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Transcript of LVEES Talk Edited
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Bess FleischmanAmulya Makkapati
Muhlenberg College Biology Department
Prevalence of Human pathogens in Ixodes scapularis ticks
Collected in the Lehigh Valley
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Blacklegged Ticks
http://bugguide.net/node/view/108373
http://www.tickencounter.org/tick_identification/deer_tick
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Winter
Fall Spring
Summer http://www.tickencounter.org/
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Lyme Disease
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Total U.S Lyme Disease Cases
1982 2004 2013
(Prior to 2014 press-release reporting likely CDC underestimate)
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Duik-Wasser et al.Am. J. Trop Med. Hyg 2012
Predictive ModelElevationClimate Variables (Temperature, Humidity)GIS-based estimates of forest fragmentation.
Field Collected DataDensity of infected nymphs per 1000 m2
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Lehigh Valley is in a transition zoneNo samples were taken from this region in this study, or any other published study
Duik-Wasser et al.Am. J. Trop Med. Hyg 2012
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LowerHudson Valley
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http://www.nsf.gov/news/special_reports/ecoinf/lyme.jsp
Conservation Biology 2003
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Borrelia burgdorferiSpirocheteLyme Disease
Borrelia miyamotoiSpirocheteTick Borne Relapsing Fever
Anaplasma phagocytophilumBacteria that live inside white blood cellsAnaplasmosis
Babesia microtiProtozoa that lives in red blood cellsBabesiosis
More than one pathogenare transmitted by blacklegged ticks!
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Babesiosis Anaplasmosis
1999 2010 1994 2010
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Our Research Questions
Are the tick-borne pathogens that are abundant in the Lower Hudson Valley already established in the Lehigh Valley?
Do the lower rates of tick-borne disease in the Lehigh Valleycorrespond with lower rates of infection in ticks?
Can we establish a baseline of the current rates of infection to inform future studies in the Lehigh Valley?
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LowerHudson Valley
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Our Study
Nine sites428 Adult and Nymph Ticks
Average of 54 ticks per site (range 11-133)
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DNAExtraction
Yittrium-ZirconiumCeramic Beads 2 mm and 0.1 mm
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Pathogen DNA detected using Real-Time PCR
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Advantages Real-Time PCRSensitive: Few false negatives for lack of detectionSpecific: Few false positives as a result of DNA contamination
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Pathogen N positive/N tested
% Positive (Our
Study)
% Positive (Hudson
Valley)
B. burgdorferi Total
Nymphs
Adults
98/428 22.9
20/111 18.0 67%
78/317 24.6 60%
Lyme Disease
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Results (Continued) Pathogen N positive/N
tested % Positive
(Our Study) % Positive
(Hudson Valley)
B. miyamotoi 2/428 0.5
Unidentified Borrelia spp. 14/428 3.3
A. phagocytophilum Total 8/428 1.9 8.5%
AP-ha 2/428 0.5
Ap-Variant 1 6/428 1.4
B. microti 2/428 0.5 1.2%
B. odocoilei 1/428 0.2
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Coinfections
Pathogens N positive/N
tested
% Positive
B. burgdorferi, A. phagocytophilum (AP-ha) 1/428 0.2
B. burgdorferi, A. phagocytophilulm (Ap- Variant 1) 1/428 0.2B. burgdorferi, B. microti 2/428 0.5
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Conclusions
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Acknowledgements• Dr. Marten Edwards• Current Members of the Edwards Lab: Laura Barbalato, Kristina Fioretti, Rachel Heist
• Dr. Luther V. Rhodes Endowment for Infectious Disease Research through the LVHN
• Lehigh Valley Hospital Scholars Program• Muhlenberg College Department of Biology • James Vaughan Fund for Summer Research• Louise Bugbee, Meaghan Bird, Katerina Pham, Sarah Richardson
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2 ul 0.12 ul
1 ul
0.25 ul
Control experiments with serial dilutions of pathogen control DNA showed that we could detect a very small number of pathogens in our tick samples.
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PCR Assays
Ticks: ITS2 Ribosomal Gene
Borrelia burgdorferi
23S Ribosomal RNA: Common to several Borrelia spp.
Outer Surface Protein A (OspA): Only found in B. burgdorferi
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PCR AssaysAnaplasma phagocytophilum
Msp2: bacterial surface proteinPositive samples were used in a sequencing reaction with a
different gene
B. microti
18S ribosomal gene sequence only found in B. microtiPositive samples were used in a sequencing reaction with a
different gene
B. miyamotoi
16S ribosomal gene sequence only found in B. miyamotoi Positive samples were used in a sequencing reaction with a
different gene
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Lyme Disease
Babesiosis
Human Granulocytic Anaplasmosis
Tick-Borne Relapsing Fever
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State/Area of residenceCalifornia 1 (0.1)Connecticut 74 (6.6)Delaware 1 (0.1)Indiana 0 (0.0)Maine 9 (0.8)Maryland 4 (0.4)Massachusetts 208 (18.5)Minnesota 73 (6.5)Nebraska 0 (0.0)New Hampshire 13 (1.2)New Jersey 166 (14.8)New York 361 (32.1)Oregon 1 (0.1)Rhode Island 73 (6.5)Tennessee 1 (0.1)Vermont 2 (0.2)Washington 0 (0.0)Wisconsin 80 (7.1)
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Three cases in Eastern PA
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http://www.cdc.gov/anaplasmosis/stats/