2013 NORTHEASTERN NATURALIST 20(2):363–371
Color and Container Size Affect Mosquito
(Aedes triseriatus) Oviposition
Gary Joseph Torrisi1,2,* and W. Wyatt Hoback3
Abstract - This investigation compared color and artificial container size as attractants
for the gravid female Aedes triseriatus (Eastern Tree-hole Mosquito) oviposition site
selection. Three colors (white, green, and black) were investigated in combination with
two container sizes (3672 ml and 12,000 ml). Presence of mosquito larvae was used to determine
percent occupancy of containers. Black containers were selected in 70% of trials,
compared to 63% for green containers and 41% for white containers. Large containers
were selected more frequently (72% of trials) than small containers (44% of trials). Results
suggest that container size may serve as an important selective cue for A. triseriatus
and may take precedence over color when gravid female mosquitoes are given the choice.
Our results provide further insight into the complexity of cues that influence mosquito
Natural tree-hole habitats serve as ovipositional sites for gravid female mosquitoes.
These habitats are common in the tropics and are also found in temperate
climates although these habitats are not as varied or abundant as those found in
the tropics (Kitching 1971). Artificial tree-holes created using plastic containers
infused with leaf litter and rainwater attract many of the same mosquito species
found in natural tree-holes including Aedes triseriatus (Say) (Eastern Tree-hole
Mosquito), which we examine in this paper (Yanoviak 2001).
Extensive research suggests that color, primarily dark, light-absorptive
backgrounds of black or brown that are similar to the linings of natural treehole
cavities serve as the main attractant for oviposition site selection for most
mosquitoes (Collins and Blackwell 2000, Frank 1986, Jones and Schreiber
1994, Yanoviak 2001). There are some exceptions where lighter-colored tropical
bromeliads are attractants to ovipositing Wyeomyia mitchelli (Theobald) and
Wyeomyia vanduzeei Dyar and Knab. In addition, pitcher plants found in sphagnum
bogs associated with temperate forests are attractive to Wyeomia smithii
Coquillett (Pitcher Plant Mosquito). Frank (1985) demonstrated that Pitcher
Plant Mosquito response to color differs dramatically from other mosquitoes in
their selection of light-colored oviposition sites.
Other cues in addition to color are used by gravid female mosquitoes in
selecting sites for the deposition of eggs. Past studies have tested variables
including: location of tree holes in illuminated gaps versus darker understory
¹Department of Entomology, University of Nebraska at Lincoln, Lincoln, NE 68508.
2Current address - 6 Jennifer Court, Saratoga Springs, NY 12866. 3Department of Biology,
Bruner Hall of Science, 905 West 25th Street, University of Nebraska at Kearney,
Kearney, NE 68849. *Corresponding author - email@example.com.
364 Northeastern Naturalist Vol. 20, No. 2
cover (Yanoviak 2001), vertical distribution of tree holes (Jones and Schreiber
1994), form and color of containers (Frank 1986), leaf-litter quality and
stability of habitats (Minakawa et al. 2005, Yanoviak 1999), light intensity
(Rodriquez-Tovar et al. 2000), and the role of conspecifics as attractants and
interspecific competitors as deterrents (Braks et al. 2004, Onyabe and Roitberg
1997, Yee et al. 2004). In addition, investigations have examined predation
(Etiam and Blaustein 2004), the influence of preexisting eggs (Allan and Kline
1998), precipitation and temperature (Alto and Juliano 2001), size and surface
area of habitats (Lester and Pike 2003, Minakawa et al. 2005) and crowding,
confinement, starvation, and infection (Zahiri and Rau 1998). These studies
point to the variety of variables that influence mosquito oviposition behavior.
Approximately seventy recognized mosquito species reside in New York
State including many species that serve as vectors of disease (NYSHD 2012).
Examples of emergence of vector-borne disease include the 1999 outbreak of
West Nile virus (WNV) in New York City leading to a subsequent epidemic
throughout the United States (Hayes and Gubler 2006). Furthermore, epidemiological
data from the Center for Disease Control (CDC) report the emergence
of dengue fever (CDC 2010a) in the Americas since the 1950s and significant
under reporting and under diagnosis of LaCrosse encephalitis (CDC 2010b)
especially in the southeastern United States. In addition, there has been an increase
in the incidence of eastern equine encephalitis (EEE) and WNV in horses
in the northeastern United States (Kurt Lutgens, Doctor of Veterinary Medicine,
Saratoga Springs, NY, pers. comm.).
Discovery of mosquito larvae in large, white vessels exposed to direct sunlight
(G.J. Torrisi, pers. observ.) seemed to be at odds with published results on
mosquito habitat selection. Based on these observations, we tested whether the
combination of color and container size serve as attractants for gravid mosquitoes.
We hypothesized that if both color and container size are cues for oviposition site
selection by gravid female mosquitoes, then large containers would be preferred
regardless of color.
Field Site Description
This investigation took place in upstate New York, in the town of Wilton
(43°18'N, 73°74'W), in a suburban housing development surrounded by parcels
of patchy temperate forest. The dominant trees of the area are Quercus rubra
L. (Red Oak), Acer rubrum L. (Red Maple), and Pinus strobes L. (White Pine).
This investigation was conducted from mid-June to mid-August in 2007, 2010,
Decaying leaf litter of oak and maple was removed from the forest floor and
cut into 1–2 cm2 sections. An equal volume of fresh leaves was stripped from
the branches of the dominant species of trees and cut into same-sized sections
and mixed with the decaying litter. The leaf litter was then covered in rainwater
2013 G.J. Torrisi and W.W. Hoback 365
collected on site and the mixture was allowed to stand for three days to promote
algal and microinvertebrate colony formations that would initially serve as the
basic nutrients for mosquito larvae.
Nine small, plastic, cylindrical containers measuring 12 x 17 x 18 cm and
nine large, plastic, cylindrical containers measuring 20 x 24 x 25 cm served as
replicates of artificial habitats. Three small and three large containers were left
white or painted green or black (Rust-Oleum® hunter green and flat black) for six
container color by size combinations per replication. Eighteen containers, three
replications, were used in each of three trials per year (n = 54/year). Thus, across
all three years, each container color and size combination was made available 27
times for ovipositing mosquitoes. Because the time of the year affected abiotic
factors including temperature and rainfall and because weather patterns across
years were different and likely to affect mosquito activity, percent occupancy of
container type was calculated by year (n = 3).
Holes were drilled into the sides of the containers half way up from the bottom
to prevent weight overload and the potential for detachment of the containers
from trees caused by heavy rainstorms or high winds. A water-repellant nylon
cord was used to attach each container by its handle to a tree at a height of 1 m
above the forest floor. Containers were placed no closer than 5 m and no further
than 10 m from one another. Containers were affixed to oak and maple trees with
a diameter greater than 20 cm. The leaf-litter mixture was placed in the bottom of
the containers covering one-half of the bottom surface area. Additional rainwater
was then added to the level of the pre-drilled holes. When rainwater was unavailable,
untreated water from a well point was substituted.
We inspected containers every 3–5 days to insure proper attachment of the
artificial containers, to replace water loss as needed, and to assess whether earlystage
mosquito larvae were present. Detection of any mosquito larvae resulted in
the container being designated as selected by a gravid female mosquito, whereas
absence of larvae resulted in the container being designated as not selected. The
number of larvae was not quantified or compared.
Detection of early-stage larvae, in any one container, initiated 21 consecutive
days of observation, record keeping, and sampling of larvae. All containers were
monitored during the 21-day period. A flashlight was used to search the bottom
of the darker containers for wrigglers. Individual late-stage larvae were removed
during each trial to identify species. Larvae were transferred to a 10-ml vial,
labeled with date, container color, and container size. Captures were then stored
in water and refrigerated. Samples were examined under magnification (400x) to
identify species. In addition, samples of late-stage larvae were reared to adult and
identified. At the conclusion of each investigation, all containers were emptied,
cleaned, and reset with leaf litter and water in the manner described above.
All collected larvae were identified as Aedes triseriatus. Across all trials and
years, 94 containers out of a possible 162 (58%) were selected by gravid females.
366 Northeastern Naturalist Vol. 20, No. 2
Percent occupancy by container size was compared using a Student’s t-test
(n = 9). Small containers were selected 44% of the time, while large containers
were selected 72% of the time (Fig. 1), which was significantly more often (twotailed,
unpaired t-test: t = 3.5, df = 8, P = 0.008). Percent occupancy by container
color was compared using an analysis of variance (n = 18). Black containers were
utilized significantly more frequently (Fig. 1) than white containers, while other
comparisons were not significant (ANOVA: F =3.8, df = 17, P = 0.047). Across
all three years (n = 3),
mosquitoes oviposited most frequently in the large green
(88% occupancy) and large black (78% occupancy) containers (Fig. 2). Small
white and small green containers were occupied least frequently (33% and 37%,
respectively). Of the 15 possible comparisons of occupancy between container
color-size combinations (Table 1), 9 comparisons were significantly different
(ANOVA: F = 18.3, df = 17, P less than 0.001).
Container size plays a significant role within the mix of complex behaviors
associated with mosquito oviposition site selection. Our finding that darker
colors influence oviposition by Eastern Tree-hole Mosquitoes is similar to the
findings of Collins and Blackwell (2000). Finding mosquito larvae in larger
containers suggests that container size also plays a role in female mosquito oviposition
Successful oviposition in a range of container sizes and colors does not
mean that all larvae would be successful. During this study, observation of late
Figure 1. Mean (± 1 S.E.) percent occupancy of Aedes triseriatus (Eastern Tree-hole
Mosquito) larvae by size and color of artificial containers over a three-year period.
2013 G.J. Torrisi and W.W. Hoback 367
instars and pupae were limited because of the short duration of each trial. Once
the presence of larvae was observed, the trials were terminated within 3 weeks.
However, we continued to rear late-stage larvae to adult stage in the laboratory
Figure 2. Mean (± 1 S.E.) percent occupancy of Aedes triseriatus (Eastern Tree-hole
Mosquito) larvae by color and size of artificial containers over a three-year period. S =
small, L = large, w = white, g = green, and b = black.
Table 1. Differences in percent of containers occupied between color-size combinations. P-values
are based on Tukey-Kramer multiple comparison test. Container size: S = small, L = large; container
color: w = white, g = green, b = black
Comparison Difference of means P-value
Lg - Sw 0.556 less than 0.001
Lg - Sg 0.519 less than 0.001
Lg - Lw 0.407 0.002
Lg - Sb 0.259 0.040
Lg - Lb 0.111 0.671
Lb - Sw 0.444 less than 0.001
Lb - Sg 0.407 0.002
Lb - Lw 0.296 0.017
Lb - Sb 0.148 0.396
Sb - Sw 0.296 0.017
Sb - Sg 0.259 0.040
Sb - Lw 0.148 0.396
Lw - Sw 0.148 0.396
Lw - Sg 0.111 0.671
Sg - Sw 0.037 0.995
368 Northeastern Naturalist Vol. 20, No. 2
for purposes of identification. In the field, on occasion, adult mosquitoes were
observed visiting the containers. This observation suggests that female mosquitoes
may have been appraising the available cues for oviposition selection (Lester
and Pike 2003). Although initially these artificial habitats were sufficient to cause
oviposition by gravid females, factors that contribute to larval development may
have been lacking. It follows that longer trial durations might have led to all
containers being selected after the more preferred large, dark containers were
colonized. Further investigation over a longer time span would provide insight
into the effects of congeners on ovipositing mosquitoes where ample containers
are available for colonization.
Past investigations suggest that deterrents to oviposition may include larval
predators, nutrient quality, competition for nutrients, presence of conspecific
eggs or larvae, competition from an alternative species having colonized the containers,
or confinement of movement for existing larvae (Allan and Kline 1998,
Braks et al. 2004, Etiam and Blaustein 2004, Jones and Schrieber 1994, Onyabe
and Rottberg 1997, Yanoviak 1999, Zahiri and Rau 1998). Many of these factors
may serve as barriers in the process of oviposition selection by gravid females.
However, no predators or evidence of larval predation were observed during this
investigation. As reported by Etiam and Blaustein (2004), there is a noticeable
response of prey to risk of predation including behavioral changes such as reduced
oviposition. This response would presume the existence of predators prior
to selection and subsequent rejection of a given container because of detection of
predator cues. However, low predator density during the onset of selection may
not be detectable by the ovipositing mosquitoes (Eitam and Blaustein 2004). This
scenario seems more likely because of the large number of containers selected
for egg deposition in this investigation. In addition, samples from each trial indicated
the same species was present in all containers and there was no evidence
of an alternate mosquito species being present in any of the containers. In one
sampling, we observed early instars along with late instars of Eastern Tree-hole
Mosquitoes in the same container.
Gravid female mosquitoes should select containers, artificial or natural, that
provide for the success of their offspring. However, previous studies have shown
that the size of the container does not necessarily correlate with successful development
of mosquito larvae. For example, predators of mosquitoes are less likely
to be found in small containers that serve as temporary habitats that are under
the threat of drying out. Larger containers serve as a more permanent habitat and
are more likely to contain mosquito predators that would reduce the abundance
of the population within the container (Service 1977, Sunahara et al. 2002).
Consequently, when selecting a suitable habitat, smaller containers are subject
to habitat instability in terms of the loss of water, where large containers hold
greater volumes of water and therefore provide a more stable habitat for female
mosquitoes but may also be subject to greater predation risk (Minakawa et al.
2005). Lester and Pike (2003) concluded that container size alone may influence
female mosquito oviposition site selection regardless of predator occupation of
2013 G.J. Torrisi and W.W. Hoback 369
Elevation and container location might also influence which species selected
the experimental containers. Placement of containers at 1 m above the forest floor
may have attracted only the Eastern Tree-hole Mosquito. Future consideration
must be given to ground-flying species and species that prefer tree-holes above
1 m from the ground.
During our study, some containers had exposure to greater periods of sunlight
because of their location closer to tree-fall gaps, while the remaining
containers were located in areas with less light under the forest understory. This
difference may have accounted for the lack of oviposition in some containers.
In support of this possibility, Yanoviak (2001) reported that artificial tree-holes
located in the understory attract more mosquitoes than artificial containers located
in tree-fall gaps.
During our experimentation, habitat stability was maintained by providing
minimum water levels throughout the trials. Higher summer temperatures deplete
water levels in containers that in turn lead to shorter development times
for mosquitoes, especially when ample food is present. In this investigation,
high summer temperatures may have resulted in fewer small containers being
colonized because of the level of water falling below stable maintenance levels
while the larger containers held adequate levels of water more consistently. In
addition, higher water temperatures hinder container selection by female mosquitoes
(Alto and Juliano 2001), perhaps forcing the female mosquitoes to continuing
searching for a more suitable container.
Container size alone does not dictate site selection. Other cues play a role in
whether a gravid female will select a particular container. Factors include leaflitter
quality, speed of decay, developmental time, water volume, lower water
temperatures, leaf-fall diversity, and nutritional yield (Barrera et al. 2006, Bently
and Day 1989, Reiskind et al. 2009, Yanoviak 1999). These factors become
influential not only in site selection but also in larval maturation to adulthood
and may actually limit oviposition even in the larger containers (Yanoviak
1999). Development may also be influenced by the effects of crowding, limiting
the mobility of the larvae and reducing container selection by subsequent
females (Yee et al. 2004).
Investigating artificial containers in and around suburban homes and patchy
forest surroundings revealed many large containers available to ovipositing mosquitoes.
Large urns, flowerpots, birdbaths, fountains, and unattended swimming
pools can all hold sufficient water and provide preferred habitats for mosquitoes.
Once these containers are established as a breeding site, conspecific eggs then
serve as additional attractant cues for gravid mosquitoes (Allan and Kline 1998,
Onyabe and Roitberg 1998). Our results suggest that large container size may
be more important than color for oviposition by gravid Eastern Tree-hole Mosquitoes
in a temperate forest setting. Results of this investigation may add to the
knowledge and applications of mosquito-control management and expand our
understanding of mosquito behavior.
370 Northeastern Naturalist Vol. 20, No. 2
The authors extend their gratitude to Apple Pools, Inc. for their initial financial support
in providing the containers and paint used in this investigation. We thank P. Torrisi
for her involvement in the proof reading and comments of the initial draft.
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