WORKSHOP:Protecting Florida from Dengue and
Chikungunya through Control of Aedes aegyptiand Aedes albopictus
June 3-4, 2014
THE FLORIDA MEDICAL ENTOMOLOGY LABORATORYUniversity of Florida IFAS
Vero Beach, Florida
Florida Medical Entomology Laboratory
Chikungunya virus
Florida Medical Entomology Laboratory
Dengue virus
Aedes aegypti Aedes albopictus
2009
2010
2011
2012
2013
DENGUE TRANSMISSION IN FLORIDA 2009-2014
DENV-2
DENV-3
Florida Medical Entomology Laboratory
DENGUE IN THE CARIBBEAN BASIN - 2013
Florida Medical Entomology Laboratory
Prevalence of Dengue: Florida vs. Singapore
Florida Medical Entomology Laboratory
0
100
200
300
400
500
600
700
800
900
2008 2009 2010 2011 2012 2013
INC
IDEN
CE/1
00
,00
0
YEAR
Singapore
Key West
Rio
79th Annual Meeting of the Florida Mosquito Control Association
2007
Jacksonville Florida
Florida Medical Entomology Laboratory
SYMPOSIUM
CHIKUNGUNYA: Is this on the horizon for Florida?
• 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)
Symposium Presentations
Florida Medical Entomology Laboratory
Florida Medical Entomology Laboratory
St. Martin (French) – 3220 8907Martinique – 27670 7088Saint Barthélemy – 7267 7018Guadeloupe – 18000 3843Dominica – 1817 2613St. Martaan (Neth) - 343 700Dominican Republic – 32500 314Anguilla – 33 205British Virgin Islands - 7 90French Guyana – 178 69Haiti – 5500 56St. Kitts – 20 39Aruba – 1 1
Chikungunya Cases
as of May, 2014
Ca. 95,000 Cases
CHIKUNGUNYA IN THE CARIBBEAN – 2013-2014
Cases/100,000
Florida Medical Entomology Laboratory
0.00
1500.00
3000.00
4500.00
6000.00
7500.00
9000.00
Cases/1
00,0
00
CHICKUNGUNYA - Caribbean
Dec 2013 - May 2014
• Dengue - 23 Cases– Travel History – Bolivia, Brazil, Cuba, Dominican Republic, Guadeloupe,
Honduras, Puerto Rico, Trinidad, Venezuela
– Florida Counties – Alachua, Broward, Clay, Hillsborough, Marion, Miami-Dade, 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 Dengue and Chikungunya -2014Imported Cases
Florida Medical Entomology Laboratory
How do we predict risks?
How do we mitigate risks?
Florida Medical Entomology Laboratory
Risk Assessment Risk Management
Surveillance Mosquito-Pathogen
Biology
Mosquito
Control Strategies
Public Health
Interventions
Florida Medical Entomology Laboratory
Mosquito Control
FL DOH
Mosquito Control and FL DOH
Beach
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 aegyptiand Aedes albopictus
June 3-4, 2014
THE FLORIDA MEDICAL ENTOMOLOGY LABORATORYUniversity 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?
Enserink. 2008. Science
Major range expansion of Aedes albopictus
occurred in the past 40 years from Asia
1985
1985
1991
1991
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
1986 & 1987
1988
1989
1990
Distribution of A. albopictus from 1986 to 1994
Modified from O’Meara et al. 1995
Lake city
Gainesville
Ocala
Leesburg
ApopkaOrlando
Kissimmee
St. Cloud
Yeehaw JunctionOkeechobee
1991
1992
1993
1994
Spread on Route 441 of A. albopictus and decline of A. aegypti
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 Spermathecae
Collection
Site
Species Mean No.
(SE) Per Coll
A.aegypti A.
albopictus
Totals
M& K Auto aegypti 73.4 (18.7) 85 0 85
(n=7) albopictus 161.0(60.0) 1 71 72
Belle Glade aegypti 67.5(2.5) 82 3 85
(n=2) albopictus 411.5(170.5) 1 61 62
Totals 169 135 304
Sperm extracted from field-collected females
and DNA amplified
Ex: Tripet et al. (2011) Am. J. Trop. Med. Hyg.
Hypothesis: Male accessory gland (MAG) products
of A. albopictus asymmetrically sterilize A. aegypti
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
Science 156: 1499-1501 (1967)
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 andsegregation of Aedes aegypti by Aedes albopictus
U
N
F
A
V
O
R
A
B
L
E
C
L
I
M
A
T
E
HABITAT
SEGREGATION
ARRIVAL &ESTABLISHMENT
1985
albopictus& aegypti
aegypti(resident)
albopictus(invader)
+
-
L ARVAL
COMPETITION
SAT YRIZ ATION
30° N
25° N
L
A
T
I
T
U
D
E
rural
suburban
urban
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
2 weeks. 4 weeks. 2 weeks 4 weeks
% H
atc
h (
SE
)
0.0
0.2
0.4
0.6
0.8
1.0
A. aegypti
A. albopictus
2001
2006
Weeks of Exposure
***
******
**
Lounibos et al. 2010 An. Entomol. Soc. Am.
Hightmax, tmean
Lowrhmin
Hig
hrh
ma
x, rh
me
an
Factor 1
-4 -2 0 2 4
Facto
r 2
-4
-2
0
2
4
MANOVA for 2001 Microclimate PCs:
Effect DF Pillai’s Trace F P
Cemetery 15,54 2.046 7.72 <0.001
Type 3,16 0.923 64.16 <0.001
A. aegypti
persists
Lounibos et al. 2010 An. Entomol. Soc. Am.
14L : 10D 21oC 10L : 14D
Assay eggs for diapause
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
0
500
1000
1500
2000
urban suburban rural
num
be
r o
f m
osq
.
WPB
BR
0
500
1000
1500
2000
urban suburban rural
num
be
r o
f m
osq.
WPB
BR
A. aegypti
A. albopictus
Habitat Segregation in South Florida Ovitraps
!
!
Ex Braks et al. 2003 J. Med Entomol
40: 785-794.
Ex Rey et al. (2006) J. Med Entomol. 43:1134-1141
Ovitrap abundances of the two species in three south Florida counties
were associated in opposite directions to compound habitat variables
quantified from aerial images
Florida Keys
2002-09
Aedes aegypti
FMEL Treeholes
1991-2003
Both species
occur throughout
the year in south
Florida
Ex Hribar
& Whiteside
(2010)
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
1
2
S1
S2
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
% S
urv
iva
l A
. a
lbo
pic
tus
Wyeomyia
Litter+
- +
-
Litter
Wyeomyia
Litter x Wyeomyia
6.68*
25.95***
4.57(ns)
F-stats
Aedes albopictus
(invasive)
Aedes triseriatus
(native)
Ecological homologs
co-occurring in eastern USA
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
2006
Days of observation
Mean
A. a
lbo
pic
tus
pe
r ti
re (
S
E)
0
10
20
30
40
50
Control
Competitor
Low Predator (2)
High Predator (8)
9 18 27
Both low and moderate-density predator treatments suppressed
colonization of tires by A. albopictus (Juliano et al. 2010 Oecologia)
0
10
20
30
40
50
CONTROL COMPETITOR LOW PRED. HIGH PRED.
Treatment
Ae
de
s a
lbo
pic
tus
eg
gs
2005
2006
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) Humans Scott et al. 2000
Uganda Rodents McClelland & Weitz 1963
Blood Meal Analyses of Wild-caught Aedes
albopictus
Location Dominant Host Reference
Brazil Humans, cows Gomes et al. 2003
Spain Humans Muňnoz et al. 2011
Thailand Humans Ponlawat & Harrington 2005
USA (Hawaii) Humans Tempelis et al. 1970
USA (Illinois) Rabbits & Rats Niebylski et al. 1994
USA (Missouri) Rabbits 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)
Jones, 1981, Physiol. Entomol. 6
Activity Rhythms of Female Aedes aegypti:
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
AFTER ~20 YR OF ABSENCE, A. AEGYPTI RECENTLY DETECTED
IN THREE SUBURBAN-RURAL SOUTH FLORIDA CEMETERIES
Part III: Will Satyr-resistant Aedes aegypti Recover in Florida?
Krigeddistribution of Ae. aegypti
0% transparency
Color intensity internally consistent within sampling period
Reiskind & Lounibos 2012
Med. Vet. Entomol.
SSix parallel collecting transects established from
the coast to inland in Palm Beach County, Florida
Six Year Comparison
Interpolated landscapes compared between sampling periods ( brown = preponderance of A. albopictus, blue = preponderance of A. aegypti)
20132006-07
0
10
20
30
40
50
60
70
80
90
100
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
% A
ed
es a
egy
pti
(n
=6 s
ites
)
Kilometer from Coast
Mean % A. aegypti in 2013
Mean % A. aegypti in 2006 & 2007
NS
NS
NS
NS
p<0.009
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 aegyptiand Aedes albopictus
June 3-4, 2014
THE FLORIDA MEDICAL ENTOMOLOGY LABORATORYUniversity 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
Water storage tanks
Production sites for Aedes aegypti
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
Integratedvector
management
Epidemiological surveillance
Communications
(COMBI)
Laboratory
Patient care
Environment
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*
Diagnostics
Integrated Vector Control
Case Management
Vaccines
Surveillance Education
Dengue Control
Primary
Prevention
Secondary
Prevention
* Courtesy CDC
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 aegyptiand Aedes albopictus
June 3-4, 2014
THE FLORIDA MEDICAL ENTOMOLOGY LABORATORYUniversity of Florida IFAS
Vero Beach, Florida
Florida Medical Entomology Laboratory
Surveillance of Aedes aegyptiand 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/ChikungunyaVector Management and Response
• Overall objective is to provide guidance to Florida mosquito control districts/programs on urban Aedesaegypti/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 peridomesticcontainer 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
Number of albos per trap
<VALUE>
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
April August
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
06/06/12 442 346 2.4
06/13/12 124 17 broken
06/20/12 321 131 7.7
06/29/12 408 238 broken
07/05/12 246 148 4.9
07/12/12 186 44 4.6
07/18/12 198 281 5.6
07/25/12 753 245 12.1
08/02/12 451 720 11.0
08/08/12 514 382 broken
08/15/12 914 465 broken
08/24/12 1252 726 4.1
08/29/12 632 156 3.3
09/05/12 401 454 broken
09/13/12 875 97 6.9
09/20/12 1142 455 2.0
10/03/12 867 252 1.0
10/10/12 309 36 3.4
10/24/12 376 193 7.3
10/31/12 168 44 19..4
11/07/12 30 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
Standard CDC
Style Light
Trap
Updraft CDC
Style Light
Trap
Updraft UV
Trap No CO2
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.
Assembling Instructions
CDC-AGO trapAutocidal gravid ovitrap
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 orleguminosae)
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.
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