VECTOR CONTROL, PEST MANAGEMENT, RESISTANCE, REPELLENTS

Field Evaluation of a Lethal Ovitrap for the Control of Aedes aegypti

(Diptera: Culicidae) in Thailand
RATANA SITHIPRASASNA, PRADITH MAHAPIBUL, CHUMNONG NOIGAMOL,
MICHAEL J. PERICH,1 BRIAN C. ZEICHNER,2 BOB BURGE,3 SARAH L. W. NORRIS,4
JAMES W. JONES,5 SONYA S. SCHLEICH,3 AND RUSSELL E. COLEMAN
Department of Entomology, U.S. Army Medical Component, Armed Forces Research Institute of Medical Sciences,
Bangkok, Thailand

J. Med. Entomol. 40(4): 455Ð462 (2003)

ABSTRACT In 1999 and 2000 we evaluated a lethal ovitrap (LO) for the control of Aedes aegypti
(L.) in three villages in Ratchaburi Province, Thailand. Two blocks of 50 houses (a minimum of 250 m
apart) served as treatment and control sites in each village, with each house in the treatment area
receiving 10 LOs. Thirty houses in the center of each treatment and control block were selected as
sampling sites, with larval and adult mosquito sampling initiated when LOs were placed. Sampling was
conducted weekly in 10 of the 30 houses at each site, with each block of 10 houses sampled every third
week. Sampling continued for 30 wk. EfÞcacy of the LO was evaluated by determining number of
containers with larvae and/or pupae per house and number of adult mosquitoes collected inside each
house. In 1999, the LO had a negligible impact on all measures of Ae. aegypti abundance that were
assessed; however, fungal contamination of insecticide-impregnated strips may have been responsible
for the low efÞcacy. In 2000, signiÞcant suppression was achieved based on changes in multiple
entomologic criteria (containers with larvae, containers with pupae, and number of adult Ae. aegypti);
however, control was not absolute and neither immature nor adult Ae. aegypti were ever eliminated
completely. We conclude that the LO can reduce adult Ae. aegypti populations in Thailand; however,
efÞcacy of the LO is lower than desired due primarily to the high number of alternative oviposition
sites. LO efÞcacy may be improved when used as part of an integrated control program that places
emphasis on reduction of adjacent larval habitats. Further studies are required to assess this issue.

KEY WORDS Aedes mosquitoes, surveillance, control, dengue, lethal ovitrap

DENGUE AND DENGUE HEMORRHAGIC fever (DHF) occur
throughout the tropical and semitropical regions of
the world (Gubler and Kuno 1997). Aedes aegypti (L.),
the primary vector of dengue viruses, is an urban
mosquito that relies on artiÞcial containers such as
ßower-pots, small cisterns, discarded tires and cans as
breeding sites (Christopher 1960, Kittayapong and
Strickman 1993). Aedes aegypti primarily feeds on hu-
mans and frequently rests in secluded locations inside
homes where conventional insecticide spraying is
minimally effective (Perich et al. 1990, 2000, Tonn et
al. 1969). Sustained control of Ae. aegypti requires a
combination of source reduction and insecticide treat-
ment (Gratz 1991, 1993, PAHO 1994). However, not
The views of the authors do not purport to represent the position
of the Department of the Army or the Department of Defense.
1 Louisiana State University, Baton Rouge, LA.
2 US Army Center for Health Promotion and Preventive Medicine,
Aberdeen Proving Ground, MD.
3 Walter Reed Army Institute of Research, Silver Spring, MD.
4 U.S. Army Medical Research Institute of Infectious Diseases, Fort
Detrick, MD.
5 E-mail: James.Jones.3@amedd.army.mil.
all breeding sites can be totally eliminated or made
mosquito-proof, and it is difÞcult to involve all mem-
bers in the community in a sustained clean-up cam-
paign. In addition, neither adulticides nor larvicides
are completely effective against Ae. aegypti. The de-
velopment of novel, effective methods for the
control of dengue vectors are therefore urgently
needed, with particular emphasis on methods that are
environmentally benign, cost-effective, and suitable
for integration into community-based control pro-
grams (Chunsuttiwat and Wasakarawa 1994, Service
1992, Swaddiwudhipong et al. 1992).
Fay and Perry (1965) and Fay and Eliason (1966)
Þrst reported the development of an ovitrap that
could be used as a tool to conduct Ae. aegypti surveil-
lance. The ovitrap has subsequently been used
throughout the world (Chadee et al. 1995, Kaul and
Geevarghese 1979, Mogi et al. 1988) and is particularly
effective in areas with low vector populations (Service
1993). Ovitraps have also proven useful for surveil-
lance of other container-inhabiting mosquitoes, such
as Ae. albopictus (Skuse) (Bellini et al. 1996, Mogi et
al. 1988).
456 JOURNAL OF MEDICAL ENTOMOLOGY
The ovitrap was Þrst used to control Ae. aegypti in
1969 in Singapore (Chan 1973). Chan et al. (1977)
subsequently modiÞed the ovitrap by incorporating an
autocidal screenÑthis modiÞed ovitrap attracted
more Ae. aegypti than did other domestic container
habitats in Þeld tests. Studies by Lok et al. (1977) and
Cheng et al. (1982) provided further evidence that
ovitraps could be used for the control of dengue vec-
tors. Zeichner and Perich (1999) modiÞed the ovitrap
by incorporating an insecticide-impregnated oviposi-
tion strip into the design. This lethal ovitrap (LO) was
evaluated against a laboratory strain of Ae. aegypti
(Zeichner and Perich 1999) and against Þeld popula-
tions of Ae. aegypti in Brazil (Perich et al. 2003). The
study in Brazil demonstrated that the LO could reduce
both sub-adult and adult populations of Ae. aegypti.
The goal of the current study was to evaluate the
efÞcacy of the LO in controlling immature and adult
Ae. aegypti populations in Thailand.
Materials and Methods
Study Site. Field tests were conducted in three
villages (designated Vil-1, Vil-2, and Vil-3) in Chom
Bung District, Ratchaburi Province, Thailand. Tests
were conducted from 7 April to 29 October in 1999 and
from 24 May to 15 December in 2000.
Study Design. A split-plot design was used, with
village as the block factor, area (treated versus control
groups) as the whole-plot factor, and sampling sched-
ule of houses as the split-plot factor. Within each of the
three villages, two groups of 50 houses each were
selected as study sites. The two groups of houses were
separated by a minimum of 250 m. One group of
houses was randomly assigned as the treated area
with the other group serving as an untreated control.
All 50 houses in the treated group received 10 LOs
each, with Þve LOs placed inside the house and Þve
outside. Ovitraps were placed in locations where they
were unlikely to be disturbed (i.e., behind furniture,
under tables, or next to walls). Thirty houses in the
center of each group of 50 houses were selected as
sampling sites. The 20 houses in each group that were
not sampled created a buffer around the sampling
siteÑthis buffer was intended to limit movement of
Ae. aegypti into the treated block of houses from sur-
rounding areas. Sampling was conducted weekly in 10
of the 30 houses (one-third of the houses in each group
were sampled each week, with all 30 houses sampled
once during a 3-wk period).
Ovitrap Design. The LO (U.S. patent number
5,983,557, 11 November 1999) used was the same black
polyethylene cup as that described by Zeichner and
Perich (1999) and Perich et al. (2003). A red-velour
paper strip (11 cm ⫻ 2.5 cm) was treated with delta-
methrin (1.0 mg active ingredient per strip) and at-
tached to the cup using a paper clip (Zeichner and
Perich 1999). This strip served as the oviposition sub-
strate. The 10 LOs in each treated house were Þlled
within 2 cm of the top with a 10% hay infusion-water
solution as described by Reiter et al. (1991). Use of the
hay infusion-water solution was intended to enhance
Vol. 40, no. 4
the attractiveness of the LOs. Ovitraps were checked
weekly, and 10% hay infusion-water added as needed
and deltamethrin-impregnated strips replaced if
found missing. Treated strips were replaced with
freshly treated strips monthly for the Þrst 8 wk of the
study in 1999; however, strips left in the ovitraps for
1-mo developed a fungal growth that physically pre-
vented female mosquitoes from directly contacting
the strips. To prevent this problem, treated strips were
replaced bi-weekly after the Þrst 8 wk of the study.
Surveillance Before Placement of Lethal Ovitraps. In
1999, a survey of larval containers was conducted from
25Ð28 January, 1Ð4 February, and 8Ð11 February in Vil-1,
Vil-2, and Vil-3, respectively. In 2000, preliminary surveys
were conducted on 27Ð31 March, 5Ð13 April, and 13Ð27
April. The types of containers, total number of contain-
ers, number of containers with water, and number of
containers with larvae or pupae were determined in 30
houses in each control and treated block in each of the
three villages. The houses sampled were the same houses
used in the actual evaluation of the LO.
Postplacement Surveillance. We conducted weekly
assessment of larval and adult mosquito populations
in the control and treated blocks of houses in each of the
three villages to evaluate the efÞcacy of the LO. Each
week, we determined (1) the total number of containers
(excluding LOs) in and around each house, (2) the
number of these containers with water, (3) the total
number of these containers with immature (larvae
and/or pupae) mosquitoes, and (4) the number of adult
mosquitoes in each house. Adult mosquitoes were col-
lected for 10 min within each house using hand-held
aspirators. Mosquitoes were transported to the Armed
Forces Institute of Medical Sciences laboratory in
Bangkok, sorted by species and sex, and enumerated.
Statistics. For the preplacement surveillance, means
of entomologic variables (number of water containers,
number of water containers with water, and number
of water containers with immature mosquitoes)
were compared for the treatment group (treated ver-
sus control) across villages by analysis of variance
(ANOVA). The Tukey multiple comparison proce-
dure (PROC ANOVA; SAS Institute 2001) was used
to separate signiÞcantly different means.
For the postplacement surveillance, the following
procedures were used. During the course of the study,
each house was sampled 10 times over the 30-wk
period. For each house by year, weekly counts for
each variable of interest were summed over the study
period. Means and standard deviations for each treat-
ment group, village and year were then calculated
based on these 30-wk cumulative totals. All analyses
were performed on these 30-wk totals for each house
instead of the weekly totals for each house. Frequency
distributions for numbers of containers per house,
containers with larvae or pupae, and adult mosquito
counts were positively skewed. LOG10 transforma-
tions were applied. After transformation, all variables
except counts for adult Ae. aegypti met assumptions of
normality and homogeneity of variance.
A mixed model ANOVA for split plot experiments
(Littell et al. 1996) was used to evaluate the main and
July 2003 SITHIPRASASNA ET AL.: FIELD EVALUATION OF A LETHAL OVITRAP 457

Table 1. Preliminary survey of water containers in treated and control blocks of three villages in Ratchaburi Province, Thailand. A
total of 30 houses each were checked for total water containers, containers with water, and containers with larvae in each control and
treated block in each village

Village 1 Village 2 Village 3

Block
Jan 1999 Mar 2000 Feb 1999 Apr 2000 Feb 1999 Apr 2000
Mean number of water containers per house (⫾SEM)
Control 24.0 (1.61) 29.9 (2.48) 23.2 (2.26) 18.6 (1.84) 22.9 (1.94) 24.7 (1.79)
Treated 24.2 (2.29) 26.7 (1.94) 23.0 (2.30) 20.8 (2.14) 26.3 (2.73) 24.2 (2.93)
Mean number of water containers per house with water (⫾SEM)
Control 16.4 (1.44) 19.9 (1.91) 13.7 (1.30) 10.6 (0.99) 17.0 (1.50) 20.0 (1.40)
Treated 16.0 (1.50) 18.2 (1.76) 13.0 (1.36) 10.5 (1.04) 13.7 (1.39) 16.9 (1.68)
Mean number of water containers per house with larvae (⫾SEM)
Control 3.6 (0.54) 0.5 (0.21) 3.7 (0.73) 0.4 (0.18) 4.2 (0.70) 1.0 (0.26)
Treated 3.5 (0.41) 2.1 (0.36)a 3.0 (0.66) 0.9 (0.23) 3.0 (0.52) 1.1 (0.42)

a
Values in the treated block are signiÞcantly different (ANOVA, with TukeyÕs mean separation procedure, P ⬍ 0.05) than corresponding
values in the control block.

interaction effects of treatment variables in the study.
Treatment group (control versus test) and weekly sam-
pling sequence of houses were treated as Þxed effects,
while village was treated as a random effect. Interactions
between treatment group and sampling sequence were
also included in the model. Data for each year were
analyzed separately. In examining the datasets for each
village there were obvious differences in the size of
mosquito populations between treatment groups across
villages. Therefore, subsequent analyses were conducted
to evaluate effects of LOs on entomologic measures for
the treatment group and sampling sequence within
each village separately. For variables that failed to nor-
malize after transformation (i.e., adult Ae. Aegypti
counts), nonparametric Wilcoxon rank-sum tests
(PROC NPAR1WAY; SAS Institute 2001) were used to
compare treatment groups overall each year as well as
treatment groups within each village each year. Unless
stated otherwise, all comparisons were made at the P ⬍
0.05 level of signiÞcance.
Results
Surveillance Before Placement of Lethal Ovitraps.
Pretreatment surveillance focused on assessing the type
and number of water containers found in each village
and whether the various containers contained immature
mosquitoes. A variety of water containers were identi-
Þed during the preliminary survey, including water jars,
ant traps, ßower pots, wash basins, cisterns, tires, coconut
husks, empty glass bottles, and tin cans. Twenty-four
percent (1,045/4,353) and 76% (3,308/4,353) of the con-
tainers were found outdoors and indoors, respectively.
Almost 45% (1,946/4,353) of all water containers were
earthen water jars of various sizes. Only 18% (185/1,045)
of the water containers found indoors were water jars,
whereas 53% (1,761/3,308) of the outdoor water con-
tainers were water jars.
Data from the preliminary survey is presented in
Table 1. Except for a higher mean number of water
containers per house with larvae in the treated block
of Village-1 (Vil-1) in 2000, there were no signiÞcant
differences between control and treated houses in any
of the three villages for any of the criteria assessed in
either 1999 or 2000.
Postplacement Surveillance, 1999. Figure 1 presents
the overall effects of the lethal ovitrap on immature
mosquito populations, while the effects of the lethal
ovitrap on adult mosquito populations is presented in
Fig. 2. The data in Figs. 1 and 2 represent mean values
recorded over each 3-wk period. In general, fewer
water containers with larvae were found in treated
blocks than in control blocks; however, absolute
(100%) control was never achieved. The LO had less
of an effect on adult mosquitoes, with a clear, sus-
tained reduction in Ae. aegypti populations only ob-
served in Vil-1 in 1999.
In 1999, there was no signiÞcant difference in the total
number of water containers or the number of containers
with water found in the treated blocks compared with
the control blocks (Table 2). Although the mean number
of containers with larvae and mean number of containers
with pupae overall were both lower in the treatment
blocks than the control blocks, these differences were
not signiÞcant. Within villages, only in Vil-1 was a sig-
niÞcantly lower (F11.56; df ⫽ 1,54; P ⫽ 0.0013) number
of containers with larvae found (Tables 2 and 3) in the
treated block (19.6 ⫾ SE ⫽ 14.9) compared with the
control block (30.6 ⫾ 16.7) (Table 2). In Vil-1 a signif-
icant (P ⫽ 0.0006) difference in the number of contain-
ers with pupae (Tables 2 and 3) was observed also, with
the treatment block having a lower mean number of
containers (6.9 ⫾ 7.6) compared with the control block
(13.1 ⫾ 9.1).
Overall, the mean number of adult Ae. aegypti mos-
quitoes in 1999 was signiÞcantly lower (Z ⫽ 3.09, P ⫽
0.0020) in the treatment blocks (21.1 ⫾ 29.0) than in
the control blocks (39.2 ⫾ 42.4) (Table 2). Although
the majority of mosquitoes were male, similar effects
were observed with both male and female mosquitoes.
Within villages, there was a signiÞcant difference be-
tween treatment groups in Vil-1 (Z ⫽ 4.6l8, P ⬍
0.0001), with the treated block having a lower mean
number of mosquitoes (26.4 ⫾ 18.8) than the control
block (62.2 ⫾ 29.7). However, differences in the num-
ber of adult Ae. aegypti found in the treated blocks of
458 JOURNAL OF MEDICAL ENTOMOLOGY
Fig. 1. Effect of the lethal ovitrap on larval container-inhabiting mosquito populations in three villages in Chom Bung
District, Ratchaburi Province, Thailand.
Vil-2 or Vil-3 compared with the control blocks were
not signiÞcant (Table 2).
In 1999, across all villages there were no signiÞcant
differences in the overall number of non-Aedes mos-
quitoes found in the treated blocks compared with the
control blocks (Tables 2 and 3). Within villages, there
was a signiÞcantly higher (F ⫽ 4.58; df ⫽ 1,54; P ⫽
0.0368) mean total number of non-Aedes mosquitoes
collected per house in the treated block (65.4 ⫾ 57.9)
than in the control block (39.4 ⫾ 29.3) of houses for
Vil-1.
Postplacement Surveillance, 2000. There were no
signiÞcant differences in 2000 between treated and
control blocks in the overall numbers of water con-
tainers or containers with water (Table 4). SigniÞcant
differences were found between treatment groups for
the number of containers with larvae (F ⫽ 63.23; df ⫽
1,174; P ⬍ 0.0001), as well as in number of containers
with pupae (F ⫽ 22.26; df ⫽ 1,4; P ⫽ 0.0092) (Tables
4 and 5). SigniÞcant differences for these variables
were also seen within each village (Tables 4 and 5).
Overall and in each village, the mean number of con-
tainers with larvae and the mean number of containers
with pupae were lower in the treated blocks compared
with the control blocks.
Vol. 40, no. 4
The mean number of total adult (male and female)
Ae. aegypti mosquitoes was signiÞcantly lower (Z ⫽
4.55, P ⬍ 0.0001) in the treatment group (12.4 ⫾ 18.5)
than in the control group (23.4 ⫾ 24.1) (Table 4).
Within each village, treated groups of houses had
signiÞcantly lower mean numbers of adult Ae. aegypti
mosquitoes than control groups. In Vil-1, the treated
group had a signiÞcantly lower (Z ⫽ 2.02, P ⫽ 0.0435)
mean number of mosquitoes (19.3 ⫾ 23. 9) than the
control group (25. 7 ⫾ 18.9). In Vil-2, the mean num-
ber of mosquitoes was also signiÞcantly lower (Z ⫽
3.59, P ⫽ 0.0003) in the treatment group (4.0 ⫾ 4.5)
than the control group (13.8 ⫾ 12.9), as was the case
in Vil-3 (Z ⫽ 2.64, P ⫽ 0.0082) (13.9 ⫾ 18.3 in the
treated group compared with 30.7 ⫾ 33.2 in the control
group).
The LO had less of an effect on adult female
Ae. aegypti than on all adults or on male adults (Table
4). Although there were fewer adult females observed
in the treated blocks in each of the three villages, these
differences were only signiÞcant in Vil-2.
No signiÞcant difference was found between treat-
ment groups in the overall number of other (non-
Aedes) mosquitoes (Table 4). However, within each
village there were signiÞcant differences between
July 2003 SITHIPRASASNA ET AL.: FIELD EVALUATION OF A LETHAL OVITRAP 459

Fig. 2. Effect of the lethal ovitrap on the number of adult Ae. aegypti collected in houses in three villages in Chom Bung
District, Ratchaburi Province, Thailand.

treated and control blocks of houses. In Vil-1, the
mean number of other species of mosquitoes was sig-
niÞcantly lower (F ⫽ 68.87; df ⫽ 1,54; P ⬍ 0.0001) in
the treated group (62.07 ⫾ 26.68) than in the control
group (188.0 ⫾ 90.3) (Table 4 and 5). In Vil-3, the
mean number of other non-Aedes mosquitoes was also
lower (F ⫽ 37.90; df ⫽ 1,54; P ⬍ 0.0001) in the treated
group (96.1 ⫾ 50.0) than in the control group (189.9 ⫾
85.1); however, in Vil-2 the mean number of other
mosquitoes was higher (F ⫽ 144.79, df ⫽ 1,54; P ⬍

Table 2. Effect of the lethal ovitrap on entomologic indices of mosquito abundance in 1999

Mean values (⫾SD) per house recorded overall and in each village
Variable Group
Overall Village 1 Village 2 Village 3
Containers with water Control 162.80 (74.60) 175.27 (72.31) 138.17 (73.15) 174.97 (74.55)
Treated 155.46 (84.96) 185.47 (94.23) 128.20 (70.95) 152.70 (80.81)
Containers with larvae Control 23.62 (16.37) 30.63 (16.67) 17.60 (14.48) 22.63 (15.67)
Treated 17.13 (12.73) 19.63 (14.91)a 12.77 (11.03) 19.00 (11.11)
Containers with pupae Control 9.31 (8.29) 13.07 (9.10) 5.60 (5.22) 9.27 (8.48)
Treated 6.56 (7.06) 6.93 (7.59)a 5.03 (5.72) 7.70 (7.68)
Total adult mosquitoes Control 131.31 (80.42) 101.57 (37.14) 161.57 (84.62) 130.80 (97.18)
Treated 137.26 (100.29) 91.90 (63.20) 197.80 (115.54) 122.07 (85.83)
Non-Aedes mosquitoes Control 92.10 (83.60) 39.37 (29.28) 142.67 (92.44) 94.27 (80.73)
Treated 116.12 (100.78) 65.43 (57.89)a 188.70 (122.08) 94.23 (65.65)
Total adult Ae. aegypti Control 39.16 (42.42) 62.20 (29.67) 18.90 (30.40) 36.37 (52.31)
Treated 21.11 (29.00)a 26.43 (18.75)a 9.07 (12.45) 27.83 (43.00)
Adult female Ae. aegypti Control 7.72 (9.10) 9.90 (6.79) 4.27 (7.42) 9.00 (11.59)
Treated 5.17 (9.85)a 6.27 (7.07)a 1.90 (3.01) 7.33 (14.89)
Adult male Ae. aegypti Control 31.43 (35.20) 52.30 (26.08) 14.63 (24.17) 27.37 (42.18)
Treated 15.94 (20.61)a 20.17 (15.04)a 7.17 (10.08) 20.50 (29.25)

a
Treated values are signiÞcantly different (P ⬍ 0.05) from control values.
460 JOURNAL OF MEDICAL ENTOMOLOGY Vol. 40, no. 4

Table 3. Mixed model ANOVA evaluation of the effect of the lethal ovitrap on three entomologic indices of mosquito abundance in
1999

Overall Village 1 Village 2 Village 3

Source
df F P df F P df F P df F P
Containers with larvae
Treatment 1,2 5.79 0.1378 1,54 11.56 0.0013 1,54 1.73 0.1943 1,54 0.61 0.4373
House sequence 2,170 1.89 0.1546 2,54 0.06 0.9406 2,54 4.65 0.0133 2,54 0.62 0.5417
Treatment*House sequence 2,170 1.73 0.1808 2,54 2.48 0.0932 2,54 0.04 0.9622 2,54 4.96 0.0105
Containers with pupae
Treatment 1,2 2.39 0.2622 1,54 13.45 0.0006 1,54 0.33 0.5695 1,54 0.34 0.5642
House sequence 1,170 3.51 0.0321 2,54 0.25 0.7783 2,54 8.33 0.0007 2,54 0.14 0.8739
Treatment*House sequence 2,170 1.09 0.3385 2,54 2.00 0.1459 2,54 0.17 0.8403 2,54 4.50 0.0156
Adult (non-Aedes) mosquitoes
Treatment 1,2 4.53 0.1670 1,54 4.58 0.0368 1,54 3.88 0.0539 1,54 0.10 0.7532
House sequence 2,170 15.85 0.0001 2,54 0.44 0.6438 2,54 80.44 0.0001 2,54 1.75 0.1827
Treatment*House sequence 2,170 0.38 0.6832 2,54 0.02 0.9848 2,54 3.30 0.0445 2,54 0.00 0.9982

0.0001) in the treated group (608.4 ⫾ 344.0) than in
the control group (113.1 ⫾ 68.8).
Discussion
Results from our study in three villages in western
Thailand suggest that the LO with an oviposition strip
treated with 1.0 mg of deltamethrin reduced natural
populations of adult Ae. aegypti. Perich et al. (2003)
reported that use of the LO in Brazil had a signiÞcant
impact on Ae. aegypti populations as measured by
positive container index, mean number of pupae/
house, and adult mosquito aspiration collections.
Perich et al. (2003) suggested that the LO could aid in
the control of Ae. aegypti when integrated into com-
munity participation programs that target removal of
containers that serve as breeding sites for this mos-
quito. This conclusion was supported by the fact that
a longer treatment period was required to impact on
vector populations (positive container index and
mean number of pupae/house) in the village with the
greatest number of containers. These “competing”
containers presumably reduced the effectiveness of
the lethal ovitrap by diverting gravid females from
visiting LOs. Perich et al. (2003) suggested the LO was
not designed to be a stand-alone devise for the control
of Ae. aegypti, but rather would be most effective when
incorporated into an integrated control program.
Data from our pretreatment surveillance demon-
strated that there were no signiÞcant differences (ex-
cept for a signiÞcantly higher number of water con-
tainers with larvae in the treated block of Vil-1 than in
the control block) between control and treated blocks
of houses before placement of the LO. Although we
did not assess adult mosquito populations during the
pretreatment surveillance, these data suggest that the
control and treated blocks of the study sites were
comparable. In addition, at no point during either year
(1999 or 2000) of the study were there signiÞcant
differences in the number of water containers in the
treated or control blocks of any of the villages.
During 1999, the LO had a minimal impact on con-
tainers with water, containers with larvae, containers
with pupae, total adult mosquito populations, and
adult populations of non-Aedes mosquitoes, with sig-
niÞcant differences noted only in Vil-1 and not in Vil-2

Table 4. Effect of the lethal ovitrap on measures of mosquito abundance in 2000

Mean values (⫾SD) per house recorded overall and in each village
Variable Group
Overall Village 1 Village 2 Village 3
Containers with water Control 168.14 (73.49) 186.00 (74.49) 126.57 (65.86) 191.87 (63.14)
Treated 168.24 (79.18) 197.70 (92.05) 139.50 (61.28) 167.53 (72.52)
Containers with larvae Control 16.76 (7.58) 17.73 (7.45) 14.77 (6.32) 17.77 (8.64)
Treated 8.47 (5.47)a 9.00 (5.56)a 8.70 (5.58)a 7.70 (5.36)a
Containers with pupae Control 5.38 (3.88) 5.97 (3.34) 4.43 (2.79) 5.73 (5.08)
Treated 2.37 (2.26)a 3.03 (2.57)a 2.37 (2.14)a 1.70 (1.88)a
Total adult mosquitoes Control 187.31 (93.05) 213.83 (81.93) 127.10 (70.49) 221.00 (96.12)
Treated 266.37 (306.66) 82.07 (25.42)a 606.73 (323.99)a 110.30 (53.04)a
Non-Aedes mosquitoes Control 163.64 (88.60) 187.97 (90.27) 113.10 (68.78) 189.87 (85.13)
Treated 255.54 (320.57) 62.07 (26.68)a 608.43 (344.01)a 96.13 (49.96)a
Total adult Ae. aegypti Control 23.38 (24.09) 25.67 (18.86) 13.80 (12.89) 30.67 (33.23)
Treated 12.39 (18.48)a 19.27 (23.89)a 4.00 (4.49)a 13.90 (18.26)a
Adult female Ae. aegypti Control 6.62 (7.77) 6.83 (6.58) 4.57 (5.19) 8.47 (10.35)
Treated 4.37 (6.88)a 6.33 (8.36) 1.57 (2.05)a 5.20 (7.63)
Adult male Ae. aegypti Control 16.76 (17.80) 18.83 (15.10) 9.23 (9.15) 22.20 (23.82)
Treated 8.02 (12.41)a 12.93 (16.54)a 2.43 (2.93)a 8.70 (11.54)a

a
SigniÞcantly different (P ⱕ 0.05).
July 2003 SITHIPRASASNA ET AL.: FIELD EVALUATION OF A LETHAL OVITRAP 461

Table 5. Mixed model ANOVA evaluation of the effect of the lethal ovitrap on three key indices of mosquito abundance in 2000

Overall Village 1 Village 2 Village 3

Variable and Effect
F df P F df P F df P F df P
Containers with larvae
Treatment 1,174 63.23 ⬍0.0001 1,54 22.71 ⬍0.0001 1,54 13.16 0.0060 1,54 28.50 ⬍0.0001
House sequence 2,174 0.76 0.4708 2,54 1.58 0.2150 2,54 0.23 0.7970 2,54 1.38 0.2602
Treatment*House sequence 2,174 0.33 0.7217 2,54 0.24 0.7888 2,54 0.07 0.9299 2,54 1.60 0.2103
Containers with pupae
Treatment 1,4 22.26 0.0092 1,54 14.58 0.0003 1,54 11.52 0.0013 1,54 27.48 0.0001
House sequence 2,170 1.37 0.2564 2,54 0.71 0.4976 2,54 0.43 0.6549 2,54 2.98 0.0594
Treatment*House sequence 2,170 0.85 0.4282 2,54 0.30 0.7387 2,54 0.76 0.4740 2,54 0.15 0.8638
Non-Aedes mosquitoes
Treatment 1,4 0.01 0.94467 1,54 68.77 ⬍0.0001 1,54 144.79 ⬍0.0001 1,54 37.90 ⬍0.0001
House sequence 2,170 1.59 0.2068 2,54 3.34 0.0430 2,54 4.43 0.0165 2,54 6.88 0.0022
Treatment*House sequence 2,170 0.17 0.8480 2,54 0.26 0.7751 2,54 0.25 0.7761 2,54 1.17 0.3191

or Vil-3. There was a signiÞcant difference in the
overall number of adult Ae. aegypti collected in treated
versus control blocks. Although fewer adults were
collected in the treated blocks of all three villages, this
difference was only signiÞcant in Vil-1.
During 2000, there were signiÞcant reductions in
containers with larvae, containers with pupae and in
total populations of all adult Ae. aegypti. However,
when sex of the adult Ae. aegypti was evaluated, the
LO had less of an effect on female mosquitoes than on
male mosquitoes, with signiÞcant reductions in female
populations only noted in Vil-2 and not in Vil-1 or
Vil-3. In contrast to the general reduction in adult
Ae. aegypti populations, the LO appeared to have no
overall impact on the number of total adult mosquitoes
collected or in the number of non-Aedes mosquitoes.
Although we would not expect that the LO would
reduce populations of non-Aedes mosquitoes, the fact
that these populations were not affected whereas
Ae. aegypti populations were reduced further supports
the argument that the LO was speciÞcally targeting
Ae. aegypti. Although there were signiÞcant differ-
ences in numbers of non-Aedes mosquitoes in each
village, in some villages there was an increase in these
mosquito populations while in others there was a de-
crease.
Although we are not certain why the performance
of the LO was signiÞcantly lower in 1999 than in 2000,
we noted fungal growth on the insecticide impreg-
nated strips during the Þrst 8 wk of the study in 1999.
To eliminate this fungal growth, we replaced the in-
secticide-impregnated strips bi-weekly rather than
monthly, beginning 8 wk after the study commenced
in 1999. This fungal growth may have affected the
efÞcacy of the LO in 1999. We did not experience this
problem in 2000.
Although we found that the LO had a signiÞcant
effect on mosquito populations, particularly in 2000,
this effect was not as long-lasting or as marked as
would be observed when applying either larvicides or
adulticides (Gratz 1991, 1993). Perich et al. (2003)
suggested that the LO would be most useful when
incorporated into an integrated control program. Our
data supports this suggestion, as control achieved in
this study was lower than desired.
Attraction of gravid Ae. aegypti away from LO to
adjacent containers is not the only possible explana-
tion for the lower than desired control achieved in this
study. Although our study design included a buffer
zone of houses with LO around the central surveil-
lance area, it is possible that adult Ae. aegypti may have
immigrated from beyond the buffer zone into the
treated areas to replace females killed by the LO.
Edman et al. (1998) reported that availability of ovi-
position sites was inversely correlated with the po-
tential for female Ae. aegypti to disperse, suggesting
that gravid females in the treated areas in this study
may have been less likely to disperse than females in
areas without LO or with fewer water containers.
Conversely, although the LO may have reduced pop-
ulations of gravid females in the treated areas, this
reduction may have led to lower rates of oviposition
in containers and corresponding decreases in density
dependent larval mortality. A decrease in larval mor-
tality may have resulted in an increased production of
adults to or near preplacement levels obfuscating ef-
fects of the LO on both immature and adult Ae. aegypti
populations (Hawley 1985, Service 1985).
The number of water containers per house in this
study was substantially higher than reported by Perich
et al. (2003) in Brazil and may have diluted the effect
of the LO. Although it remains to be determined how
signiÞcant a role the LO could play in an integrated
control program, the results of our study provide ev-
idence that the LO can reduce Ae. aegypti populations.
Active community participation in a clean-up program
has been regarded as an effective method of reducing
populations of Ae. aegypti in Thailand (Chunsuttiwat
and Wasakarawa 1994, Eamchan et al. 1989, Swaddi-
wudhipong et al. 1992). Cheng et al. (1982) reported
that the use of a modiÞed ovitrap resulted in a decrease
in the Breteau index of 36% in the control area in
Houston, TX compared with a nearly 500% increase in
the nonovitrap area; however, use of the ovitrap was
accompanied by removal of all other breeding habi-
tats. Incorporation of the LO into a clean-up program
would presumably result in more rapid and a greater
level of control than clean-up alone, particularly if
adjacent water containers were eliminated; however,
further studies will be required to conÞrm this.
462 JOURNAL OF MEDICAL ENTOMOLOGY
Acknowledgments
Funding for this project was provided by the Military
Infectious Diseases Research Program of the U.S. Army Med-
ical Research and Materiel Command, Fort Detrick, MD.
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Received for publication 11 March 2002; accepted 27 Janu-
ary 2003.