WORKSHOP: Protecting Florida from Dengue and Chikungunya through Control of Aedes aegypti and Aedes albopictus June 3-4, 2014 THE FLORIDA MEDICAL ENTOMOLOGY LABORATORY University of Florida IFAS Vero Beach, Florida

Florida Medical Entomology Laboratory

Aedes aegypti

Dengue virus

Aedes albopictus

Chikungunya virus Florida Medical Entomology Laboratory

DENGUE TRANSMISSION IN FLORIDA 2009-2014

2009 2010 2011 2012 2013

DENV-3 DENV-2

Florida Medical Entomology Laboratory

DENGUE IN THE CARIBBEAN BASIN - 2013 Florida Medical Entomology Laboratory

Prevalence of Dengue: Florida vs. Singapore 900

800

Singapore

INCIDENCE/100,000

700

Key West

600

Rio

500 400 300 200 100 0 2008

2009

2010

2011

2012

YEAR

Florida Medical Entomology Laboratory

2013

79th Annual Meeting of the Florida Mosquito Control Association 2007 Jacksonville Florida

SYMPOSIUM CHIKUNGUNYA: Is this on the horizon for Florida?

Florida Medical Entomology Laboratory

Symposium Presentations • Chikungunya outbreak in the Indian Ocean (Walter Tabachnick)

• Chikungunya on an island off the coast of Kenya: impact on the health of the global community and potential development of an early warning system in the U. S. (Ken Linthicum) • Chikungunya in Florida: what to expect if it arrives (Jon Day) • Florida response capabilities to a Florida Chikungunya outbreak (C. Roxanne Connelly)

Florida Medical Entomology Laboratory

CHIKUNGUNYA IN THE CARIBBEAN – 2013-2014

Chikungunya Cases as of May, 2014 Ca. 95,000 Cases Cases/100,000 St. Martin (French) – 3220 Martinique – 27670 Saint Barthélemy – 7267 Guadeloupe – 18000 Dominica – 1817 St. Martaan (Neth) - 343 Dominican Republic – 32500 Anguilla – 33 British Virgin Islands - 7 French Guyana – 178 Haiti – 5500 St. Kitts – 20 Aruba – 1 Florida Medical Entomology Laboratory

8907 7088 7018 3843 2613 700 314 205 90 69

56 39 1

CHICKUNGUNYA - Caribbean Dec 2013 - May 2014 9000.00

Cases/100,000

7500.00 6000.00 4500.00 3000.00 1500.00 0.00

Florida Medical Entomology Laboratory

Florida Dengue and Chikungunya -2014 Imported Cases

• Dengue - 23 Cases – Travel History – Bolivia, Brazil, Cuba, Dominican Republic, Guadeloupe, Honduras, Puerto Rico, Trinidad, Venezuela – Florida Counties – Alachua, Broward, Clay, Hillsborough, Marion, MiamiDade, Orange, Osceola, Seminole – DENV Serotypes – 1(5), 2(6), 3(1), 4(3), 1&2(1)

• Chikungunya – 10 Cases – Travel History - Dominica, Dominican Republic, Haiti (6), Martinique (2). – Florida Counties - Hillsborough (2), Miami-Dade (5), Palm Beach, Pasco.

Florida Medical Entomology Laboratory

How do we predict risks? How do we mitigate risks? Surveillance

Mosquito Mosquito-Pathogen Control Strategies Biology

Risk Assessment

Risk Management

Public Health Interventions Florida Medical Entomology Laboratory

Beach

Mosquito Control FL DOH Mosquito Control and FL DOH

Florida Medical Entomology Laboratory

Attendees represent • ca. 75% of Florida’s population • ca. 90% of the total of Florida Mosquito Control budgets • 32 Florida Counties • 25 FL MCD’s • 17 FL DOH

WORKSHOP: Protecting Florida from Dengue and Chikungunya through Control of Aedes aegypti and Aedes albopictus June 3-4, 2014 THE FLORIDA MEDICAL ENTOMOLOGY LABORATORY University of Florida IFAS Vero Beach, Florida

Florida Medical Entomology Laboratory

Natural History and Ecology of Aedes aegypti and Aedes albopictus with Special Reference to Florida

P Lounibos, Univ. of Florida, Florida Medical Entomology Lab

Part I: Invasion Biology & Competitive Displacement

Part II: Natural History & Ecology of Life Stages in Florida Part III: A Resurgence of A. aegypti in Florida?

Major range expansion of Aedes albopictus occurred in the past 40 years from Asia 1991

1985

1991

1985

Enserink. 2008. Science

These two species both frequent container habitats provided by humans and, as adults, mate in association with host-seeking

Decline of A. aegypti after the arrival of A. albopictus in Florida Distribution of A. albopictus from 1986 to 1994 1988 1986 & 1987

1990 1989

1994

1992

1991

1993

Spread on Route 441 of A. albopictus and decline of A. aegypti Lake city Gainesville Ocala Leesburg

Apopka Orlando Kissimmee St. Cloud Yeehaw Junction Okeechobee

Modified from O’Meara et al. 1995

Surveillance in south Florida cemeteries since 1990 documented rapid displacement and led to testing hypotheses of proposed mechanisms

Patterns of exclusion and co-existence in south Florida cemeteries (modified ex Lounibos et al. 2010 An. Entomol. Soc. Am.)

Larval densities, leaf litter, and species were manipulated in experiments in tires and plastic cups in Vero Beach, Florida and Rio de Janeiro, Brazil

Auto salvage yards harbor sympatric A. albopictus and A. aegypti in south Florida

Evidence of interspecific mating among field-collected females Sperm from

Collection Site M& K Auto

Species aegypti

Mean No. (SE) Per Coll 73.4 (18.7)

(n=7)

albopictus

Belle Glade (n=2)

Totals

85

A. albopictus 0

161.0(60.0)

1

71

72

aegypti

67.5(2.5)

82

3

85

albopictus

411.5(170.5)

1

61

62

169

135

304

Totals

A.aegypti

Spermathecae

85

Ex: Tripet et al. (2011) Am. J. Trop. Med. Hyg.

Sperm extracted from field-collected females and DNA amplified

Hypothesis: Male accessory gland (MAG) products of A. albopictus asymmetrically sterilize A. aegypti

Science 156: 1499-1501 (1967)

Experiment: 1. Inject MAG homogenates from conspecific or heterospecific males into unmated females of both species. 2. Allow post-injection females access to conspecific males in cages. 3. Assess insemination rates by spermathecal dissections

Results (Tripet et al. [2011] AJTMH 85) confirmed that the sterilizing effects of MAG extracts are asymmetric in their interspecific effects

Factors contributing to competitive displacement and segregation of Aedes aegypti by Aedes albopictus

U N F A V O R A B L E

LARVAL COMPETITION

rural

SATYRIZ ATION

+

albopictus (invader)

C L I M A T E

HABITAT SEGREGATION

aegypti

albopictus & aegypti

(resident)

suburban

-

30° N

urban 1985

ARRIVAL & ESTABLISHMENT

L A T I T U D E

25° N

Natural History & Ecology of Life Stages in Florida • Eggs:

A. albopictus

A. aegypti

Desiccation-resistance of aedine eggs influences what habitats may be colonized. All species shown (3 included as ‘forest’) belong to the subgenus Stegomyia. Numbers refer to different geographic strains. (ex: Sota & Mogi 1992. Oecologia 90:353-358)

Aedes eggs, laid on tongue depressors

Depressors with eggs of each species in a screened vase

1.0

2001

A. aegypti A. albopictus

0.8

% Hatch (SE)

2006 0.6

***

***

0.4

*** **

0.2

0.0 2 weeks.

4 weeks.

2 weeks

4 weeks

Weeks of Exposure Lounibos et al. 2010 An. Entomol. Soc. Am.

MANOVA for 2001 Microclimate PCs: Effect

DF

Pillai’s Trace

F

P

Cemetery 15,54

2.046

7.72

<0.001

Type

0.923

64.16

<0.001

Factor 2

High rhmax, rhmean

3,16 4

2

0 A. aegypti persists

-2

-4 -4 Low rhmin

-2

0

Factor 1

2

High 4 tmax, tmean

Lounibos et al. 2010 An. Entomol. Soc. Am.

14L : 10D

21oC

Assay eggs for diapause

10L : 14D

Short-day (diapause) eggs survive south Florida winter better than long-day (non-diapause) eggs Ex Lounibos et al. 2011 JAMCA 27: 433-436

Quiescence and Diapause in Aedine Eggs • Quiescence = inactivity owing to unfavorable environment - flooding, especially with deoxygenated water, stimulates hatch - conditioning influences hatchability Diapause eggs will not hatch with flooding alone, and require reactivation (=diapause termination) - obligate state in univoltine species - facultative in temperate, multivoltine species

Habitat Segregation in South Florida Ovitraps number of mosq.

2000

A. aegypti 1500 WPB BR

1000 500

!

0 urban

suburban

rural

number of mosq.

2000

A. albopictus 1500 WPB BR

1000 500

!

0 urban

suburban

rural

Ex Braks et al. 2003 J. Med Entomol 40: 785-794.

Ovitrap abundances of the two species in three south Florida counties were associated in opposite directions to compound habitat variables quantified from aerial images

Ex Rey et al. (2006) J. Med Entomol. 43:1134-1141

Florida Keys 2002-09

Aedes aegypti

Ex Hribar & Whiteside (2010)

Both species occur throughout the year in south Florida

FMEL Treeholes 1991-2003 Ex Lounibos & Escher (2008)

Natural History & Ecology of Life Stages in Florida Aquatic Stages:

Container Habitats Producing A. aegypti Pupae in the Florida Keys

Ex Hribar & Whiteside (2010) Stud Dipteralog 17:237-251

Ex Lounibos et al. (2003) Ecol. Entomol. 28

0.8

% Survival A. albopictus

0.7

F-stats Litter Wyeomyia Litter x Wyeomyia

6.68* 25.95*** 4.57(ns)

0.6 0.5 0.4 0.3 0.2 0.1 S2

0

+

1

Wyeomyia

S1

-

2

-

Litter

+

Ecological homologs co-occurring in eastern USA

Aedes albopictus (invasive)

Aedes triseriatus (native)

Dipterous predators in Florida treeholes preferentially consume A. albopictus over A. triseriatus

T. rutilus IV consuming Aedes sp. IV instar

C. appendiculata IV consuming Aedes sp. II

Prediction – multigeneration field experiment – (open to immigration):

C. appendiculata will reduce the invasability of naturally exposed tires by A. albopictus and facilitate coexistence with A. triseriatus

Containers & Setting: Golf cart tires with sieved tire water and 2.0 g oak leaves, arranged in groups of four at eight random sites in hammock Treatments (4): - control (no added macroinvertebrates) - 100 I A. triseriatus - 100 I A. triseriatus + 2 IV C. appendiculata - 100 I A. triseriatus + 8 IV C. appendiculata Census every nine days, adding supplementary A. triseriatus Is and C. appendiculata IVs

Repeat in summers of 2005 & 2006

Mean A. albopictus per tire ( SE)

50

2006

Control Competitor Low Predator (2) High Predator (8)

40

30

20

10

0 9

18

27

Days of observation Both low and moderate-density predator treatments suppressed colonization of tires by A. albopictus (Juliano et al. 2010 Oecologia)

50 2005 2006

Aedes albopictus eggs

40

30

20

10

0 CONTROL

COMPETITOR

LOW PRED.

HIGH PRED.

Treatment

Juliano et al. 2010 Oecologia 162

Natural History & Ecology of Life Stages in Florida

Adults:

What are the bloodmeal hosts of these species in nature?

Blood Meal Analyses of Wild-caught Aedes aegypti Location

Dominant Host

Reference

Kenya

Reptiles

McClelland & Weitz 1963

Thailand

Humans

Ponlawat & Harrington 2005

USA (Hawaii) Humans

Tempelis et al. 1970

USA (PR)

Scott et al. 2000

Uganda

Humans Rodents

McClelland & Weitz 1963

Blood Meal Analyses of Wild-caught Aedes albopictus Location Brazil

Dominant Host Humans, cows

Spain

Humans

Muňnoz et al. 2011

Thailand

Humans

Ponlawat & Harrington 2005

USA (Hawaii) Humans

Reference Gomes et al. 2003

Tempelis et al. 1970

USA (Illinois) Rabbits & Rats USA (Missouri) Rabbits

Niebylski et al. 1994

Savage et al. 1993

Evidence for Multiple Blood Meals per Gonotrophic Cycle Based on Dissections of Biting Females % of A. albopictus with eggs: Thailand 19% (n=2341)

Singapore 8% (n=631)

Kenya data from Trpis & Hausermann 1986

Thai data from Gould et al. (1970); Singapore from Chan (1971)

Activity Rhythms of Female Aedes aegypti:

Jones, 1981, Physiol. Entomol. 6

Dispersal of gravid A. aegypti, marked with Rb, in Rio de Janeiro

Ex Honorio et al. 2003 Mem. Inst. Osw. Cruz 98

Dispersal of gravid A. albopictus, marked with Rb, in Rio de Janeiro

Ex Honorio et al. 2003 Mem. Inst. Osw. Cruz 98

In experiments in outdoor cages, Rey & O’Connell (2014) found both skip oviposition and laying eggs on the water surface to be more common in A. aegypti than A. albopictus

Part III: Will Satyr-resistant Aedes aegypti Recover in Florida?

AFTER ~20 YR OF ABSENCE, A. AEGYPTI RECENTLY DETECTED IN THREE SUBURBAN-RURAL SOUTH FLORIDA CEMETERIES

SSix parallel collecting transects established from the coast to inland in Palm Beach County, Florida

Kriged distribution of Ae. aegypti

0% transparency

Color intensity internally consistent within sampling period Reiskind & Lounibos 2012 Med. Vet. Entomol.

Six Year Comparison

2006-07

2013

Interpolated landscapes compared between sampling periods ( brown = preponderance of A. albopictus, blue = preponderance of A. aegypti)

100

NS

NS

% Aedes aegypti (n=6 sites)

90

NS

Mean % A. aegypti in 2013

80 p<0.009

70

Mean % A. aegypti in 2006 & 2007

60 50 40 30 20 NS

10 0 0

1

2

3

4

5

6 7 8 9 10 11 12 13 14 15 Kilometer from Coast

Paired T-tests on proportion A. aegypti (df=5)

Courtesy M. Reiskind NCSU

Selected References

Christophers SR. 1960. Aedes aegypti (L.) The Yellow Fever Mosquito. Its Life History, Bionomics and Structure. Cambridge University Press, UK. Hawley WA. 1988. The biology of Aedes albopictus. Journal of the American Mosquito Control Association 4 (Supplement #1): 1-39. Silver, JB. 2008. Mosquito Ecology. (3rd edition). Springer, NY

WORKSHOP: Protecting Florida from Dengue and Chikungunya through Control of Aedes aegypti and Aedes albopictus June 3-4, 2014 THE FLORIDA MEDICAL ENTOMOLOGY LABORATORY University of Florida IFAS Vero Beach, Florida

Florida Medical Entomology Laboratory

Development of pilot programs for dengue prevention

Gary G. Clark, Ph.D. Mosquito and Fly Research Unit CMAVE, ARS, USDA Gainesville, Florida

Presentation topics  World Health Organization (WHO) recommendations for Integrated Vector Management (IVM)  Methods of vector control  CDC evaluations  COMBI (COMmunication for Behavioral Impact)  Status of dengue vaccines  Improved vector control tools

WHO/HTM/NTD/DEN/2009.1

Integrated Vector Management (IVM)* • • • • •

Advocacy, social mobilization and legislation Collaboration within health and other sectors (vector-borne disease control programs) Integration of non-chemical and chemical vector control methods Evidence-based decision-making (adapt interventions to local vector ecology) Capacity building (train staff to manage IVM programs based on the local situation)

* Promoted by the World Health Organization

Methods of vector control* 1. Environmental management (actions to control immature Aedes aegypti). • Improved water supply and water storage systems • Mosquito-proof water-storage containers • Solid waste disposal • Street cleansing • Building structures (remove roof gutters) * Promoted by the World Health Organization

Methods of vector control cont. 2. Chemical control: larvicides (complementary to environmental management and recommended by WHO) • Organophosphates- Temephos* and Pirimiphos-methyl • Insect Growth Regulators- Diflubenzuron, Methoprene*, Novaluron, and Pyriproxyfen* • Biopesticides- Bacillus thuringiensis var. israelensis* and Spinosad * WHO-approved to treat potable water

Production sites for Aedes aegypti

Water storage tanks

Production sites for Aedes aegypti

Discarded tires

Methods of vector control cont. 2. Chemical control: adulticides (applied as residual surface treatments or as space treatments; perifocal treatment = effects of both a larvicide and an adulticide) • Insecticide susceptibility, droplet size, application rate, and indoor penetration (open doors and windows) are crucial to efficacy • Ground applications should be focused on areas where people congregate (high density housing, schools, etc.)

Adulticides cont.

• • •

In emergencies, treatments should be carried out every 2-3 days for 10 days Space sprays recommended for emergency use during an ongoing epidemic* With resources, emergency space spraying can be initiated at the same time that source reduction and larviciding are intensified

“* No well-documented example of the effectiveness of this approach interrupting an epidemic.” WHO, 2009.

Insecticides used as cold aerosols or thermal fogs • Organophosphates- Fenitrothion, Malathion, and Pirimiphos-methyl • Pyrethroids- Bioresmethrin, Cyfluthrin, Cypermethrin, Cyphenothrin, Deltamethrin, D-Phenothrin, Etofenprox, Lambda-Cyhalothrin, Permethrin and Resmethrin

CDC evaluations of emergency control tools in Puerto Rico • • •

•

•

Ground ULV applications versus Aedes aegypti C-130 (Hercules transporter) with USAF Reserve Unit from Columbus, OH US Navy (DVECC = NECE) with PAU-9 from Jacksonville Naval Air Station, FL Wild mosquitoes susceptible to naled (Dibrom 14) and the insecticide reached the ground but did not penetrate houses Limited, transitory impact on wild population

Ground ULV application in San Juan, Puerto Rico

Ground ULV application

WHO/CDS/CPE/PVC/2001.1

Aerial application of naled with USAF’s C-130 over San Juan, Puerto Rico

US Navy’s PAU-9 unit (no longer in the DoD inventory)

Aerial application of naled with PAU-9 over San Juan, Puerto Rico

Indoor application with thermal fog unit in Cuba

Indoor application with portable ULV unit in Panama City, Panama

Biological Control Introduction of organisms that prey upon, parasitize, compete to reduce populations of the target species. • Larvivorous (native species) fish used to eliminate larvae from large containers used to store potable water in many countries. • Predatory copepods have been effective in eliminating dengue vectors in Viet Nam for several years.

Individual and household protection • Clothing for protection against dengue vectors • Repellents (DEET, IR 3535 or picaridin) for the skin or clothing • Household insecticide aerosol products, mosquito coils, or insecticide vaporizers reduce biting activity • Window and door screens and air conditioning also reduce biting activity

Communication for Behavioral Impact (COMBI) • COMBI is a methodology to plan communication and social mobilization activities in support of a public health program • COMBI focuses on measuring changes in specific behaviors, not just changes in knowledge or attitudes If a dengue program is not working very well, communications and COMBI will not be successful!

COMBI Guide

WHO/CDS/WMC/2004.2

The 3 COMBI principles 1. Identify key, specific behavioral objectives linked to dengue prevention and control objectives 2. Conduct a situational analysis to define the behavioral and communication objectives as part of the formative research activities 3. Develop a strategic communications (COMBI) plan that includes evaluation of behavior change

Epidemiological surveillance

Integrated

Communications

vector management

(COMBI)

Laboratory

Environment

Patient care Panamerican Health Organization

Dengue vaccines •

High levels of country interest – prevention and vaccines

•

Low levels of awareness about dengue vaccine development in the “vaccine community”

•

Effective vaccine must be tetravalent (4 viruses)

•

A number of candidates – ‘strong pipeline’ in different stages of evaluation

Dengue vaccines • Field testing of an attenuated tetravalent vaccine currently underway • 5 companies soon to enter human trials • No licensed vaccine at present - some projections for a licensed vaccine in 2015 • Vector control continues to be the key to dengue prevention and will still be needed (e.g., YF and JE vaccines)

Towards improved vector control tools* • Insecticide-treated materials (ITMs)

• Deployed as bed nets, window curtains,

•

and long-lasting insecticidal fabric covers for domestic water storage containers can reduce vector densities to low levels Curtains were well-accepted in Mexico and Venezuela by the community and reinforced for controlling mosquitoes and household pests

* Not sufficiently field-tested to be recommended by the WHO as public health interventions

Improved vector control tools cont. • Lethal ovitraps 1. Ovitraps used for surveillance have an insecticide incorporated on the oviposition substrate (= lethal ovitrap) 2. Autocidal trap allows oviposition but prevents adult emergence 3. Sticky trap “traps” females that land on it • Population densities reduced with large numbers of frequently-changed traps. Success in Singapore and Brazil.

Increasing Concern at WHO • Published “Global Strategy for Dengue Prevention and Control (2012-2020)” • Challenges: • Surveillance/diagnostics • Case management • How to sustain vector control (need political will) • Post-vaccine availability (still need vector control)

Provided to workshop participants

From the “WHO Global Strategy for Dengue Prevention and Control 2012-2020*” “An effective response is based: (1)on well-developed contingency plans (2)that are broadly disseminated (3)and thoroughly understood (4) and pre-tested before an epidemic.” * Page 13

The Tool Box* Integrated Vector Control

Vaccines

Primary Prevention

Dengue Control Case Management

Diagnostics

Surveillance Education * Courtesy CDC

Secondary Prevention

Children’s Museum in San Juan, Puerto Rico First museum in the Caribbean specifically designed for children (established 1993)  Hands-on exhibit with live mosquitoes in typical larval habitats  Provided annual training for facilitators  Developed video describing dengue and prevention  Modified exhibit in 1997 to create a laboratory  Funded by San Juan Rotary and Rotary Intl. Continuing strong in 2013; 24,000 visitors! 

Children’s Museum Exhibit

WORKSHOP: Protecting Florida from Dengue and Chikungunya through Control of Aedes aegypti and Aedes albopictus June 3-4, 2014 THE FLORIDA MEDICAL ENTOMOLOGY LABORATORY University of Florida IFAS Vero Beach, Florida

Florida Medical Entomology Laboratory

Surveillance of Aedes aegypti and Ae. albopictus Daniel L. Kline Mosquito and Fly Research Unit USDA-ARS, CMAVE Gainesville, FL

Why Surveillance? • Entomological surveillance is used to determine changes in the geographical distribution and density of the vector, evaluate control programs, obtain relative measurements of the vector population over time and facilitate appropriate and timely decisions regarding interventions. It may also serve to identify areas of high-density infestation or periods of population increase. A number of methods are available; the objective of this talk is to direct participants towards known resources.

Peridomestic Mosquito Surveillance in Florida • Surveillance of peridomestic mosquito species has NOT been a focus of most Florida Mosquito Control Districts/Programs. • However, there are literature references, websites, and other developing resources to guide the development of a successful peridomestic surveillance program.

Resources • Focks, D.A. 2003. A review of entomological sampling methods and indicators for dengue vectors, WHO TDR/IDE/Den/03.1 • 2014. Website set up by Rutgers University with lessons learned during a 5 year USDA sponsored Area Wide Asian Tiger Mosquito program. • March 2014: The Governing Board of the Florida Mosquito Control Association created an ad hoc working group to address Aedes aegypti/albopictus as it relates to possible dengue and chikungunya virus transmission in Florida.

Ad hoc committee on Dengue/Chikungunya Vector Management and Response • Overall objective is to provide guidance to Florida mosquito control districts/programs on urban Aedes aegypti/albopictus management. • Working group is being co-chaired by Jim Cilek and FMCA President, Neil Wilkinson. Four broad categories will be addressed by the group: surveillance, control, education, and administrative/agency response. • Current committee members are Jim McNelly, Dan Kline, Roxanne Connelly, Peter Connelly, Chris Lesser and Joel Jacobson.

Surveillance Standard Operating Procedures Have Been Sent Out for Review

• SOP 1---Peridomestic container survey • SOP 2---Ovijar survey procedure • SOP 3---Peridomestic adult surveillance

Standard Operating Procedure (SOP-1): Peridomestic Container Survey • Overall Methodology – Initially, grid area or neighborhood to aid in surveying and documenting location of containers and prioritize areas for control. – Identify “super producers,” i.e. sites with a multitude of potential production sites such as cemeteries, salvage yards, boat yards, etc. – Coordinate with other entities with related responsibilities (e.g. code enforcement). Some of these entities are already actively tracking sites, such as illegal tire dump sites.

Specific Methods for peridomestic container and immature surveillance • House to House Surveys: determine if, what type, and number of containers per residential unit contain larvae and pupae to get quantitative abundance data in neighborhoods. • Pupal Surveys: (# pupae per type of container or # pupae per house). Pupal productivity in containers is the most important parameter to determine because this is a closer estimate of adult production from that habitat.

Container Surveys

Numbers of Containers per house April

August Number of albos per trap 1.006565213 - 5 5.000000001 - 10 10.00000001 - 15 15.00000001 - 20 20.00000001 - 25 25.00000001 - 30 30.00000001 - 40 40.00000001 - 50 50.00000001 - 100

Site receiving education only

Quantitative Indices • Baseline determination of immature abundance. • Provide a tool to track population changes over time • Examples – Container Index: # positive containers/total containers surveyed – Breteau Index: # positive containers/100 houses surveyed

Larval Surveillance

Larval Surveys (1)

Look at container preference/productivity

(2)

Rural to urban differences

Standard Operating Procedure (SOP-2): Ovijar Survey Procedure • Jars/ovitraps---Many types available. Filled ca. ¾ full with water or infusion; seed germination paper or wooden tongue depressor “roughened” to facilitate egg laying. • Placement---Rutgers ATM website recommends 1-4/city block; no less than 30 m apart so that they do not interact with each other. Place in shady spots. • Sampling frequency---check at least once per week. • Evaluation---count eggs (identify to species if you can) or alternatively hatch eggs and rear to 4th instar larvae and identify using the taxonomic key of Darsie and Morris (2003) • Operational note: In Italy, Carrieri et al. (2012) estimated that 44 Ae. albopictus eggs/ovijar was the epidemic threshold for chikungunya outbreaks.

Egg Surveillance

Ovitrapping

Tiger Tubes

Tiger Tubes

Comparison of 3 Ovitraps Tiger Tube

LBJ

Green Vase

top

162

106

228

no top

493

442

316

TOTAL

655

548

514

Photos by Pete Obenauer

DATE

HEATED

UNHEATED

Δ⁰F

06/06/12 06/13/12 06/20/12 06/29/12 07/05/12 07/12/12 07/18/12 07/25/12 08/02/12 08/08/12 08/15/12 08/24/12 08/29/12 09/05/12 09/13/12 09/20/12 10/03/12 10/10/12 10/24/12 10/31/12 11/07/12

442 124 321 408 246 186 198 753 451 514 914 1252 632 401 875 1142 867 309 376 168 30

346 17 131 238 148 44 281 245 720 382 465 726 156 454 97 455 252 36 193 44 4

2.4 broken 7.7 broken 4.9 4.6 5.6 12.1 11.0 broken broken 4.1 3.3 broken 6.9 2.0 1.0 3.4 7.3 19..4 15.2

TOTAL

10,601

5,434

SOP 3: Adult Mosquito Surveillance • Sticky ovitrap survey • BG Sentinel(BGS) trap survey

Adult Surveillance

Landing/Biting Counts • A technique which has fallen into some disfavor due to the risk of exposing individuals to disease agents.

WHAT KIND OF TRAPS SHOULD BE USED?

NEW JERSEY LIGHT TRAP

CDC TYPE TRAPS

Updraft UV Trap No CO2

Standard CDC Updraft CDC Style Light Style Light Trap Trap

Aedes albopictus is not readily attracted to traditional light traps (Chan 1985a, 1985b).

Sticky Ovitrap Surveys • Used in Australia by Scott Ritchie – Operational Note: Threshold >2 females/ovitrap indicated increased risk of dengue transmission with Ae. aegypti in Australia (Ritchie et al. 2004).

• CDC Autocidal Gravid Ovitrap (AGO) – Uses a black 5-gal bucket with a sticky sheet inside a tube in the bucket. – AGO sticky traps can be left in the field without replacing the adhesive for 6-8 weeks. Traps are currently not commercially available.

CDC-AGO trap Autocidal gravid ovitrap Assembling Instructions

Trap components A. ¾” black polypropylene netting (Industrial Netting, Minneapolis, Minnesota, USA) covering the entrance of the trap to exclude the entry of larger debris or organisms B. 3.8 l black, polyethylene cylinder that serves as the trap entrance (12.8 cm diameter) and capture chamber C. Sticky surface covering the interior of the capture chamber that is made of a black styrene cylinder (16 cm diameter); the inner surface is coated with 155 g / m2 of a non-setting, polybutylene adhesive (32UVR, Atlantic Paste & Glue Co. Inc. Brooklyn, New York, USA) D. Screen barrier at the bottom of the capture chamber to prevent adult mosquitoes from reaching the infusion or prevents any mosquito emerging from the infusion to escape from the trap E. Black pail lid F. Black polyethylene pail (5 gal.) G. Micro-drainage holes to allow excess infusion to drain from the trap H. 10 l of water I. 30 g hay packet (do not use alfalfa or leguminosae)

Place the trap in a place where you can comfortably inspect its contents

Take the rubber band and top net off

Pick the trapped mosquitoes using a needle or forceps and place each specimen on a paper towel – by species (e.g., Aedes aegypti, Culex spp.)

Usually, there is no need to remove other insects (if there are too many, change the sticky board). Mosquitoes need to be removed so you do not count them twice

With practice, you will be able to separate mosquitoes by species and sex. You may want to use magnifiers if needed.

Put the net and rubber band back on and place the trap where it was.

Adult Surveillance • BioGents Sentinel Trap Survey – Current commercial version (available through BioQuip) has a white outside casing; will adequately assess adult populations when baited with BG lure. – An experimental version of the same trap design with black outside casing and white top was better than the commercial all white casing for attracting Ae. aegypti.

Adult Survey With BGS Traps • Traps should be placed: – In shaded areas, not in direct sunlight – Away from other dark objects (especially if using black housing) – Near resting areas for mosquitoes – Out of areas subject to heavy winds – Traps in Florida will likely require a rain shelter – Collection size can be increased by using the BG Lure, BG-Lure + Octenol, CO2 alone or in combination with the other lures, but do NOT use octenol alone

BioGents Sentinel Trap (BGS)

Optional Lures • BG-Lure---a dispenser which releases a defined combination of lactic acid, ammonia, and fatty acids, all substances that are found on human skin. Originally developed for Aedes (Stegomyia) aegypti. • CO2 can be added to the air stream as an additional attractant.