Control of Aedes Breeding Sites With a Dual-action Insecticidal Coating is Effective in Reducing Dengue Transmission: Cluster-randomised Controlled Trial Study
Phase: NA
Conditions: Dengue, Vector Borne Diseases, Arbovirus Infections, Zika, Chikungunya Fever
Sponsor: Rocio Cardenas Sanchez

 
 
 
 
Control of Aedes breeding sites with a dual-action 
insecticidal coating is effective in reducing Dengue 
transmission: Cluster-randomised Controlled Trial 
Study 
 
 
 
 
Study Protocol 
 
 
 
 
Ubication of study: Metropolitan Area of Cucuta, Colombia 
 
 
Approval of the protocol: 11 March 2019 
Update: 12 February 2024 
 
 
 
 
 
 
 

 
Study Protocol 
Place of study 
The base-line study was conducted within 2.5 months, the intervention 
was applied within 3 months and the follow up study with final evaluation 
was within 12 months, in 2019–22 in Metropolitan Cucuta in the North-
East of Colombia, including the city of Cucuta (629,414), and two adjacent 
municipalities Villa del Rosario (93,735) and Los Patios (81,411 
inhabitants) located in the Norte de Santander state [1]. The climate is 
warm and dry, with a mean annual temperature of 26.7 °C and annual 
rainfall of 806 mm. There are two rainy seasons from the end of March to 
the beginning of June and from the end of September to the beginning of 
December. DZC are endemic, with occasional epidemic outbreaks [2]. 
Study design and sample size to baseline study and evaluation  
This is design study for a large cluster randomized trial on the impact of 
treating water containers with a protective paint. The sample size was 
calculated for detection of a 50% reduction in the House Index with > 99% 
power at 5% significance level. Given a baseline HI of 30%, an intra-
cluster correlation coefficient (ICC) of 0.01 and a cluster size of 2,000 
households it was found that a minimum of 12 clusters per study arm was 
needed [3]. The same result but with a lower power (68%) was found for a 
baseline DZC incidence of 3% and a 50% reduction in the intervention 
arm. A large number of households per cluster was needed as Ae. 
aegypti vectors are day-biters [4,5,6] and it was assumed that many 
household members stay within their neighbourhoods during the day 
(mainly housewives, small children and school children when the school 
is close to their house) where they are at risk to get infected. All 
estimations were done through The Shiny CRT Calculator which is a web-
based app to determine sample size and power for cluster trials [7]. 
Sampling procedure for household survey and entomological inspections 
On the basis of available maps, the city was stratified into high, middle 
and low endemic/infested areas for Ae. aegypti (strata) using the 
surveillance information from SIVIGILA of the preceding year as well as 
information obtained from interviews with the vector control 
coordinators in Cúcuta. By this way, 24 high and medium endemic areas 
(clusters) were selected in Cucuta (16 clusters), Los patios (4 clusters) and 
Villa del Rosario (4 clusters). Figure 1 shows the selected clusters in 

Cucuta to be sampled by staff. For the baseline study they were not yet 
paired and allocated to the intervention and control arm as this was 
planned to do with the information resulting from the baseline study 
enabling us to find pairs of clusters with similar characteristics. The 
following characteristic should be similar in each pair of clusters: (I) High 
and middle level endemicity for Aedes-borne diseases; (II) Similar socio-
economic conditions; (III) Similar types of the built environment: houses, 
schools, industry, health centers, workshops, churches and stores; (IV) 
Similar characteristics of public spaces. Within each cluster, all blocks of 
houses were inspected including public spaces. For the 
entomological/socio-demographic household survey, a proportion of 10% 
of 2000 houses was sampled in this study, resulting in 200 houses in each 
cluster. This number was rounded to 250 by convenience. Therefore, the 
study targeted 6000 households (250 households in each cluster) using a 
systematic random sampling method. For this we first estimated the 
sampling interval by dividing the total number of households in each 
cluster (around 2000) by the number of sample households (250), 
resulting in a sampling interval of 8. All households located within each 
cluster were mapped and a randomly defined starting point was selected 
and from there every 8th household was visited. Clear replacement rules 
were given to the interviewers and the compliance was checked in sample 
of families. 
Vector control staff and trained interviewers participated in the survey. 
They were divided into “couples” of one vector inspector and one 
interviewer. Each couple was assigned one cluster where they had to visit 
the sample houses. The interviewer did the interview with the head of 
household and simultaneously the inspector (local vector control staff) 
did the inspection of water containers both of them filling a form. 
Data collection 
The baseline surveys included a household questionnaire and 
entomological survey (see Additional files). The instruments were 
discussed in a participatory workshop with experts and pre-tested among 
the members of vector control staff in the three municipalities leading to 
some modifications. The first instrument was the questionnaire which 
was applied by trained interviewers and the other part was the form for 
the entomological inspections (larvae/pupae survey) which was applied 
by the vector control staff. As this is part of the national vector control 
programme only verbal consent of the house owner was obtained to check 
their water containers. An extensive training was conducted by the 
research team before starting the field work. 

Household questionnaire 
Demographic and health data was collected through face-to-face 
interviews using a standard questionnaire that included both structured 
and semi- structured questions. Household questionnaire was adapted 
from published research in Colombia [8]. The questions referred to socio-
demographic parameters (number of people living in the house, age, sex, 
and educational level), self-reported DZC disease acquired during the last 
months, population mobility during the day, usage of water in tanks and 
willingness to accept the intervention trial (see Additional file). 
Entomological survey 
The standard entomological survey form was adapted following the 
guidelines of Standard Operational Procedures (SOPs) by WHO [9]. The 
questions were adapted to local conditions and survey requirements (see 
Additional file). The following data were recorded from each household: 
total number of containers (potential breeding sites), number of mosquito 
larvae positive and negative containers (any species), pupae count per 
container, container type, and other container characteristics (if they were 
covered or uncovered, outdoors or indoors). 
Dengue surveillance and case definition 
Data of the national surveillance system were obtained from SIVIGILA 
(the national health surveillance system), aggregated by year and setting 
(study areas) over the study period. Notified dengue cases including those 
classified as dengue fever (DF) and severe dengue, relying on a clinical 
case definition or lab confirmed or hospitalized patients. This study used 
annual population data (from the National Institute of Statistics-DANE; 
[10]) for calculating the incidence rate. 
Ethical considerations 
The household questionnaire was only applied to adults who provided 
information related to the purpose of this study. No child or adolescent 
below the age of 18 was interviewed in this study. All people participating 
in the study were informed in local language through the study 
information sheet in a written and oral way. They were asked to sign the 
informed consent form. All participants were informed that their 
participation was voluntary and that their responses remained 
anonymous, therefore the study used numbers which replaced the names 
of individuals and codes which replaced the address of house. Before 

examining the domestic and peri-domestic water-holding containers, the 
field team requested permission to enter the house, did the inspection and 
collected entomological and sociodemographic data. 
The study received approval from local health authorities in Cucuta and 
Norte de Santander and the study protocol was approved by the ethical 
committee of the Albert–Ludwigs-Universität (application number 
141/19) in Freiburg, Germany  
Fig. 1  
 
Selection of clusters in Cucuta 
Map shows the 16 clusters selected in Cucuta which were visited by 
interviewers and vector control staff 

Fig. 2 
 
A concrete ground tank for washing purpose 
Concrete ground tanks for washing (lavadero) are made of cement or ceramic tiles 
with the shape of a rectangle or square and variable sizes 
 
References 
1. Wilson AL, Courtenay O, Kelly-Hope LA, Scott TW, Takken W, Torr 
SJ, Lindsay SW. The importance of vector control for the control 
and elimination of vector-borne diseases. PLoS Negl Trop Dis. 
2020;14(1):e0007831. https://doi.org/10.1371/journal.pntd.00078
31. 
2. Wolbers M, Kleinschmidt I, Simmons CP, Donnelly CA. 
Considerations in the design of clinical trials to test novel 
entomological approaches to dengue control. PLoS Negl Trop Dis. 
2012;6(11):e1937. 

3. DANE. Censo Nacional de Población y vivienda 
2018. https://geoportal.dane.gov.co/geovisores/sociedad/cnpv-
2018/. Accessed 30 Apr 2021. 
4. Instituto Departamental de Salud de Norte de Santander (INS 
Norte de Santander). 2016. Análisis de Situación de Salud con el 
Modelo de Determinantes Sociales. Coordinación de vigilancia en 
Salud Pública. Fecha de consulta: 4 de abril de 2018. Available from 
en: http://salasituacionalidsnds.weebly.com/uploads/1/0/7/1/1071
4324/asisdepartamental-2016-nortedesantander.pdf. Accessed 19 
Mar 2018. 
5. The Shiny CRT Calculator: Power and Sample size for Cluster 
Randomised Trials. 
(2022). https://clusterrcts.shinyapps.io/rshinyapp/. Accessed 20 
June 2022. 
6. Wong J, Astete H, Morrison AC, Scott TW. Sampling considerations 
for designing Aedes aegypti (Diptera:Culicidae) oviposition studies 
in Iquitos, Peru: Substrate preference, diurnal periodicity, and 
gonotrophic cycle length. J Med Entomol. 2011;48(1):45–52. 
7. Yasuno M, Tonn RJ. A study of biting habits of Aedes aegypti in 
Bangkok. Thailand Bull World Health Organ. 1970;43(2):319–25. 
8. Chadee DD. Landing periodicity of the mosquito Aedes aegypti in 
Trinidad in relation to the timing of insecticidal space-spraying. 
Med Vet Entomol. 1988;2(2):189–92. 
9. Hemming K, Kasza J, Hooper R, Forbes A, Taljaard M. A tutorial on 
sample size calculation for multiple-period cluster randomized 
parallel, cross-over and stepped-wedge trials using the Shiny CRT 
Calculator. Int J Epidemiol. 2020;49(3):979–95. 
10. Higuera-Mendieta DR, Cortés-Corrales S, Quintero J, González-
Uribe C. KAP Surveys and Dengue Control in Colombia: 
Disentangling the Effect of Sociodemographic Factors Using 
Multiple Correspondence Analysis. PLoS Negl Trop Dis.  
11. Special Programme for Research and Training in Tropical Diseases 
(TDR), World Health Organization. Operational guide for assessing 
the productivity of Aedes aegypti breeding 
sites. http://www.who.int/tdr/publications/documents/sop-pupal-
surveys.pdf]. http://www.who.int/tdr/publications/documents/sop
-pupal-surveys.pdf. Accessed 20 Jul 2018. 

12. DANE. Geovisor CNPV 2018: Geoportal DANE GEOPORTAL 
DANE. https://geoportal.dane.gov.co/geovisores/territorio/analisis
-cnpv-2018/?lt=4.646075&lg=-74.088605&z=20. Accessed 15 Dec 
2021. 
13. World Health Organization. Comprehensive Guidelines for 
Prevention and Control of Dengue and Dengue Haemorrhagic 
Fever. Revised an. World Health Organization, Regional Office for 
South-East Asia 2011. Available online 
at: http://origin.searo.who.int/entity/vector_borne_tropical_disea
ses/documents/SEAROTPS60/en/. 
 
 
 
 
 
 
 
 

Part B: Practices on dengue and water use 
Household questionnaire 
Form for extraction of socio-demographic data 
We are evaluating a new strategy against dengue. If you allow me, I will ask a few short questions that will not take up 
much of your time. If you feel uncomfortable with any question, you cannot answer or end the interview whenever you 
want. 
House code: _________________________  
 
 
        Cluster code: ____ 
1. ¿ Could you give us the following details about the people who live in your house, including yourself in the 
following table?  
  
Person 
Age 
Sex 
1=F 
2=M 
3=O 
Education 
Primary 
school=1 
High school= 2 
University= 3  
No= 4 
During this year, 
did you or anyone 
in your household 
have DZC? 
1=Dengue 2=Zika 
3=Chikunguna 4=No 
Which 
month?  
Did you go 
to the 
doctor or 
health 
unit? 
1=Yes  2=No 
 
During the day, where 
do you spend most of 
the time (7 am -5 pm)? 
1=Inside neighborhood, 
2=Outside neighborhood 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
11 
 
 
 
 
 
 
 
12 
 
 
 
 
 
 
 
 
 
Mark the correct answer 
Part C: Willingness of the community to receive the insecticide coating 
 
 
Thanks for your participation 
2. Do you apply some type of vector control in your laundry tank to prevent dengue? 
  1= Use of fish   2 = Addition of chlorine   3= Use of tank lid    4=Washing and brushing    5= None       6=I don’t know    
 
3. Has the health service applied any type of powder or tablet insecticide in your laundry tank this year? 
1= Yes            2= No          3= I don’t know 
 
4. Which deposits do you use for water consumption? 
1= Ground tank             2=Tap water           3=Other deposits, specify____________           4=None 
 
5. Do you know where mosquitoes lay their eggs or where they breed? 
1=Clean water      2=Dirty water (e.g. puddle)        3= Grass or pasture        4= I don’t know 
 
6. ¿ which of these media would you like to receive information about dengue? (Multiple choices) 
1=TV 2=Radio 3=Cell phone 4=internet 5=Newspaper 6=Flyer 6=Visit to your home by a technician 7=Loudspeakers 8=I don't know 
 
 
7. Would you like to apply freely a transparent coating as a control method in your laundry tank? 
1 = Yes      2=No      3= I don’t know    
Part A: Household sociodemographic characteristics 

SECCIÓN B: Prácticas sobre el dengue y uso del agua 
 
 
(Spanish version) 
Estamos evaluando una nueva estrategia contra el dengue. Si me permite le realizaré unas preguntas cortas que no le 
quitarán mucho tiempo. Si usted se siente incómodo con alguna pregunta, puede no contestar o terminar la entrevista 
cuando lo desee. 
Código casa: _________________________  
 
 
Código conglomerado: ____ 
8. ¿Podría darnos los siguientes detalles sobre las personas que viven en la casa, incluyéndose en la siguiente 
tabla?  
  
Persona Edad 
Sexo 
1=F 
2=M 
3=O 
Educación 
Primaria=1 
Bachillerato= 2 
Universidad= 3  
No= 4 
¿En este año ud. 
 o alguien de su 
hogar tuvo DZC?  
1=Dengue 2=Zika 
3=Chikunguna 4=No 
¿En 
qué 
Mes?  
¿Asistió al 
médico o 
unidad de 
salud? 
1=SI  2=No 
 
¿Durante el día, donde 
permanece la mayor 
parte del tiempo (7 am -
5 pm)? 1=En mi hogar o 
barrio, 2=Fuera del barrio 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
11 
 
 
 
 
 
 
 
12 
 
 
 
 
 
 
 
 
 
Marque la respuesta correcta 
SECCIÓN C: Disposición de la comunidad para recibir el recubrimiento insecticida 
 
 
Gracias por participar 
9. ¿Usted aplica algún tipo de control en su tanque de lavadero para prevenirse del dengue? 
1= Uso peces    2 = Adición de cloro    3= Uso tapa tanque      4=Lavado y cepillado   5= Ninguno    6=No sé    
 
10. ¿En este año el servicio de salud ha aplicado algún insecticida tipo polvo o pastilla en su tanque de lavadero? 
1= Si            2= No          3= No sé 
 
11. ¿Cuáles de los depósitos usa para el consumo de agua? 
1= Tanque bajo         2=Agua de la llave         3=Otros depósitos, especifique                                     4=Ninguno 
 
12. ¿Sabe usted en dónde ponen los mosquitos sus huevos o dónde se crían? 
1=Agua limpia          2=Agua sucia (charco)   3= Monte o Pasto            4= No sé 
 
13. ¿Con cuál de estos medios le gustaría recibir información sobre el dengue?  (opción multiple) 
1=TV     2=Radio    3=Celular    4=Internet    5=periódico    6=Volante   6=Visita a su casa por técnico   7=Perifoneo   8=No sé 
  
14. ¿Le gustaría que pongamos una pintura transparente como método de control de forma 
gratuita en su tanque de lavadero? 
1 = Si      2=No      3=No sé   
SECCIÓN A: Características Sociodemográficas del hogar 

 
Entomological survey 
Form for extraction of entomological data 
House code: ________        Cluster: _____                 Inspector: _______________ 
                          Date: ___/____/_____       
Building type: House: _   Car store: _ Health unit: _  Church: _  Grocery store: _  School: _  Restaurant: _  other: _       Public space:__ 
# 
*Water tanks 
Other water containers  
* Details of each water tank 
Entomological 
indices 
Measures 
 
Tanque bajo 
Storage tanks 
Elevated tanks 
buckets 
Cans / small containers / Jars 
Flowerpots 
Tires 
Sumps / sewers 
Bottles (with water) 
Gasoline bottles (Pimpinas) 
Natural breeding site 
Other: 
Other: 
Volume capacity (litres) 
Water type (1=Watertap, 
2=Rain, 3=other) 
Tank location 
(1=Inside, 0=Outside) 
Shading  
(1=Yes, 2=partly, 3=No) 
Covered tank 
(1=Yes, 2=partly, 3=No) 
Larvae 
 
Pupae 
count 
Length (centimeters) 
Width (centimeters) 
Height (centimeters) 
Concrete tanks 
Plastic tanks 
Tiled tanks 
Metallic tanks 
Positive (+),  
Negative (-) 
 Total number 
Each water tank has its row (Water tank such as bottles etc. can be put together) 
1 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
11 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
12 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
13 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
15 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
16 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
17 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
18 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Comment: __________________________________________________________________________________ 

 
 
(Spanish version) 
Código casa: ________        Conglomerado: _____                 Inspector: _______________ 
                          Fecha: ___/____/_____       
Tipo de edificación: Casa: _   Taller: _ Unidad de salud: _  iglesia: _ Tienda: _  Colegio: _  Restaurante: _  otro: _       Espacio Público:__ 
No. 
*Tanques de agua 
Otros depósitos con agua 
*Detalles de cada tanque de 
agua 
Índices 
entomológicos 
Medidas 
SITIOS 
Tanque bajo 
Tanque almacenamiento 
Tanque elevado 
baldes 
Latas / Envases / Jarras 
Floreros 
Neumáticos (Con o sin agua) 
Sumideros/alcantarillas 
Botellas (Con agua) 
Galones ( Pimpinas) 
Criaderos naturales 
Otro: 
Otro: 
Capacidad en volumen (litros) 
Tipo de agua 
(1=la llave, 2=Lluvia, 3=otro) 
Ubicación del tanque 
(1=Adentro, 0=Fuera) 
Sombreado 
(1=Sí, 2=parcialmente, 3=No) 
Cubierto 
(Si=1, parcial= 2, No=3) 
Larva 
 
Conteo 
de 
pupas 
Longitud 
Anchura 
Altura 
Cemento 
Plástico 
Baldosa 
Toneles metálicos 
Positivo (+),  
Negativo (-) 
 Número total 
Cada tanque de agua tiene su fila (Depósito de agua como botellas etc.. pueden ser puestos 
1 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
11 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
12 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
13 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
15 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
16 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
17 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
18 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Observación: __________________________________________________________________________________