant V, Cabrera A, Schmehl D, Sullivan J, Huang ZY, Ellis JD (2015) An analysis of the pollen collected by honey bees (Apis mellifera L.) in developed areas. Electronic conference proceedings (abstract), annual meeting of the Entomological Society of America Southwestern Branch. Tulsa, OK.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Walsh E, Rangel J (2015) The effect of in-hive miticides on honey bee (Apis mellifera) queen retinue response and queen mandibular pheromone. Electronic conference proceedings (abstract), annual meeting of the Entomological Society of America Southwestern Branch. Tulsa, OK.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Fisher II, A, Rangel J (2015) Honey bee (Apis mellifera) drone reproductive health. Electronic conference proceedings (abstract), annual meeting of the Entomological Society of America Southwestern Branch. Tulsa, OK.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Rangel J, Lau P, Bryant V, Cabrera A, Schmehl D, Sullivan J, Huang ZY, Ellis JD (2015) Pesticides found in pollen and nectar collected by honey bees in urban environments. Invited Symposium presentation. Electronic conference proceedings (abstract), Entomological Society of America annual meeting. Minneapolis, MN.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Lau P, Rangel J, Bryant V, Cabrera A, Schmehl D, Sullivan J, Huang ZY, Ellis JD (2015) Analysis of pollen collected by honey bees, Apis mellifera, in developed areas. Invited Symposium presentation. Electronic conference proceedings (abstract), Entomological Society of America annual meeting. Minneapolis, MN.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Rangel J, Giresi M, Pinto M, Baum K, Rubink W, Coulson R, Johnston JS (2015) Africanization of a feral honey bee (Apis mellifera) population in South Texas: Does a decade make a difference? Electronic conference proceedings (abstract), Entomological Society of America annual meeting. Minneapolis, MN.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Walsh E, Rangel J (2015) The effects of in-hive miticides on honey bee (Apis mellifera) queen retinue response and mandibular pheromones. Electronic conference proceedings (abstract), Entomological Society of America annual meeting. Minneapolis, MN.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Fisher II, A, Rangel J, Hoffmann C (2015) The Effects of Crop Protection Pesticides and Chemicals on Honey Bee (Apis mellifera). Electronic conference proceedings (abstract), Entomological Society of America annual meeting. Minneapolis, MN.
Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Rangel J, Baum K, Coulson R, Johnston JS, Traver B (2015) Prevalence of Nosema species in a feral honey bee population: A 20-year survey. Invited Symposium presentation. Electronic conference proceedings (abstract), Apimondia International Beekeeping Congress. Daejeon, South Korea.

Progress 11/13/13 to 09/30/14

Outputs
Target Audience: The target audiences for this proposed work includes the following: Scientific community interested in the fields of honey bee biology, apiculture, social insects, animal behavior, behavioral ecology, toxicology The beekeeping community in the state of Texas and in the United States The farming community that is directly or indirectly involved in using honey bees for crop pollination, or those who own or operate farms where pesticides might be used to control crop pests The students at Texas A&M University who will be involved in designing, carrying out, analyzing, and presenting experiments as well as taking courses relavant/related to this grant The general public, as an increase in honey bee health will help increase food security in the country Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? We were able to participate in the following professional and training events: Annual and Regional meetings of the Entomological Society of America (ESA) American Bee Research Conference Graduate Research Forum, Department of Entomology, Texas A&M University How have the results been disseminated to communities of interest? The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented oral talks or posters at at least one scientific conference per year, in one of the following conferences: Annual meeting of the Entomological Society of America (ESA) American Bee Research Conference Biannual meeting of the International Union for the Study of Social Insects Annual meeting of the Southwestern Branch of ESA Animal Behavior Society International Society of Behavioral Ecology The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented oral talks or posters at least one beekeeping meeting per year to share our results with the beekeeping community, at: Texas Beekeepers Association Summer Clinic or Annual Convention American Beekeeping Federation Local, county beekeepers association in TX Heartland Beekeepers Association or any State or Regional Beekeeping conference The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects will attend and present oral talks or posters at least one university-wide conference or meeting per year to share our results with the local academic/student community, which can be any combination of the following conferences: Texas A&M University Department of Entomology's Annual Graduate Research Forum Texas A&M University Department of Entomology's Seminar Series Ecological Symposium, Texas A&M University Student Research Week, Texas A&M University The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects attended and presented in at least one community outreach event to share our results with the public and communicate our work through outreach activities to our local community, which can be any combination of the following conferences: Brazos Valley Worldfest Brazos Valley Museum of Natural History's Boom Days Texas A&M University's College of Agricultural and Life Sciences Annual Tailgate What do you plan to do during the next reporting period to accomplish the goals? We will change knowledge of the students and postdocs involved in the projects through: Development, execution, and delivery of research project Completion of graduate level courses at Texas A&M University that will help increase the knowledge of studnets in the field of Entomology and Apiculture Biweekly laboratory meetings with all members of the Rangel Lab to assess progress in all objectives Instruction of Honey Bee Biology, and Honey Bee Management courses at the undergraduate level at Texas A&M University Development and execution of several scientific techniques/tools to accomplish research goals We will evaluate the outputs involved in the projects by developing and executing the following experimental procedures: Insect source. We will use Italian honey bee colonies and queens (Apis mellifera Ligustica) for all experiments, to be purchased from nearby producers in the city of Navasota, TX. We will label all queens to ensure they do not get inadvertently replaced by Africanized bee stocks. All queens heading source larvae for grafting will be super-sisters to each other. To create super-sister queens, all queens will be daughters of the same mother who will be single-drone inseminated (SDI), following the technique described by Laidlaw (1977), which has been used in similar studies Grafting. Experimental queens will be raised by transferring young larvae from a colony to cells in another queenless colony to initiate queen-rearing behavior, a common procedure known as "grafting" Observation colonies. We will create medium-sized honey bee colonies from larger colonies that live in nearby bee yards, all headed by artificially inseminated super-sister Italian queens. Genotyping. To calculate queen mating frequency we will collect emerging brood from a queen's colony and extract DNA from their thoraces using DNeasyTM kits. The DNA samples will be screened at 6 highly polymorphic microsatellite loci Spermatozoa viability analysis. We will analyze spermatozoa viability following a protocol similar to that described by Collins and Donoghue (1999). This will be done in collaboration with the Theriogenology Laboratory directors at Texas A&M University. Statistical analysis. To test the effects of the queen-rearing environment on the morphological characteristics of queens, we will conduct non-parametric Kruskal-Wallis and ANOVA tests to analyze data Drone rearing. We will use plastic frames sold commercially with hexagonal cell templates to rear drones in laboratory conditions. We will evaluate our progress in the following general ways. We hope to be successful in grafting queens for all experiments. We also hope that drone rearing will occur early in each year's spring so that they can be produced early in the year, not only for drone quality measurements, but also to enable successful mating frequency of queens. We also hope that RNA can be extracted in large numbers to create the microRNA and mRNA libraries for the fertility fingerprint. We will produce the following: The PI, postdoctoral research fellow, graduate student(s) and undergraduate student(s) involved in these projects will produce at least two of the following tangible products per year to share our results with the public: Scientific articles in peer-reviewed scientific journals Articles in Trade magazines such as American Bee Journal, Bee Culture Columns in the Texas Beekeepers Association Journal Blogs and posts in social media sites such as the Facebook page of the TAMU honey bee lab, or its website Oral presentations, posters, and outreach events outlined above

Impacts
What was accomplished under these goals? We submitted for publication four manuscripts, as follows: Prevalence of Nosema species in a feral honey bee population: A 20-year survey Juliana RANGEL1 *, Kristen BAUM2, William L. RUBINK3, Robert N. COULSON4, J. Spencer JOHNSTON1 and Brenna E. TRAVER5 1Department of Entomology, Texas A&M University, 2475 TAMU, College Station, Texas 77843-2475 2Department of Integrative Biology, Oklahoma State University, 501 Life Sciences West, Stillwater, Oklahoma 74078 3P.O. Box 2686, Edinburg, Texas 78540 4Knowledge Engineering Laboratory, Department of Entomology, Texas A&M University, College Station, Texas 77843-2475 5Department of Biology, Penn State Schuylkill, 200 University Drive, Schuylkill Haven, PA 17972 Keywords: Apis mellifera / Africanized feral honey bees / Nosema apis / Nosema ceranae / qPCR Abstract - Nosema spp. are microsporidian pathogens of honey bees that cause nosemosis, a disease implicated in colony losses worldwide. Few studies have measured Nosema spp. levels in feral honey bees. We evaluated the presence and infection intensity of Nosema apis and N. ceranae in a feral Africanized honey bee population in south Texas from 1991-2001 and in 2013. Overall, less than 6% of samples had Nosema spp. spores. Nosema apis was only found in samples from 1991 to 1995. Conversely, N. ceranae was found every year examined, ranging from 16.7% infection in 1991 to 85.7% in 2013. There were no effects of temperature or rainfall on either infection over time. This suggests that feral honey bees are relatively free of Nosema spp. compared to managed colonies. More studies on the incidence of Nosema spp. in feral honey bee populations are needed. Africanization of a feral honey bee (Apis mellifera) population in South Texas: Does a decade make a difference? Juliana Rangel,1* Melissa Giresi,2 M. Alice Pinto,3 Kristen A. Baum,4 William L. Rubink,5 Robert N. Coulson,6 J. Spencer Johnston1 1Department of Entomology, Texas A&M University, 2475 TAMU, College Station, Texas 77843-2475 2Department of Biology, Texas A&M University, 3258 TAMU, College Station, Texas 77843-3258 3Mountain Research Centre (CIMO), Polytechnic Institute of Bragança, Campus de Sta. Apolónia, Apartado 1172, 5301-855 Bragança, Portugal 4Department of Zoology, Oklahoma State University, 501 Life Sciences West, Stillwater, Oklahoma 74078 5P.O. Box 2686, Edinburg, Texas 78540 6Knowledge Engineering Laboratory, Department of Entomology, Texas A&M University, College Station, Texas 77843-2475 SUMMARY The arrival to the United States of the Africanized honey bee (AHB), a hybrid between European honey bees and the African honey bee Apis mellifera scutellata, is a remarkable model for the study of biological invasions. This immigration has created an opportunity to study the dynamics of secondary contact of honey bee subspecies from African and European lineages in a feral population in South Texas. An 11-year survey (1991-2001) of this population showed that mitochondrial haplotype frequencies changed drastically over time from a resident population of eastern and western European maternal ancestry, to a population dominated by African haplotypes. A subsequent study of the nuclear genome showed that the Africanization process included bidirectional gene flow between European and Africanized honey bees, giving rise to a new panmictic mixture of A. m. scutellata- and European-derived genes. In this study, we examined gene flow patterns in the same population 23 years after the first hybridization event occurred. Of 89 tree cavities known to shelter colonies from earlier surveys, 28 were inhabited in 2013. Of these, 25 colonies were of A. m. scutellata maternal ancestry, and 3 colonies were of western European maternal ancestry. No colonies of eastern European maternal ancestry were detected, although they were present in the earlier samples. Nuclear DNA revealed an increase in the introgression of A. m. scutellata-derived genes into the population. Our results suggest this feral population is an admixed swarm with a now greater presence of African-derived nuclear and mitochondrial genetic composition. Keywords: Apis mellifera; feral honey bees; Africanized honey bee; hybridization; introgression; microsatellites; mitochondrial DNA; nuclear DNA; spatial distribution The role of β-ocimene in regulating foraging behavior of the honey bee, Apis mellifera RONG MA1*, ULRICH G. MUELLER,1 JULIANA RANGEL2 1Section of Integrative Biology, The University of Texas at Austin, 1 University Station #C0990 Austin, TX 78712 2Department of Entomology, Texas A&M University 315 Minnie Belle Heep, 2475 TAMU College Station, TX 77843 Abstract--The larval pheromone (E)-β-ocimene (EBO) inhibits worker ovary development and accelerates the behavioral transition from nursing to foraging in the honey bee, Apis mellifera. Because EBO diffuses throughout the hive, it is hypothesized that worker honey bees can use it to indirectly assess brood abundance. Here, we test whether EBO plays a role in the regulation of worker foraging activity. We exposed colonies to a pulse of either synthetic ocimene or a paraffin oil control, and then recorded the number of returning pollen and non-pollen foragers. The effect of pheromone treatment on pollen foraging was highest directly after pheromone exposure and returned to baseline after three hours. Exposure to synthetic ocimene significantly increased overall foraging activity, thus providing evidence that honey bee foragers assess and respond to airborne larval odor cues such as ocimene to modulate foraging behavior. Endopolyploidy changes with age-related polyethism in the honey bee, Apis mellifera Juliana Rangel1*, Kim Strauss1, Kaileah Seedorf1, Carl E. Hjelmen1 and J. Spencer Johnston1 1 Department of Entomology, Texas A&M University, College Station, Texas 77843 Abstract Honey bees (Apis mellifera) exhibit age polyethism, whereby female workers assume increasingly complex colony tasks as they age. While changes in DNA methylation accompany age polyethism, other DNA modifications accompanying age polyethism are less known. Changes in endopolyploidy (DNA amplification in the absence of cell division) with increased larval age are typical in many insect cells and are essential in adults for creating larger cells, more copies of essential loci, or greater storage capacity in secretory cells. However, changes in endopolyploidy with increased adult worker age and polyethism are unstudied. In this study, we examined endopolyploidy in honey bee workers ranging in age from newly emerged up to 55 days old. We found a nonsignificant increase in ploidy levels with age (P < 0.1) in the most highly endopolyploid secretory cells, the Malpighian tubules. All other cell types decreased ploidy levels with age. Endopolyploidy decreased the least amount (nonsignificant) in neural (brain) cells and the stinger (P < 0.1). There was a significant reduction of endopolyploidy with age in leg (P < 0.05) and thoracic (P < 0.001) muscles. Ploidy in thoracic muscle dropped from an average of 0.5 rounds of replication in newly emerged workers to essentially no rounds of replication (0.125) in the oldest workers. Ploidy reduction in flight muscle cells is likely due to the production of G1 (2C) nuclei by amitotic division in the multinucleate striated flight muscles that are essential to foragers, the oldest workers. We suggest that ploidy is constrained by the shape, size and makeup of the multinucleate striated muscle cells. Furthermore, the presence of multiple 2C nuclei might be optimal for cell function, while higher ploidy levels might be a dead-end strategy of some aging adult tissues, likely used to increase cell size and storage capacity in secretory cells. Key words: Age polyethism, Apis mellifera, endopolyploidy, flow cytometry, honey bees

Publications

Type: Journal Articles Status: Awaiting Publication Year Published: 2015 Citation: The effects of miticides on the mating health of honey bee (Apis mellifera L.) queens. Journal of Apicultural Research
Type: Journal Articles Status: Accepted Year Published: 2015 Citation: Endopolyploidy changes with age-related polyethism in the honey bee, Apis mellifera. PLoS ONE
Type: Journal Articles Status: Submitted Year Published: 2015 Citation: Honey bee (Apis mellifera) queen reproductive potential affects queen mandibular gland pheromone composition and worker retinue response. Journal of Chemical Ecology
Type: Journal Articles Status: Published Year Published: 2014 Citation: Optimal colony fissioning in social insects: testing an inclusive fitness model with honey bees. Insectes Sociaux. Voted Best Paper in 2013 by the Editorial Board. DDOI: 10.1007/s00040-013-0309-3.
Type: Journal Articles Status: Submitted Year Published: 2015 Citation: The role of ?-ocimene in regulating foraging behavior of the honey bee, Apis mellifera. Apidologie.
Type: Journal Articles Status: Submitted Year Published: 2015 Citation: In-hive miticides and their effect on queen supersedure and colony growth in the honey bee (Apis mellifera L.). Apidologie. In review.
Type: Other Status: Published Year Published: 2014 Citation: Newsletter. J Rangel. 2014. Texas Beekeepers Association Bimonthly Column. 6 columns per year.
Type: Journal Articles Status: Submitted Year Published: 2015 Citation: Africanization of a feral honey bee (Apis mellifera) population in South Texas: Does a decade make a difference? JULIANA RANGEL,1* MELISSA GIRESI,2 M. ALICE PINTO,3 KRISTEN A. BAUM,4 WILLIAM L. RUBINK,5 ROBERT N. COULSON,6 J. SPENCER JOHNSTON1 Proceedings of the Royal Society of London Biological Sciences. Submitted
Type: Journal Articles Status: Submitted Year Published: 2015 Citation: Prevalence of Nosema species in a feral honey bee population: A 20-year survey Juliana RANGEL1 *, Kristen BAUM2, William L. RUBINK3, Robert N. COULSON4, J. Spencer JOHNSTON1 and Brenna E. TRAVER5. Apidologie. Submitted.