Phylogenetic study of Aedes albopictus and Aedes koreicus (Diptera, Culicidae) origin, two invasive mosquito species in Belgium

1
Phylogenetic study of Aedes albopictus and Aedes
koreicus (Diptera, Culicidae) origin, two
invasive mosquito species in Belgium
Raharimalala F.N.1 , 2, Boukraa S.1 , Bawin T.1,
Zimmer J.-Y.1, Francis F.1 1Functional and
Evolutionary Entomology University of Liege
(GxABT) Belgium 2Institut Pasteur of
Madagascar Email entomologie.gembloux_at_ulg.ac.be
Introduction In Belgium, presence of the three
invasive species of Aedes were mentioned until
2009 Aedes (Stegomya) albopictus (Skuse, 1894),
Aedes japonicus japonicus (Theobald, 1901) and
Aedes koreicus (Edwards, 1917) (Versteir et al.,
2009, 2012). Ae. albopictus was detected for the
first time in 2000 for Belgium (Schaffner et al.,
2004). Then, it was absent during 12 years until
investigator field in July 2013. One male has
been caught in Oost-Vlaanderen Province, in his
older site (Boukraa et al., 2013). Larvae of Ae.
koreicus were also caught in abundance in
Maasmechelen village. Despite an intensive
investigation, Ae. japonicus was not seen.
Investigation study of the probable origin of
these two species with their phylogenetics was
conducted. Nested PCR with three mitochondrial
loci were used cytochrome-oxydase subunit 1
nicotinamide adenine dinucleotide dehydrogenase
subunit 4 and nicotinamide adenine dinucleotide
dehydrogenase subunit 5 for evaluation of the
relationships in the phylogenetics origin of
these two species.
Material and methods
Types of breeding sites for larvae collections
The CO2-baited traps Mosquito Magnet Liberty
Plus (MMLP) for adults captures
DNA extracted Nested PCR with specifics primers
Results
Concatenation of COI-ND5 (919bp) gives a profile
very closed to Aedes albopictus USA
Percentage bootstrap supports (1000 replicates)
superior to 50 are given at each node.
Branch lengths represent estimated substitutions
per site.
Figure 2- Concatenated COI-ND4 genes of Aedes
koreicus compared with Aedes japonicus sp
sequences by the maximum likelihood method based
on the Jukes-Cantor model .
Percentage bootstrap supports (1000 replicates)
superior to 50 are given at each node. Branch
lengths represent estimated substitutions per
site. Aedes notoscriptus (KF034773.1) was used as
out-group K1-K4 Aedes koreicus in July 2013 for
Belgium.
Aedes aegypti (AF390098.2) was used as out-group
Recovered of monophyletic grouping of Aedes
japonicus sp and Aedes koreicus.
Figure 1- Concatenated COI-ND5 genes of Aedes
albopictus by the maximum likelihood method based
on the Jukes-Cantor model .
Discussion
This study suggests that the probable origin of
Ae. abopictus could be the United States.
Phylogeny result confirms the investigation
conducted from the proprietor of the firm and
allowed us to suggest that the probable
introduction way for Ae. albopictus in Belgium
was the importation of used tires via USA. For
Ae. koreicus, data deposited in GenBank about
this invasive specie was very poor until today
and has not allowed us to study its origin from
the phylogenetic study. Nevertheless, research
allowed us to confirm the theory of the closed
relatives of Aedes japonicus sp and Aedes
koreicus.
Conclusions
Discovery of these two invasive species suggests
a significant risk of their establishment in
Belgium and requires serious prophylactic
measures. Knowledge of their origin is a crucial
key element in the management of their invasion -
development and should allow better tag actions
that must be taken. Undeniably, these data will
be explored (epidemiology, epidemic) and the
development of a monitoring in the probable
entrance doors.
Key words Culicidae, Aedes, invasive,
phylogenetic study, Belgium
References Schaffner F, Van Bortel W, Coosemans
M First record of Aedes (Stegomyia) albopictus
in Belgium. J Am Mosq Control Assoc 2004,
20201-203. Boukraa S, Raharimalala FN, Zimmer
JY, Schaffner F, Bawin T, Haubruge E, Francis F
Reintroduction of the invasive mosquito species
Aedes albopictus in Belgium in July 2013.
Parasite 2013, 2054. Versteirt V, Schaffner F,
Garros C, Dekoninck W, Coosemans M, Van Bortel W
Introduction and establishment of the exotic
mosquito species Aedes japonicus japonicus
(Diptera Culicidae) in Belgium. J Med Entomol
2009, 461464-1467. Versteirt V, De Clercq EM,
Fonseca DM, Pecor J, Schaffner F, Coosemans M,
Van Bortel W Bionomics of the established exotic
mosquito species Aedes koreicus in Belgium,
Europe. J Med Entomol 2012, 491226-1232.    
This work was supported by Subside Federal for
Research (grant R.DIVE.05558-J-F), University of
Liege (ULg) to FNR.