Acta Tropica 229 (2022) 106339

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Acta Tropica
journal homepage: www.elsevier.com/locate/actatropica

Water quality characteristics of breeding habitats in relation to the density

of Aedes aegypti and Aedes albopictus in domestic settings in Gampaha
district of Sri Lanka
Rasika Dalpadado a, b, Deepika Amarasinghe b, *, Nayana Gunathilaka c
a
Regional Director of Health Services Office, Gampaha District, Gampaha, Sri Lanka
b
Department of Zoology and Environmental Management, Faculty of Science, University of Kelaniya, Dalugama, Sri Lanka
c
Department of Parasitology, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka

A R T I C L E I N F O A B S T R A C T

Keywords: In spite of the unavailability of measures to target dengue control in human populations, the promising way of
Water quality index combating the disease is by controlling vector mosquito larvae and their breeding habitats. Water quality
Mosquito characteristics of the breeding habitats of dengue vectors are among the crucial parameters that determine the
Breeding
female mosquito’s oviposition and breeding. Therefore, a cross-sectional study was conducted to investigate the
Dengue vectors
habitat characteristics of dengue vector mosquitoes by assessing the water quality of positive breeding habitats
using the weighted arithmetic water quality index (WQI). The present study was conducted in domestic areas of
the selected medical officer of health areas of the Gampaha district, Sri Lanka, from 2017 to 2019. Water quality
characteristics of breeding habituations were measured and analyzed. The relationship between water quality
parameters in breeding habitats with the occurrence of mosquito species was assessed using Kruskal-Wallis one-
way analysis of variance, followed by pairwise comparison using Dunn’s test and Mann-Whitney U test at a 5%
level of significance. This study revealed that the temperature of Aedes mosquito breeding water ranged between
25.3 and 39.8 ◦ C, and bred at temperatures as high as 39.8 ◦ C in discarded receptacles. The results indicated that
Ae. aegypti was prominent in alkaline water ranging between 7.5 and 8.5 pH, whereas Ae. albopictus was
abundant in water with a pH range of 6.5–7.5. Both species of Aedes inhabited waters of low turbidity and TDS
level. Ae. aegypti and Ae. albopictus immatures were prominent in water where TDS levels ranged between 250
and 350 ppm. The mean conductivity in the mosquito breeding water was recorded as 228.3 ± 63.9 µs/cm. The
study revealed that Aedes mosquitoes could breed in water with a mean dissolved oxygen level of 6.9 ± 0.7 mg/L,
ranging between 6.35 ± 1.09 mg/L and 7.28 ± 0.26 mg/L. The water quality indices were calculated for the
eight previously identified breeding habitat categories of Aedes mosquitoes.

1. Introduction management and water storage practices (Waewwab et al., 2019) as

Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus are the
two most significant vector mosquitoes of several arboviral diseases of
humans. Among them, dengue and dengue haemorrhagic fever together
has become an important public health problem causing significant so­
cioeconomic pressures worldwide, notably in tropical and subtropical
areas (WHO, 2017). After the first reported case in 1962, Sri Lanka has
experienced periodic epidemics once in a few years from the 1990s to
2008. The worst epidemic was observed in 2017, with 186,000 sus­
pected dengue cases and 440 deaths (Liyanage et al., 2021). The risk of
the extent of the disease increases with poor environmental
these mosquitoes prefer to breed in man-made artificial water containers
in and around human habitation.
Both Ae. aegypti and Ae. albopictus carry high vectorial competency
for dengue viruses. In the South-East Asia region, Ae. aegypti is consid­
ered the principal epidemic vector of dengue viruses in urban cities,
while Ae. albopictus has been recognized as the secondary vector of
Dengue which is also crucial in maintaining the viruses (Lambrechts
et al., 2010). The virus can infect female mosquitoes of these two species
during their blood-feeding process from an infected person. Once
infected, the mosquito can retain the virus throughout its adult life
(Bhatt et al., 2013). Climatic change, deforestation, travel, and trade

* Corresponding author.
E-mail address: deepika@kln.ac.lk (D. Amarasinghe).

https://doi.org/10.1016/j.actatropica.2022.106339
Received 4 November 2021; Received in revised form 26 January 2022; Accepted 29 January 2022
Available online 31 January 2022
0001-706X/© 2022 Elsevier B.V. All rights reserved.
R. Dalpadado et al.
have forced the dengue mosquitoes to spread, adapt and survive in a
range of extended areas in the globe. These mosquitoes prefer to breed in
natural and artificial water-holding stagnant habitats that include clear
water in domestic and semi-domestic environments (Awang et al., 2020;
Madzlan et al., 2016; Chaterrgy et al., 2015).
Aedes mosquitoes follow visual and olfactory cues to select appro­
priate water containers and then the chemical and physical properties of
water for choosing it for their oviposition (Texler et al., 1998). Breeding
water quality is critical in determining whether female mosquitoes de­
posit eggs and if the following immature stages successfully grow to
adulthood. Even though numerous studies have been focused only on
the types of key breeding sites for dengue mosquitoes, knowledge on the
breeding ecology comprising physical, biological, and chemical char­
acteristics of the breeding habitat is vital in identifying preferences for
breeding sites and developing successful vector control measures against
dengue outbreaks around the globe. As the immature mosquito stages
are confined to aquatic habitat, physicochemical characteristics of the
breeding water have a significant effect on the growth, development,
and survival of mosquitoes, and hence it affects the transmission and
magnitude of Dengue (Mereta et al., 2013; Emidi et al., 2017). Only a
few studies have been done in Sri Lanka (Amarasinghe and Dalpadado,
2014) and even globally (Gopalakrishnan et al., 2013; Madzlan et al.,
2016) determine the relationship between the occurrences of dengue
vectors and physicochemical parameters of their breeding habitats. The
ecological parameters influencing mosquito life cycle and biology help
facilitate the successful implementation of the vector control pro­
grammes. Therefore, the current research was conducted for the period
of April 2017 to December 2019 to determine the water quality
Fig. 1. The map of Gampaha District showing selected MOH areas for the larval surveillance.
2
Acta Tropica 229 (2022) 106339
characteristics of varieties of Aedes breeding habitats and to arrive at
water quality indices of Aedes breeding sites in selected areas in Sri
Lanka.
2. Methods
2.1. Study design and area selection
The present study was conducted as a cross-sectional study in the
Gampaha district, the Western province of Sri Lanka. Gampaha has been
the main contributor to dengue reported with increasing frequency in Sri
Lanka over the last few decades. Approximately 47–55% of dengue cases
have been reported in the previous five years in the Western Province of
Sri Lanka, the country’s most populated province, representing 28.7% of
the country’s whole population. The second-highest prevalence of
dengue cases was observed in the Gampaha District since 2010 despite
all the efforts taken to control the disease. Above the average of 13,000
dengue cases were reported annually from 2015 to 2019 in the district of
Gampaha, with approximately 32,000 reported cases only during the
year 2017 with around 40 deaths (Epidemiology Unit, 2018). The Dis­
trict of Gampaha is also said to have the highest residential population in
Sri Lanka, with an area of 1387 km2 (Alahakoon et al., 2021).
Geographic areas inside towns and cities were described as urban. In
contrast, rural areas located outside towns and cities and these areas
were usually less developed with significant land cover under agricul­
ture and natural vegetation. Areas with mixed characteristics were
considered sub-urban (Ndenga et al., 2017); the Gampaha district is
divided into fifteen Medical Officer of Health (MOH) areas for the
R. Dalpadado et al.
convenience of the operation of administrators. The present study was
conducted in the following six MOH areas (Fig. 1); Negombo (7◦ 12′
60.00′′ N: 79◦ 49′ 59.99′′ E) and Wattala (6◦ 58′ 40.44′′ N, 79◦ 53′ 23.28′′ E)
MOH areas as urban; Attanagalla (7◦ 05′ 60.00′′ N: 80◦ 06′ 60.00′′ E) and
Gampaha (7◦ 5′ 22.92′′ N, 79◦ 59′ 33′′ E) MOH areas as sub-urban; Dompe
(6◦ 56′ 25.42′′ N: 80◦ 4′ 37.91′′ E) and Divulapitiya (7◦ 2′ 30.30′′ N,
80◦ 01′ 65′′ E) MOH areas as rural MOH areas.
2.2. Entomological surveillance
To assess the water quality characteristics of dengue vector breeding
places, water samples were collected from the pre-identified eight
different breeding categories, namely A/C refrigerator trays, water
storage containers, concrete slabs, low roof gutters, tube wells, dis­
carded items, temporary removals, and natural breeding places; leaf
axils, bamboo stumps, tree holes, where Aedes larvae were found. During
the surveillance, twenty-five house premises were randomly selected,
house to house (indoor) and peri-domestic area mosquito larval sam­
pling was done, and analysis of mosquito breeding water quality pa­
rameters was performed in-situ. The distance between households
ranged from 5 to 100 m, and verbal consent was obtained from all the
households before the inspection. Each of the selected MOH areas was
visited every other month for 30 months, from April 2017 to December
2019.
During larval surveillance, habitat was examined visually for the
presence of dengue vector immatures. Then, the larvae and pupae were
captured using a pipette, dipper, or scoop based on the breeding habi­
tat’s characteristics and water availability following the guidelines
specified by the World Health Organization and National Dengue Con­
trol Unit Sri Lanka (WHO 2016; National Dengue Control Unit Sri Lanka
2016, 2019). Then larval counts were recorded along with the container
types and volume of breeding water. A maximum of ten larvae from each
site was transferred to labeled plastic bottles containing 70% alcohol,
and they were brought to the laboratory for species identification.
2.3. Water quality characterization
A water sample (20 mL) was collected from each breeding habitat
that was positive for Aedes larvae. Seven abiotic variables, temperature,
hydrogen ion concentration (pH), conductivity, Total Dissolved Solids
(TDS), turbidity, salinity, and Dissolved Oxygen (DO), were measured in
situ. Temperature (portable meter, Hach SenSION TM), pH (portable
meter, Hach SenSION TM), and DO (digital meter EUTECH Dowp 300/
02 K) and conductivity, TDS, and salinity were measured (Hach Sen­
SION TM multi-probe meter). Further, the clarity and color of water
collected from the breeding habitats were recorded.
2.4. Calculation of the water quality index (WQI)
The quality of Aedes mosquito breeding water was assessed by
calculating the water quality index (WQI) based on five factors; pH,
Total Dissolved Solids (TDS), turbidity, conductivity, and dissolved ox­
ygen levels in each breeding habitat using a weighted arithmetic water
quality index according to Brown et al., (1972) described below.
WQI ¼ ΣQiWi / ΣWi
Where,
Qi = Quality rating of nth parameter
Wi = Unit weight of nth parameter
The Qi for each of the water quality parameters was calculated as
follows,
Qi ¼ 100 [(Vo – Vi) / (Si – Vi)]
Where,
Vo = Estimated mean value of the nth water quality parameter in
each breeding habitat category
Si = The water quality standard value for Sri Lanka of the nth
parameter (Table 1; Sri Lanka standards for potable water (Sri Lanka
3
Acta Tropica 229 (2022) 106339
Table 1
Indicator for larval density level.
Density level Mean number of larvae per 250 mL
Low < 10.00
Moderate 10.00- 20.00
High > 20.00
Standards, 1983)
Vi = Ideal value of the nth parameter in pure water according to the
Sri Lankan standards
The Wi each water quality parameter is calculated as follows,
Wi ¼ K/Si
Where,
K = proportionality constant, which can also be calculated by using
the following equation
K¼ 1/ Σ(1/Si)
The quality in the Aedes mosquito breeding habitat was specified as
Excellent (0–25), Good (26–50), Poor (51–75), Very poor (75–100) and
contaminated water (>100), according to the water quality index scale
described by Brown et al., (1972).
2.5. Data analysis
The statistical analysis was carried out using IBM SPSS 26 statistical
software. Chi-square (χ2) analysis was performed to determine whether
there was any significant difference in the distribution of the mosquito
larvae across different breeding habitat type categories with water
quality parameters. The densities of immature stages (larvae and pupae)
were calculated as a number of immature stages collected over 250 mL
of water in the breeding habitat. The relationship between water quality
parameters in breeding habitats with the occurrence of mosquito species
was assessed using Kruskal-Wallis one-way analysis of variance, fol­
lowed by pairwise comparison using Dunn’s test and Mann-Whitney U
test at a 5% level of significance. A P value < 0.05 was taken as statis­
tically significant.
3. Results
3.1. Distribution of breeding habitats and vector density
A total of 4812 potential breeding sites were investigated at 2250
randomly selected premises during the study. Of them, 15.7% of sites (n
= 755) were found positive for Aedes larvae, and a total of 18,476 Aedes
larvae and pupae were collected from eight preidentified breeding
habitats categories. Aedes albopictus (77.9%; n = 589 sites) was recorded
as the dominant species, followed by Ae. aegypti (22.1%; n = 166 sites).
Aedes larvae identified from each site were varied between Ae. albopictus
and Ae. aegypti (Table 2). The mean density of Ae.aegypti immatures was
highest in concrete slabs (27.4±14.2), followed by water storage con­
tainers (22.2 ± 10.7) and temporary removal items (20.2 ± 10.7); and
they were not seen in natural breeding places or roof gutters throughout
the study. Ae.albopictus mosquitoes were detected in all the eight
breeding type categories with the greatest mean immature density in
concrete slabs (33.5± 20.9), roof gutters (31.8 ± 14.6) and natural
breeding places (22.0 ± 8.5). The least mean immature density for Ae.
albopictus was reported in indoor breeding places such as air condi­
tioners (A/C) and refrigerator trays (13.2 ± 6.4).
Table 3 shows that the immature density of mosquito larvae of most
breeding sites is moderate; Ae. aegypti (45.8%), Ae. albopictus (64.9%),
or low; Ae. aegypti (43.4%), Ae.albopictus (25.5%). The highest number
of Ae. aegypti positive containers recorded was water storage containers
(4.8%), followed by concrete slabs at 4.2%, while that value for Ae.
albopictus was highest in temporary removal items (3.7%). Although
discarded receptacles were identified as one of the most frequently
found breeding habitats (Ae.aegypti; 28.3%, n = 47, Ae.
R. Dalpadado et al. Acta Tropica 229 (2022) 106339

Table 2
Mean density ± SD (range in parenthesis) of immature stage across eight habitat categories for Ae. aegypti and Ae. albopictus.
Mosquito AC/ Refrigerator Concrete slabs Discarded Roof gutters Natural Temporary Tube wells Water storage
species trays breeding places removable items containers
Aedes aegypti 17.3± 5.2 27.4±14.2 14.9 ± 6.6 0.0 0.0 20.2 ± 10.7 14.3 ± 5.1 (6.0- 22.2 ± 10.7 (3.0-
(8.0–26.7) (4.4–80.0) (2.0–32.0) (8.0–56.0) 19.0) 60.0)
Aedes 13.2 ± 6.4 33.5 ± 20.9 17.6 ± 6.9 31.8 ± 14.6 22.0 ± 8.5 20.6 ± 8.7 13.9 ± 5.8 17.5 ± 10.0
albopictus (1.7–23.0) (2.0–72.0) (3.0–60.0) (12.0–55.0) (8.0–45.0) (4.0–60.0) (8.0–23.5) (8.0–48.0)

Table 3
Frequency of occurrence of Ae. aegypti and Ae. albopictus containers across different habitat categories according to the level of larval density.
Habitat Frequency of occurrence
Aedes aegypti Immature density Aedes albopictus Immature density
Low density (< 10.0) Moderate density High Density (> Low density (< 10.0) Moderate density High Density (>
(10.0–20.0) 20.0) (10.0–20.0) 20.0)
A/C Refregirator trays; C1 5.4% (n = 9) 1.8% (n = 3) 0.6% (n = 1) 0.5% (n = 3) 5.1% (n = 30) 0.2% (n = 1)
Concrete slabs; C2 6.0% (n = 10) 4.8% (n = 8) 4.2% (n = 7) - 2.2% (n = 13) 1.0% (n = 6)
Discarded items; C3 15.7% (n = 26) 12.0% (n = 20) 0.6% (n = 1) 8.7% (n = 51) 21.1% (n = 124) 1.0% (n = 6)
Gutters; C4 - - - 1.0% (n = 6) 2.0%(n = 12) 1.0% (n = 6)
Natural breeding places; - - - 3.7% (n = 22) 11.7% (n = 69) 1.5% (n = 9)
C5
Temporary removable 6.0% (n = 10) 9.6% (n = 16) 0.6% (n = 1) 7.3% (n = 43) 15.1% (n = 89) 3.7% (n = 22)
items; C6
Tubewells; C7 6.6% (n = 11) 1.8% (n = 3) - 1.2% (n = 7) 1.7% (n = 10) -
Water storage containers; 3.6% (n = 6) 43.4% 15.7% (n = 26) 45.8% 4.8% (n = 8) 10.8% 3.1% (n = 18) 25.5% 5.9% (n = 35) 64.9%(n 1.2%(n = 7) 9.7%
C8 Total (n = 72) (n = 76) (n = 18) (n = 150) = 382) (n = 57)

Note: n Values in the parenthesis include: (Number of positive containers of each breeding habitat for Ae.aegypti or Ae.albopictus / Total number of positive containers
for Ae. aegypti (n = 166)or Ae. albopictus (n = 589))*100.
*n represents the number of Aedes positive breeding places in each category inspected.

albopictus;30.7%, n = 181) for both species of mosquitoes, mostly their
densities were low or moderate in discarded receptacles (Ae. aegypti;
low; 15.7%, moderate; 12.0%, high; 0.6%, Ae.albopictus: low; 8.7%,
moderate; 21.1%, high; 1.0%). Water storage containers (frequency
prevalence; 24.1%, n = 40) were identified second prevalent breeding
place for Ae. aegypti followed by temporary removals (frequency
prevalence;16.1%, n = 27) while, temporary removal items (26.1%, n =
154) and natural breeding habitats (17%, n = 100) were identified as the
most frequently found breeding habitats for Ae. albopictus in the study
area (Fig. 2). Around 90.7% (n = 685) of Aedes positive containers were
outdoors, and only 9.3% (n = 70) were found indoors, such as refrig­
erator trays and A/Cs. The study found that the outdoor mosquito

Fig. 2. Percentage prevalence of Ae.aegypti and Ae. albopictus in study sites of Gampaha district.

4
R. Dalpadado et al. Acta Tropica 229 (2022) 106339

breeding localities had a higher immature mean density than inside
houses (outdoor; 20.0 ± 10.5, indoor; 17.3 ± 9.5). However, Ae. aegypti
indoor breeding habitats had a higher mean density than outdoors
(outdoor; 20.4 ± 10.2, indoor; 19.3 ± 12.2). The results of the Mann-
Whitney U test also revealed that the density of the immature was
significantly different across indoor and outdoor localities in the study
area (Mann-Whitney U test; U = 27,857, P = 0.025, P < 0.05).
3.2. Water quality characteristics of breeding habitat for Aedes mosquito
Water quality characteristics of Aedes mosquito species breeding
sites in this study are shown in Table 4 and Fig. 3. The water temperature
of Aedes mosquito breeding sites was ranged between 25.3 and 39.8 ◦ C,
and mosquitoes were reported to breed at temperatures as high as 39.8
◦
C in discarded receptacles (Table 4; Fig. 3a). The temperature range
between 30.0 and 32.0 ◦ C has been described as the optimum temper­
ature for both Ae. aegypti and Ae. albopictus, with around 75% of
mosquitoes being found within this range. The water pH in the different
breeding habitats of Aedes mosquitoes ranged from acidic to alkaline pH;
5.6 to 8.7 within the study period. The results indicate that immature
stages of Aedes prefer slightly alkaline pH as 65% (n = 485) of Aedes
larvae were detected at pH above 7. Ae. aegypti was prominent in
alkaline water ranging between 7.5 and 8.5 pH, whereas Ae. albopictus
was abundant in water with a pH range of 6.5–7.5 (Fig. 3b). Results
indicate that both species of Aedes inhabited waters of low TDS and
turbidity level (Fig. 3c and d). The frequency distribution of Ae. aegypti
and Ae. albopictus immatures with TDS level was almost identical. The
vectors were prominent in water, where TDS levels ranged between 250
and 350 ppm (Ae. aegypti; 70.5% Ae. albopictus; 71.5%). However, Ae.
aegypti had a lower range of TDS level (162 - 649 ppm) compared to Ae.
albopictus since secondary vector Ae. albpictus was found in water with a
darker appearance, ranging from 86 to 1002 ppm with a higher level of
degrading organic matter (Fig. 3c). The mean conductivity in the mos­
quito breeding water recorded at 228.3 ± 63.9 µs/cm, and it was ranged
between 195.2 ± 35.2 µs/cm and 438.3 ± 76.7 µs/cm. The lowest
conductivity value was recorded in natural breeding places (72 µs/cm)
while highest at roof gutters (578 µs/cm) (Fig. 3e). The results also
revealed that Aedes mosquitoes could breed in water with a mean dis­
solved oxygen level of 6.9 ± 0.7 mg/L, ranging between 6.35 ± 1.09 mg/
L and 7.28 ± 0.26 mg/L. They were also found in breeding in water with
2.62 mg/L, lower dissolved oxygen level in discarded items, and water
with a higher dissolved oxygen level of 8.92 mg/L in water storage
containers. About 90% (n = 676) of Aedes aquatic stages were inhibited
in water with higher dissolved oxygen levels (6.5–8.5 mg/L). (Fig. 3f)
Kruskal –Wallis statistics (P < 0.05 at 95% of significance) followed
by pairwise comparisons by Dunn’s multiple comparison tests, corrected
with the Bonferroni correction demonstrated that all estimated water
quality parameters varied significantly across different breeding cate­
gories of Aedes species during the study period (Table 5, Fig. 4). Ac­
cording to the Kruskal–Wallis statistics, there were significant variations
on the occurrence of both Ae. aegypti and Ae. albopictus larvae with
temperature (H = 5.487, df = 1, P= 0.019) and dissolved oxygen level
(H = 11.248, df = 1, P= 0.001) in breeding habitats. In addition, sig­
nificant variations were also indicated with pH (H = 1.566, df = 1, P=
0.211), total dissolved solids (H = 0.633, df = 1, P= 0.426), turbidity
(H = 2.068, df = 1, P= 0.150) and conductivity level of breeding habitat
categories (H = 0.210, df = 1, P= 0.647). Except for pH, all water
quality parameters denoted significant variations among the breeding
habitat categories encountered indoor and outdoor settings (Table 6).
Most breeding sites are positive for Ae. aegypti (87.4%; n = 145) and Ae.
albopitcus (88.8%; n = 523) were clear (non-opaque) in nature (Fig. 5).
The color of the breeding water in the examined containers were non
opaque (86.7%; n = 144, 86.9%; n = 512), light yellow (7.2%; n = 12,
1.1%; n = 7), light brown (4.8%; n = 8, 8.1%; n = 48) and dark brown
(1.2%; n = 2, 3.7%; n = 22) respectively for Ae. aegypti and Ae. albopitcus
(Fig. 6). The Chi-square test suggests both appearances of water
(χ 2=23.321, df=3, P=<0.001) and the color of water (χ 2=2.151, df=2,
P=0.034) positive for Aedes were significantly varied with the species
encountered.
Table 7 shows the density distribution of immature stages of Aedes
mosquitoes in different breeding habitats, classified according to the
condition of breeding water. Ae. aegypti mosquito immature mean
density is highest in concrete slabs (Overall; 27.4±14.2, non-opaque
water; 30.6 ± 10.2, turbid water; 17.3 ± 16.9) followed by water stor­
age containers (Overall; 22.2 ± 10.7, non-opaque water; 23.1 ± 8.4,
turbid water; 10.7 ± 4.7) while the immature mean density of Ae.
albopitcus was also found to be highest concrete slabs (Overall; 33.5±
20.9, non-opaque water; 42.1 ± 17.6, turbid water; 16.9 ± 5.3, highly
turbid water; 4.0 ± 0.0) followed by (Overall; 31.8 ± 14.6, non-opaque
water; 51.0 ± 2.6, turbid water; 30.0 ± 12.5, highly turbid water; 23.7 ±
11.1). Even though both Ae. aegypti and Ae. albopitcus mosquitoes found
to be different breed types of breeding habitats, the productivity of the
habitat in terms of the density distribution of immature were signifi­
cantly varied in the Gampaha district with habitat categories (Kruskal-
Wallis test; H = 63.027, df = 7, P = 0.000, P < 0.05) and with the
condition of breeding water ((Kruskal-Wallis test; H = 40.285, df = 2, P
= 0.000, P < 0.05). Kruskal-Wallis test followed by pairwise compari­
sons by Dunn’s multiple comparison test showed the density of larvae in
non-opaque and turbid water (H = 161.187, df = 2, P = 0.000, P <
0.05) and non-opaque and highly turbid water (H = 139.557, df = 2, P

Table 4
Water quality parameters of different breeding habitats of Aedes spp.
Habitat
A/C Concrete Discarded Gutters Natural Temporary Tube wells Water Average
Refrigerator slabs items places removable storage
trays items containers
Temperature/ Mean 33.5 ± 2.3 32.8 ± 1.1 30.6 ± 1.3 30.4 ± 0.9 29.4 ± 1.3 30.6 ± 1.0 29.0 ± 1.3 30.6 ± 1.1 30.6±1.6
◦
C
Range 27.5- 36.5 29.5–34.5 26.5- 39.8 27.6–32.1 25.4- 32.0 28.0- 33.0 25.4 – 31.5 25.30- 32.8 25.3–39.8
pH Mean 7.0 ±0.1 7.4 ±0.4 7.1 ±0.4 6.9 ±0.3 6.9 ±0.4 7.1 ±0.3 7.0 ±0.2 7.0 ±0.3 7.0 ± 0.4
Range 6.8- 7.2 6.9 – 8.4 5.6- 8.7 5.9 – 7.6 5.9 - 8.4 5.9- 8.2 6.5–7.1 6.0 – 7.9 5.6 - 8.7
TDS /ppm Mean 352 ± 107 342 ± 92 251 ± 73 729 ±149 250 ±52 235 ±33 201 ± 30 243 ± 95 270 ±116
Range 169 - 714 156 – 645 145 – 705 413 - 1002 142- 527 164- 416 142- 271 86 -795 86 -1002
Turbidity/ Mean 5.30 ±0.98 5.63 ±0.96 4.53 8.64 ±1.54 4.49 ±0.60 4.34 ±0.71 3.70 ±0.93 3.98 ±1.40 4.62 ±1.25
NTU ±0.91
Range 2.95- 8.26 3.12- 7.88 1.02 - 5.62- 12.20 2.83 - 6.95 2.00 - 7.69 1.28 – 4.98 0.85- 10.07 0.85–12.20
11.69
Conductivity/ Mean 259.1 266.2 218.2 438.3±76.7 225.5±35.8 204.9±29.9 195.2±35.2 225.6±63.1 228.3
µs/cm ±58.3 ±64.7 ±49.9 ±63.9
Range 118- 429 201- 511 96- 492 293- 578 72- 346 116- 365 76- 242 82- 469 72- 578
DO/ mg/L Mean 6.35±1.09 6.92±0.38 6.86±0.78 7.24±0.64 6.83±0.75 6.96±0.64 7.28±0.26 6.77±0.64 6.87 ±0.73
Range 3.02–7.86 6.09- 7.91 2.62- 7.20 5.26- 7.94 3.59- 7.88 3.59- 7.98 6.94- 7.96 4.26- 8.92 2.62 -8.92

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R. Dalpadado et al. Acta Tropica 229 (2022) 106339

Fig. 3. Frequency distribution of water quality parameters: (a) temperature; (b) pH; (c) total dissolved solids; (d) Turbidity; (e) conductivity; (f) Dissolved Oxygen
level for Aedes mosquitoes.

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R. Dalpadado et al. Acta Tropica 229 (2022) 106339

Table 5 3.3. Water quality index

Asymptotic significance of water quality characteristics across different
breeding habitats of Aedes mosquitoes. The water quality indices were calculated for the eight previously
Physicochemical Test Degree of Asymptotic Sig. (2- identified breeding habitat categories of Aedes mosquitoes in the Gam­
Parameter Statistic Freedom sided test) paha district (Fig. 6). The results illustrate WQI values ranged between
Temperature/ ◦ C 232.050a 7 0.000 47.5 and 66.5 (Good to Poor) among Aedes positive breeding habitats in
pH 64.255a 7 0.000
the Gampaha district where tube wells and water storage containers
TDS /ppm 226.519a 7 0.000
Turbidity/ NTU 224.285a 7 0.000 with lowest WQI’s (47.5 and 50.4), respectively, while roof gutters with
Conductivity/ µs/cm 183.206a 7 0.000 the highest WQI (66.5). It was noted highest immature density for both
DO/ mg/L 45.325a 7 0.000 Ae.aegypti and Ae.albopictus were observed in breeding water with
a. The test statistic is adjusted forties. higher WQI with and poor potable water standard (Ae.aegypti; Concrete
slabs; larval density 27.4 ± 14.2, WQI; 64.9, Ae.albopictus; Gutters;
= 0.027, P < 0.05) varied significantly. In contrast, the larval densities larval density 31.8 ± 14.6, WQI; 66.5). A strong positive correlation was
in turbid and highly turbid water are the same (H = 21.629, df = 2, P = noted between Aedes immature density and WQI calculated at 95%
0.100, P > 0.05). The results of the Mann-Whitney U test also revealed confidence level (R=0.58, F=8.3, P = 0.000) where Aedes larval density
that the density of the immature was the same across the two species increases with increasing WQI in the Gampaha district.
(Mann-Whitney U test; U = 52,689, P = 0.124, P > 0.05).

Fig. 4. Box-plot (median and quartiles) of the Kruskal-Wallis rank-sum test showing the variation of the distribution of water quality parameters (a) temperature; (b)
pH; (c) total dissolved solids; (d) Turbidity; (e) conductivity (f) Dissolved Oxygen level across different breeding habitats for Aedes mosquitoes followed by Dunn’s
posthoc test, P < 0.05. Breeding sites that share the same letters were not significantly different following pairwise comparisons by Dunn’s multiple comparison tests.

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R. Dalpadado et al. Acta Tropica 229 (2022) 106339

Table 6
Asymptotic significance of physicochemical characteristics across different breeding habitats and indoor-outdoor localities for Aedes mosquitoes.
Physicochemical Parameter Number of samples Across species Across locality(indoor/outdoor)
Test Statistic Degree of Freedom Asymptotic Sig Test Statistic Degree of Freedom Asymptotic Sig.
Temperature/◦ C n = 755 5.487a, 1 0.019 66.184a 1 0.000
pH n = 755 1.566a 1 0.211 0.362 a
1 0.547
TDS /ppm n = 755 0.633a 1 0.426 21.277 a
1 0.000
Turbidity/ NTU n = 755 2.097a 1 0.148 7.126 a
1 0.008
Conductivity/ µs/cm n = 755 0.210a 1 0.647 10.542a 1 0.001
DO/ mg/L n = 755 11.24a 1 0.001 14.752a 1 0.000

a. The test statistic is adjusted forties.

Fig. 5. Percentage distribution of positive containers for Ae. aegypti and Ae. albopictus appearance and condition of breeding water.

4. Discussion
Understanding the local epidemiology and ecology of the vector is
essential for the prevention and control of dengue. Many variables affect
the magnitude of dengue epidemics, either directly or indirectly. Water
quality is a principal factor in the dengue vector breeding habitat, which
determines the oviposition success of female mosquitoes and larval
development survival of the juvenile stages up to adulthood (Oyewole
et al., 2009; Romeo et al., 2005). Female Mosquitoes, Ae. aegypti and Ae.
albopictus lay their eggs and then mature into larvae and pupae in do­
mestic and peridomestic water storage containers, rain-breeding items
such as a wide variety of discarded receptacles, tyres, temporary
removable items, covering polythenes, concrete slabs, gutters and nat­
ural water retention structures such as tree holes, leaf axils and bamboo
stumps (National Dengue Control Unit, 2016). In the first part of the
current study, immature vector prevalence and density were evaluated
in preidentified most common breeding habitat categories in Sri Lanka.
discarded non-reusable items, water storage items and temporary re­
movals, were identified as the most frequently found breeding habitats
for Ae. aegypti mosquitoes. In contrast, discarded non-reusable items,
temporary removals and natural breeding places were identified as the
most frequently identified breeding habitats for Ae. albopictus mosqui­
toes. While frequency prevalence of habitat categories is widely used in
vector control interventions in Sri Lanka, it only indicates the presence
or absence of mosquito larvae in a particular habitat, not the relative
significance or accurate picture of a specific vector’s abundance in each
breeding site category. The majority of previous studies conducted in Sri
Lanka had also focused exclusively on the removal of discarded re­
ceptacles by solid waste management as the primary vector control
strategy by assessing the availability of breeding sites for dengue pre­
vention and management (Surendran et al., 2021; Louis et al., 2016;
Abeyewickreme et al. 2012), However, the current study examined
larval productivity in each habitat category, which may be utilized in
conjunction with frequency distribution and habitat preference to
identify key breeding sites in each area and design an effective man­
agement approach. In the current research, concrete slabs were found as
breeding sites with the greatest larval productivity for both Ae. aegypti
and Ae. albopictus. Even if the number of positive concrete slabs are
smaller due to higher larval density, they must be recognized as a crucial
breeding habitat in dengue management. The District of Gampaha is the
most populous district with the third-highest urban population in Sri
Lanka. The district is situated near Colombo district, the economic hub

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R. Dalpadado et al. Acta Tropica 229 (2022) 106339

Fig. 6. Water quality index of different breeding categories for Aedes mosquitoes in Gampaha District Sri Lanka.

Table 7
Mean density of immature stage across different habitat categories for Ae. aegypti and Ae. albopictus in Gampaha.
Habitat category Mean Density of immature/ mean number positive per dip
Aedes aegypti Aedes albopictus
Clear Turbid Highly Overall mean Clear Turbid Highly turbid Overall mean
turbid
A/C Refrigerator 17.5 ± 5.4 15.0 ± 0.0 NA 17.3± 5.2 16.7± 3.1 7.5 ± 2.2 (1.7 - 3.0 ± 0.4 (2.7- 13.2 ±6.4
trays (8.0–26.7) (8.0–26.7) (9.0- 23.0) 18.0) 3.3) (1.7–23.0)
Concrete slabs 30.6 ± 10.2 17.3 ± 16.9 NA 27.4±14.2 42.1 ± 17.6 16.9 ± 5.3 4.0 ± 0.0 33.5 ± 20.9
(7.2- 80.0) (4.4 - 48.0) (4.4–80.0) (18.0- 72.0) (2.0–40.0) (2.0–72.0)
Discarded 17.0 ± 5.0 8.00 ± 3.0 2.5 ± 0.5 14.9 ± 6.6 18.6 ± 6.2 5.3 ± 2.1 5.5 ± 2.1 17.6 ± 6.9
(8.0–32.0) (2.7- 20.0) (2.0 - 2.7) (2.0–32.0) (6.0- 60.0) (3.0–10.0) (4.0–7.0) (3.0–60.0)
Gutter NA NA NA NA 51. 0 ± 2.6 30.0 ± 12.5 23.7 ± 11.1 31.8 ± 14.6
(48.0- 55.0) (12.0–44.0) (12.0–45.0) (12.0–55.0)
Natural NA NA NA NA 22.2 ± 8.7 20.4 ± 4.2 NA 22.0 ± 8.5
(8.0- 45.0) (12.0–25.0) (8.0–45.0)
Temporary 20.5 ± 6.8 13.0 ±0.0 NA 20.2 ± 10.7 20.8 ± 8.6 15.8 ±6.9 (4.0- NA 20.6 ± 8.7
removable items (8.0–56.0) (8.0–56.0) (6.0- 60.0) 30.0) (4.0–60.0)
Tube wells 14.3 ± 3.1 (6.0- NA NA 14.3 ± 5.1 (6.0- 13.9 ± 5.8 NA NA 13.9 ± 5.8
19.0) 19.0) (8.0- 23.5) (8.0–23.5)
Water storage 23.1 ± 8.4 (7.0- 10.7 ±4.7 NA 22.2 ± 10.7 18.2 ± 7.7 5.3 ± 2.5 (3.8 - NA 17.5 ± 10.0
containers 60.0) (3.0- 15.2) (3.0- 60.0) (5.0- 48.0) 14.0) (8.0–48.0)

Note: Values in the parenthesis include: Mean larval density ± SD (Minimum-Maximum) for each habitat category for Aedes aegypti and Aedes abopictus, and highest
recorded values were highlighted for each category of breeding water.

of Sri Lanka and It comprises five important industrial zones of the
nation. Therefore, numerous infrastructure development projects are
occurring inside the area. Most of the building construction sites are
dynamic ecosystems; varying stages of development may enable the
sites to be suitable for Aedes reproduction; when possible, habitats are
continually generated or not eliminated.
The present research recognized these unfinished building con­
structions with mostly unreachable concrete slabs and floors as one of
the attractive and prolific breeding habitats for Aedes mosquitoes which
demand the urgent attention of the governing authorities of dengue
management. Similar results were also reported in some studies con­
ducted in other countries such as Singapore (Liang et al., 2018) and
China (Liu et al., 2021). Since temporary water collections are retained
for months in these concrete slabs, long-lasting larvicides such as
temephos sand granules (3, 4 months) or larval growth regulator (6–8
weeks) can be applied to prevent mosquito breeding under the proper
monitoring of local health authorities (National Dengue Control Unit,
2016).
According to the present research, temporary removal items are also
a major breeding environment in dengue control. Temporary removals
in all three environmental setups in the Gampaha district had the
maximum number of detected breeding sites for Ae. aegypti, and they
were also recognized as one of the most preferred habitats for Aedes
mosquitoes, with the district’s third-highest larval productivity when
considering primary dengue vector Ae. aegypti. Proper storage of these
items under shade will help reduce the dengue burden to a great extent

9
R. Dalpadado et al.
in the District of Gampaha since they represent about 30% of positive
breeding sites of the district.
The present research also highlights the necessity of preventing
mosquito breeding in Gampaha district roof gutters, identified as one of
the most favoured breeding locations for Aedes mosquitoes with high
larval production. Gutters were often overlooked owing to the diffi­
culties of accessing them during most household and government con­
trol measures, and the importance of gutters was typically
underestimated. As a result, a suitable public awareness campaign
should be created to highlight the relevance of these highly productive
larval breeding sites to the general population and the need for frequent
cleaning of these cryptic breeding locations in the management of
dengue fever within the area. Gutter mesh of the proper size may also be
put over the gutters to prevent mosquito breeding and restrict the
entrance of leaves and debris into the gutters. If they are unreachable for
regular cleaning, long-lasting larvicide can be applied with proper
monitoring. Simultaneously, laws must be strengthened to propose new
rules on the use of gutters in structures and eliminate any unnecessary
roof gutters. Other recognised key breeding sites, such as water storage
items and natural breeding places with higher habitat preference,
immature productivity, and higher availability of positive containers in
the field, should also receive greater attention similar to solid waste
management to manage dengue in the Gampaha district effectively.
Before reaching adulthood, the life cycle of a mosquito passes
through several phases, and environmental factors influence each phase
in various ways. The success of completion of each stage is determined
by both internal and external factors. The characteristics of aqueous
larval habitat also play an important role as it can provide nutrients and
a suitable environment that can increase the survival rate of the larvae
(Madzlan et al., 2016). According to Thangamathi et al. (2014), both
Aedes species favor similar breeding habitats in turbidity, pH, alkalinity,
chloride, and phosphate concentrations.
The larvae have a wide range of temperature tolerance. The tem­
perature is critical in the proper growth of mosquito immatures since
they are poikilothermic. Apart from other variables like food and min­
eral concentrations, the temperature is the primary determinant of
larval growth and development (Gopalakrishnan et al., 2013). Increased
water temperature accelerates the growth of mosquito immatures but
reduces their size (Gubler, 2011), whereas the same increases in tem­
perature generate fewer adult mosquitos (Chatterjee et al., 2015). In the
current research, the optimum temperature range for Ae. aegypti and Ae.
albopictus mosquitos in the Gampaha region was determined to be
30.0–32.0 ◦ C, with around 75% of Aedes mosquitos detected in this ideal
temperature range. Aedes larvae were not found in gutters when the
temperature reached higher over 33 ◦ C, mainly when the gutters were
built of steel. They’ve primarily been found in plastic gutters in the
shade, where the temperature did not rise over the recommended level.
As a result, using steel roof gutters may assist in reducing the dengue
load since they typically heat up, preventing mosquito breeding.
Furthermore, the life cycle durability of Aedes species at various tem­
peratures gives crucial information for determining the consequences of
seasonal changes. As a result, this information may be integrated with
other secondary details to develop a monitoring system for the early
identification of dengue fever cases in the area.
The current investigation showed that Aedes sp. exploits various
breeding habitats with varying physicochemical features. It is most
likely due to the Aedes species’ great environmental adaptability. Our
results revealed that pH levels influenced Aedes immatures’ presence,
and above 65% of Aedes mosquitoes were prominent in breeding water
with 7–8 pH. They also survived in breeding water, where the pH range
within 5.6–8.7. Few other studies also implied that a wide range of pH
levels from 6.5 to 8.0 is suitable for Aedes mosquito survival and
breeding (Waewwab et al., 2019; Umar and Donpedro, 2008). According
to Umar and Donpedro (2008), the pH of the breeding water affects the
osmoregulation and oxygen transport mechanisms in mosquitoes.
Increased pH, salinity, and alkalinity lead to quicker mosquito growth
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Acta Tropica 229 (2022) 106339
larvae up to 9.0 pH. After pH reaches the substantial alkalinity range, pH
inhibits the further growth of Aedes larvae at the field level. So, at the
household level, basic actions like common salt may be used to reduce
Aedes breeding. According to Mukhopadhyay et al. (2010), adding
common salt to Ae. aegypti breeding sites might have a good impact,
especially in areas lacking monitoring and supervision. As a result, in
control programs where breeding habitat reduction is impossible, pH
modification (pH above the range of 8, 9) may be used to manage Aedes
mosquitoes. According to Mukhopadhyay et al. (2010), adding common
salt to probable breeding sites of Ae. aegypti may provide positive out­
comes, particularly in locations where monitoring and supervisory
procedures are inadequate. Therefore, pH manipulation (pH above the
range of 8, 9) may be utilized to manage Aedes mosquitoes in control
programmes where breeding habitat reduction is impossible.
Overgaard et al. (2017) stated that Aedes aegypti immature infesta­
tion was positively associated with total dissolved solids but negatively
associated with dissolved oxygen. Contrary to this, the present study
resulted in higher mosquito larval density when the TDS and DO levels
were higher. Mosquito larvae are metapneustic, meaning they breathe
via their posterior spiracles and breath on mostly ambient oxygen
(Clements, 1992). The dissolved oxygen level, on the other hand, in­
fluences the number, density, and habitat productivity of Aedes larvae
by indicating the degree of pollution in the water. Because Aedes
mosquitos prefer clear water, they are usually inhibited in water with
higher dissolved oxygen levels, and oxygen deficit has an impact on
larval numbers and productivity in Aedes breeding sites. In general,
Aedes mosquitos like to spawn in clean water.
The results of the current study also suggested Aedes larvae are
preferred to breed in habitats with clear and non-opaque breeding
water. Ae. aegypti was rarely reported in breeding water with highly
turbid, brown-colored musky water. Only in discarded receptacles Ae.
aegypti was reported in highly turbid water even with lesser larval
density. Comply with the physical nature of the breeding water, the
results of the present study highlighted both the Aedes vectors inhabited
in water with lower turbidity and TDS levels. However, it was noted Ae.
aegypti had a lower TDS range compared to Ae. albopictus. Ae. aegypti
favours clear water with little dissolved solids and turbidity, but Ae.
albopictus was also reported murky water with high organic matter
dissolved solids and turbidity including natural breeding places such as
tree holes and leaf axils as well as in gutters with higher organic debris.
Agreeing with Montgomery and Ritchie, 2002 Ae. aegypti, in general,
needs less food for oviposition than Ae. albopictus, making it prefer to
nest in less murky water. TDS, total hardness, conductivity, fluoride,
chloride, phosphate, sodium, and potassium concentrations, as well as
the dissolved oxygen content of water, all play a role in mosquito
breeding site selection in addition to these physical factors. Therefore,
when planning controlling programmes for Ae. aegypti, based on the
above interventions, vector control activities must be targeted on stag­
nant clear /clear water collecting breeding habitats rather than highly
polluted container breeding habitats especially in urban areas, Ae.
aegypti was prominent.
However, targeting the control program for Ae. albopictus all possible
turbid breeding places should also consider with clear stagnant water
collections since they can survive in broad turbidity and TDS range.
When comparing the (WQI) of Aedes breeding habitats to potable water
standards in Sri Lanka, it was discovered that Aedes mosquitoes breed in
poor water. The greater dissolved solid concentration in breeding en­
vironments was the significant cause. TDS also indicate the availability
of food for growing larvae. Despite the fact that Aedes mosquitos cannot
grow at higher turbidity levels, their survival is dependent on the
availability of food and organic detritus in certain environments. In
comparison to potable water, they are more plentiful in breeding sites
with low water quality but not in extremely contaminated water bodies.
They may, however, thrive in environments with drinkable water, such
as tube wells and water storage containers. The results of the study
comply with a similar study conducted in India about the water quality
R. Dalpadado et al.
of Aedes breeding habitats (Jasmine and Robin, 2020).
5. Conclusion
Controlling Aedes mosquitoes in key breeding habitats such as con­
crete slabs, water storage items, temporary removals and gutters is
critical for dengue outbreak prevention. Study findings highlighted the
importance of vector control interventions targeting stagnant and clear
water collecting breeding habitats rather than highly polluted container
breeding habitats especially in urban areas, where Ae. aegypti is
prominent.
Ethics approval and consent to participate
Verbal consents were taken from house owners where required
Consent for publication
Not relevant
Availability of data and materials
The data supporting the conclusions of this study are included in this
article
Funding
Not relevant
CRediT authorship contribution statement
Rasika Dalpadado: Visualization, Data curation, Formal analysis,
Writing – original draft. Deepika Amarasinghe: Visualization, Writing
– review & editing. Nayana Gunathilaka: Visualization, Data curation,
Formal analysis, Writing – review & editing.
Declaration of Competing Interest
We the authors certify here that we have no conflicts of interest
regarding publication of manuscript titled “Water quality characteristics
of breeding habitats in relation to the density of dengue vector
mosquitoes, Aedes aegypti and Aedes albopictus, indoor and outdoor do­
mestic settings in Gampaha District; Western Province of Sri Lanka’
Acknowledgments
Not relevant
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