Source: CLEMSON UNIVERSITY submitted to
BIODIVERSITY OF INSECTS OF MEDICAL AND VETERINARY IMPORTANCE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0230188
Grant No.
(N/A)
Project No.
SC-1700433
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jul 1, 2012
Project End Date
Sep 30, 2016
Grant Year
(N/A)
Project Director
Adler, P.
Recipient Organization
CLEMSON UNIVERSITY
(N/A)
CLEMSON,SC 29634
Performing Department
Agricultural & Environmental Sciences
Non Technical Summary
More than 13,000 species of biting flies in the world feed on blood from vertebrate animals, and often transmit disease agents to humans and domestic and wild animals. Threats to human and animal welfare, such as economic losses and diseases caused by bloodsucking flies, provide motivation and justification for biodiversity research, with the ultimate goal of eliminating the risk. However, only the adult females directly pose a risk. The males and the immature stages of biting flies perform key ecological roles in the environment, such as pollination and organic matter processing. A tradeoff thus exists between controlling the problematic female flies and conserving the beneficial life stages. The project aims to contribute fundamental scientific knowledge in the areas of community ecology, host-parasite relationships, and the processes underlying the diversification of life, while also ensuring the public a level of protection from biting flies and fly-borne diseases. Methods include trapping and other collecting techniques, cytogenetic and molecular analyses, surveys of natural enemies living in or on the flies, ecological characterization of breeding habitats, and development of predictive models that will aid in identifying future pest problems. The project promotes the idea of biomonitoring for biting flies and vector-borne diseases, while also providing a first-line of protection for citizens, domestic animals, and wildlife. It encourages the involvement of agencies and the education of the public. Because the research is directed at blood-feeding flies, nearly all of which breed in water, the project also addresses environmental health issues through water-quality characterization and modeling of the breeding habitats. Potential impacts are both conceptual and practical and would be shared with the scientific and public communities. A vigorous program of presenting findings as publications and oral and written reports and seminars will ensure dissemination of information to a broad community of researchers, teachers, and extension specialists. Training of students, both undergraduate and graduate, would provide the mechanism for continued research and education in medical and veterinary entomology. Results from the research would be conveyed to appropriate personnel in agencies responsible for implementing solutions.
Animal Health Component
100%
Research Effort Categories
Basic
70%
Applied
20%
Developmental
10%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3123110107010%
3123110108010%
3123110111010%
3123110113020%
7213110107010%
7213110108010%
7213110111010%
7213110113020%
Knowledge Area
312 - External Parasites and Pests of Animals; 721 - Insects and Other Pests Affecting Humans;

Subject Of Investigation
3110 - Insects;

Field Of Science
1130 - Entomology and acarology; 1070 - Ecology; 1110 - Parasitology; 1080 - Genetics;
Goals / Objectives
The primary objective of this project is to discover, describe, and interpret the rich biodiversity of blood-feeding flies in South Carolina and other regions harboring species that potentially affect the economy and the human and animal health and welfare in the region. The specific objectives are 1) to discover and describe the biodiversity of flies of medical and veterinary importance and their associated symbiotes (e.g., natural enemies) at scales ranging from the genome to the ecological community; 2) to interpret the phylogenetic and ecological relationships of flies of medical and veterinary importance and their associated symbiotes; and 3) to develop solutions to problems caused by biting flies. New species, cytogenetic forms, and molecular forms discovered through surveys and biomonitoring will be described and, where appropriate, formally named. Symbiotic organisms identified in surveys of biting flies will be described, if new, and their potential as natural enemies explored. Ecological models and phylogenetic relationships of biting flies and their associated symbiotic organisms will be produced from the data collected in surveys, with emphasis on predictive models, such as predicting the factors that control pest distributions. The process of identifying and providing solutions for problems will form a feedback loop: Biting fly problems will be identified through surveillance and biomonitoring, and problems that come to light from citizens and agencies will provide the impetus for focused surveys and biomonitoring. Overall, the proposed research will address the growing concerns posed by biting flies of medical and veterinary importance.
Project Methods
To discover and describe biodiversity of biting flies, various trapping techniques will be used for collecting adult biting flies. Immature biting flies will be collected by hand and with nets from pools, ponds, tree holes, streams and rivers, and pupae will be reared to adults. These samples also will provide the material to assay for symbiotic organisms such as bacteria and fungi. Morphospecies of biting flies will be screened for hidden biodiversity, such as sibling species, cytoforms, and molecular forms, through cytogenetic and molecular analyses. To provide a predictive framework for interpreting biological phenomena of biting flies, a phylogenetic analyis will be conducted for selected groups of biting flies, using standard cladistic methodology and appropriate outgroups. Characters will be obtained from morphological, chromosomal, and molecular sources, and independent and combined analyses will be evaluated with the goal of obtaining most parsimonious trees. Surveys of biting flies will be accompanied by the collection of ecological data from the habitats of the immature stages, and various statistical analyses will be performed on the ecological data sets to establish predictive models. Investigations of natural enemies of biting flies, especially parasites and pathogens, will focus on documenting host associations and determining host specificity. Biting-fly problems will be addressed both preventatively and reactively. Surveys of biting flies, investigations of their bionomics and life histories, and studies of their symbiotic organisms (potential natural enemies) will provide baseline data for species that pose a health or economic threat. Specific problems that are brought to light will be investigated and potential solutions and recommendations will be provided.

Progress 07/01/12 to 09/30/16

Outputs
Target Audience:The target audience reached included members of the scientific community, undergraduate and graduate students, and public health officials and citizens in the state, nation, and world. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project continued to provide an education and training center for students and scientists from throughout the world to acquire new knowledge, skills, and techniques about the genetics, systematics, and ecology of biting flies. In addition to providing training in medical-veterinary entomology for Clemson University students, one professor fromIstanbul University (Turkey) and two students (1 doctoral student, 1 undergraduate student) from Mahasarakham University (Thailand) were hosted for a total of 14 months of specialized training. How have the results been disseminated to communities of interest?Project results were disseminated through numerous peer-reviewed journal publications; research presentations at professional meetings and universities throughout the world (e.g., Colombia, Russia, Thailand); a website; and many phone calls, e-mails, and in-person responses to colleagues, the public, the press, and students. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? IMPACT: To aid pest management and the control of vector-borne diseases, research was focused on understanding the fundamental biology and underlying genetics of pests and vectors of blood-feeding flies by discovering species, elucidating the ecology of each species, and determining the process by which some species become pests or vectors. Consequently, 14 new species and 32 chromosomal and molecular forms of blood-feeding flies were discovered throughout the world, indicating a far richer fauna than previously appreciated. The genetic mechanisms, such as chromosomal rearrangements, responsible for generating biodiversity were explored, and the mechanisms for becoming pests and vectors were elucidated. For instance, in the case of black flies, the historical process of becoming a vector of disease organisms involved the evolution of repetitive blood feeding, augmented by dispersal from the cool, mountainous, northern ancestral habitat into lowland and tropical areas where multiple generations of flies could be achieved and where the availability of potential hosts, including humans, increased. The process of attaining pest status, although accomplished via different pathways, always involves the ability to achieve large populations. Factors that promote large populations, such as impounding and polluting water in the breeding habitats, were identified. Objective 1: to discover and describe the biodiversity of flies of medical and veterinary importance and their associated symbiotes (e.g., natural enemies) at scales ranging from the genome to the ecological community. By exploring the genome of morphologically well-known species of blood-feeding flies, several dozen new species and genetic varieties were discovered throughout the world, including Europe, North Africa, South America, Southeast Asia, and the United States. The habitats and general ecological relationships of a number of these species and variants were elucidated. The discovery of these new species and genetic forms provides a database for recognizing and predicting pest and vector problems, not only in the areas where they were discovered but potentially in areas where they might become introduced. Southeast Asia, in particular, was identified as a biodiversity hotspot for blood-feeding flies. Objective 2: to interpret the phylogenetic and ecological relationships of flies of medical and veterinary importance and their associated symbiotes. By establishing phylogenetic relationships of species in numerous groups of blood-feeding flies, insights were gained into mechanisms of diversification, including speciation. In one large, widespread species complex of economically important biting flies, the Simulium cholodkovskii complex, chromosomal and genomic restructuring phenomena included the differential expression of individual chromosomal rearrangements (i.e., as fixed, polymorphic, or sex linked) in different species, species-specific differentiation of sex chromosomes, and reciprocal translocations of chromosome arms. Although rather rare, translocations were documented in about 12 independent lineages of the family Simuliidae. At least six of these lineages were associated with subsequent speciation events, suggesting that translocations have facilitated adaptive radiations. This work provides the basis for future comparative genomic research to elucidate the evolution of blood feeding in biting flies, and consequently, to provide a platform for understanding the development of pest and vector species. Objective 3: to develop solutions to problems caused by biting flies. Procedures were outlined for developing pest-management plans for blood-feeding flies. This work focused on endangered birds that experience fitness losses when they are attacked by swarms of blood-feeding flies. Solutions to the pest problem were developed with the recognition that multiple management actions are possible, including, for example, suppression of the pest population via biological control or relocation of the endangered species. A decision-making framework was outlined as a procedural blueprint that can be used to determine how to advance conservation of endangered species when there are competing objectives, such as the need to ensure the viability of an endangered species while also preserving the ecological function of native blood-feeding flies.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Sirin, �. D. & P. H. Adler. 2015. Chromosomal and morphological taxonomy of a new species of black fly in the genus Metacnephia (Diptera: Simuliidae) from Western Anatolia, Turkey. Zootaxa 4048: 269-280.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Lehnert, M. S., C. E. Beard, P. D. Gerard, K. G. Kornev & P. H. Adler. 2016. Structure of the lepidopteran proboscis in relation to feeding guild. Journal of Morphology 277: 167-182.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adler, P. H., T. K�delov�, M. K�dela, G. Seitz & A. Ignjatovi?-?upina. 2016. Cryptic biodiversity and the origins of pest status revealed in the macrogenome of Simulium colombaschense (Diptera: Simuliidae), historys most destructive black fly. PLoS ONE 11(1): 1-25 e0147673. Doi: 10.1371/ journal.pone.0147673
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adler, P. H., A. Yildirim, Z. Onder, G. T. Tasci, O. Duzlu, M. O. Arslan, A. Ciloglu, B. Sari, N. Parmaksizoglu & A. Inci. 2016. Rearrangement hotspots in the sex chromosome of the Palearctic black fly Simulium bergi (Diptera: Simuliidae). Comparative Cytogenetics 10: 295-310. Doi: 10.3897/CompCytogen.v10i2.8855
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adler, P. H., B. Belqat, J. Garrido Gonz�lez, A. J. P�rez & G. Seitz. 2016. Chromosomal relationships of Simulium armoricanum and its undescribed sister species in the Simulium vernum species group (Diptera: Simuliidae). Zootaxa 4137: 211-222.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adler, P. H., O. Yadamsuren & W. S. Procunier. 2016. Chromosomal translocations in black flies (Diptera: Simuliidae)facilitators of adaptive radiation? PLoS ONE 11 (6): e0158272. 23 pp. Doi: 10.1371/ journal.pone.0158272
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adler, P. H., H. Takaoka, M. Sofian-Azirun, V. L. Low, Z. Yacob, C. D. Chen, K. W. Lau & X. D. Pham. 2016. Vietnam, a hotspot for chromosomal diversity and cryptic species in black flies (Diptera: Simuliidae). PLoS ONE 11(10): e0163881. 29 pp. Doi: 10.1371/journal.pone.0163881
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Adler, P. H. 2016. Pathways to becoming pests and vectors: lessons from the Simuliidae. In: Arthropod-borne infectious diseases and arthropods as disease agents in human and animal health. Nova Acta Leopoldina (Neue Folge) 411: 23-32.


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

Outputs
Target Audience:The target audience reached during the past year included the scientific community of arthropod systematists, medical and veterinary entomologists, conservation biologists, and public health workers; the public, especially citizens affected by pest problems caused by biting flies; and undergraduate and graduate students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project continued to provide a training center for students and scientists from throughout the world to acquire new knowledge, skills, and techniques about the genetics, systematics, and ecology of biting flies. In this reporting period, a one-month workshop in insect cytogenetics was offered to a doctoral student from the Institute of Tropical Medicine, Belgium. How have the results been disseminated to communities of interest?Results of the project were disseminated through 4 peer-reviewed journal publications; 9 research presentations at regional, national, and international professional meetings and universities, including invited and keynote addresses (Germany, Turkey); presentations to the public, including one for a South Carolina Trust and one for an Arizona Rotary Club; a web site; and many routine phone, e-mail, and in-person responses to colleagues, the public, and students regarding information related to blood-feeding flies. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? IMPACT: Blood-feeding flies, because they breach the circulatory systems of animals, represent not only a physical stress often resulting in economic losses in the human and livestock sectors, but also a tremendous risk of diseases from pathogens and parasites transmitted while the flies are feeding. The diversity of blood-feeding flies--more than 13,000 species--presents a challenge to the prediction and control of the pest and disease problems they cause. Yet, an understanding of this biodiversity is directly related to the success of managing and controlling the problems. Emphasis was placed on revealing this biodiversity, especially the biodiversity that cannot be seen under a stereomicroscope but exists at the genetic level and often reveals different species with different ecological relationships, including tendency toward pest status and competency for transmitting agents of disease. An integrated approach was used, employing various microscopic, cytogenetic, and molecular techniques. Consequently, more than 10 new species were discovered and two were described formally. A recurring pattern in these studies is that a blood-feeding fly that has long been considered a single species by the scientific community is actually a composite of two or more species, not all of which are pests or vectors of disease organisms. History's most economically devastating black fly, for instance, consists of five genetically distinct entities, at least two of which are distinct species and only two of which have any medical or veterinary significance. Objective 1: to discover and describe the biodiversity of flies of medical and veterinary importance and their associated symbiotes (e.g., natural enemies) at scales ranging from the genome to the ecological community. Multiple new species were discovered in various parts of the world, including the United States, Colombia, Portugal, Turkey, and Vietnam. A world perspective of the biodiversity of biting flies allows general patterns of species diversity and pest and vector problems to be discovered. A compendium of all cytogenetic and chromosomal studies of the biting-fly family Simuliidae over the past 100 years was completed and published, providing an entry point for researchers worldwide to understand the current state of knowledge and to initiate novel research from an established base. Objective 2: to interpret the phylogenetic and ecological relationships of flies of medical and veterinary importance and their associated symbiotes. The phylogenetic relationships of several economically and ecologically important members of the genus Simulium were resolved. The most significant outcome of this work was an understanding of the major pathways by which species of black flies become economic pests of humans and domesticated animals. The phylogenetic inferences indicate that the acquisition of an ability to colonize large rivers, particularly in one large lineage of black flies, enhanced the probability that large populations of blood-feeding flies could be achieved. These large populations potentially translate into pest problems. The most serious pests, historically and presently, have been large-river species. Objective 3: to develop solutions to problems caused by biting flies. Two approaches to developing novel solutions to problems caused by blood-feeding flies, and possible means of disrupting the enabling traits, were investigated. One approach involved the identification of species traits, such as structural and behavioral characteristics, that enable certain species to become pests. For example, colonization of large rivers is one of the principal pathways by which black flies reach pest status. Identification of the features that allow these flies to exploit large rivers would allow the genes that code for these traits, such as strong adhesive silk, to be identified. Once the enabling features have been linked to the associated genes, the possibility of disrupting their expression could provide an alternative and proactive approach to pest management and vector control. The second approach involved understanding the mechanisms of fluid uptake via the tiny mouthparts of blood-feeding flies by investigating the larger mouthparts of butterflies as a model, with the goal of transferring an understanding of the physical mechanisms and constraints of fluid uptake by butterflies to blood-feeding by biting flies. Results from the investigation of butterflies indicate that the general mouthpart architecture promotes fluid uptake from droplets and films, based on principles such as capillarity, and that these principles are probably relevant for all fluid-feeding insects. The diversity of liquid foods, such as floral nectar and animal blood, suggests that specific structural adaptations for fluid uptake selected for by details of these different food sources, such as flower structure, are overlain on the fundamental structural organization of all mouthparts adapted for fluid feeding.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Adler, P. H., A. Inci, A. Yildirim, O. Duzlu, J. W. McCreadie, M. K�dela, A. Khazeni, T. Br�derov�, G. Seitz, H. Takaoka, Y. Otsuka & J. Bass. 2015. Are black flies of the subgenus Wilhelmia (Diptera: Simuliidae) multiple species or a single geographical generalist? Insights from the macrogenome. Biological Journal of the Linnean Society 114: 163-183.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Adler, P. H., M. Cherairia, S. F. Arigue, B. Samraoui & B. Belqat. 2015. Cryptic biodiversity in the cytogenome of bird-biting black flies in North Africa. Medical and Veterinary Entomology 29: 276-289.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Adler, P. H. & �. D. Sirin. 2015. A new species of black fly in the Prosimulium hirtipes (Fries) group (Diptera: Simuliidae) from Western Turkey. Proceedings of the Entomological Society of Washington 117: 85-94. Doi: 10.4289/0013-8797.117.2.85
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Adler, P. H. & R. W. Crosskey. 2015. Cytotaxonomy of the Simuliidae (Diptera): a systematic and bibliographic conspectus. Zootaxa 3975: 1-139.


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: The target audience reached during the past year included the scientific community of medical entomologists, public health workers, and conservation biologists; the public, particularly the citizens affected by pest problems caused by biting flies; and students from high school to graduate school. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Students and scientists from throughout the world routinely visit the laboratory of the project director to learn techniques, skills, and concepts related to the genetics, systematics, and ecology of biting flies. During this reporting period, atraining session of 1 monthwas provided for a scientist from Zentrum für Agrarlandschaftsforschung (Germany). How have the results been disseminated to communities of interest? Project results during this reporting period have been disseminated through four publications in peer-reviewed journals; 17 research presentations at regional, national, and international professional meetings; and many examples of direct transfer via e-mails and phone calls in response toinformation requests fromcolleagues, students, and the public. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? IMPACT: The cytogenomics, molecular biology, and ecology of biting flies in various regions of the world were studied to provide a general awareness of possible pest problems and vector threats. A world inventory of black flies, available freely on the internet, was maintained and updated. The inventory gives the names, classification, and distributions of all known black flies of the world, providing scientists, public health workers, and interested members of the public immediate access to information on black flies for any area of the world. An emerging threat from biting flies was investigated and involved the impact of biting flies, especially black flies, on endangered species. Intensive trapping and modelling of adult flies on the landscape and at nests of whooping cranes implicated at least one species of blackfly as a problem for the endangered cranes. The model now can be used to help guide the management of this critically endangered species. To begin understanding blood feeding by biting flies, the structure and function of the mouthparts of a nonblood-feeding group of insects, butterflies, were studied. The butterfly proboscis provides not only a beginning model for understanding blood feeding in biting flies, but also a model for designing medical probes capable of acquiring and transporting minute volumes of fluid. Objective 1: to discover and describe the biodiversity of flies of medical and veterinary importance and their associated symbiotes (e.g., natural enemies) at scales ranging from the genome to the ecological community. Intensive sampling of biting flies was conducted through international collaborations in Algeria, Armenia, Turkey, and the United States. Black flies in northeastern Algeria were sampled at 31 sites, revealing eight nominal species and species complexes in three genera among more than 31,000 specimens. The most abundant and ubiquitous species, representing nearly 80% of collected specimens, occupied the widest range of habitats, and prospered in habitats with anthropogenic influences. These species present perhaps the greatest pest and vector threats because they are adapted to breeding in streams and rivers in highly populated areas. Cytogenomic analysis of more than 500 larval black flies from 19 sites in Armenia and Turkey revealed five species of the genus Prosimulium, including two cytoforms and one species new to science. Objective 2: to interpret the phylogenetic and ecological relationships of flies of medical and veterinary importance and their associated symbiotes. Phylogenetic relationships were resolved for a number of species of mammal-feeding and bird-feeding black flies, such as flies inthe genus Prosimulium and the subgenus Eusimulium. Understanding these relationships provides insights into how and why some species of black flies become pests while others do not. Objective 3: to develop solutions to problems caused by biting flies. This objective involved two novel approaches to finding solutions for biting fly problems. One approach involved the butterfly proboscis as an initial model for understanding uptake of blood by biting flies. The butterfly proboscis has been treated historically as a sealed tube with fluid uptake restricted to the apex. This hypothesis was tested, and revealed that fluid uptake can occur along the length of the proboscis,but theinsects can control fluid entry by behaviorally straightening or flexing their proboscis. A second approach involved collaborative efforts with ecological modelers and conservation biologists to understand the nesting failure of a reintroduced population of the critically endangeredwhooping crane. The collaboration produced hierarchical models for daily nest survival. The modelling efforts and empirical data suggest that nest survival is negatively related to the abundance of at least one species of black fly. Thus, biting flies must be considered in management programs for whooping crane populations if breeding is to be successful.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Converse, S. J., J. A. Royle, P. H. Adler, R. P. Urbanek & J. A. Barzen. 2013. A hierarchical nest survival model integrating incomplete temporally varying covariates. Ecology and Evolution 3: 4439-4447. doi: 10.1002/ece3.822
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Cherairia, M., P. H. Adler & B. Samraoui. 2014. Biodiversity and bionomics of the black flies (Diptera: Simuliidae) of northeastern Algeria. Zootaxa 3796: 166-174.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Adler, P. H. & �. ?irin. 2014. Cytotaxonomy of the Prosimulium (Diptera: Simuliidae) of Western Asia. Zoological Journal of the Linnean Society 171: 753-768.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Kwauk, K., D. K. Hasegawa, M. S. Lehnert, C. E. Beard, P. D. Gerard, K. G. Kornev & P. H. Adler. 2014. Drinking with an unsealed tube: fluid uptake along the butterfly proboscis. Annals of the Entomological Society of America 107: 886-892.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Adler, P. H. & G. Seitz. 2014. Chromosomal characteristics and evolutionary relationships of the Palearctic black fly Simulium carthusiense (Diptera: Simuliidae). European Journal of Entomology 111: 469-474.


Progress 01/01/13 to 09/30/13

Outputs
Target Audience: The research and teaching efforts associated with this projectreached (1) the worldwide scientific community, which further disseminated the information via citations in additional publications and media; (2) the general public (local, national, international), especially via web-based information on insects of medical-veterinary importance; (3) the pest-management community who uses the information to improve control programs for insects of medical-veterinary importance; and (4) national and international students and colleagues interested in gaining knowledge of insects of medical-veterinary concern by attending one- to three-month long workshops offered in the research laboratory. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Training Activities. Study and research opportunities were offered by the Principal Investigator on the project to international students and scientists. The objective was to enhance, train,and expand the cooperative global network of scientists conducting research on bloodfeeding flies. The study and research curriculum was designed, directed, and taught by the Principal Investigator in his laboratory, and involved training in genetics, systematics, fundamental biology and ecology, and biodiversity of bloodfeeding flies. Six participants attended for one to three months from Algeria, Colombia, and Slovakia. Professional Development. The following conferences were attended to exchange and acquire new information related to biting flies of medical-veterinary importance: Annual Meeting, North American Black Fly Association, Athens, GA. Freshwater Animal Diversity Assessment Meeting, Ostend, Belgium. International Conference on Biodiversity and Integrated Pest Management, Manado, North Sulawesi, Indonesia. Annual Member’s Meeting of the International Crane Foundation, Baraboo, WI. How have the results been disseminated to communities of interest? Several research and educational oral presentations related to the projectwere given to the scientific community and the public: Adler, P. H. & E. W. Gray. 2013. Populations of black flies on the landscape of an endangered species. 11th Annual Meeting, North American Black Fly Association. Athens, GA. Adler, P. H. & D. C. Currie. 2013. Diptera: Simuliidae. 3rd Freshwater Animal Diversity Assessment (FADA) Meeting, Ostend, Belgium. Adler, P. H. 2013. Biodiversity and integrated pest management of biting flies: taking a bite out of the conflict. 1st International Conference on Biodiversity and Integrated Pest Management: Working Together for a Sustainable Future. Manado, North Sulawesi, Indonesia. Adler, P. H. 2013. Biology of black flies. Annual Member’s Meeting of the International Crane Foundation. Baraboo, WI. McCreadie, J. W. & P. H. Adler. 2013. The North American larval black fly database. 11th Annual Meeting, North American Black Fly Association. Athens, GA. Williams, R., J. W. McCreadie, P. H. Adler & S. Stutsman. 2013. Gamma and taxonomic diversity of preimaginal black flies (Diptera: Simuliidae) of the mid and eastern United States. 11th Annual Meeting, North American Black Fly Association. Athens, GA. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Bloodfeeding flies, such as mosquitoes and black flies, transmit the agents of dozens of diseases of humans and domestic animals, and as nuisance pests they cause enormous economic losses. However, only the female flies of most species create these risks. The males and larvae perform key functional roles in the environment, such as pollination and organic matter processing that provides food for other organisms. Thus, a tradeoff exists between controlling the problematic female flies and conserving the beneficial life stages. The research associated with this project contributes fundamental scientific knowledge in areas of ecology and biodiversityon a global scale, including insights into the genetic and environmental factors that drive pest status, while also facilitating theprotection of public and animal health. Objective 1) To discover and describe the biodiversity of flies of medical and veterinary importance and their associated symbiotes (e.g., natural enemies) at scales ranging from the genome to the ecological community: Surveys of biting flies, particularly the Simuliidae,were conducted in a number of areas of the world, from the southeastern United States to Europe, North Africa, Japan, and western Asia. All of these surveys involved morphological and cytogenetic analyses to detect genomic and organismal biodiversity. These surveyswere conducted by (1) on-site visits (southeastern United States), (2) scientists in other parts of the world (e.g., multiple European countries, Japan, Malaysia, Turkey) who conducted surveys and sent the material to the laboratory of the Principal Investigator for analysis, and (3) students from other countries (Algeria, Iran, Slovakia, Colombia) who conducted surveys in their countries and then visited the laboratory of the Principal Investigator, with their survey material. to learn techniques and skills for cytogenomic and taxonomic analyses. The application of cytogenomic and morphological analyses to the material revealed a wealth of new biting-fly biodiversity, including more than 10 new species and numerous chromosomally distinct forms (cytoforms). The discovery of multiple species of biting flies in what previously were considered single species is remarkable. Some of these putativespecies have been serious medical and veterinary concerns for years, with pest management applied as though a single species had been involved. The analysesrevealed that not all of the newly discovered species within what previously were considered single species are actually pests. Thepractical implication is that pest management now can be more precisely targeted against the specific pest species.Black flies from oceanic islands were used to test the utility of a macrogenomic (chromosomal) approach for determining geographic origins of insular populations. The test case used a black fly from Okinawa Island and comparative populations throughout the mainland of Asia. During the analysis, new species of black flies were discovered. The island population was shown to be chromosomally unique and to have originated from the main islands of Japan rather than from the Asian mainland or fromnearbyTaiwan. The macrogenomic approach applied to biting flies illustrates its potential for defining source areas of invasive species of flies that might pose medical and veterinary risks. Objective 2) To interpret the phylogenetic and ecological relationships of flies of medical and veterinary importance and their associated symbiotes: The discovery of new biting-fly biodiversity provides the first step in recognizing and understandingits implications and significancefor medical-veterinary entomology and pest management. To provide a database of these new biodiversity discoveries, integrated with existing knowledge, a world inventory of black flies was placed on the web, with the intent of updating it annually. For the 2013 update, all informally and formally named species, cytoforms, and molecular forms of black flies and their geographic areas of occurrence were included. Researchers throughout the world now canuse this database to guide their ecological and phylogenetic work. All biodiversity discovered under Objective 1 was also analyzed phylogenetically, largely using cytogenomic information. Several thousand larvae representing multiple species groups in three subgenera of black flies were analyzed cytogenetically to provide characters for phylogenetic inference. Analyses also were conducted using chromosomal outgroups so that actual phylogenies, rather than unrooted dendrograms, could be obtained. From these phylogenetic analyses, insights were gained into the evolution and development of pest status. For example, of the five cytoforms discovered in the notorious Golubatz fly of Europe, only two have pest status and they are the two that are most closely related in the phylogeny. Phylogenetic analysis also provided insight into the evolution of the characters responsible for driving pest status, especially the ability to achieve extraordinarily high levels of abundance. Objective3) To develop solutions to problems caused by biting flies: We used trichomycete fungi, which live in the digestive tracts of arthropods, to begin understanding the potentialfor using symbiotic organisms from one hostspeciesas possible agents of infection in nontypical host species. Specifically, we investigated the ability ofa midgut trichomycete fungus (Harpella sp.) that typically lives in larval black flies to colonize larval mosquitoes, which would represent novel hosts. Mosquitoes have never been documented to harbor these midgut fungi. To test the suitability of mosquito midguts for hosting these fungi, we exposed them to trichospores of Harpella obtained from wild larval black flies. The prevalence of fungi in the mosquito midguts and in the controls was recorded. The spores were ingested by mosquito larvae of three species and exhibited growth in the midguts of two species.In the various replicates, up to two-thirds of larval mosquitoes were colonized by the Harpellafungi. In light of the ability of the fungi to colonize larval mosquitoes, we suspect that environmental factors might be more important than physiology in explaining why mosquitoes lack midgut trichomycetes in nature. Larval mosquitoes tend to inhabit still water, while larval black flies are found exclusively in running water.The results suggest that symbiotes of larval black flies can be transferred to nontypical (mosquito) hosts. Although midgut fungi in larval black flies are believed to be commensalisitc, they also are known to become parasitic in adult black flies. Thus, the study provides a model for working with potential biological control agents for biting flies. The study also provides insight into the physiology of the insect gut in relation to the digestive tract as an environment for other living organisms (symbiotes).

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Khazeni, A., P. H. Adler, Z. Telmadareiiy, M. A. Oshaghi, H. Vatandoost, S. M. Abtahi & A. Lofti. 2013. The black flies (Diptera: Simuliidae) of Iran. Zootaxa 3694: 67-74.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Adler, P. H., Y. T. Huang, W. K. Reeves, S. K. Kim, Y. Otsuka & H. Takaoka. 2013. Macrogenomic evidence for the origin of the black fly Simulium suzukii (Diptera: Simuliidae) on Okinawa Island, Japan. PLOS ONE 8(8): e70765. doi:10.1371/journal.pone.0070765
  • Type: Websites Status: Published Year Published: 2013 Citation: Adler, P. H. & R. W. Crosskey. 2013. World blackflies (Diptera: Simuliidae): a comprehensive revision of the taxonomic and geographical inventory [2013]. 120 pp. http://www.clemson.edu/cafls/departments/esps/biomia/pdfs/blackflyinventory.pdf


Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: This project was initiated during the current reporting period, and was focused on research to expand knowledge of the diversity of insects that influence the health and welfare of humans, domestic animals, and wildlife. A network of international collaborations was developed to monitor insects of medical and veterinary importance and to determine the factors that influence their geographic distributions, dispersal, and variations in pest status. Selected species of bloodfeeding flies with wide distributions and strong dispersal abilities, such as the capacity to colonize remote islands, were investigated as models for understanding the factors that drive dispersal and distribution. Integrated approaches were used to probe the underlying genetic structure of these model insects. The influence of bloodfeeding flies on endangered wildlife was investigated to provide more effective means of managing the endangered species. The work was disseminated by routine correspondence with the scientific community and the public, by presentations at professional meetings, by reports to environmental and conservation agencies, through a university web site centered around the research, and by incorporating the information in classes taught by the lead investigator. PARTICIPANTS: The principal investigator (P. H. Adler) designed or co-designed and directed the research and conducted laboratory and field research, data analysis, presentation of results at professional meetings, and preparation of publications. C. E. Beard (Research Technologist) provided technical support in all phases of the project and conducted research on natural enemies of medically important flies. Collaborations were in place with numerous colleagues worldwide, the U.S. Fish & Wildlife Service, and the International Crane Foundation. TARGET AUDIENCES: The beneficiaries of this research included researchers and students worldwide, (2) citizens of South Carolina, whose health and welfare was addressed by investigations of insects of medical and veterinary importance, and (3) policy makers involved in government health and environmental programs, who received information related to insects of medical and veterinary importance. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Threats from invasive insects of medical and veterinary importance are increasing, especially with greater globalization and climate change. The ability to proactively address these threats is based on a sound understanding of organisms, from the genetic to the community level. Macrogenomic profiles in the giant chromosomes of remote-island black flies were compared with those of mainland populations on two continents, enabling the discovery of the mainland source from which the island populations were derived. The success of this test case demonstrated that the macrogenomic approach provides a powerful tool that can be used to determine the geographic area from which an invasive pest species arrived. Similar analyses of black flies with wide geographic distributions revealed multiple, heretofore unrecognized forms and species with more restricted distributions. Within species, the analyses also identified areas of high and low levels of macrogenomic variation, typically involving a reduction in variation in populations toward at least part of the periphery of the distribution. These patterns probably reflect the length of time the populations have been in the area, with less variation in more recently colonized areas. The project provided insights into movements of invasive bloodfeeding insects, which can be used as the basis for improved detection and management of these medically important insects.

Publications

  • No publications reported this period