RATANA SITHIPRASASNA, PRADITH MAHAPIBUL, CHUMNONG NOIGAMOL,
MICHAEL J. PERICH,1BRIAN C. ZEICHNER,2BOB BURGE,3SARAH L. W. NORRIS,4
JAMES W. JONES,5 SONYA S. SCHLEICH,3AND RUSSELL E. COLEMAN
(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. Ef\u00decacy 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 ofAe. aegypti abundance that were assessed; however, fungal contamination of insecticide-impregnated strips may have been responsible for the low ef\u00decacy. In 2000, signi\u00decant suppression was achieved based on changes in multiple entomologic criteria (containers with larvae, containers with pupae, and number of adultAe. aegypti); however, control was not absolute and neither immature nor adultAe. aegypti were ever eliminated completely. We conclude that the LO can reduce adultAe. aegypti populations in Thailand; however, ef\u00decacy of the LO is lower than desired due primarily to the high number of alternative oviposition sites. LO ef\u00decacy 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.
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 arti\u00decial containers such as \u00dfower-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 ofAe. aegypti requires a combination of source reduction and insecticide treat- ment (Gratz 1991, 1993, PAHO 1994). However, not
all breeding sites can be totally eliminated or made mosquito-proof, and it is dif\u00decult to involve all mem- bers in the community in a sustained clean-up cam- paign. In addition, neither adulticides nor larvicides are completely effective againstAe. 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) \u00derst reported the development of an ovitrap that could be used as a tool to conductAe. 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 asAe. albopictus (Skuse) (Bellini et al. 1996, Mogi et al. 1988).
The ovitrap was \u00derst used to controlAe. aegypti in 1969 in Singapore (Chan 1973). Chan et al. (1977) subsequently modi\u00deedthe ovitrap by incorporating an autocidal screen\u00d1this modi\u00deed ovitrap attracted moreAe. aegypti than did other domestic container habitats in \u00deeld 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) modi\u00deed the ovitrap by incorporating an insecticide-impregnated oviposi- tion strip into the design. This lethal ovitrap (LO) was evaluated against a laboratory strain ofAe. aegypti (Zeichner and Perich 1999) and against \u00deeld popula- tions ofAe. aegypti in Brazil (Perich et al. 2003). The study inBrazil demonstrated that the LO could reduce both sub-adult and adult populations ofAe. aegypti. The goal of the current study was to evaluate the ef\u00decacy of the LO in controlling immature and adult
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.
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 \u00deve LOs placed inside the house and \u00deve 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 site\u00d1this buffer was intended to limit movement of
rounding areas. Sampling was conducted weekly in 10 of the 30 houses (one-third of the houses in eachgroup were sampled each week, with all 30 houses sampled once during a 3-wk period).
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\ue000 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 \u00delled 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
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 \u00derst 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 \u00derst 8 wk of the study.
1999, a survey of larval containers was conducted from 25\u00d028 January, 1\u00d0 4 February, and 8 \u00d011 February in Vil-1, Vil-2, and Vil-3, respectively. In2000, preliminary surveys were conducted on 27\u00d031 March, 5\u00d013 April, and 13\u00d027 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.
assessment of larval and adult mosquito populations in the control and treatedblocks of houses in each of the three villages to evaluate the ef\u00decacy 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.
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 signi\u00decantly 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 eachhouse. 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 adultAe. aegypti met assumptions of normality and homogeneity of variance.
interaction effects of treatment variables in the study. Treatment group (control versus test) and weekly sam- pling sequence of houses were treated as \u00dexed 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., adultAe. 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 theP\ue001 0.05 level of signi\u00decance.
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- \u00deed during the preliminary survey, including water jars, ant traps, \u00dfower 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 earthenwater 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 signi\u00decant differences between control and treated houses in any
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 inAe. aegypti populations only ob- served in Vil-1 in 1999.
In 1999, there was no signi\u00decant 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 (Table2). Althoughthe 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 signi\u00decant. Within villages, only in Vil-1 was a sig- ni\u00decantly lower (F11.56; df\ue002 1,54;P\ue002 0.0013) number of containers with larvae found (Tables 2 and 3) in the treated block (19.6\ue003 SE\ue002 14.9) compared with the control block (30.6\ue003 16.7) (Table 2). In Vil-1 a signif- icant (P\ue002 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\ue003 7.6) compared with the control block (13.1\ue003 9.1).
Overall, the mean number of adultAe. aegypti mos- quitoes in 1999 was signi\u00decantly lower (Z\ue002 3.09,P\ue002 0.0020) in the treatment blocks (21.1\ue003 29.0) than in the control blocks (39.2\ue003 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 signi\u00decant difference be- tween treatment groups in Vil-1 (Z\ue002 4.6l8,P\ue001 0.0001), with the treated block having a lower mean number of mosquitoes (26.4\ue003 18.8) than the control block (62.2\ue003 29.7). However, differences in the num- ber of adultAe. aegypti found in the treated blocks of
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
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