Student Engagement and Vector Biology Research at Western Carolina University

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1 Student Engagement and Vector Biology Research at Western Carolina University Brian Byrd and Bruce Harrison Environmental Health Sciences Program College of Health and Human Sciences Cullowhee, NC 28723

2 Cullowhee, NC Western Carolina University

3 My Role at WCU 9 Month Faculty (Env. Health Science) Teaching (Undergraduate): Medical Entomology, Epidemiology, Etiology of Infectious Diseases, Freshman Seminar Research: La Crosse Encephalitis Eco Epidemiology Molecular Assays for Mosquito ID Service Intramural Extramural

4 Student Engagement Undergraduate Research Experiences Service Learning Engaged Teaching

5 La Crosse Encephalitis Eco epidemiology of Disease (LACv Infection) Vertical Distribution Studies Molecular ID of Mosquitoes Spatial Epidemiology of LACE CDC CDC Byrd

6 La Crosse Encephalitis: Mosquito-Borne Disease in our Backyard Haddow AD and Odoi A, The incidence risk, clustering, and clinical presentation of La Crosse virus infections in the eastern United States, PLoS One Jul 3;4(7):e6145.

7 Molecular Identification (rdna ITS2) [L] [1] [2] [3] [4] [5] [6] [7] [8] Ae. albopictus Ae. triseriatus Ae. japonicus 600 bp [L] Lane No. 1 Ae. aegypti 2 Ae. albopictus 3 Ae. bahamensis 4 Ae. triseriatus 5 Ae. mediovittatus 6 Ae. japonicus 7 Ae. atropalpus 8 Negative Control Genus species 100 bp L 100 bp Ladder (Invitrogen) PRIMERS: CP-P1A gtggatcctgtgaactgcaggacacatg CP-P1B gtgtcgacatgcttaaatttagggggta *

8 Aedes atlanticus and Ae. tormentor Current Problem: Morphologically Indistinguishable as Adult Females A Aedes atlanticus B Aedes tormentor

9 ITS2 Seqs Distinguish the Two Species and Validate a Novel Morphological Character Blind 015 H01 Blind 005 C01 ATLANTICUS-CBS-2-2 ATLANTICUS-CBS-2-1 Blind 001 A01 Blind 003 B01 Blind 013 G01 TORMENTOR-CBS-1-2 TORMENTOR-CBS-1-1 M13F Blind 011 F01 Blind 002 A02 Blind 009 E01 Blind 004 B Hpy188I Sequence Recognition Site 5 TCNGA 3 3 AGNCT 5 Hpy188I Digest (Predicted) Ae. atlanticus Ae. tormentor Aedes atlanticus Aedes tormentor NEBcutter V2.0

10 Hpy88I Restriction Digest Assay Aedes atlanticus ITS2 PCR Product Digested ITS2 Aedes atlanticus (Undigested ITS2) Aedes atlanticus (Digested ITS2) Aedes tormentor ITS2 PCR Product Digested ITS2 * Aedes tormentor (Undigested ITS2) Aedes tormentor (Digested ITS2) Agilent 2100 BioAnalyzer electropherograms demonstrate successful Hpy188I digestion of ITS2 PCR products. Post-digest products match predicted sizes. *This additional peak is due to intraindividual sequence heterogeneity (common near the 5 end of ITS2) in Aedes tormentor.

11 Vertical Distribution Studies CDC CDC Byrd Image (HOPE)

12 Trap Type Studies Determine the best trap type to collect adult container inhabiting mosquitoes. Aedes triseriatus Aedes albopictus Aedes japonicus Mosquitoes Mosquitoes Mosquitoes BGS CDC FP Gravid 0 BGS CDC FP Gravid 0 BGS Gravid FP CDC Gonotrophic Differences: BG-Sentinel, CDC, FP (Host Seeking) Gravid (Gravid)

13 Student Engagement: Teaching ENVH 430: Medical Entomology ENVH 470: Epidemiology ENVH 499: Undergraduate Research ENVH 493: Special Topics Molecular ID of Pathogens

14 Medical Entomology ENVH 430: Medical Entomology 3 hour lecture Tuesday and Thursday (1 2:15) 1 hour lab Tuesday (2:30 4:30)

15 Medical Entomology Lecture Lab

16 Medical Entomology Collection

17 Medical Entomology: Fall 2011 Hobcaw Barony, Georgetown SC Mosquitoes (Found at HB) Aedes albopictus Aedes vexans Anopheles crucians s.l. Anopheles quadrimaculatus s.l. Culiseta melanura Culex erraticus Culex restuans Culex territans Culex salinarius Culex nigripalpus Ochlerotatus fulvus pallens Ochlerotatus atlanticus Ochlerotatus infirmatus Ochlerotatus taeniorhynchus Ochlerotatus tormentor Ochlerotatus sollicitans Ochlerotatus c. canadensis Psorophora ciliata Psorophora columbiae Psorophora ferox Uranotaenia sapphirina

18

19

20 New Building: August (Research Lab, Insectary, Teaching Lab)

21 Lab Capacities Microscopy/Mosquito ID Dissecting Scopes (90X) Adult and Larval Identification Molecular Capabilities DNA Amplification (PCR) Cloning Cell Culture LACv Sequencing GIS Forensic Science Program ArcGIS Collection Traps CDC Light Traps Gravid Traps BG Sentinels

22

23 Prep/Autoclave Room 80 C Ultralow Freezer Dishwasher Gel Documentation System Chemical Hood Autoclave

24 Teaching Lab (Microbiology/Medical Entomology)

25 Arthropod Containment Lab (Insectary)

26 Undergraduate Research Group (Previous Members) Alan Goggins Mike Singleton Jamie Wyatt Samantha Kunze Charlie Sither Erin Gymburch Billy Wiseman

27 Acknowledgements Resources: WCU Faculty Research Grant NC Biotechnology Center WCU Quality Enhancement Program NC DENR WCU Honors College WCU College of Health and Human Sciences Collaborators: Dr. Bruce Harrison (WCU) Dr. Gideon Wasserberg (UNC-G) Forensic Science Program (WCU) Natural Resource Program (WCU) Dr. Sheri Denslow (Mission Hospital) Dr. Carl Williams (NC DHHS) Brian Byrd, PhD, MSPH Environmental Health Sciences College of Health and Human Sciences Cullowhee, NC (828)

28 Who s your Redneck Daddy? Bruce Harrison Affiliate Professor WCU

29

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