Division of Environmental Health Department of Public Health and Social Services Guam Mosquito Laboratory

Threat of Mosquito-borne Diseases to Guam Introduction and Spread of Mosquito-borne Disease Mission of the Mosquito Surveillance and Control Program Guam Mosquito Laboratory BioSecurity Level Presentation Outline

Threat of Mosquito-borne Diseases to Guam Guam’s economy, U.S. military presence, tourism, and geographic location have led to the entry of millions of items and people into the island from all over the world Many of these goods and travelers originate from areas where mosquito-borne diseases are endemic Infected passengers and mosquitoes, and mosquito carrying ships and planes, can enter Guam Hot and humid climate of Guam provides ideal habitat for mosquitoes to breed and populate Thus, the risk of introducing and spreading mosquito- borne diseases on Guam is high

Threat of Mosquito-borne Diseases to Guam (Cont.) Mosquito-borne diseases include: Dengue Fever: >100 million cases worldwide each year Malaria: Kills about 660,000 people annually Japanese Encephalitis: Causes inflammation of the brain membrane Zika Virus and Chikungunya: Emerging mosquito-borne diseases Guam is free of endemic mosquito-borne diseases Most recent tests of captured mosquitoes showed no infection All documented cases post-WWII were imported Importation of infected mosquito Can enter island via ship or plane with imported goods and arriving passengers Arrival of infected person Person infected elsewhere arrives to Guam Importation of certain mosquito species can increase the threat to Guam Some mosquito species are more efficient than others in transmitting the disease

Introduction and Spread of Mosquito-Borne Diseases Outbreak from an infected mosquito Infected mosquito entering island can bite and transmit disease to local resident or visitor Infected mosquito may transmit virus to its offspring Outbreak from an infected person Non-infected mosquito can bite an arriving visitor or returning resident who is infected with the disease Now infected mosquito can subsequently bite another person to transmit the disease Importation of certain mosquito species Aedes aegypti mosquito is an efficient transmitter of dengue fever Eradicated on Guam post WW-II by the military Can be imported into the island via ship or plane and become re- established Establishment of the disease As more mosquitoes get infected, the greater the potential for the spread of the disease in the population leading to an outbreak If infected mosquitoes are not eradicated, the disease may become established

Mosquito Surveillance and Control Program (MSCP) MSCP of the Division of Environmental Health, DPHSS seeks to prevent the introduction and spread of mosquito-borne disease on Guam MSCP will conduct mosquito surveillance Collect and identify mosquito species on Guam Gather data on mosquito species, location and density Establish monitoring sites (i.e., ports of entry) Test mosquitoes for infection MSCP will conduct mosquito control Determine triggers to initiate investigation, control measures, etc. Conduct outbreak investigations and control activities Determine pesticide resistance MSCP will conduct mosquito education Promote mosquito education and awareness Conduct outreach and campaigns

Guam’s Need for a Mosquito Laboratory To conduct mosquito surveillance, control, and education activities, MSCP will require space, equipment, personnel and community support DOI CIP grant ($3 million) to fund DEH Office Expansion Project Additional office space House MSCP staff, store equipment and materials Mosquito lab Identify species, test for infection and determine pesticide resistance Training room Provide onsite training and education on mosquito control Food screening lab Project to be constructed in Dededo, adjacent to the Northern Community Health Center

The building will consist of 34 rooms total 15 rooms for mosquito laboratory Lab safety and security utmost importance Protecting staff and citizens from lab hazards Limited access to building Restricted access to lab Internal and external security Received assistance/guidance from CDC Constructed to meet certification standards for structure Biosafety Level (BSL) of a lab describes the degree of protection for containing biological agents BSL-4 > BSL-3 > BSL-2 > BSL-1 Pathogens are identified by the CDC as to which biosafety level they belong to Majority of our lab rooms will be BSL-2; few rooms will be BSL-3 Project Information

AgentsPractice Primary Barriers and Safety Equipment Facilities (Secondary Barriers) BSL-1 Not known to consistently cause diseases in healthy adults Standard microbiological practices No primary barriers required PPE; laboratory coats and gloves; eye and face protection, as needed Laboratory sink and bench required BSL-2 Agents associated with human diseases and posts moderate hazards Includes bacteria and viruses that cause only mild disease to humans, or are difficult to contract via aerosal in a lab setting BSL-1 practice plus: Limited access Biohazard warning signs “Sharps” precautions Biosafety manual defining any needed waste decontamination or medical surveillance policies Primary Barriers: BSCs or other physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materials PPE: Laboratory coats, gloves, face and eye protection, as needed BSL-1 plus: Autoclave available BSL-3 Indigenous or exotic agents that may cause serious or potentially lethal disease Includes various bacteria, viruses, and parasites that can cause severe to fatal diseases in humans, but for which treatment exists BSL-2 practice plus: Controlled access Decontamination of all waste Decontamination of laboratory clothing before laundering Primary Barriers: BSCs or other physical containment devices used for all open manipulations of agents PPE: protective laboratory clothing, gloves, face, eye, and respiratory protection, as needed BSL-2 plus: Physical separation from access corridors Self-closing, double door access Exhausted air not recirculated Negative air flow into laboratory Entry through airlock or anteroom Hand washing sink near laboratory exit BSL-4 Dangerous/exotic agents which post high individual risk of aerosol-transmitted laboratory infections, Causes severe to fatal disease in humans for which vaccines or other treatments are not available BSL-3 practice plus: Clothing change before entering Shower on exit All material decontamination on exit from facility Primary Barriers: All procedures conducted in Class III BSCs or Class I or II BSCs in combination with full-body, air- supplied, positive pressure suit BSL-3 plus: Separate building or isolated zone Dedicated supply and exhaust, vacuum, and decontamination systems Other requirements outlined in text Summary of Biosafety Levels 1-4

BSL for specific Pathogens PathogensBSL-2BSL-3 Dengue fever virusX Chikungunya virusX(recommended) Japanese encephalitis virus X(recommended) MalariaX West Nile virusX Yellow fever virusX Zika virusX Note: For Chikungunya and Japanese encephalitis viruses, a BSL-3 is recommended but not required

Conclusion Guam’s economy, location, and climate may lead to the importation and the establishment of mosquito- borne disease(s) MSCP seeks to prevent the introduction and spread of mosquito-borne disease MSCP requires a dedicated laboratory to implement mosquito surveillance, control, and education A mosquito lab can prevent and control mosquito- borne diseases on Guam Proposed mosquito lab will be constructed to meet the physical standards to contain biohazards

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Table 3: Food Lab Pathogens and BSL Category Laboratory Information (Cont.) PathogensBSL-2BSL-3 Bacillus cereus X Campylobacter jejuni X If high concentrations or high production volumes are used or when there is high potential for aerosol production Clostridium botulinum X If high concentrations or high production volumes are used or when there is high potential for aerosol production Clostridium perfringens X E.coli 0157:H7X Hepatitis AX Listeria monocytogenesX If high concentrations or high production volumes are used or when there is high potential for aerosol production Norwalk and Norwalk-like viral agent If used for research work (co-cultivation, replication studies, or manipulation involving concentrated virus) only If used for virus production activities SalmonellaX ShigellaX Staphylococcus aureusX

Primary Barriers Biological Safety Cabinets (BSC) are enclosed, ventilated laboratory workspace BSC Class II Type A2 will be used Vertical, laminar flow ventilated cabinets with an open-front Mass airflow is re-circulated within work space Uses HEPA-filter for both re-circulated air and exhaust air; HEPA filters are effective for trapping particulates and infectious agents Provides personnel, environment, and product protection Insects will be contained in cages and placed inside environmental chambers Laboratory Information (Cont.) BSC Class II Type A2Environmental Chambers

Secondary Barriers Laboratory Facilities Design Access is limited to lab staff Double doors for main entrance/exits Laboratory portion is separated by additional doors Self-closing doors with locks are provided BSL-3 rooms and insectaries are separated from unrestricted traffic flow An anteroom is provided for the insectary rooms BSL-3 rooms have an anteroom and are equipped with a negative airflow into the rooms; ducted air ventilation system that circulates air from “clean” area to “potentially contaminated” areas is also provided No windows to the outside are provided for lab portion Exhaust vents and vacuum lines are protected with HEPA filters Laboratory Information (Cont.)

Mosquito Sorting and Identifying Insecticide Chemical Preparations Insecticide Resistance Testing Polymerase Chain Reaction (PCR) Food and water screening and sampling Types of Tests/Procedures Performed

Autoclaves An autoclave is a strong, pressurized, steam-heated vessel used for sterilization Guam Mosquito Lab will have two (2): One (1) pass-thru autoclave from BSL-3 rooms to sterilization room One (1) back up autoclave Disinfection and Decontamination Pass-thru autoclaveAutoclave

Facilities are designed to follow BSL-2 and BSL-3 specifications: Rooms sealable for decontamination Surfaces of walls, floors, and ceilings are smooth, impermeable durable, chemical resistant, and easily cleanable and decontaminated; all penetrations sealed Handwashing and eye wash stations are provided in each room and located near exits Detailed protocols and strategies will be established regarding disinfection and decontamination once laboratory is set up Disinfection and Decontamination (Cont.)