UNCLASSIFIED Public Health Command - Pacific Leptospirosis Surveillance by Filtration Sampling and...

UNCLASSIFIED

UNCLASSIFIEDPublic Health Command - Pacific

Leptospirosis Surveillance by Filtration Sampling and Next GenerationSequencing in Okinawa, Japan

CPT Sean Kluckman, P.E.

CPT Sean Kluckman, [email protected] 1 of 16

HUMANENVIRONMENT

ANIMAL

ONE HEALTH

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UNCLASSIFIEDPublic Health Command - Pacific AS OF: Slide 2

OVERVIEW

• Public Health Command – Pacific • Leptospirosis Overview• Leptospirosis Case Study

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UNCLASSIFIEDPublic Health Command - Pacific AS OF: Slide 308SEPT2015

Public Health Command-Pacific

Human HealthMedical Entomology

Preventive Medicine Doctors

Animal HealthVeterinary Care

Environmental HealthEngineering ServicesLaboratory Services

Leptospira Outbreak

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Leptospirosis

Victoriano et al. BMC Infectious Diseases, 2009

• Multiple strains of disease-causing Leptospira that are endemic to all tropical and subtropical regions.

• Outbreaks have been linked to heavy rainfall events.

• Transmitted through contact with water, moist soil and vegetation contaminated with the urine of infected animals.

• Symptoms for humans and dogs, resulting in fever, kidney failure and can cause death if untreated.

SuspectedHost

(Mongoose)

Infected Military

Working Dog

Infected Military

Personnel

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Leptospirosis Case Study

Trigger Event – September 2014• A long hot dry period followed by heavy rainfall

occurred prior to a water training exercise.

• Marines present with the symptoms of “Leptospirosis”.

• Loss of a Marine Military Working Dog (MWD) with presumptive Leptospirosis diagnosis.

Navy Times, 03 NOV 2014

Stripes Japan, 09 OCT 2014

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Initial Response

United States Air Force• Collects Water/Tissue

Samples from Nuisance Wildlife (MONGOOSE)

United States Army• Analyzes Water and Tissue

Samples• Conduct Site Evaluation

United States Marine Corps• Provided medical oversight

OKINAWA

Presumptive

Samples

CAMP ZAMA

Team

Dispatc

hed

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UNCLASSIFIEDPublic Health Command - Pacific AS OF: Slide 8


Problems Encountered

Raw water was sediment laden and clogged the instruments.

Polymerase Chain Reaction (PCR) Sequencing showed that all samples were negative for Leptospira.

Military Working Dogs do not show symptoms of being infected until the late stages of the infection.

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Human Health

Animal Health Environmental Health

Leptospira Outbreak

Response TeamEntomologistsMicrobiologistsEngineersVeterinariansDoctors

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Turbidity Drinking Water

0.3 NTUs

Raw Water 800 NTUs

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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

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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

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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.

Max. Normal Precipitation

http://weatherspark.com/history/33303/2014/Okinawa-Okinawa-Prefecture-Japan

2014 Potential Predictive Environmental Cue

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Leptospirosis Case StudyConclusions

Worked as an integrated team to identify and provide solutions to the Leptospirosis outbreak.

Develop a water filtration device and incorporated Next Generation Sequencing to evaluate samples.

Recommendations apply to similar training environments around the Pacific.

Provided recommendations to allow training to continue.

ChemoprophylaxisVaccination of MWDsEducation and Awareness

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Questions

HUMANENVIRONMENT

ANIMAL

ONE HEALTH

UNCLASSIFIED Public Health Command - Pacific Leptospirosis Surveillance by Filtration Sampling and...

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