Vol. 35, no.

2 Journal of Vector Ecology 277

Bionomics of Aedes aegypti subpopulations (Diptera: Culicidae) from Argentina

Marta Gladys Grech, Francisco Ludueña-Almeida, and Walter Ricardo Almirón
Centro de Investigaciones Entomológicas de Córdoba, Edificio de Investigaciones Biológicas y Tecnológicas, Facultad de
Ciencias Exactas, Físicas y Naturales, Universidad Nacional de Córdoba. Av. Vélez Sarsfield 1611 (X5016GCA) Ciudad
Universitaria, Córdoba, Argentina

Received 24 November 2009; Accepted 5 May 2010

ABSTRACT: Differences in biological features of immature and adult Aedes aegypti, as well as variability in vector competence,
seem consistent with the existence of genetic variation among subpopulations and adaptation to local conditions. This
work aims to compare the bionomics of four Ae. aegypti subpopulations derived from different geographical regions reared
under temperate conditions. Life statistics of three Ae. aegypti subpopulations from the provinces of Córdoba, Salta, and
Misiones were studied based on horizontal life tables. The Rockefeller strain was used as a control. The development time
required to complete the larva and pupa stages varied from 6.91 to 7.95 and 1.87 to 2.41 days, respectively. Significant
differences were found in mean larval development time between the Córdoba and Orán subpopulations. The larva-pupa
development time was similar in all the subpopulations. However, survival values varied significantly between the Orán
and San Javier subpopulations. The proportion of emergent males did not differ from females within each subpopulation
nor among them. Adult longevity was similar among the subpopulations. The average number of eggs laid by each female
was significantly different. The Rockefeller strain laid a significantly greater number of eggs (463.99 eggs/female) than the
rest of the subpopulations. Moreover, differences in the demographic growth parameter Ro were detected among the four
subpopulations. The differences obtained in larval development time, larva-pupa survival values, and net reproductive rates
among the subpopulations might reflect underlying genetic differences as a result of colonization from different regions that
probably involve adaptations to local conditions. Journal of Vector Ecology 35 (2): 277-285. 2010.

Keyword Index: Aedes aegypti, subpopulations, biological variation, life statistics, Argentina.

INTRODUCTION and south (La Pampa province) (Rossi et al. 2006), in

Aedes aegypti (L.) has long been studied because of
its importance as a vector of viral diseases such as dengue
and urban yellow fever (Christophers 1960, Gubler
1988, Clements 1992, PAHO 1995). Governments and
health organizations have invested considerable effort in
eradicating Ae. aegypti from the Americas. Eradication
programs initiated in 1915 achieved the apparent
elimination of Ae. aegypti from most of the region by
1970, utilizing vertically-structured programs (PAHO
1994, Gubler and Clark 1994). By 1964, these mosquitoes
were considered eradicated from Argentina (Kerr et al.
1964, Bejarano 1979, Gubler 1988, Nathan 1993). The first
re-infestation of Argentina was reported in 1986 in the
provinces of Formosa and Misiones, and in 1991 Ae. aegypti
was detected in Buenos Aires province. In 1995, the Ministry
of Health of Córdoba province reported the presence
of Ae. aegypti and confirmed the establishment of this
mosquito in Córdoba city (Carcavallo and Martínez 1968,
Bejarano 1979, Campos 1993, Campos and Maciá 1996,
Almirón and Ludueña-Almeida 1998, Coto et al. 2003).
The re-infestation in Argentina might have entered the
northwestern provinces from Bolivia and the northeastern
provinces from Paraguay and Brazil (Rondán-Dueñas et al.
2009). In Argentina, the current geographical distribution
of Ae. aegypti is wider than before its eradication (Curto
et al. 2002). The distribution has expanded toward the
west (Mendoza province) (Domínguez and Lagos 2001)
comparison to the historical distribution.
Between April and November 1997, 19 cases of dengue
fever by DEN-2 serotype were detected in the localities
of Orán, Salvador Maza, Güemes, and Tartagal (Salta
province) in northwestern Argentina (Avilés et al. 1999).
In 1998, the first epidemic of dengue was recorded in Salta
province with 359 confirmed cases by the same serotype
and it was thought most likely to be linked to an under-
reported outbreak of DEN-2 in Bolivia in 1996 and 1997
(Gianella et al. 1998, Van der Stuyft et al. 1998). During
2000, a second epidemic of DEN-1 serotype was recorded in
Argentina, affecting the provinces of Misiones and Formosa
(in the northeast). Both the 1998 and 2000 epidemics were
associated with outbreaks in the neighboring countries of
Bolivia, Brazil and Paraguay (Avilés et al. 2003). During
2002, 214 cases by DEN-1 were identified in Salta province,
and the DEN-3 serotype was recorded for the first time
in Misiones province. The emergence of DEN-3 in Salta
province in 2003 (co-circulating with DEN-1 and DEN-
2) was shortly followed in 2004 by significant outbreaks
of DEN-3 there and in the neighboring provinces of Jujuy
and southwestern region of Formosa (National Ministry
of Health 2006). During the summer-autumn of 2009,
an unprecedented epidemic of DEN-1 affected the north-
western provinces reaching central areas of the country,
with autochthonous cases recorded for the first time in
provinces such as Catamarca, Chaco, Córdoba, Corrientes,
Buenos Aires, Santa Fe, Santiago del Estero, and Tucumán
278 Journal of Vector Ecology December 2010

(National Ministry of Health 2009).

According to Dègallier et al. (1988), the biological
features of Ae. aegypti larva and adults seem to vary
according to local conditions. Based on genetic studies
of Ae. aegypti subpopulations from Argentina using
molecular markers, great differences were detected in
haplotype compositions between different regions of the
country (de Sousa et al. 2000, 2001, Rondán-Dueñas et
al. 2009). Tejerina et al. (2009a) reported differences in
life statistics between subpopulations of Ae. aegypti from
four localities from the north-east of Misiones province
(Argentina). Moreover, variability in vector competence of
these mosquitoes in several subpopulations has also been
demonstrated (Beerntsen et al. 2000).
At present, the best option to prevent epidemic
outbreaks and reduce dengue transmission is to control
the vector populations. Since there is autochthonous
transmission of dengue virus and also the introduction
of cases of dengue and yellow fever from neighboring
countries (Bolivia, Brazil, Paraguay), it is necessary to
study the ecological features of Ae. aegypti populations to
understand the dynamics of viral diseases and be able to
implement effective control programs. Therefore, the aim
of this work was to compare life statistics of four Ae. aegypti
subpopulations derived from different geographical regions
(the provinces of Córdoba (Córdoba city), Salta (San Ramón
de la Nueva Orán city -hereafter Orán), and Misiones (San
Javier city)), reared under temperate conditions, because
reinfestation is moving south. The Rockefeller strain
provided by the CDC of Puerto Rico was used as a control Figure 1. Map of Argentina showing location of collection
for comparison with a strain highly adapted to laboratory sites. Or = Orán city (Salta Province), SJ = San Javier
conditions. city (Misiones Province), Co = Córdoba city (Córdoba
Province).
MATERIALS AND METHODS
Annual average cumulative rainfall for Orán is 1,067
Aedes aegypti subpopulations were from the cities of mm and the mean maximum, average, and minimum
San Javier, Orán, and Córdoba (Figure 1). San Javier city temperatures are 27.7°, 21.2°, and 16.4° C, respectively
(27º52’ S; 55º08’ W) is located in the southeastern region (National Meteorological Service 2009). The National
of the subtropical province of Misiones, along the banks Coordination for Vector Control of the National Ministry
of the Uruguay River opposite the Brazilian city of Porto of Health recorded entomological data from Orán city,
Xavier. Annual average cumulative rainfall for San Javier is showing HI and BI of 6.46% and 8.9, 14.28% and 29.27, and
1,948 mm and the mean maximum, average, and minimum 22.49% and 46.26 during February 2003, 2004, and 2005
temperatures are 27.6° C, 21.5° C, and 16.6° C, respectively. respectively (Estallo et al. 2008). No specific information
Entomological surveillance data were estimated for about Ae. aegypti breeding containers is available for the
Misiones province (Colonia Delicia city) in an area in northwestern region of Argentina.
which a dengue outbreak took place. The House index (HI) Córdoba city (31º22’ S; 64º12’ W), in the central-
was 51% and the Breteau index (BI) 106, recorded during western area of Córdoba province, is the main city and
2000 (Masuh et al. 2003). In the city of Posadas, capital of capital of the province, with a temperate climate. Annual
Misiones Province, breeding sites found in the homes were average cumulative rainfall for Córdoba city is 869.9 mm.
buckets of 10 to 20 liters, tires, and containers up to one liter The mean maximum, average, and minimum temperatures
(Tejerina et al 2009b). Pupal surveys conducted in Formosa are 24.03°, 17.28°, and 11.21° C, respectively (National
province (Clorinda city, northeastern Argentina) during Meteorological Service 2009). Entomological surveys were
the fall and spring 2007 showed that large tanks used for performed in Córdoba city during the summer months
drinking water storage were both the most abundant and (December-February) of 1997-2000 and during February
the most productive types of container (Garelli et al. 2009). 2004, 2005, and 2009, in order to determine traditional
Orán (23º08’ S; 64º20’ W) is an endemic dengue area larval indices. The infestation values during 1997-1998 were
in the northeastern region of the subtropical province of HI = 23%, CI = 16%, and BI = 47, but during 1999-2000
Salta, 270 km north of Salta, capital city of the province. the infestation values were higher (HI = 47%, CI = 24%,
Vol. 35, no. 2 Journal of Vector Ecology 279
and BI = 106). During 2004, 2005, and 2009, the HI, CI,
and BI were 33%, 7%, and 77, 29%, 13%, and 70, and 25%,
7%, and 34, respectively (Almirón and Asis 2003, Estallo et
al. 2009). Larvae and pupae of Ae. aegypti were found in a
great variety of containers, with discarded tires, buckets,
cans, and tins being those most commonly used as breeding
sites according to entomological surveys performed in six
neighborhoods of Córdoba (Almirón and Asis 2003).
Aedes aegypti used in this study were obtained from
laboratory colonies established from field immature stages
collected in the cities of San Javier, Orán, and Córdoba; mass
rearing techniques by Gerberg et al. (1994) and Domínguez
et al. (2000) were followed. The source of colony material
came from a large number of households and containers
from each city. The material collected was transferred to
Córdoba city, where the colonies from each locality were
developed simultaneously in the laboratory of the Córdoba
Entomological Research Center (National University of
Córdoba).
Larvae and pupae collected from artificial containers
were laboratory reared. Larvae were kept in 750 ml water-
filled trays and fed with liver powder (0.25 mg/larva/day).
Water surface was daily cleaned with filter paper to avoid
complications due to fungus and bacteria development.
Pupae were put in plastic flasks and then placed in
cardboard entomological cages (30 cm diameter, 50 cm
high). The emerging adults were provided with 10% sugar
solution soaked in cotton wick on 70-ml plastic flasks.
Females were offered a restrained mouse for blood meals
for two h twice a week. For oviposition, plastic flasks with
filter paper on the inside wall and dechlorinated water were
placed inside the cages. Eggs remained on moist filter paper
for at least four to five days to ensure embryogenesis, and
then were air-dried to store until the experiment started.
When there were enough eggs from each locality, they
were immersed in water for hatching. Four cohorts of 30 1st
instar larvae were established and followed simultaneously
for each subpopulation; experimental material was the
first generation progeny from individuals that were field-
collected as larvae. Each cohort was kept in a 750 ml water-
filled tray, the larvae fed with liver powder and monitored
daily, as previously described. Life statistics were estimated
by horizontal life tables from these cohorts.
Aedes aegypti mass rearing was conducted in Córdoba
City, in the laboratory of the Córdoba Entomological
Research Center (National University of Córdoba),
between November 2006 and March 2007. During this
period, daily mean temperatures ranged from 18.5° to 28°
C, with a mean value of 22.9° C (mean maximum = 24.7°
C; mean minimum = 21.1° C). The mean relative humidity
was 59.1% and the natural photoperiod was approximately
13 h of light.
The number of days spent in each preimaginal life stage
and numbers of survivors in each stage were recorded daily.
Survival was expressed as the percentage of individuals
that reached the next instar/stage. The sex ratio of emerged
adults was also estimated (Gómez et al. 1977).
Adults that emerged from each cohort were placed in a
cardboard cage (an average of ten males and eight females
emerged per cohort). The number of living and dead males
and females was recorded daily. Adults were provided
with a 10% sugar solution daily. Females were offered a
restrained mouse for blood meals twice a week, starting
two days after the first female of the cohort emerged. Once
the mouse was removed from the cage, a plastic flask with
filter paper and dechlorinated water was placed inside the
cage and replaced daily. Male and female longevity as well
as fecundity were estimated. Daily fecundity was expressed
as the number of eggs laid/female/day.
Survival data ( l x ) and daily fecundity of females ( m x )
were used to calculate mean net reproductive rate (Ro) for  
each subpopulation. Net reproductive rate was expressed
as R 0 = ∑ l x m x , the summation of mean fecundity at
age x i times the probability of survival to stage x i . R 0
expresses the per-generation growth rate of the population
as the total number of female eggs a mosquito lays during
her lifetime ( R 0 › 1.0 the population increases in size, R 0 =
1.0 no increase in population size and R 0 ‹ 1.0 population
growth is declining) (Carey 1993, Service 1993, Irvin et al.
2004).
The development time and survival in each preimaginal
stage, adult longevity, oviposition time, and fecundity were
analyzed with ANOVA followed by the Duncan’s Test to
detect significant differences and determine whether these
life statistics varied among the four subpopulations studied.
Sex ratio in each cohort was analyzed by the Student t-test
for differences of proportions (Steel and Torrie 1988). Sex
ratio among subpopulations was analyzed by ANOVA,
following arcsin data transformation. All the data were
checked for normality. Mean net reproductive rates were
estimated by jackknife analysis of lxmx life table data and
compared by using ANOVA followed by the Duncan’s test.
The jackknife method recombines the original data by
omitting one of the n replicates from the jackknife sample
and calculates pseudovalues of the parameter of interest for
each recombining of the data. These new estimates form
a set of numbers from which mean values and variances
can be calculated and compared statistically. The jackknife
method is well-suited for estimating variance for population
growth statistics (Miller 1974, Meyer et al. 1986, Shao and
Tu 1995, Krebs 1999).
RESULTS
Significant differences were found in the mean larval
development time between Córdoba (6.91 days) and Orán
(7.95 days) subpopulations (F=8.90; d.f=(3;12); P=0.0022)
(Table 1). The Córdoba subpopulation had a slightly lower
development time for instars 1+2 (2.41 days), analyzed
together, which differed significantly from the San Javier
and Orán subpopulations (F=3.86; d.f.=(3;12); P=0.0383)
(Table 1). The longest development time for the 4th instar
(3.21 days) was recorded for the Orán subpopulation
(F=5.28; d.f=(3;12); P=0.0149), differing significantly from
the San Javier subpopulation and the Rockefeller strain
(Table 1).
280 Journal of Vector Ecology December 2010
Pupal development time did not differ significantly
among the four subpopulations, ranging between 1.87 and
2.41 days (F=2.94; d.f.=(3;12); P=0.0761) (Table 1). Larvae
and pupae completed their development in approximately
9.5 days, with no significant differences detected among the
four subpopulations (F=1.71; d.f.=(3;12); P=0.2175) (Table
1).
The lowest survival was observed for the Orán
subpopulation, for the larval stage (73.3%) as well as for the
larval and pupal stages analyzed together (59.2%), differing
significantly from the San Javier subpopulation (95% and
85.8%, respectively) [F=4.94; d.f=(3;12); P=0.0185; F=7.59;
d.f=(3;12); P=0.0042] (Table 2). The survival recorded
for the pupal stage varied between 80.2 and 91.7% but
no significant differences were observed among the 4
subpopulations (F=1.67; d.f=(3;12); P=0.2268) (Table
2). Pupal survival was higher than larval survival in the
subpopulations of Córdoba, Orán, and the Rockefeller
strain.
No significant differences were detected between
the mean number of males and females that emerged in
each subpopulation (Table 3). Although no significant
differences were found in the sex ratio from 1:1, more
males were recorded in the four subpopulations (F=0.37;
d.f.=(3;12); P=0.7748) (Tables 2 and 3).
Mean male and female longevity varied from 15.7 days
(Orán) to 33.8 days (Rockefeller strain), and 35.7 days (San
Javier) to 43.8 days (Orán), respectively (Table 4). Longevity
of adults was similar among the four subpopulations (Table
4), for both females (F=0.19; d.f.=(3;12); P=0.8995) and
males (F=2.87; d.f.=(3;12); P=0.0804). Females lived longer
than males in all subpopulations. The greatest oviposition
time was recorded for females from the subpopulation
of Córdoba (31.3 days), followed in decreasing order by
females from the Orán (29.5 days) and San Javier (29.1
days) subpopulations, and the Rockefeller strain (26.74
days) (Table 4), but no significant differences were detected
(F=0.12; d.f.=(3;12); P=0.9495).
Significant differences were found in fecundity
values (F=11.48; d.f=(3;12); P=0.0008) (Figure 2), with
Figure 2. Mean of Aedes aegypti fecundity for four subpopulations studied. SJ = San Javier, CO =
Córdoba, OR = Orán, ROC = Rockefeller.
the Rockefeller strain females showing the highest daily
fecundity (10.9 eggs/female/day) and fecundity values as
well as total number of eggs laid. In decreasing order follow
the subpopulations of San Javier (2.6 eggs/female/day),
Córdoba (2.0 eggs/female/day), and Orán (1.4 eggs/female/
day) (Table 4).
Mean net reproductive rate was highest for the
Rockefeller strain ( R 0 = 134.22), differing significantly
from the San Javier ( R 0 = 27.04), and the group
formed by the Córdoba ( R 0 = 9.71) and Orán ( R 0 =
10.33) subpopulations. Differences in the demographic
growth parameter R 0 were detected among the four
subpopulations (F = 380.26; d.f. = 3;12); P=0.0001) (Table
4).
DISCUSSION
Larval development times obtained in this study
(among 6.91 and 7.95) were lower than the 9.65 days at
20° C reported in an earlier study (Bar-Zeev 1958) under
laboratory conditions. Domínguez et al. (2000) reported
that under temperate semi-natural conditions (Córdoba
city), larval development time was 8.9 days at 22.1° C.
However, under field conditions (26° C) in the temperate
city of La Plata (Buenos Aires province, Argentina), Maciá
(2006) obtained a larval development time that varied from
four to 42 days for both sexes, with a mean of 8.3 days
and 10.5 days for males and females, respectively. Under
subtropical semi-natural conditions in Misiones province, a
larval development time of 5.8 days at 25.6° C was reported
by Tejerina et al. (2009a). Temperatures lower than 20º
C, even for a few hours a day, interfere in the normal
development of larvae (Bejarano 1956).
Pupal development time obtained in this study was
lower than the three days reported by Bar-Zeev (1958) and
Domínguez et al. (2000). Tejerina et al. (2009a) also found
a pupal development time of 2.5 days in the subtropical
province of Misiones.
The differences obtained here for the larval development
time of Córdoba and Orán subpopulations may reflect
Vol. 35, no. 2 Journal of Vector Ecology 281
underlying genetic differences that probably imply local
life history differentiation. The Córdoba subpopulation had
shorter development times, probably because it is better
adapted to local environmental conditions in Córdoba city,
where the duration of favorable temperatures permitting
survival of adults and immature stages is shorter than in
subtropical cities. Isozyme analyses conducted after the
range expansion of Aedes albopictus in the United States
were interpreted as indicating rapid local differentiation
(Black et al. 1988). Rondán-Dueñas et al. (2009) reported
different haplotype composition among the different
geographical regions in Argentina. Moreover, high levels
of genetic polymorphism were detected in Argentinian Ae.
aegypti subpopulations, which suggests different origins of
different subpopulations (de Sousa et al. 2000). Biber et al.
(2006) found resistance values for cohorts from Córdoba
similar to those observed for cohorts from Yacuiba
(southern Bolivia, bordering with the NW of Argentina).
Larval and pupal survival values were close to 100% in
all subpopulations from Misiones province, including the
San Javier subpopulation (Tejerina et al. 2009a). However,
the same subpopulation reared here in Córdoba city under
laboratory conditions has shown survival values close to
86%. This probably reflects that the San Javier subpopulation
is well-adapted to local environmental conditions. Larval
survival value obtained here was higher than the 76.75%
recorded by Domínguez et al. (2000) for Ae. aegypti reared
under semi-natural conditions during summer in Córdoba
city, but pupal survival was similar (96.4%).
The differences obtained in larval-pupal survival
between Orán and San Javier subpopulations, both
from subtropical regions, probably reflect differences in
haplotype composition as a result of colonization from
different regions of South America and also an adaptation
to local conditions if appropriate selection existed.
Previous studies reported that subpopulations of NW and
NE Argentina were genetically differentiated, probably
because they have received different samples of the gene
pool of the species from Bolivia and Brazil, respectively,
where Ae. aegypti was established much earlier than in
Argentina (de Sousa et al. 2001). The current gene pool of
the subpopulations of NW Argentina may be the result of
remaining populations that survived the last eradication
campaign and of a recent colonizing event that may have
occurred in the northwestern provinces from Bolivia,
whereas the genetic similarity between the subpopulations
of NE Argentina, south Brazil, Paraguay, and Uruguay
suggest the transportation of mosquito vectors between
cities due to international travel and commerce (Rondán-
Dueñas et al. 2009).
As in this study, a sex ratio of 1:1 was also reported in
Misiones province (Tejerina et al. 2009 a) and for Ae. aegypti
adults emerging from larvitraps in the temperate province
of Buenos Aires (central-western Argentina) (Campos and
Maciá 1996, Maciá 2006). However, a subpopulation from
Córdoba city had a slight to high preponderance of females,
but only when mosquitoes were reared during the summer
months (Domínguez et al. 2000).
Female longevity varied significantly among
subpopulations from different localities of Misiones
province, appearing to be longer in subpopulations where
insecticide application is more frequent (Tejerina et al.
2009a). Females from the San Javier subpopulation (eastern
Misiones province) reared in Misiones province showed
less longevity (11.45 days) (Tejerina et al. 2009a) than the
same subpopulations reared in Córdoba city (20.46 days).
Under laboratory conditions, longevity was highly variable,
with a minimum of one day and a maximum of 76 days for
both females and males, showing the genetic plasticity of
Ae. aegypti and its potential to survive when environmental
conditions are favorable (Muir and Kay 1998, Fouque et al.
2006).
The mean number of eggs laid by Ae. aegypti females
under natural conditions (La Plata city, Buenos Aires
province) averaged 59.8 eggs/female, which were reared
as larvae at low densities (Maciá 2006). Daily fecundity
obtained in this study was lower than the mean value of 4.5
eggs/female/day for the San Javier subpopulation, reported
by Tejerina et al. (2009a).
The fecundity values reported in this study revealed
that, in subpopulations from NE and NW of Argentina,
there is a general trend that separates both subtropical
region subpopulations. The reproduction pattern observed
for Ae. aegypti populations from tropical and subtropical
regions indicates that reproduction occurs during the whole
year and their abundance may be either associated with
rainfall regimens or not (Sheppard et al. 1969, Moore et al.
1978, Chadee 1991, 1992, Barrera et al. 1997, Micieli and
Campos 2003). In temperate regions, winter temperatures
affect mosquito abundance. A discontinuous reproduction
pattern was reported in Ae. aegypti populations from Buenos
Aires province as a result of low winter temperatures, and
its abundance may be limited mainly due to temperature
(Campos and Maciá 1996, Schweigmann et al. 2002). A
similar reproduction pattern was reported for Córdoba
province, and the abundance was associated with both
temperature and rainfall regimens (Avilés et al. 1997,
Domínguez et al. 2000). In northern Argentina, however,
the reproduction of Ae. aegypti populations showed
different patterns. High temperatures allow reproduction
during the whole year in Salta and Misiones provinces
(Micieli and Campos 2003). However, for Resistencia city
(Chaco province, subtropical region) the reproduction
pattern was similar to that observed in temperate regions
(Stein et al. 2002).
Coinciding with survival and fecundity values reported
in this study, the San Javier strain had the highest mean
net reproductive rate compared with Orán and Córdoba
subpopulations (Table 4). Once again there is a trend that
separates central and northwestern from northeastern
subpopulations. Irvin et al. (2004) reported that the mean
net reproductive rate was higher for the nontransformed
laboratory Ae. aegypti Orlando strain (Ro = 87.23) when
comparing results with the other transformed mosquito
strains with Ro between 25.26 and 47.72.
Environmental factors as well as genetic composition
282 Journal of Vector Ecology December 2010

Table 1. Mean and standard deviation of Aedes aegypti larval and pupal development time for four subpopulations studied.
Four replicates were performed for each subpopulation.
San Javier Córdoba Orán Rockefeller p-value
Larva 1+2 2.72 ± 0.11a 2.41 ± 0.09b 2.71 ± 0.16a 2.46± 0.25ab 0.0383
Larva 3 2.15 ± 0.30a 1.75 ± 0.28a 2.03 ± 0.29a 2.23 ± 0.30a 0.0875
Development Larva 4 2.61 ± 0.21a 2.75± 0.24ab 3.21 ± 0.56b 2.23 ± 0.29a 0.0149
time (days) Total larva 7.48 ± 0.29a 6.91 ± 0.18b 7.95 ± 0.50a 7.02± 0.22ab 0.0022
Pupa 2.10 ± 0.33a 2.41 ± 0.22a 1.87 ± 0.29a 2.25 ± 0.33a 0.0761
Larva+Pupa 9.57 ± 0.25a 9.31 ± 0.21a 9.83 ± 0.59a 9.40 ± 0.14a 0.2175
Different letters among columns indicate significant differences (p < 0.05)..

Table 2. Mean and standard deviation of Aedes aegypti larval and pupal survival, number and sex ratio of adults that emerged
in four subpopulations studied. Four replicates were performed for each subpopulation.
San Javier Córdoba Orán Rockefeller p-value
Larva 95 ± 5.77c 88.33 ± 6.38bc 73.33 ± 13.33a 79.16 ± 6.87ab 0.0185
Immature
stage survival Pupa 90.37 ± 5.66a 91.66 ± 6.07a 80.21 ± 6.25a 90.27 ± 12.78a 0.2268
(%)
Larva+Pupa 85.83 ± 7.39c 80.83 ± 5.69bc 59.16 ± 13.44a 70.83 ± 5.01ab 0.0042

No. of adults Male 10.75 ± 3.95a 11.5 ± 3.79a 8.25 ± 2.99a 9.75 ± 1.89a 0.5459
emerged Female 9.5 ± 2.08a 7.75 ± 2.06a 7.75 ± 1.50a 7.5 ± 1.00a 0.367
Male 0.52 ± 0.16a 0.59 ± 0.15a 0.5 ± 0.1a 0.56 ± 0.8a 0.7748
Sex Ratio (%)
Female 0.48 ± 0.16a 0.41 ± 0.15a 0.5 ± 0.15a 0.44 ± 0.8a 0.7748
Different letters among columns indicate significant differences (p < 0.05).

Table 3. Differences between the mean number of males and females that emerged in each subpopulation. Four replicates
were performed for each subpopulation.
Aedes aegypti subpopulations
San Javier Córdoba Orán Rockefeller
Male emerged 10.75± 3.95a 11.50± 3.79a 8,252.99a 9.75± 1.89a
Female emerged 9.5 ± 2.08a 7.75± 2.06a 7.75± 1.50a 7.50± 1.00a
p-value 0.7007 0.2530 0.7306 0.2152
Different letters among columns indicate significant differences (p < 0.05).

Table 4. Mean and standard deviation of Aedes aegypti male and female longevity, oviposition time, daily fecundity, total
number of eggs collected and net reproductive rate (Ro) for four subpopulations studied. Four replicates were performed
for each subpopulation.
Aedes aegypti subpopulations
San Javier Córdoba Orán Rockefeller p-value
Male 20.46 ± 6.55a 22.63 ± 2.01a 15.69 ± 9.72a 33.47 ± 13.16a 0.0804
Longevity (days)
Female 35.65 ± 21.56a 43.22 ± 13.13a 43.83 ± 23.74a 42.99 ± 6.38a 0.8995
Oviposition time (days) 29.14 ± 8.57a 31.28 ± 2.93a 29.48 ± 19.47a 26.74 ± 4.80a 0.9495
Daily fecundity 2.58 ± 1.37a 2.00 ± 0.88a 1.42 ± 0.64a 10.88 ± 2.37b 0.0001
Total no. of eggs 854.5 ± 644.82a 515.75 ± 316.16a 296.00 ± 211.92a 2122.00 ± 347.08b 0.0002
Net reproductive rate 27.04b 9.71a 10.33a 134.22c 0.0001
Different letters among columns indicate significant differences (p < 0.05).
Vol. 35, no. 2 Journal of Vector Ecology 283
exert a strong influence on the response of mosquito
subpopulations of the same species under different
environmental conditions. The results of our study clearly
show differences in Ae. aegypti life statistics among the
subpopulations studied. We observed differences between
temperate and subtropical subpopulations when they were
reared in a common temperate environment, and these
differences may reflect underlying genetic differences that
probably imply adaptation to local conditions. Therefore,
it would be interesting to repeat this experiment with the
same subpopulations under subtropical conditions and see
how the Córdoba strain performs under the conditions
under which the Orán and San Javier strains normally exist.
The subpopulation from the temperate region
(Córdoba) presented a shorter development time than
subtropical region subpopulations (Orán and San Javier).
In the temperate climate of Córdoba province, this may
contribute to the vector competence of Ae. aegypti since
autochthonous dengue cases were reported for the first
time in this province in 2009. It is also important to note the
differences in larval-pupal survival reported here between
the subtropical region subpopulations, one genetically
associated with subpopulations from Bolivia and the
other with subpopulations from Brazil. The Aedes aegypti
subpopulation from Orán (northwestern Argentina)
showed the longest larval development time, the lowest
percentage of larva and pupa survival, and the lowest values
of fecundity in a temperate region. However, the vector
competence of this subpopulation in a subtropical region
is still quite good since several dengue epidemics have been
recorded in Orán.
Acknowledgments
The authors thank Iliana Martinez for her helpful
comments and suggestions that improved the quality
of this manuscript. This work was supported by grants
from FONCYT (Fondo para la Investigación Científica
y Tecnológica), CONICET (Consejo Nacional de
Investigaciones Científicas y Técnicas), SECYT (Secretaría
de Ciencia y Tecnología), and UNC (Universidad Nacional
de Córdoba). WRA is a scientific member of CONICET,
Argentina. MGG is a scholarship holder of CONICET,
Argentina.
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