Lymes on the rise………
The Increase of Lymes in Minnesota
 A local family physician inquiring about Lymes
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disease.
Physician noted an increase of patients experiencing
ring shaped rashes, headaches, and fever chills which
point to Lymes.
Rigid diagnosis was not made as conformation
requires an diagnostic assay.
The physician expressed concern that the Minnesota
Department of Natural Resources maybe inaccurate.
Duluth area people might need to be better informed
before planning outdoor activities.
symptomology
Contracting Lymes Disease
 The bite of an infected nymph or adult tick can only transmit
Lyme disease after it attaches to the host for 36 – 48 hours
 the generation time of B. Burgdorferi: ~ 12 hours
 Spirochetes waiting in the mid-gut of the tick begin to multiply as
the tick starts its blood meal then migrate to the salivary glands
where they are discharged into the host with the tick's saliva.
 It takes several hours before a large enough infectious dose of
spirochetes can penetrate the new host.
 Most incidences of Lyme disease occur from May through
September because the nymphal stage of the tick is responsible
for most cases
 They are too small to be easily detectable
 Their bites cause much milder pain than an adult tick.
Lymes Disease
Symptomology of Lymes Disease
 Lyme is a multisystem inflammatory disease
 affects the skin in the early stage
 and then spreads to the joints, nervous system and may
eventually affect the organ systems.
The symptoms are divided into three stages, each stage
being more advanced than the previous.
Symptomology
Stage 1 –
Early localized infection
 Skin Rash Around Tick Bite
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(75%)
Mild Illness
Flu-like Symptoms
Fevers
Chills
Swollen Lymph Nodes
Headache
Stiff Neck
Muscle Fatigue
 Initial sign and symptoms of
Lyme disease include an
appearance of a rash, which has
the appearance of being solid
red or bull’s eye rash with flu
like symptoms. These symptoms
may appear up to a month after
infection.
 Within weeks of stage one
Symptomology
Stage 2
– Early Disseminated Infection
 Neurological Problems
 Meningitis
 Encephalitis
symptoms, the bacterium
may spread to other parts of
the body through the
bloodstream.
 infected individuals may
start having acute
neurological problems
such as Bell's Palsy. *15%
of untreated patients
progress to this stage.
 Meningitis, as well as
mild Encephalitis is
common at this stage,
leading to memory loss,
sleep disturbances, or
changes in mood or
affect.
Symptomology
Stage 3 –
Late Persistent Infection
 Polyneuropathy
 Pain and Numbness
 Cognitive Impairment
 Arthritis
 Acrodermatitis chronica
atrophicans (primarily in
Europe)
 The third and the most severe
stage may occur months to years
after initial infection. Symptoms
at this stage are primarily
rheumatologic and neurologic,
and mostly untreatable.
Resources:
http://www.nature.com/nrmicro/journal/v3/n2/full/nrmicro1086.html
3 stages of Lyme Disease
 Stage 1: Early Localized (previous slides)
 Stage 2: Early Disseminated
 Weeks to Months
 Affects skin, joints, nervous system, heart
 Stage 3: Late Persistent
 Months to Years
 Damage to joints, nerves, brain
Stage 3
 Also Known as:
 Post-Lyme Disease Syndrome
 Chronic Lyme Disease
 Occurs 6 months after treatment
Possible Cause
 If antibiotics are not received quickly, then the body
may build up antibodies to B. b. These antibodies may
later attack healthy tissues.
 Multiple studies hint at the autoimmune process
behind Lyme Disease but nothing is known.
Diagnosis
 Clinical Evidence
 Enzyme Linked Immunosorbent Assay
 Western Blotting
 Indirect Immunofluorescence
 Detect antibodies
 Lack s sensitivity and specificity
 PCR
Treatment
 Short Term Treatment
 Amoxicillin
 Probenecid
 Ceftriaxone
 Doxycycline
 Tetracycline
 Chronic Antibiotics
 Penicillin
 Erythromycin
 Tetracyline
Our tick collection methods
Materials:
• Light-colored bed sheet
• Hand held GPS devices
• Sealable plastic
containers
Methods:
• We began by stopping at
the banks of pre-selected
locations listed in the
GPS.
• We separated into
groups of about two and
while holding the sheet
open, we swept the
ground.
• After dragging for 100
yards we examined the
sheet in hopes of finding
black-legged ticks.
Our Results
• We continued collecting over
two days for a total of eight
stops.
• We found six ticks.
• Some observations were made
about the general environment
in which the ticks were found,
however, very little information
was recorded.
• All of the ticks found were in
their nymph stage and were not
much larger than the point of a
pencil.
• Our sampling was done during
the mid summer season rather
than the late spring or fall, this
may be why so few ticks were
collected.
Project title
Michael Cherry| Dr. Ben Clarke| Bridges and Pathways
Program 2010
 What
criteria and procedures should be used when
Tick
Collection
collecting ticks in the ‘wild’?
 Where ticks
are located
Project
Overview
 Survey techniques should be consistent
 Use of standardized equipment
 Integrity of Data Collection Criteria and methods of
recording
Where are ticks located in the
midwest?
 Near or in deciduous forests
 Dry to mesic (moderate moisture) forests
 Alfisol-type soils of sandy or loam-sand textures
overlying sedimentary rock
 Along ecotones (boundaries between varying
ecosystems)
Alfisol Soils
 Alfisols are a soil order in USDA soil taxonomy.
Alfisols form in semiarid to humid areas, typically
under a hardwood forest cover. They have a clayenriched subsoil and relatively high native fertility.
Alfisol from
East and
Tennessee.
"Alf" refers to Aluminium
(Al)
Iron (Fe).
http://www.uamont.edu/facultyweb/francis/soilprofiles/so
Where are ticks not located in the
midwest?
 Near coniferous forests
 Grasslands
 Wet and very damp forests
 Acidic soils of low fertility and a clay soil texture
 Precambrian bedrock
Construction of dragging
 70 x 90cm piece of light-colored corduroy
equipment
 1cm diameter wooden dowel along leading edge
 Steel rod sewn into trailing edge for added weight
 Rope or wood handle to allow front edge of ‘sheet’ to
contact vegetation
Collection Techniques from
 Same individual should perform the dragging for
Schulze/jordan
consistency
 Speed, height
 Collected hourly from 0600 – 2100 hours
 100m long transects, collect every 10m to prevent
ticks from falling off
 Ramdomize dragging order, direction of travel,
location within each transect (left, right, center) to
minimize repeated dragging of same areas
 Minimize dew collection by drying on running
pickup truck hood
 To minimize effects of moisture on tick collection
 For each sampling event (each hour) (use of Weather
Statistics
Station):
 Ambient Temperature
 Ambient Humidity
 Place Weather Station in center of area, 1m above
vegetation height
 Litter temperature and humidity (digital
thermometer)
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At least 2.5cm below leaf litter surface but not into the soil
 Use of repeatedanalysis
measures of analysis of variance
Statistical
 Use of multiple linear regression to examine relationships
 Use separate regressions for each species
 Tolerance value of 0.1 is acceptable
 Calculate temp gradient: subtract hourly litter temp from
ambient
 Calculate humidity gradient: subtract litter H from
ambient H
 Use Pearson correlation and linear regression to
determine relationships between calculated gradients
and numbers of questing ticks
 Consistency regarding the gathering of ticks and in
Conclusion
measuring and recording statistics are critical to
success
 Meteorologically
Works
CitedMediated Diurnal Questing of
Ixodes Scapularis and Amblyomma Americanum
Nymphs – Terry Schulze and Robert Jordan Journal
of Medical Entomology (2003)
 Distribution of the Common Tick, Ixodes ricinus, in
different vegetation types in Southern Sweden –
Anders Lindstrom and Thomas Jaenson, Journal of
Medical Entomology (2003)
Possible Outcomes
o Lymes disease occurances have been on the rise in
the US partially due to climate change and
expansion in tick population, but also due to
increased human/tick interaction in fringe areas near
cities. By identifying specific ecotomes required for
the complex life cycles of ticks and boriela, we can
possibly eliminate some of those factors in these
fringe areas possibly reducing the incidence of
disease being transferred to humans
GIS Modeling of the Interrelationship
of Ticks, Mice, and Ecotones
o Collecting ticks while recording specific geographical
data
 Soil/vegetation/elevation/etc.
o Analyze ticks to detect the presence of Borriela
bacteria
o Analyze the Data searching for patterns of Borriela in
relation to the specific ecotone in which the tick was
found
By: Nina M. Holz
Borrelia burgdorferi
Complete genome
Characteristics
 Borrelia burgdorferi are
 spiral shaped microorganism (spirochetes)
 Gram-negative
 Motile
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On the average, seven periplasmic flagella are located at each cell
end, and these flagella overlap at the central region of the cell.
 inhabit the gut of deer ticks (Ixodes scapularis).
 Named after American scientist Willy Burgdorfer who in
1982 isolated and determined the bacterium.
 Lymes disease is the most common tick-borne disease in
the US.
Characteristics
 Chemo-organotropic
 derive energy by metabolizing organic molecules.
 Microaerophilic
 they require oxygen to survive, but only at low concentrations.
Too much or lack of oxygen can be harmful to them.
 Catalase negative
 they are unable to break down hydrogen peroxide.
 They cannot synthesize their own amino acids, fatty acids,
enzyme cofactors, and nucleotides.
Characteristics
 Optimal growth temperature is 34° to 37°C
 Generation time of 11 to 12 hours at 35°C.
 Their life cycle is directly linked to their host, and they depend on
arthropod vectors (ticks) to infect mammals.
 They reside in infected mammals as well as in the gut of ticks.
 They multiply when there is enough blood ingested by the tick to
support bacterial growth.
Genome structure
DNA Molecule Summary
Chromosomal Gene Summary
 18 DNA molecules
 853 genes
 28.5%
 793 protein coding genes
G+C
chromosomal bases
 63.1% linear DNA bases
 502 genes assigned a role, 51
genes not assigned a role
 102 hypothetical genes
Cultivation
 The organism's small genome lacks the genes that are
necessary for the synthesis of amino acids, fatty acids,
enzyme cofactors, and nucleotides.
 In order for these spirochetes to be cultured, a very
special type of medium is used called BarbourStoenner-Kelly (BSK) medium.
BSK =Barbour-Stoenner-Kelly
 Contains serum-supplemented mammalian tissue-culture
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that enables the bacterium to reproduce without ever
having the need to synthesize too many biological
compounds.
Contains over thirteen ingredients in a rabbit serum base.
Optimal temperature for growth =32-35oC
A microaerobic environment.
Even under optimal conditions, the generation time is slow,
about 12-24 hours.
BSK solidified with 1.3% agarose allows the production of
colonies from single organisms.
Some Basic Background Information
Regarding Ticks and Lymes Disease
 The tick that passes enables animals to contract Lymes
Disease is called: Ixodes scapularis.
 The ticks generally feed on a wide variety of small mammals,
birds, and reptiles, but prefer to feed on white-footed mice
(Peromyscus leucopus), which are the important reservoir
of infection in nature.
 Adults prefer to feed and mate on white-tailed deer .
 The infection is a bacteria called Borrelia burgdorferi. The
ticks usually get infected from feeding on a infected host.
 Not all humans that get bite by ticks get Lymes diesease
 Only ticks that are infected with the bacteria can give Lymes
to humans
 This is generally after a prolonged amount of time feeding on
the human where they can also get Lymes disease
Internal Anatomy of a Tick
 Shown to the left
is the internal
anatomy of a
tick. The picture
shows the major
areas involved in
feeding and
reproducing in
the tick.
Early Onset Symptoms of Lymes
Disease
 Within 1-4 weeks of being bitten by an
infected tick, most people will
experience some symptoms of Lyme
disease.
 A circular, expanding rash (called
erythema migrans) at the site of the bite
develops in about 70%-80% of cases.
 Some people report flu-like symptoms
at this stage, including fever, chills,
headaches, fatigue, swollen lymph
nodes, joint pain, and muscle aches.”
(WebMD)
Mid-Stage Onset Symptoms of
Lymes Disease
 If the disease is not detected and treated in its
early stages, it can extend to more areas of the
body, affecting the joints, heart, and nervous
system (about 1-4 months after the initial bite).
 Additional rashes may occur, and there may be
intermittent periods of pain and weakness in
the arms or legs.
 Facial-muscle paralysis (Bell's palsy),
headaches, and poor memory are other
symptoms at this stage, along with a rapid
heartbeat and some loss of control of facial
muscles.” (WebMD)
Late-Stage Onset Symptoms of
Lymes Disease
 This is the most serious stage of the disease,
when treatment was either not successful or
never started (usually occurring many months
after the initial bite).
 Joint inflammation (arthritis), typically in the
knees, becomes apparent, and may become
chronic.
 The nervous system can develop abnormal
sensation because of disease of peripheral
nerves (peripheral neuropathy), and
confusion.
 Heart problems are less common, but can
include inflammation of the heart muscle.”
(WebMD)
Current Treatments for Lymes
Disease
 Most Lyme disease is curable with
antibiotics, particularly when the
infection is diagnosed and treated
early.
 Doxycycline, amoxicillin, and
cefuroxime are the drugs of choice
most of the time for early illness.
 Later stages might require longerterm, intravenous antibiotics, such
as ceftriaxone.” (WebMD)
Vaccines made for Lymes Disease
 Currently, there is no human vaccine for
Lyme disease.
 The one that did exist -- LYMErix -- is no
longer available.
 Originally approved by the FDA in 1998 to
help prevent the disease, the vaccine was
pulled from the market by the
manufacturer in 2002 due to poor sales.
 There was concern that the vaccine could
trigger arthritis problems, although the
FDA never found evidence that the
vaccine was dangerous.” (WebMD)
Possible Ideas for a Future Vaccine
for Lymes Disease
 Since the Salp15 protein is the protein
that enables the bacteria to enter the
host initially undetected, learning
more about the protein anatomy and
morphology would be beneficial.
 If there is some way to possibly
degrade the protein so that the
bacteria is exposed and becomes
vulnerable, I think this would allow
the host’s body to fight off the
bacteria faster and more efficiently
with hopefully less damage to the
body.