UNCLASSIFIE D UNCLASSIFIE D Public Health Command - Pacific Leptospirosis Surveillance by Filtration Sampling and Next Generation Sequencing in Okinawa, Japan CPT Sean Kluckman, P.E. CPT Sean Kluckman, [email protected]1 of 16 HUMAN ENVIRONMENT ANIMAL ONE HEALTH
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UNCLASSIFIED Public Health Command - Pacific Leptospirosis Surveillance by Filtration Sampling and Next Generation Sequencing in Okinawa, Japan CPT Sean.
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UNCLASSIFIED
UNCLASSIFIEDPublic Health Command - Pacific
Leptospirosis Surveillance by Filtration Sampling and Next GenerationSequencing in Okinawa, Japan
UNCLASSIFIEDPublic Health Command - Pacific AS OF: Slide 1008SEPT2015
Leptospirosis Case Study
Turbidity Drinking Water
0.3 NTUs
Raw Water 800 NTUs
UNCLASSIFIED
UNCLASSIFIEDPublic Health Command - Pacific AS OF: Slide 118SEPT2015
Leptospirosis Case Study
Water Source
PumpFilter
Waste
Option 1: Envirocheck Method Option 2: Innovative Disk Filter System
DNA Extraction
Water Source
Trap Filter(10/2µm pore)
Analytical Filter (0.45µm pore)
Waste
DirectDNA Extraction
Pros- Established method- Commercially available
Cons- High equipment cost- High consumable cost- Requires electricity- Clogs easily
Pros- Low cost- Light weight- No electricity- Rugged- Easy to Use
Cons- Undeveloped- Untested
100mL
140 ml syringe for delivery of 100ml of sample
UNCLASSIFIED
UNCLASSIFIEDPublic Health Command - Pacific AS OF: Slide 1208SEPT2015
Leptospirosis Case Study
Next Generation Sequencing16S rDNA Amplicon Sequencing
Conventional Polymerase Chain Reaction (PCR)
• Rapid ID of specific threat• Prior sample knowledge• All Samples Negative (Possibly Due to Sensitivity)
• Rapid ID of ALL threats• No prior sample knowledge• 3 variations of pathogenic Leptospira
UNCLASSIFIED
UNCLASSIFIEDPublic Health Command - Pacific AS OF: Slide 1308SEPT2015
Leptospirosis Case Study
Recommendations
Administer Leptospirosis vaccine for all Military Working Dogs.
Continue monthly sampling to try and determine a correlation between Leptospira detection and environmental conditions.
Develop a formal kit for filtration-based water sampling.
The daily number of hourly observed precipitation reports during the last 12 months (2014), color coded according to precipitation type, and stacked in order of severity. From the bottom up, the categories are thunderstorms (orange); heavy, moderate, and light snow (dark to light blue); heavy, moderate, and light rain (dark to light green); and drizzle (lightest green). Not all categories are necessarily present in this particular graph. The faint shaded areas indicate climate normals. The bar at the top of the graph is green if any precipitation was observed that day and white otherwise.