Human Bloodfeeding by the Recently Introduced Mosquito, Aedes japonicus japonicus, and Public Health Implications

Abstract

Knowledge of the host-feeding behavior and extent of interactions with human hosts are important in evaluating the role and vector potential of invasive mosquitoes in transmission of native arboviruses. We collected blood-engorged females of the recently established exotic species Aedes japonicus japonicus from sites in New Jersey during 2000 to 2007 and identified the sources of vertebrate blood meals by sequencing portions of the cytochrome b gene of mitochondrial DNA. Over 1/3 (36%, n = 36) of the engorged mosquitoes acquired blood meals from humans. Other mammalian hosts included white-tailed deer (53%), fallow deer (5%), horse (3%), and Virginia opossum (3%). No avian, amphibian, reptilian, or mixed blood meals were identified. Our detection of a comparatively high prevalence of human bloodfeeding in Ae. j. japonicus in association with its local abundance, vector competence, and repeated detection of West Nile virus from field-collected specimens illustrates the potential for this invasive mosquito to serve as a "bridge" vector in transmission of West Nile and other mosquito-borne viruses in North America.

Full text

SCIENTIFIC NOTE
HUMAN BLOODFEEDING BY THE RECENTLY INTRODUCED
MOSQUITO, AEDES JAPONICUS JAPONICUS, AND PUBLIC
HEALTH IMPLICATIONS
GOUDARZ MOLAEI,
1
ARY FARAJOLLAHI,
2,3
JAMESINA J. SCOTT,
4
RANDY GAUGLER,
2
AND
THEODORE G. ANDREADIS
1
ABSTRACT. Knowledge of the host-feeding behavior and extent of interactions with human hosts are
important in evaluating the role and vector potential of invasive mosquitoes in transmission of native
arboviruses. We collected blood-engorged females of the recently established exotic species Aedes japonicus
japonicus from sites in New Jersey during 2000 to 2007 and identified the sources of vertebrate blood meals
by sequencing portions of the cytochrome bgene of mitochondrial DNA. Over 1/3 (36%,n536) of the
engorged mosquitoes acquired blood meals from humans. Other mammalian hosts included white-tailed deer
(53%), fallow deer (5%), horse (3%), and Virginia opossum (3%). No avian, amphibian, reptilian, or mixed
blood meals were identified. Our detection of a comparatively high prevalence of human bloodfeeding in Ae.
j. japonicus in association with its local abundance, vector competence, and repeated detection of West Nile
virus from field-collected specimens illustrates the potential for this invasive mosquito to serve as a ‘‘bridge’’
vector in transmission of West Nile and other mosquito-borne viruses in North America.
KEY WORDS Aedes japonicus japonicus, bloodfeeding behavior, West Nile virus, arboviruses
The global spread of exotic mosquito species as
nuisance pests and potential vectors of diseases
poses profound impacts on human health and
creates serious challenges for public health and
mosquito control agencies. Aedes japonicus japo-
nicus (Theobald), an invasive container habitat
species native to Japan, Korea, and Eastern
China (Tanaka et al. 1979), was first discovered
in the northeastern USA in 1998 (Peyton et al.
1999) and has rapidly spread throughout much of
eastern North America and southern Canada
(Andreadis et al. 2001, Fonseca et al. 2001,
Sardelis and Turell 2001, Falco et al. 2002,
Harrison et al. 2002, Oliver et al. 2003, Joy
2004, Reeves and Korecki 2004, Roppo et al.
2004, Young et al. 2004, Caldwell et al. 2005,
Gallitano et al. 2005, Gray et al. 2005, Joy and
Sullivan 2005, Larish and Savage 2005, Sames
and Pehling 2005, Holman et al. 2006, Saenz et al.
2006, Thielman and Hunter 2006, Bevins 2007,
Morris et al. 2007, Hughes et al. 2008).
Laboratory studies have shown that Ae. j.
japonicus is a highly efficient vector of West Nile
(WN) and St. Louis encephalitis viruses and a
moderately efficient vector of eastern equine
encephalitis and La Crosse viruses (Sardelis and
Turell 2001; Sardelis et al. 2002a, 2002b; Sardelis
et al. 2003). Furthermore, WN virus has been
detected in field-collected Ae. j. japonicus from at
least 9 different states in the northeastern and
north central USA (New Hampshire, Massachu-
setts, Rhode Island, Connecticut, New York,
New Jersey, Pennsylvania, Ohio, and Indiana)
(CDC 2009), implicating this species as a
‘‘bridge’’ vector to humans.
Studies on the host-feeding behavior of Ae. j.
japonicus are needed to define its feeding prefer-
ences in areas where it may serve as a potential
vector of WN and other mosquito-borne viruses.
Laboratory studies with native Asian populations
indicate that this species readily feeds on chickens
and mice but not on reptiles or amphibians
(Miyagi 1972, Tanaka et al. 1979). However,
blood meal analyses of field-collected mosquitoes
in New York (Apperson et al. 2004) and
Connecticut (Molaei et al. 2008) indicate that
Ae. j. japonicus acquire blood meals exclusively
from mammalian hosts, including humans. The
present study examined the host-feeding habits of
Ae. j. japonicus to extend knowledge of the
behavioral ecology of this mosquito in other
areas of northeastern USA and to further
elucidate its role and vector potential in arbovirus
transmission.
Bloodfed mosquitoes were collected from a
variety of localities in rural and suburban New
Jersey including farms, wood lots, waste tire and
automobile disposal sites, residential backyards,
and an animal park during 2000 to 2007 using
New Jersey light trap collections (JW Hock
Company, Gainesville, FL), grass infusion–baited
1
The Connecticut Agricultural Experiment Station,
123 Huntington Street, P.O. Box 1106, New Haven, CT
06511.
2
Center for Vector Biology, Rutgers University, 180
Jones Avenue, New Brunswick, NJ 08901.
3
Mercer County Mosquito Control, 300 Scotch
Road, West Trenton, NJ 08628.
4
Lake County Vector Control District, P.O. Box 310,
410 Esplanade, Lakeport, CA 95453.
Journal of the American Mosquito Control Association, 25(2):210–214, 2009
Copyright
E
2009 by The American Mosquito Control Association, Inc.
210

gravid traps (BioQuip Products, Inc., Rancho
Dominguez, CA), carbon dioxide–baited Centers
for Disease Control and Prevention (CDC) and
encephalitis virus surveillance light traps (Bio-
Quip), and aspiration from resting sites by using a
backpack aspirator (BioQuip). Engorged mos-
quitoes were identified to species level on a chill
table with the aid of a stereomicroscope using
descriptive keys (Tanaka et al. 1979). Individual
mosquitoes were dissected on microscope slides
using flame-sterilized scalpels and forceps or
disposable razor blades, and DNA was isolated
from the abdominal contents of engorged mos-
quitoes using DNA-zol BD (Molecular Research
Center, Cincinnati, OH) according to the manu-
facturer’s recommendation with some modifica-
tions as described elsewhere (Scott 2003; Molaei
et al. 2006, 2008). Isolated DNA from the
mosquito blood meals served as DNA templates
in subsequent polymerase chain reaction (PCR)
with primers based on cytochrome bsequences of
avian and mammalian species using previously
described thermal-cycling conditions (Scott 2003;
Molaei et al. 2006, 2008). The GeneAmp PCR
system 9700 (Applied Biosystems, Foster City,
CA) was used to perform PCR reactions and
sequenced directly in cycle sequencing reactions
using the sequencer, 3730xl DNA analyzer
(Applied Biosystems) at the Keck Sequencing
Facility, Yale University, New Haven, CT, or
using ABI PrismH3100 automated capillary
genetic analyzer (Applied Biosystems) at the
Biotechnology Center for Agriculture and the
Environment, Rutgers University. Sequences
were analyzed and annotated by using Chroma-
sPro version 1.22 or 2.2.6 (Technelysium Pty
Ltd., Tewantin, Australia) and identified by
comparison to the GenBank DNA sequence
database (NCBI 2008).
Analysis of the vertebrate blood meal sources
for Ae. j. japonicus identified 5 mammalian hosts
(Table 1). White-tailed deer, Odocoileus virginia-
nus (Zimmermann), was the most frequently
identified host (52.8%of total), followed by
human, Homo sapiens L. (36.1%). Specimens
with human-derived blood meals were collected
by aspiration (92.3%,n512) and a gravid trap
(7.7%), from woodlots (69.2%), waste tire dis-
posal sites (15.4%), and residential backyards
(15.4%). Other mammalian species identified
included horse, Equus caballus (L.), and Virginia
opossum, Didelphis virginiana (Kerr). Two mos-
quitoes collected within a captive wildlife facility
had acquired blood meals from fallow deer,
Dama dama L. This small facility (ca. 5 acres)
houses a group of 15 fallow deer that are allowed
to roam freely within the enclosure. No avian,
amphibian, reptilian, or mixed blood meals were
identified.
Examination of the host-feeding behavior of
mosquitoes is vital to understanding their vecto-
rial capacity in a new distribution range, partic-
ularly in instances where humans may be the
preferred host and the mosquito species are
capable of transmitting viruses that circulate
annually and cause human disease. Our blood
meal analysis of Ae. j. japonicus identified
exclusively mammalian-derived blood meals with
a high prevalence of human feeding that was
notably greater than that reported for other
mammalophilic mosquito species in the north-
eastern USA (Apperson et al. 2002, 2004; Molaei
et al. 2009). This finding, in concert with the
detection of WN virus from field-collected
females throughout New Jersey in 8 of the last
9 years (n524 WN virus positive pools) (CDC
2008), supports a likely bridge vector role for this
species in transmission of WN virus to humans in
the region that has not been previously recog-
nized. Our results also corroborate the reported
attraction of Ae. j. japonicus to human bait
stations and collection of indoor biting females in
suburban and rural environs in Connecticut and
New Jersey (Andreadis et al. 2001, Scott 2003).
Our results with populations in New Jersey are
further consistent with the report of exclusive
mammalian-derived blood meals from Ae. j.
japonicus in New York and Connecticut, where
98%and 67%of mammalian-derived blood meals
were from white-tailed deer, respectively (Apper-
son et al. 2004, Molaei et al. 2008). The
prevalence of white-tailed deer as hosts for Ae.
j. japonicus and other mammalophilic mosquitoes
in the present and the most recent studies is likely
Table 1. Number and percentage of mammalian-derived blood meals identified from Aedes japonicus japonicus
collected in New Jersey, 2000–07.
Vertebrate host species No. %total
Trap type
GT
1
NJ LT CO
2
LT ASP
White-tailed deer, Odocoileus virginianus 19 52.8 8 3 3 5
Human, Homo sapiens 13 36.1 1 — — 12
Fallow deer, Dama dama 2 5.5 — 2 — —
Horse, Equus caballus 1 2.8 1 — — —
Virginia opossum, Didelphis virginiana 1 2.8 — — 1 —
Total 36 100.0 10 5 4 17
1
GT, gravid trap; NJ LT, New Jersey light trap; CO
2
LT, CO
2
-baited light trap; ASP, aspirator.
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a function of deer abundance in the northeastern
USA (Molaei et al. 2006, 2008).
The absence of avian-derived blood meals from
Ae. j. japonicus in this and other studies from the
region is enigmatic. Laboratory observations
indicate that Ae. j. japonicus readily feeds on
birds in addition to mammalian species (Miyagi
1972, Scott 2003), and vector competency studies
in the USA use chickens as the blood meal source
for this mosquito (Sardelis and Turell 2001;
Sardelis et al. 2002a, 2002b; Sardelis et al.
2003). Furthermore, a free mating colony of Ae.
j. japonicus established since 2000 at Rutgers
University has been exclusively maintained by
feeding on bobwhite quail, Colinus virginianus
(L.) (Williges et al. 2008). The absence of avian-
derived blood meals from field-collected mosqui-
toes could be due to the relatively small numbers
and inherent experimental bias in collecting
engorged mosquitoes. Samples analyzed in our
study were collected at ground level, which may
not harbor a large number of Ae. j. japonicus that
feed on birds in canopies and rest near hosts
(Scott 2003). Efforts in collecting this mosquito
species in the canopy in New Jersey have been
unsuccessful; however, Andreadis and Armstrong
(2007) reported the collection of a relatively small
number (16.1%,n5535) of Ae. j. japonicus in
CO
2
-baited CDC light traps in tree canopies
(7.6 m) when compared with light traps placed at
ground level (1.5 m) in Connecticut.
Lack of avian feeding is also problematic with
regard to the acquisition of WN virus by this
species, since birds are viewed as the principal
reservoir and amplifying hosts. Serological evidence
of WN virus infection has been noted in white-
tailed deer populations in New Jersey (Farajollahi
et al. 2004) and Iowa (Santaella et al. 2005);
however, it is not known whether they develop a
sufficient viremia to infect mosquitoes and contrib-
ute to local transmission cycles. Alternatively, WN
virus could be acquired from other mammals, such
as eastern chipmunk, Tamias striatus (L.) (Platt et
al. 2007), and fox squirrels, Sciurus niger L. (Root
et al. 2006, Platt et al. 2008), for example, which
have been shown to develop serum titers sufficient
to infect mosquitoes in the laboratory.
Successful establishment, rapid range expan-
sion, abundance, repeated isolation of WN virus
in nature, and vector competency at rates
comparable to other susceptible bridge vectors
in concert with pronounced bloodfeedings on
human hosts are consistent with the view that Ae.
j. japonicus likely plays a role as a bridge vector in
transmission of this virus and conceivably a
number of other mosquito-borne viruses to
humans and other mammals in northeastern
and other regions of the USA.
We are grateful to Wayne Crans for his insights
and encouragement and Michael Thomas, John
Shepard, and Melanie J. Raubeson for technical
assistance. We thank Linda McCuiston for
identification assistance and pooling of mosquito
specimens, along with Maureen Musarra, Thom-
as L. Scott, Jennifer Gruener, and John Phelps.
We also thank the superintendents and staff of
Bergen County Mosquito Control (Peter Plu-
chino, Jimmy Bartlett, and Warren Staudinger),
Burlington County Mosquito Control (Dominic
Chappine and Tom Verna), Cumberland County
Mosquito Control (Heather Lomberk and Doug
Abdil), Sussex County Division of Mosquito
Control (John Holick and Marta Iwaseczko),
and the Warren County Mosquito Extermination
Commission (Christine Musa, Abie Musa, Bob
Duryea, Sara May, Teresa Duckworth, Katy
Parise, Heather Buckley, and Veronica Ronnie
Galbraith), and Jessie Sebbo for assistance in
collecting specimens. Funding for this research
was provided in part by Laboratory Capacity for
Infectious Diseases Cooperative Agreement
Number U50/CCU6806-01-1 from the Centers
for Disease Control and Prevention, US Depart-
ment of Agriculture (USDA) Specific Coopera-
tive Agreement Number 58-6615-1-218, USDA-
administered Hatch funds CONH00768 to the
Connecticut Agricultural Experiment Station.
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