1. MOLECULAR DIFFERENCES RESULTING FROM LARVAL GROWTH CONDITIONS IN AEDES AEGYPTI DAVID PRICE DISEASE VECTOR MOLECULAR BIOLOGY LAB NMSU PI IMMO HANSEN

2. Background  Aedes aegypti is an important disease vector  Relatively safe model for malaria Plasmodium gallinaceum  Existing Reference Genome

3. Aedes/Dengue Situation http://www.healthmap.org/dengue/index.php

4. Small vs Large Mosquitoes  Larval conditions affect adult insect  Crowding, food availability -> Changes in adult size, time to reach adulthood, immunity, reproductive success, energy stores, life span, fecundity on first blood meal, vectorial capacity

5. Fat Body

6. Bloodfeeding  Purdue Medical Entomology PBM- Post Blood Meal NBF- Not Blood Fed

7. Hypothesis  Significant metabolomic, transcriptional and metabolic differences exist in the fat body between small and large mosquitoes, before and after blood meal.

8. Methods  Metabolic CO 2 production  Metabolomic GC/MS, LC/MS, LC/MS/MS analysis  Transcriptomic RNA-Seq approach using Illumina sequencing

9. Metabolomics  Questions:  Map metabolic pathway up/down regulation after blood meal Price et al, unpublished data

10. Metabolic  Questions:  Do small and large mosquitoes burn energy at the same rate?  Are the substrates being used for energy the same?  In conjunction with other work: Is there a difference in how far energy stores go?

11. Metabolic Price et al, unpublished data, Sable Systems

12. Transcriptomic  Questions:  Determine changes in gene expression between mosquitoes raised under different nutrient conditions  What patterns of expression are present?

15. Conclusions  Used metabolomic, metabolic, and transcriptional descriptive work to identify differences between small and large mosquitoes; pre and post bloodmeal.  Immune regulation is curious  Results of small vs large vectorial capacity experiments have been conflicting  Signs of autophagy  No apparent metabolic difference

16. Hansen Lab/Collaborators  Alexander Ulanov – University of Chicago, Urbana  Wayne Van Voorhies - NMSU molecular Biology  NIAID/NIH

17. Bloodfeeding  Pre and Post Bloodmeal  Bloodfeeding is required for egg production in Ae. Aegypti.  During blood feeding, body weight more than doubles in a minute.  Three day process, bloodmeal to eggs (Ave. 55)  Steinwascher, American Midland Naturalist. Vol 112 No 1.  Metabolic peak at 24 hours  Gray and Bradley, Parasitology. Feb 2006.

18. Fat Body Physiology  Function similar to a liver and fat tissue  Most metabolic pathways  Energy - Fat and Glycogen stores  Nutrient stores  Synthesizes and supplies yolk proteins for vitellogenesis to the ovaries

19. Metabolic  Experiment in planning  Larger groups of mosquitoes over course of 4 or 5 days to determine RQ  RQ is ratio of CO 2 produced to O 2 consumed 1 Carbohydrate 0.9 Protein 0.7 Lipid

20. Vector Control Methods Using Mosquitoes  Sterile insect technique  Release of sterile males to reduce population numbers and interrupt disease transmission  Population replacement  Upregulation of immune system components typically leads to increased fitness cost  Increased fitness cost in the wild could lead to less fit mosquito  Population reduction  Change the average size of a mosquito and what will happen to a wild population

21. Future  Sequence Small and Large mosquito transcriptomes, pre and post bloodmeal  Connect transcriptional data to metabolomic Enzymes to metabolites – KEGG  Identify differences which may be important for nutrient accumulation and explore use in vector control  Infection response  Dengue, plasmodium

23. unfed 24 h PBM 48 h PBM PBM- Post Blood Meal NBF- Not Blood Fed