BAMA Article Assignment 7/30/13

1. Preventing the spread of infectious diseases through the endosymbiotic relationships with Wolbachia By Lois Crozier BAMA Teacher Johnsonburg Area School District, Johnsonburg, PA BAMA Article Assignment 7/30/13

2. Article: Doudoumis, V., Alma, U., Askoy, E., Abd-Alla, A. M. M., Tsiamis, G., Brelsfoard, C., Aksoy, S., & Bourtzis, K. (2013). Tsetse-wolbachia symbiosis: Comes of age and has great potential for pest and disease control. Journal of Invertebrate Pathology, 112, S94-S103. doi: PMID: 22835476 Tsetse-Wolbchia symbiosis: Comes of age and has great potential for pest and disease control

3. Tsetse fly Order: DipteraFamily: GlossinidaeGenus: Glossina Vector in the transmission of Trypanosomiasis the cause of African Sleeping Sickness

5. thread-like parasitic nematodes (roundworms) transmitted by arthropod vectors microfilariae infect vector arthropods, in which they mature to larvae adult worms inhabit specific tissues where they mate and produce microfilariae, thread-like larvae. microFilariae ! What are they?

7. 60 million people live in regions where the parasite is located. • African Animal Trypanosomiasis is the biggest problem in preventing the increase of livestock populations • African Animal Trypanosomiasisis the leading cause of hunger and poverty The Cost of Trypanosomiasis

8. Current vector control is not sustainable or environmentally friendly. Urgent need for pesticide free, environmentally friendly method of vector control The Point?

9. http://www.rochester.edu/college/bio/labs/WerrenLab/WerrenLab-WolbachiaBiology.htmlhttp://www.rochester.edu/college/bio/labs/WerrenLab/WerrenLab-WolbachiaBiology.html http://www.who.int/mediacentre/factsheets/fs259/en/ http://emedicine.medscape.com/article/214581-overview

10. 1. Wigglesworth glossinidia Found in 2 distinct populations in the insect: Intracellular population in the specialized epithelial cells in the end gut Plays a role in providing essential vitamins and minerals absent from a blood diet Extracellular population in female milk secretion Method of transmission to the intrauterine larvae through mothers milk. An absence of Wigglesworth g. results in female sterility Wigglesworth is heterozygous in the natural population influences host juvenile larvae development of immune system 2.Sodalis glossinidius Present in all lab populations and heterozygous in natural populations Distributed in many tissues including haemolymph, salivary gland, milk gland, an midgut where it lives close to where trypanosomes develop. Passed to intrauterine larva through mothers milk 3 Symbiotic Bacteria in the Tsetse fly

11. 3. Wolbachiapipiens • Obligatory endosymbiont • (lives within the organism and either the endosymbiont or the host cannot survive without the other) • maternally transmitted • Found in reproductive tissue of insect

12. Parthenogenesis( female reproduction without fertilization ) • Feminization ( male killing, male zygotes don’t survive ) • Cytoplasmic Incompatibility (CI) • Most common • Can be unidirectional or bidirectional • Results in a reduction in the population of the host Effects of Wolbachiapipiens in Host

13. Incompatible Insect Technique (IIT) • IITcauses sterilitydue to the presence of a Wolbachiastrain genetically capable of inducing cytoplasmic incompatibility. • IIT Has been done successfully in the lab and in the field Another method to stop transmission

14. Wolbachia-induced CIcan control populations of agricultural pests and disease vectors • might someday play a role in spreading or replacing desired phenotypes (because infected females have a reproductive advantage of passing their DNA onto the offspring ) • Methods for inducing the transfer of desired genes have yet to be worked out fully. Incompatible Insect Technique (IIT)

15. Infected Uninfected Offspringwith withWolbachiaWolbachia No Yes Yes Yes No Higher frequency in the Population Proportional to the number of infected males Yes Yes Yes

16. vector genus of tsetse flies is Glossinia • http://www.who.int/mediacentre/factsheets/fs259/en/ • Using 16S rRNAbased PCRanalysis • 3,750 flies from 12 species from 10 African countries were tested • Found multiple cytoplasmic strains and Wolbachia gene insertions on the Glossinia chromosomes ( host DNA) GlossiniaandWolbachia

17. 3 Wolbachia genes were found on the host DNA of Glossiniam. morsitans in natural and lab populations Horizontal Gene TransferandWolbachia • 1. 16S rRNA(used in classifying and identifying bacterial species) • 2. fbpA(ferric iron binding protein-encoding gene or fibronetin binding protein) • 3. wsp(Wolbachia surface protein)

18. led to the identification of at least 2 horizontal gene transfer events 52% of Wolbachia genome transferred in one event and 47% of Wolbachia genome being transferred in the other event. • Whole Genome Sequence of Glossiniam.morsitans

19. The HGT of Wolbachiagenome can provide clues into the evolution and transfer of endosymbiont genes into multicellular organisms aiding in the development of Wolbachiaas a transport device to get desired genes into vectors such as the tsetse fly to stop the spread of African Trypanosomiasisand other diseases spread by insects. Possibilities and Hope

20. disease produced by a filarial worm depends on the tissue locations preferred by adults and type of microfilariae • lymphatic filariasis • disease is caused by the presence of worms in the regional lymphatic vessels • and by the host response to the worms and worm products Diseases

21. Elephantiasis of leg caused by chronic infection with the filarial nematode Wuchereriabancroftispread by a mosquito vector • Onchocercavolvulus is a filarial worm that is transmitted to humans by blackflies. • Mature worms live in the subcutaneous tissues and produce microfilariae that migrate through the skin and connective tissues. • do not move through the circulatory system • 3-4 months after infection, form nodules on outer layer of connective tissue which turns into calcifications Other Filarial Nematode infections

23. HAT and AAT are serious problems in Africa and in North America someday, as climates change. Insecticides are not environmentally responsible or sustainable, neither are baiting or releasing sterilized male tsetse flies. Wolbachia has the potential to rid the world of this sickness and others, one fly at a time, one generation at a time. Wouldn’t that be nice. Summary

24. 16S ribosomal RNAis a component of prokaryotic ribosomes having 1,542 nucleotides used in reconstructing phylogenies multiple sequences of 16S rRNA can exist within a single bacterium What is 16s rRNA?

25. Carl R.Woese, an American microbiologist and biophysicist along with George E. Fox, a researcher at the University of Houston, were well known for their defining work in identifying a new domain in classifying organisms, Archaea. • by phylogenetic taxonomy of 16S rRNAstemming from their introduction of the lowest common ancestor (a progenote). • comparative analysis of ribosomal RNA • which they believed was the earliest genetic resemblance to life on Earth today. 16s rRNA(continued)

26. PCR applications of 16S rRNA highly conservative primer binding sites making PCR easier ( very few differences in primer sites for the gene) Has hyper variable regions in its gene sequence making gene sequences making it a very species specific tool used to classify bacteria Used in medical microbiology due to it being fast and inexpensive, compared to using phenotypes to identify bacteria Originally used to classify bacteria now used to identify new species of bacteria http://en.wikipedia.org BACK