Vol. 41, no.

1 Journal of Vector Ecology 1

Effects of temperature on the life cycle, expansion, and dispersion of Aedes aegypti
(Diptera: Culicidae) in three cities in Paraiba, Brazil
Rafael A. Marinho1*, Eduardo B. Beserra1, Maria A. Bezerra-Gusmão¹, Valbia de S. Porto1, Ricardo A. Olinda2,
and Carlos A. C. dos Santos3

Entomology Laboratory, Nucleus for Systematic and Bioecology of Insects, Department of Biology, Universidade Estadual da Paraíba,
¹

Campina Grande, Paraiba, Brazil, rafa_paraibano@hotmail.com

2
Department of Statistics, Universidade Estadual da Paraíba, Campina Grande, Paraiba, Brazil
3
Unity of Atmospheric Sciences, Center of Technology and Natural Resources, Federal University of Campina Grande, Paraíba, Brazil

Received 9 April 2015; Accepted 27 September 2015

ABSTRACT: The mosquito Aedes aegypti is the primary vector of dengue and is common throughout tropical and subtropical regions. Its
distribution is modulated by environmental factors, such as temperature. This study aimed to evaluate the influence of temperature on the
life cycle and expansion of Ae. aegypti populations in the cities of Campina Grande, João Pessoa, and Patos. Samples of Ae. aegypti were
collected in the three cities and raised in the laboratory. We assessed the life cycles of the three Ae. aegypti populations under six constant
temperatures (16, 22, 28, 33, 36, and 39° C), selected on the basis of historical temperature tendencies of each city. We also used existing
climate data to calculate projected temperature increases for all three areas. Our results suggest that Campina Grande, João Pessoa, and
Patos will experience, respectively, maximum temperature increases of 0.030° C/year, 0.069° C/year, and 0.061° C/year, and minimum
temperature increases of 0.019° C/year, -0.047° C/year, and -0.086° C/year. These projected increases will result in temperatures favorable
to the Ae. aegypti life cycle, causing rapid population growth. Therefore, Ae. aegypti populations are likely to expand in the mesoregions
represented by these cities. Journal of Vector Ecology 41 (1): 1-10. 2016.

Keyword Index: Aedes aegypti, vector biological cycle, intra-population variation, development, temperature tendency.

INTRODUCTION that regulate aspects of mosquito ecobiology in natural and urban

The mosquito Aedes aegypti is the primary vector of dengue
fever and an extremely synanthropic insect due to its anthropophilic
nature and specific reproductive needs (Lambrechts and Failloux
2012). It is abundant in neotropical regions, where environmental
factors (e.g., rainfall, temperature, and relative humidity) favor its
life cycle (Eisen et al. 2014). The global distribution of Ae. aegypti
is strongly influenced by climatic factors, with temperatures of
10° C or lower limiting larval development and adult survival
(Slosek 1986, Hopp and Foley 2001). Optimal temperatures for
development, longevity, and fecundity are between 22° C and 32°
C (Beserra et al. 2009). With higher temperatures in the favorable
survival range of Ae. aegypti, egg-laying time decreases, causing an
increase in egg number (Costa et al. 2010). Moreover, the extrinsic
incubation period of the dengue virus is reduced, resulting in
higher rates of viral transmission (Focks et al. 2000, Hopp and
Foley 2001). According to estimates by Bhatt et al. (2013), 390
million dengue virus infections occur annually, including close to
100 million cases of dengue disease.
Vector-borne diseases, such as dengue, may be particularly
sensitive to both periodic fluctuations and sustained changes
in global and local climates because the vectors themselves
are temperature- and moisture-dependent (Thai and Anders
2011). As the effect of temperature on Ae. aegypti life cycles are
substantially established (Bar-Zeev 1958, Keirans and Fay 1968,
Rueda et al. 1990, Kamimura et al. 2002, Mohammed and Chadee
2011, Richardson et al. 2011), recent studies (Williams et al. 2014,
Campbell et al. 2015) focus on the investigation of the interaction
between life cycle and global climate changes and future scenarios
ecosystems that favor the transmission of dengue.
Based on current understanding of the close relationship
between climate and the life cycle of Ae. aegypti, a projected
temperature elevation of 2° C by the end of the 21st century will
likely broaden the distribution of dengue worldwide (Donalísio
and Glasser 2002). Furthermore, each of the last three decades has
been successively warmer than any decade since 1850. In tropical
regions of South America where Ae. aegypti are particularly
prevalent both maximum and minimum temperatures have risen
in recent decades, corresponding to the worldwide trend.
Based on these previous findings, we hypothesized that the
rise in temperature will favor vector dispersal and geographical
expansion. In this study we aimed to (1) estimate increasing
temperature trends over the next 18 years (until 2032) for the
cities of Campina Grande, João Pessoa, and Patos; (2) evaluate
the effects of temperature on the Ae. aegypti life cycle; and (3)
investigate whether estimates of temperature increase in the three
cities will favor Ae. aegypti dispersion and expansion.
MATERIALS AND METHODS
Study site
The assessment was conducted for 12 months (January, 2012
to February, 2013) and samples of Ae. aegypti populations were
collected from Campina Grande (Bodocongó neighborhood -
07°13′14.92″S, 035°55′1.32″W), João Pessoa (Castelo Branco -
07°08′11″S, 034°51′9.33″W), and Patos (Jatobá - 07°02′40′′S,
037°16′16′′W). These cities were selected because they are
located in different mesoregions (Agreste, Mata, and Sertão da
2 Journal of Vector Ecology June 2016
Paraíba, respectively) that vary in thermal characteristics (Figure
1). Population samples of Ae. aegypti were obtained from eggs
collected in oviposition traps that were installed in 50 residences,
one per block, for each city.
Based on the Köppen-Geiger climatic classification, the three
cities are characterized as follows: (1) Campina Grande has a
moderate tropical climate, with a spring/summer dry season and
an autumn/winter wet season. Maximum summer and winter
temperatures are 30 and 18° C, respectively. Minimum summer
and winter temperatures are 20 and 15° C, respectively. The
annual relative humidity is between 75–82%. (2) João Pessoa has
a hot and humid intertropical climate, with an average annual
temperature of 26.1º C. Maximum temperatures (around 30.5° C)
occur in February and minimum temperatures (around 20.5° C)
occur in August. The annual relative humidity is approximately
89%. (3) Patos has a hot semi-arid climate, with the hottest month
being December (average minimum and maximum temperatures
of 20.4 and 32.7° C, respectively) and the coldest month being July
(average minimum and maximum temperatures of 17.1 and 28.1°
C, respectively). The annual relative humidity is between 40% and
60%.
Aedes aegypti maintenance in the laboratory
Rearing and bioassays were carried out at the Entomology
Laboratory of the Center of Systematic and Insect Bioecology
of Paraíba State University. Rearing was conducted in
environmentally controlled rooms, with the temperature set to
26 ± 2° C, photoperiod to 12:12 hours light-dark, and relative
humidity to 70%, the latter being verified using a thermo-
hygrometer dry bulb and wet. Rearing procedures followed those
described by Beserra et al. (2009).
We attempted to minimize interference between different
Aedes species in our study area with the following procedure. Eggs
from each block were submerged in separate containers filled
with water and allowed to hatch. Upon reaching the 4th instar,
larvae were fixed on a slide for species identification under the
microscope. The study species (Ae. aegypti) was differentiated from
the sympatric Ae. albopictus based on morphological characters
of the eighth abdominal segment, following the identification
key presented by Consoli and Lourenço de Oliveira (1994). We
successfully identified 30% of the larvae in each container.
Calculation of temperature tendencies
To set appropriate temperatures in the laboratory, we
calculated temperature tendencies for each study region using
historical (1975 to 2011) annual temperature data, which were
obtained from the National Institute of Meteorology. RClimdex
software 1.9.0, based on the R statistical package, was used for index
calculation. RClimdex calculates all 27 indexes recommended
by Climate Change Initiative/Climate and Ocean: Variability,
Predictability, and Change (CCI/CLIVAR) Expert Team for
Climate Change Detection Monitoring and Indexes (ETCCDMI)
(Orlowsky and Seneviratne 2012). The program is able to adjust
for errors in the database by replacing all faulty data with -99.9,
an internal format recognized by R. These errors include negative
daily precipitation values and maximum daily temperatures lower
than the minimum daily temperatures.
Upon obtaining annual temperature tendencies, we multiplied
the values by 10 to obtain decadal tendencies. Starting from 2012,
we projected mean maximum and minimum temperatures for two
decades, until 2032. Based on the decadal ones, we were able to
linearly calibrate the temperatures of the three cities. Temperatures
between the estimated minimum temperature (16° C) and the
maximum temperature (39° C) were calibrated at differences of 6,
5, and 3° C, resulting in constant temperature ranges of 16, 22, 28,
33, 36, and 39° C. These temperatures were used to evaluate the
cycle life and thermal requirements for Ae. aegypti development.
Although the effects of constant temperatures on Ae. aegypti are
well established (Delatte et al. 2009, Richardson et al. 2011), we
chose not to alter the selected temperatures to better compare our
results with previous studies.
Figure 1. Map of Paraíba, Brazil, showing the
three cities (João Pessoa, Patos, and Campina
Grande) sampled in this study.
Vol. 41, no. 1 Journal of Vector Ecology 3
Life cycle of Ae. aegypti and developmental thermal
requirements
For each of the 50 population samples per city, life cycles
were assessed through the first generation (F1), considering the six
different temperatures (see previous section), 12-h photoperiod,
and 70% relative humidity. Evaluation of the incubation period
was performed from the first five ovipositions of each repetition,
which were immersed in 500 ml disposable cups filled with
distilled water. Water temperatures ranged from 15.0 to 15.7° C,
21.0 to 21.7° C, 27.2 to 27.5° C, 32.0 to 32.8° C, 34.9 to 35.5° C, and
38.1 to 38.7° C for the temperatures conditions of 16, 22, 28, 33,
36, and 39° C, respectively. To assess the length of the larval stage,
20 larvae/cup were used per repetition. Assessments were made
daily; for each temperature, we recorded the egg development
periods and mortality, larval and pupal stages, larval emergence,
and adult longevity and fecundity.
Statistical analysis
The experiment was completely randomized in a factorial 3
× 6 design (18 treatments). Independent variables consisted of
six temperatures (16, 22, 28, 33, 36, and 39° C), three populations
(Campina Grande, João Pessoa, and Patos), and five replications.
Group differences and any significant interactions between
independent variables were examined using ANOVA. Dependent
biological variables were checked for homogeneity of variance
using the Box-Cox test (Box and Cox 1964) and the Hartley test
(Hartley 1950). Subsequently, treatment averages were compared
using Tukey’s test. All analyses were performed in SAS/STAT
(2003). Significance was set at p < 0.05.
Linear regression equations y1 = ai + bit were estimated
between the inverse of development time (1/D response variable,
yt in days) and studied temperatures (dependent variable, t in
°C). The base temperature (Tb) was estimated as the relationship
between the intercept and the linear coefficient of the equation
(Tb = -ai/bi) when development time = 0 (0 = ai + bit). The
thermal constant (K), which is expressed as degrees/day (GD),
was calculated as the inverse of the linear coefficient (K = 1/bi)
(Santana et al. 2010).
In order to estimate the annual number of generations, we
used biological data from the temperature setting that resulted in
the shortest life cycle. The likely generation number (NG) of insects
in the field was estimated based on their thermal requirements
and on the typical temperatures in their area of origin (obtained
from INMET). Specifically, we used the following equation: NG
= T(tc-tb)/K, where T is the time considered in a year, tc is the
average monthly temperatures of the study areas, K is the thermal
constant, and tb is the base temperature.
RESULTS
Maximum and minimum temperature tendencies
Due to missing temperature data in some years, we selected
only the period from 1994 to 2011 (18 years) for analysis (Figure
2). We found no significant linear increases in the maximum
and minimum temperature tendencies of our study regions (p >
0.05). However, we noted that only Campina Grande exhibited a
trend of increasing minimum temperature, while the minimum
temperatures for the other two cities tended to decrease (Figure
2). We attribute the lack of significance to the short-time series
used in the analysis (only 18 years of data), but we do not consider
the amount of data available to have compromised the methods
applied in this study.
Based on existing data, maximum temperatures in João
Pessoa tended to increase at approximately 0.069° C/year, while
minimum temperatures tended to decrease at -0.047° C/year.
Projected maximum and minimum temperatures for 2032 were
32.8° C and 18.8° C, respectively. Maximum and minimum
temperatures in Campina Grande tended to increase at rates
of 0.030° C/year and 0.019° C/year, respectively, resulting in
projected 2032 temperatures of 33.7 °C (maximum) 16.6 °C
(minimum). Patos exhibited an increasing trend of 0.061 °C/year
for maximum temperature and a decreasing trend of -0.0869°
C/year for minimum temperature. Projected 2032 temperatures
in Patos were calculated to be 39.1 °C (maximum) and 16.8°
C (minimum). The minimum (16° C) and maximum (39° C)
projected temperatures were used as the reference for determining
the six temperatures used in our rearing experiments.
Ae. aegypti aquatic life cycle at six constant temperatures
We found no significant interaction between population
origin and temperature on egg viability (p > 0.05), showing that
independently of vector collection point and temperature, egg
viability did not differ between population samples. Observed
viabilities were generally high: the greatest percentage of viable
eggs (85.2%) was in João Pessoa, while the greatest percentages
of viable larvae 87.7% and pupae (92.6%) were in Patos (Table 1).
Temperature affected every stage of the Ae. aegypti life cycle
similarly in all three populations: the length of developmental
stages was inversely proportional to the temperature increase.
However, the egg stage and the overall life cycle (egg to adult) of the
populations differed significantly in the intensity of their response
to temperature (Tables 2 and 3). No embryonic development or
survival of the larval stages occurred at 39° C (water temperature
37.8 to 38.5° C), and so this temperature was excluded from
statistical analysis.
There was a significant interaction between population origin
and temperature on the Ae. aegypti embryonic development
period (Campina Grande p = 0.0490; Patos p = 0.0366), except in
João Pessoa, which ranged from 10.4 days at 16° C to 2.59 days at
36° C. In general, Ae. aegypti from João Pessoa had the shortest
embryonic development (5.03 days), whereas mosquitoes from
Patos (5.92 days) and Campina Grande (5.86 days) did not differ
significantly from each other (Table 2). No significant differences
were found in the duration of larval and pupal stages among
population samples at any temperature. However, the length of
the full life cycle (egg to adult) differed significantly among the
populations at temperatures between 16° C and 22° C (Campina
Grande p = 0.0100, João Pessoa p = 0.0047, Patos p = 0.0082) ;
specifically, the Campina Grande population required more
days to develop. In general, Ae. aegypti from João Pessoa had the
shortest development period of 18.3 days (Table 3).
Adult longevity and number of eggs per Ae. aegypti female
Longevity and female fecundity (measured by egg number)
were significantly affected by the interaction between population
and temperature. The temperature extremes of 16 and 36° C were
4 Journal of Vector Ecology June 2016

Figure 2. Trends of maximum and minimum temperatures for Campina Grande (a), João Pessoa (b), and Patos (c), based on the INMET
historical data series.
Vol. 41, no. 1 Journal of Vector Ecology 5
Table 1. Average viability (% ± SD) of egg, larval, and pupal stages across five temperatures, in Aedes aegypti (Linnaeus, 1762) populations
from the cities of Campina Grande, João Pessoa, and Patos (Paraíba, Brazil). Insects were reared in a 12:12 h light:dark photoperiod.
Viability (%)
Population
Eggns Larvans Pupans
Campina Grande 81.8 ± 14.57 78.2 ± 1.97 91.2 ± 1.79
João Pessoa 85.2 ± 14.38 80.0 ± 5.74 91.0 ± 2.37
Patos 82.59 ± 6.23 87.7 ± 4.59 92.6 ± 0.71
Overall mean 83.2 81.9 91.6
CV 7.09 11.5 7.79
ns
Not significant (ANOVA; p ≤ 0.05).

Table 2. Embryonic developmental duration (days) of Aedes aegypti from Campina Grande, João Pessoa, and Patos. Insects were reared
in a 12:12 h light:dark photoperiod under five temperatures. Data are presented as means ± SD. Original means are listed; data were log-
transformed for statistical analysis. Means followed by the same lowercase letters in the rows and the same capital letters in the columns
are not significantly different (Tukey’s test; p < 0.05).
Embryonic development (days)
16°C 22°C 28°C 33°C 36°C Overall mean
Campina Grande 13.4 ± 0.47aA 7.68 ± 0.59 bA 3.35 ± 0.62 cA 2.70 ± 0.41 cB 3.66 ± 0.59 cA 5.86 ± 3.95 A

João Pessoa 10.4 ± 0.59 aB 5.16 ± 0.30 bB 3.15 ± 0.40 cA 2.61 ± 0.64 cB 3.48 ± 0.32 cA 5.03 ± 301 B

Patos 11.5 ± 0.66 aB 7.92 ± 0.65 bA 3.32 ± 0.48 cdA 4.23 ± 0.43 cA 2.59 ± 0.36 dA 5.92 ± 3.46 A
CV = 9.32%

Table 3. Duration (days) of larval and pupal stages, as well as full development (egg to adult emergence) in Aedes aegypti populations
from Campina Grande, João Pessoa, and Patos. Insects were reared in a 12:12 h light:dark photoperiod under five temperatures. Means
followed by the same lowercase letters in the rows and the same capital letters in the columns are not significantly different (Tukey’s test;
p <0.05).
Temperature
Population 16° C 22° C 28° C 33° C 36° C Overall Mean
(15.0–15.7°C) 2
(21.0–21.7°C) (27.2–27.5°C) (32.0–32.8°C) (34.9–35.5°C)
Larval stage
Campina Grande 20.51 ± 1.45 aA 13.40 ± 0.54 bA 6.16 ± 0.63 cdA 6.70 ± 0.37 cA 5.17 ± 0.79 dA 10.39 ± 6.0 A
João Pessoa 19.86 ± 0.70 aA 12.49 ± 0.49 bA 7.20 ± 0.46 cA 6.12 ± 0.72 cdA 5.26 ± 0.35 dA 10.19 ± 5.58 A
Patos 20.19 ± 1.01 aA 12.12 ± 0.49 bA 7.25 ± 0.30 cA 7.12 ± 0.38 cA 5.43 ± 0.97 dA 10.42 ± 5.51 A
CV = 6.90%
Pupal stage
Campina Grande 6.97 ± 1.53 aA 3.68 ± 0.70 bA 2.08 ± 0.48 cA 2.51 ± 0.33 bcA 1.29 ± 0.30 dA 3.30 ± 2.16 A
João Pessoa 5.95 ± 0.74 aA
3.25 ± 0.50 bA
2.16 ± 0.69 bcA
2.69 ± 0.27 bA
1.50 ± 0.39 cA
3.11 ± 1.64 A
Patos 7.06 ± 1.61 aA 3.38 ± 0.91 bA 2.51 ± 0.34 bcA 1.88 ± 0.16 cA 1.30 ± 0.18 cA 3.22 ± 2.21 A
CV = 23.31%
Full development (egg to adult)
Campina Grande 41.42 ± 2.56 aA 24.76 ± 1.06 bA 11.59 ± 0.57 cA 11.92 ± 0.43 cA 10.13 ± 1.29 dA 19.96 ± 12.2 A
João Pessoa 36.28 ± 1.63 aB
20.91 ± 0.54 bB
12.52 ± 0.49 cA
11.43 ± 0.64 cdA
10.67 ± 1.40 dA
18.36 ± 9.93 B
Patos 38.82 ± 1.46a AB 23.42 ± 1.46 bAB 13.08 ± 0.60 cA 13.24 ± 0.46 cA 9.33 ± 0.90 dA 19.58 ± 10.9 A
CV = 6.51%
6 Journal of Vector Ecology June 2016

Table 4. Longevity (days) of adults and number of eggs/female in Aedes aegypti populations from Campina Grande, João Pessoa, and
Patos at five temperatures and with a 12:12 h light:dark photoperiod. Means followed by the same lowercase letters in the rows and
the same capital letters in the columns are not significantly different (Tukey’s test; p < 0.05). Original means are listed; data were log-
transformed for statistical analysis.
Temperature
Population
16°C 22°C 28°C 33°C 36°C Overall mean
Longevity of males
Campina Grande 10.89 ± 3.83 bA 29.26 ± 2.03 aAB 21.94 ± 3.32 abB 14.75 ± 5.11 bB 8.32 ± 1.50 bA 17.04 ± 8.40 B
João Pessoa 15.37 ± 2.64 cA
35.55 ± 3.41 aA
32.06 ± 2.26 aA
23.18 ± 3.38 bA
8.72 ± 1.43 cA
22.98 ± 10.5 A
Patos 10.86 ± 1.38 bA 27.59 ± 4.16 aB 31.05 ± 3.92 aA 23.54 ± 1.19 aA 10.53 ± 2.13 bA 20.71 ± 9.1 A
CV = 14.88%
Longevity of females
Campina Grande 12.93 ± 2.44 bcA
25.44 ± 4.13 aB
23.30 ± 2.16 aB 15.05 ± 3.33 bB 7.10 ± 1.45 cA 16.76 ± 7.39 C
João Pessoa 14.76 ± 2.74 cA 34.71 ± 1.88 aA 30.08 ± 3.95 aA 22.16 ± 1.98 bA 10.53 ± 1.68 cA 22.45 ± 9.56 A
Patos 10.22 ± 2.73 bA
26.56 ± 1.56 aB
27.45 ± 2.05 aAB
22.64 ± 2.11 aA
9.49 ± 3.77 cA
19.27 ± 8.38 B
CV = 13.72%
Number of eggs/female
Campina Grande 3.50 ± 1.44 dA
79.22 ± 11.2 aB
64.42 ± 8.65 abB 40.4 ± 11.7 bA 16.12 ± 8.83 cA 40.73 ± 30.1 C
João Pessoa 2.75 ± 1.51 dA 161.5 ± 7.34 bA 262.3 ± 19.7 aA 51.3 ± 10.5 cA 30.57 ± 14.1 cA 101.6 ± 99.8 A
Patos 1.46 ± 0.70 dA
126 ± 23.3 aAB
181.3 ± 43.1 aA
52.92 ± 10.8 bA
22.27 ± 8.73 cA
76.79 ± 71.53 B
CV = 21.50%

Table 5. Results of regression analyses on the development time of Aedes aegypti populations from Campina Grande, João Pessoa, and
Patos. Insects were raised in a 12:12 light:dark photoperiod. Base temperatures (tb), thermal constants (K), intercepts (a), and slopes (b)
are shown.

Tb (°C) K (GD) a ± SEM b ± SEM R²

Population
Egg stage
Campina Grande 9.92 74.6 -0.13286 ± 0.11663 0.01339 ± 0.00417 0.77
João Pessoa 4.32 90.9 -0.04758 ± 0.13102 0.01099 ± 0.00469 0.64
Patos 10.35 73.3 -0.14111±0.10571 0.01363 ± 0.00378 0.81
Larval stage
Campina Grande 9.41 139.8 -0.06734 ± 0.03843 0.00715 ± 0.00137 0.90
João Pessoa 9.45 140.8 -0.06713 ± 0.01274 0.00710 ± 0.00045 0.98
Patos 8.31 156.9 -0.05298 ± 0.02273 0.00637 ± 0.00081 0.95
Pupal stage
Campina Grande 11.1 38.2 -0.29193 ± 0.21694 0.02612 ± 0.00776 0.79
João Pessoa 7.31 49.8 -0.14683 ± 0.17590 0.02006 ± 0.00629 0.77
Patos 11.9 35.4 -0.33578 ± 0.12554 0.02824 ± 0.00449 0.92
Full development (egg to adult)
Campina Grande 9.57 261.0 -0.03667 ± 0.01830 0.00383 ± 0.00065 0.91
João Pessoa 7.30 290.6 -0.02513 ± 0.01259 0.00344 ± 0.00045 0.95
Patos 9.65 264.5 -0.03648 ± 0.01688 0.00378 ± 0.00060 0.92
Vol. 41, no. 1 Journal of Vector Ecology 7
Table 6. Number of annual generations estimated for laboratory and field populations of Aedes aegypti in Campina Grande, João Pessoa,
and Patos.
Laboratory Annual Generations
Population
16º C 22º C 28º C
Campina Grande 9.0 17.3 25.7
João Pessoa 10.9 18.4 26.0
Patos 8.7 17.0 25.3
highly unfavorable to adult longevity in all populations. In Patos,
temperatures between 22 and 33° C were favorable to both male
and female longevity, with no significant difference between them.
However, in Campina Grande and João Pessoa, male and female
longevity were significantly reduced at 33° C (Campina Grande
pfemale < 0.0001; pmale < 0.0001)/( João Pessoa pfemale < 0.0001; pmale <
0.0001) compared with 22 and 28° C, respectively. In general, João
Pessoa males and females were longer-lived than mosquitoes from
the other two cities (Table 4).
The temperature extremes of 16 and 36° C decreased the
number of eggs per female considerably, with 16° C resulting
in the lowest fecundity, between 1.46 eggs/Patos female and
3.5 eggs/Campina Grande female. The largest number of eggs/
female (average = 262.3) was observed at 28° C in João Pessoa.
Considering the overall average of all populations across all
temperatures, the females of João Pessoa exhibited high fecundity
(101.6 eggs/female).
Developmental thermal requirements and estimation of Ae.
aegypti annual generation number
The development speed of Ae. aegypti responded significantly
to an increase in temperature. The estimated values for Tb and (K)
in Campina Grande, João Pessoa, and Patos populations varied
between immature development stages (egg, larval, pupal) and the
full life cycle.
In the João Pessoa sample, we observed the lowest Tb for
viable egg (4.32° C) and pupal (7.31° C) stages, as well as for
the full egg to adult cycle (7.30° C), whereas the lowest Tb for a
viable larval stage occurred in the Patos sample (8.31° C). These
data suggest that the João Pessoa and Patos populations have
higher tolerances to reduced environment temperature (Table 5).
However, we also note that Patos mosquitoes exhibited the highest
Tb (11.9° C) across all stages and populations.
Respectively, K values obtained from egg, larval, and pupal
stages were as follows: 74.6, 139.8, and 38.2 GD for Campina
Grande; 90.9, 140.8, and 49.8 GD for João Pessoa; 73.3, 156.9, and
35.4 for Patos. As the thermal constant is the sum of favorable
temperatures for mosquito development, 261.0 GD (Campina
Grande), 290.6 GD (João Pessoa), and 264.5 GD (Patos) were
required to complete the full Ae. aegypti life cycle in each of the
three cities.
The slope of the regression equations for each life stage
represents developmental speed, where a steeper slope indicates
a higher development speed. The Patos population exhibited the
steepest slopes in egg (b = 0.01363) and pupal (b = 0.02824) stages.
Field
Annual
33º C 36º C Average o C
Generations
32.7 37.0 23.6 23.6
32.2 36.0 24.7 24.7
32.2 36.3 28.0 28.0
For the larval stage, the Campina Grande population exhibited
the fastest development (b = 0.00715) compared with the other
two populations (b = 0.00637 for Patos and b = 0.00710 for João
Pessoa. When examining the full life cycle (egg to adult), the
Campina Grande population exhibited faster development (b =
0.00383), followed by the João Pessoa population (b = 0.00378).
Based on the thermal requirements for Ae. aegypti
development and laboratory temperatures, we found that the
number of annual generations varies between 8.7 (at 16° C) to 37
(at 36 °C). On the basis of these results and data on the typical
climate in each region, we observed that the number of insect
generations is higher in Patos (28.0) than in Campina Grande
(23.6) or João Pessoa (24.7) (Table 6). Therefore, increase in the
Ae. aegypti population in Patos is likely to be faster than that in
the other two cities.
DISCUSSION
Climate tendencies and potential effects on vector expansion
Climate tendencies estimated for the regions of Campina
Grande, João Pessoa, and Patos are similar to patterns described
in the 5th IPCC-AR5 report, which found a progressive
temperature increase over the past 100 years, caused primarily
by anthropogenic activity but also by climate variability. Other
states in the northeastern region of Brazil are also experiencing
a trend of increasing temperature. For example, cities such as
Teresina, Piaui, and Sao Luís, Maranhão are projected to have an
average increase of approximately 0.014° C per year, causing a rise
in temperature of 0.62° C by 2050 and 1.34° C by 2100 (Santos
et al. 2010). Collectively, our results combined with previous data
suggest that neotropical regions will tend to grow hotter over the
years. On a worldwide scale, the IPCC affirms this trend of warmer
climates and predicts that 2016–2035 will experience an increase
of 0.3 to 0.7° C in global mean surface temperature, relative to
1986–2005.
Our projected temperature increases for Campina Grande
(33.7° C) and João Pessoa (32.6° C) by 2032 were similar to the
30.4 to 34.8° C projections of the IPCC (2013). Because these
temperature conditions are favorable to Ae. aegypti development
(Tables 1–6), future climatic changes will likely encourage the
spread and expansion of Ae. aegypti. Patos is a notable exception,
however, because the city’s estimated 2032 temperature of 39.1
°C is too hot for Ae. aegypti. Overall, as regional climates shift to
become warmer, the resultant changes in Ae. aegypti populations
(Vargas et al. 2010) should also influence the distribution of
8 Journal of Vector Ecology June 2016
dengue (Campbell et al. 2015).
Effect of temperature on the Ae. aegypti life cycle
Most of the temperatures selected for this study allowed Ae.
aegypti populations to fully develop without causing deleterious
effects. The sole exception was 39° C, which suppressed
embryonic development and led to larval death within hours of
hatching. The negative effect of the 39° C condition is consistent
with the influence of temperature on culicid vector development
(Mohammed and Chadee 2011, Zequi and Lopes 2012) and
suggests that 39° C approaches the lethal temperature (Mourya et
al. 2004, Aghdam et al. 2009, Couret et al. 2014) for Ae. aegypti.
According to Eisen et al. (2014), the relationship between water
temperature and development rate is similar for eggs, larvae, and
pupae. At temperatures favorable to the life cycle, insects not
only complete their development but do so more quickly, which
may enhance vector competence for arboviruses (Muturi et al.
2012). Above the optimal temperature, development rates remain
relatively stable and may decrease slightly until temperatures reach
an upper limit, at which point development drops dramatically.
This upper limit occurs at ∼38 to 42° C (Eisen et al. 2014),
corroborating our findings. Despite the lack of full life cycle data
at 39° C, the fact that some larvae still hatched indicates a possible
physiological adaptive response to high water temperatures (37.9
to 38.5° C under the 39° C condition).
According to Beserra et al. (2009), development time and
temperature are inversely related, and the optimum growth
range is between 22 and 32° C. Insects that develop within this
optimum range experience improvements in other intrinsic
characteristics, such as starvation resistance (Padmanabha et
al. 2011). Temperature-influenced variation in development
and related characteristics have been reported in Ae. aegypti
populations originating from several cities in the Agreste and
Sertão da Paraíba mesoregions, with each population differing in
ways that reflected eco-climatic adaptations to their home regions
(Beserra et al. 2006).
The mean viability of egg, larval, and pupal stages for all
population samples was high (above 80%), corroborating previous
reports (Beserra et al. 2006, Tejerina et al. 2009). However, we
should note that due to differences in experimental conditions
such as variation in feed, insect density, amount of water, insect
strain, and equipment, comparisons across multiple studies may
not always be reliable and should be considered with caution
when the objective is to define the best ecological parameters of
the vector (Walsh et al. 2011).
The temperature extremes of 16 and 36° C considerably
decreased adult longevity and female fecundity, similar to the
findings of Costa et al. (2010). Twice as many female Ae. aegypti
survived when subjected to 25 vs 35° C, resulting in 40% more
eggs produced. According to Carrington (2013a) that evaluated
temperature fluctuations (DTR) in an Ae. aegypti population of
Thailand, females from the 35° C constant treatment were the only
ones to successfully take a blood meal. The mean number of eggs
laid was 25.5 eggs/female. At 35° C, with small fluctuations, no
females were interested in feeding on human blood. At 16° C the
authors reported that the reproduction had failed. Males tended to
live longer than females at most temperatures. This pattern may be
due to the intensity of oviposition, which is energetically expensive
and likely to reduce longevity. Our longevity results are similar
to a previous study showing reduced longevity with increasing
temperatures and significantly higher mortality for females than
for males (Beserra et al. 2009). However, studies in Ae. albopictus
found that females tended to live longer than males (Löwenberg
and Navarro-Silva 2004, Delatte et al. 2009).
Thermal requirements and number of annual generations
The Campina Grande, João Pessoa, and Patos populations
differed in thermal requirements at all stages of the life cycle. The
João Pessoa population possessed the lowest base temperatures
required for successful development, reflecting their ability to
tolerate gradual temperature reduction to the threshold at which
development ceases. In general, variation in Tb and K values may
indicate adaptability to adverse environmental conditions that
require an immediate response to temperature changes. The rapid
proportional development between instars following rising water
temperatures is an example of such an immediate response.
Tb and K values of all developmental stages were generally
within previously reported ranges (Beserra et al. 2006), regardless
of population origin. One exception was our K value for the pupal
stage, which was higher at 35 GD than the range of 19.6–32.1
GD reported by Beserra et al. (2006). Beserra et al. (2009) have
also found tb and K within the ranges above. However, a later
study by Beserra et al. (2009) reported observed Tb and K that
were generally lower than those reported in this study, indicating
considerable variation in these values. A very low value of Tb for
Ae. aegypti was probably caused by physiological differences of
each stage when exposed to stressful environmental conditions.
One major caveat in our study is the use of constant
temperatures in the laboratory. Some authors (Carrington et al.
2013a) have suggested that such conditions may not accurately
reflect the variable environmental conditions in the field. For
example, estimates of degree/days will differ under constant
versus fluctuating temperatures. Carrington et al. (2013b) also
noted that in their Thailand Ae. aegypti population, they were
unable to find a significant difference in developmental stages
based on K values calculated with constant temperature, while
differences emerged under fluctuating temperatures. While we
acknowledge this issue, we were able to obtain significant results
using constant temperatures, suggesting that the importance of
using natural temperature variation may depend on the specific
Ae. aegypti population.
Our study strongly suggests that future thermal conditions
of the three examined cities will result in Ae. aegypti population
growth, as well as increases in dispersion and expansion.
Specifically, the optimal temperature (22 to 36° C) for Ae.
aegypti development falls within both the current and projected
temperature ranges for Campina Grande (max. = 33.7° C and
min. = 16.6° C), João Pessoa (max. = 32.8° C and min. = 18.8° C),
and Patos (max. = 39.1° C and min. = 16.8° C). These favorable
temperatures resulted in a larger number of generations per
year, most notably in Patos, and increased development speed in
Campina Grande and João Pessoa. Both factors contributed to an
Ae. aegypti population spike by shortening the time required for
completing a full life cycle and reproducing.
To conclude, our results showed that local populations of
Ae. aegypti were able to complete the cycle of life in extreme
Vol. 41, no. 1 Journal of Vector Ecology 9

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Delatte, H., G. Gimonneau, A.Triboire, and E.D. Fontenille.
2009. Influence of temperature on immature development,
Thanks to the Municipal Health Department of Campina
survival, longevity, fecundity, and gonotrophic cycles of Aedes
Grande, João Pessoa, and Patos, and to Paraíba State University
albopictus, vector of chikungunya and dengue in the Indian
for supporting this research.
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