Discover the Microbes Within! The Wolbachia Project

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Discover the Microbes Within! The Wolbachia
Project
Real-world Research in the Classroom
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Welcome to Woods Hole!April 11, 2008
This project was one of the most fascinating
things I have ever done in the research area. I
really enjoyed the idea that we, as high school
students, are part of a big research project
which is going to benefit future generations of
scientists. Maggie Pakalska, 3/5/08, Junior,
Lake Region HS, Maine
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Fact Most high impact scientific research is the
product of interdiscipilinary teams Then
Participation in a community of biologists is
essential to teach effectively what it really
means to be a scientist
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Deb
Laurel
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The Marine Biological Laboratory
Charles Whitman 1st Director of the MBL
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• The Marine Biological Laboratory
• International center for research, education,
  and training in biology
• Founded in 1888
• Oldest independent marine laboratory in the
  Americas
• 55 Nobel Laureates
• 275 year round staff
• 1500 summer personnel
• Bay Paul Center Microbial diversity, evolution,
  ecology, and genomics

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Why Are WeHere?
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To bring real-world research to your classroom
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2007 Binghamton HS
To engage high school students in nature and
discovery-based science
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To enhance student interest in science spanning
bio-diversity to bio-informatics
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Some interesting outcomes
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To encourage nationwide participation in the
collection of new data
Rochester, NY
Ithaca, NY
-Falmouth, MA
-Woods Hole, MA
Bronx, NY
Loudon County, VA
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Discover the Microbes Within!The Wolbachia
Project

• 1,000,000,000,000,000,000 insects on the planet
• Insects present on earth for 500 MY
• 85 of all animal species are insects (1-30
  million species)
• 20 of all insect species harbor heritable
  symbionts called Wolbachia
• Scientists cant discover them all on their own.
• YOU ARE OUR BIGGEST ASSET TO SCIENTIFIC DISCOVERY
  AND REAL RESEARCH!

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What is symbiosis?

• The living together of dissimilar organisms (de
  Bary 1859)
• Often for mutual benefit, but also parasitism and
  commensalism
• Endosymbiosis is when one organism lives and
  replicates inside another one

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Microbial Endosymbiosis In Eukaryotic Evolution
2.0 Bya
Origin of mitochondria
Human Microbiome
1.5 Bya
Animals and land plants evolve in microbial soup
0.5 Bya
Today
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SYMBIOSIS QUIZ
What of the cells in your body are human?
10 30
60 90
Raise your hand to answer
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Vertically-transmitted(i.e., Inherited)
Symbionts
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Insect egg with symbiotic bacteria
Insect egg
Credit Michael Clark Seth Bordenstein
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Ways that vertically transmitted microbes can
increase in frequency

• Increase host survival reproduction (mutualism)
  
• Very common
• Why might vertical transmission be associated
  with mutualistic effects on hosts?
• Most famous cases are the lineages leading to
  organelles
• Mitochondria evolved from the alpha-Proteobacteria
  about 2 billion years ago
• Chloroplasts evolved from cyanobacteria about 1
  billion years ago

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Beneficial microbes in animal hosts
Aphids-Buchnera
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Buchnera
Aphid eggs (Uroleucon ambrosiae)
A. Mira
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Must heritable symbionts always be beneficial?
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Reproductive Parasites
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Asymmetric (Uniparental) Inheritance
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Wolbachia are Infectious Widowmakers!
Male-Killing
Parthenogenesis
Feminization
Reproductive Parasitism each of these
reproductive distortions leads to more infected
females in the insect host species
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Cytoplasmic Incompatibility (CI)
CI
X
x

Wolbachia-infected offspring
x

Uninfected offspring
x

Wolbachia-infected offspring
x

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Wolbachia Master Manipulators
Courtesy of Bandi Sacchi
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Imagine
physicians across the United Statesdocumenting
similar events. In each case, an unfertilized egg
in a woman had spontaneously begun to develop,
ultimately producing a healthy female baby. One
young researcher, who had analyzed the timing and
locales of the virgin births, suggested a
spreading infection might be causing the
incidents. The Federal Centers for Disease
Control and Prevention in Atlanta quickly
dismissed the idea, calling it "ridiculous." Seve
ral months later came a well-publicized report in
the Journal of the American Medical Association
concluding that the number of infertile couples
was rising rapidly worldwide. The international
uproar intensified when physicians began to
observe another reproductive curiosity Some
newborns that were genetically male appeared to
be female. One week, the New England Journal of
Medicine and the National Enquirer ran articles
with the headline, "Is this the end of mankind,
or just men?" Science fiction? Definitely. For
many insect species and other arthropods,
however, the truth can be as strange as fiction
when bacteria known as Wolbachia are around.
By JOHN TRAVIS Undesirable Sex
Partners Bacteria manipulate reproduction
ofinsects and other species
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The Wolbachia Pandemic
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The Wolbachia Pandemic
Insects (at least 20 !)
Filarial nematodes
Crustaceans
(family Onchocercidae)
Chelicerates
2-6 million insect species are infected with
Wolbachia!!
Arthropods
Nematodes
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Number of scientific papers with Wolbachia in
title
PCR detection of Wolbachia in 20 of all insects
Nobel prize awarded for PCR
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(No Transcript)
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How do YOU discover the Wolbachia within?
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Integrated Set of Lab ExercisesFrom Organisms to
Molecules and Back!
Lab 1 - Insect Identification (Biodiversity)
Lab 2 - Isolation of Insect and Wolbachia DNA
(Molecular Biology)
Lab 3 - PCR of Wolbachia 16S rDNA (Molecular
Biology)
Lab 4 - Presence/Absence of Amplicon (Molecular
Biology)
Lab 5 - Evolutionary Sequence Analysis
(Bioinformatics)
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Features of These Labs

• Original Research in Lab Exercises

• YOU can make new discoveries

• Integrates across Science
• Biodiversity -gt Molecular -gt Evolution

• Integrates Science and Technology

• Covers Emerging Fields
• Microbial diversity, Bioinformatics,
  Molecular Phylogeny

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Discover the Microbes Within The Wolbachia
Project Lab 1 Insect Identification
Hemiptera aphids 67,500 species
Coleoptera beetles 370,000 species
Collembola spring tails 6,000 species
Diptera flies 120,000 species
Lepidoptera butterflies, moths, skippers 140,000
species
Orthoptera grasshoppers, crickets, katydids
17,000 species
Odonata dragonflies, damselflies 5,000 species
Dermaptera earwigs 1,200 species
Hymenoptera bees, wasps, ants 108,000 species
Isoptera termites 1,900 species
Dictypotera cockroaches and mantids 6,000
species
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EXTRACT DNA FROM YOUR SAMPLES WHY?
Discover the Microbes Within The Wolbachia
Project Lab 2 DNA Extraction
Infected sample () Uninfected Sample
(-) Insect Insect Mitochondria
Mitochondria WOLBACHIA -
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Discover the Microbes Within The Wolbachia
Project Lab 3 Polymerase Chain Reaction
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Discover the Microbes Within The Wolbachia
Project Lab 4 Gel Electrophoresis
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Discover the Microbes Within The Wolbachia
Project Lab 4 Gel Electrophoresis





















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Discover the Microbes Within The Wolbachia
Project Lab 5 Sequence Analysis

• NCBI
• BLAST
• No programming skills required!

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When It all comes together, it looks like this!
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High School
MBL
ATGCGC
Primers, Insect controls,
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Where Does Your Data Go?
To
Consequences

• Class reports
• Peers, family, teachers, research scientists
• Online database (website repository for your
  data)

• New discoveries
• Collaborations between research scientists and
  high schools
• Summer envisionships
• Professional meetings
• Professional societies
• Journal publications

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How Important Are Wolbachia?
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Wolbachia spurs Nasonia speciation
Interspecific cross (male x female)
N. giraulti (G)
Infected
Hybrid Offspring
Cured
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Human Health Wolbachia may help control the
transmission of arthropod-borne diseases
(Malaria, Dengue fever, Filariasis,
Trypanosomiasis, West Nile, Chagas)
Does Not Transmit Disease
Transmits Disease
Reinfect vector with transgenic symbiont
Isolate and culture symbiont
Transform symbiont
Anti-pathogen gene
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Drive via Cytoplasmic Incompatibility
Male
Female
Progeny
X
None (Incompatible)
X
X
X
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Population Replacement
Transmits Disease
Does Not Transmit Disease
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Human Health Bill and Melinda Gates Foundation
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Wolbachia are Chemotherapy Targets for Curing
River Blindness Elephantiasis Caused By
Filarial Nematodes
Onchocerca volvlulus
Untreated
11 months post-treatment
Horeauf et al, 2003
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Web Resource - http//www.eol.org/taxa/17123196
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http//discover.mbl.edu
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Discover the Microbes Within!

• 1,000,000,000,000,000,000 insects on the planet
• 85 of all animal species are insects (1-30
  million species)
• 20 of all insect species harbor Wolbachia
• Scientists cant discover them all on their own.
• YOU ARE OUR BIGGEST ASSET TO SCIENTIFIC DISCOVERY
  AND REAL RESEARCH!

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The Wolbachia Community (San Juan 2006)
Help Discover the Microbes Within!
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Thank you - Assessment!
Consent Forms
Questionnaires
EDC Center for Science Education
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Do Good Science and Discover!