Medical and Veterinary Entomology (2018) 32, 145–154 doi: 10.1111/mve.

12272

EDITOR’S CHOICE

A report on the mosquitoes of mainland Åland,

southwestern Finland and revised list of Finnish
mosquitoes
C. L. C U L V E R W E L L 1,2
1
Medicum, Department of Virology, University of Helsinki, Helsinki, Finland and 2 The Natural History Museum, London, U.K.

Abstract. To successfully implement surveillance or control strategies for mosquitoes,

up-to-date knowledge of regional species composition is vital. The last report regarding
mosquitoes (Diptera: Culicidae) in the Åland archipelago, southwestern Finland listed
19 species (Utrio, 1979). To determine the current species diversity, one collection
trip was made to mainland Åland in 2015 and three in 2016. Mosquitoes (n = 3286)
were collected as both adult and immature life stages from 88 collections within 29
1-km2 areas. Fifteen of the 19 previously reported species were obtained, leaving the
current status of four species uncertain. At least 11 species previously not reported from
Åland, but confirmed on the Finnish mainland, were collected. Aedes geminus Peus
was identified based on examination of the gonostylus, and represents a new species
distribution for Finland. Anopheles maculipennis s.s. Meigen was confirmed from
cytochrome c oxidase subunit I (COI) and internal transcribed spacer 2 (ITS2) sequence
data and is reinstated on the list of Finnish species, along with Ochlerotatus sticticus
(Meigen). Dahliana geniculata (Olivier) was found in two locations, in 2 months,
indicating that there is an established population in Åland. The present data confirm
that at least 27 species inhabit mainland Åland, rising to 31 when historical data are
included. The Finnish mosquito fauna is increased from 38 to 41 species.
Key words. Aedes geminus, Anopheles maculipennis, Dahliana geniculata, Ochlerota-
tus sticticus, Mosquitoes, Åland, Finland.

Introduction is potential for more southerly species to become established

on the Åland Islands that are not yet established on the Finnish
The most fundamental knowledge in mosquito biology is the mainland. From a surveillance point of view, any differences in
species composition and the distribution of species in a given species composition that occur between the Finnish mainland,
area. This information is key for control initiatives, and forms a the Åland Islands, and southern Sweden should be known.
building block for all further research. While some species lists In 1979, 19 species of mosquitoes were reported from Åland,
are well maintained, others for many countries or regions are out representing half of the 39 species that were then recog-
of date, based on incomplete data or missing altogether. nized from the whole of Finland (Utrio, 1979). These species
Mainland Åland is the largest of over 6500 islands in the are representatives of the genera Aedes Meigen, Anopheles
autonomous archipelago of Åland in the Gulf of Bothnia, Meigen, Culex Linnaeus, Culiseta Felt and Ochlerotatus Lynch
situated between Finland and Sweden. Officially part of Finland, Arribálzaga (following Reinert et al., 2009). Scant information
Åland forms the most southerly and westerly points of the was presented regarding the dates or locations of these collec-
country (N 59∘ , E 19∘ ), and is a busy area for ferries taking tions, and the methods were presented very generally, omitting
passengers and vehicles between ports in Finland, Sweden and much useful information. At the time, Utrio stated that ‘the
Estonia. The climate is milder than in mainland Finland, so there present picture of the distribution of the species is still far from

Correspondence: C. L. Culverwell, E326, University of Helsinki, Medicum, Department of Virology, Haartmaninkatu 3, Helsinki 00014, Finland.
Tel.: +358 2941 911; Fax: +358 29 41 26491; E-mail: [email protected]

© 2017 The Royal Entomological Society 145

146 C. L. Culverwell

adequate … ’, a comment that applied to his research on the during 2016 to determine seasonal variation of species within
entire country’s mosquito species, not just those restricted to habitats (Table 1).
Åland, and which still applies. Adult mosquitoes were collected using either an Improved
Since 1979, the list of Finnish mosquitoes has had a couple of Prokopack Aspirator with regular mesh cups or a Mosquito
revisions, bringing the number of species recognized from 39 to Magnet® Patriot (John W. Hock Company, Gainesville, FL,
40 with the addition of Dahliana geniculata (Olivier) (Itämies, U.S.A.) baited with octenol, or caught while biting using a
1981), and then to 38 with the removal of Anopheles maculipen- 50-mL Falcon™ tube. Unbaited CDC Light traps were set up
nis s.s. Meigen and Ochlerotatus sticticus (Meigen) based on in July and September 2016, but were not successful because of
a literature review (Huldén & Huldén, 2014). In comparison, poor weather conditions. Adult mosquitoes were frozen briefly
the list of Swedish mosquitoes was recently updated to include at −20 ∘ C, identified to species, then either stored at −20 ∘ C
50 species (Lundström et al., 2013; Statens Veterinärmedicin- in 80% ethanol (EtOH) for DNA extraction or mounted onto
ska Anstalt, 2016) of which at least three are confirmed from cardboard points and pinned for morphological examination.
locations compatible with Åland that are not currently recorded Immature life stages were collected from ground water habi-
from Finland (An. maculipennis s.s., Aedes rossicus Dolbeškin, tats, including puddles, road, field and woodland ditches,
Gorickaja & Mitrofanova and Oc. sticticus). streams, lake margins and flooded pools, using a larval dipper
The climate in Finland has warmed marginally since Utrio and/or a fine-meshed aquarium net. Tree cavities were searched
collected mosquitoes in the lead up to his publication in 1979 using a turkey baster and plastic tubing.
(Pirinen et al., 2012). This trend is predicted to continue in Fourth-instar larvae were processed by being either: (a)
forthcoming years (European Centre for Disease Prevention dropped into sub-boiling water, stored in 80% EtOH, then
and Control, 2012), and has the potential to result in the slide-mounted for morphological examination (see below), or
ranges of many native and invasive species being altered. (b) transferred to individual rearing vials and link reared, retain-
This increases the need to know where mosquito species are ing larval and pupal exuviae in 80% EtOH. Adults were frozen
distributed, as a warming climate will also expand the ranges for approximately 24 h after emergence to allow time for the cuticle
mosquito-borne arboviruses that affect humans, several of which to sclerotize and for male genitalia to undergo rotation. These
are known from Finland, including but not limited to Sindbis adults were then identified and either mounted on points or
virus (Sane et al., 2012), Inkoo virus (Brummer-Korvenkontio frozen at −20 ∘ C.
et al., 1973; Putkuri et al., 2016) and Chatanga virus (Putkuri
et al., 2016).
As results from extensive collections have not been published
Morphological identification and slide preparation
since the early 1980s, there is a need to update the records for
Finnish mosquitoes with contemporary data. This study, as part
Morphological identifications were made using a combination
of a larger project, aimed to ascertain initially which mosquito
of keys for adult females, fourth-instar larvae and male genitalia
species currently inhabit mainland Åland, and subsequently
(Utrio, 1976; Stojanovich & Scott, 1995; Becker et al., 2010). In
which species inhabit the Finnish mainland.
many cases, slide preparations were required to reveal necessary
morphological characters. Larval and pupal exuviae were placed
into Cellosolve™ (2-ethoxyethanol) (Thermo Fisher Scientific,
Methods
Waltham, MA, U.S.A.) for approximately 10 min then arranged
dorsal side up in Euparal (ANSCO Laboratories, Manchester,
Mosquito collections, field sites and processing
U.K.) on a microscope slide. Male abdomens, including the
Mosquitoes (n = 3286) were collected from mainland Åland genitalia, were clipped between abdominal segments VI and VII,
on 30 August 2015, and 14–17 May, 6–9 July and 6–9 and then processed using the same protocol as for fourth-instar
September 2016. In total, 88 collections were made across the larvae, as follows. Specimens were cleared using 1% NaOH at
island. As a consequence of time restrictions, the collection sites 55 ∘ C for 2 h, washed in ddH2 O, covered in Cellosolve™ for
loosely followed roads, for ease of access, and aimed to cover 1 h, washed in ddH2 0, covered in clove oil for 1 h and then
as many habitat types as possible. Efforts were also made to dissected/arranged dorsal side up in Euparal. All slides were
cover as large a geographical area as possible and to visit each then dried in an oven for at least 8 weeks at 55 ∘ C.
of the 10 municipalities on the mainland. Each unique field
collection (using a single collection method in a set time period)
was assigned a collection number starting with the country prefix Molecular identifications
‘FI’. Mosquitoes from each collection were numbered with
this country and collection code as well as a unique specimen Selected members of the An. maculipennis complex
number based on the number of individuals obtained. (n = 15/33) were distinguished using sequences from two
In 2015, only one collection of biting adults was made, gene regions: partial mitochondrial cytochrome c oxidase sub-
whereas in 2016 a combination of adult (n = 29) and immature unit I (COI) and ribosomal internal transcribed spacer 2 (ITS2).
(n = 58) collections were made simultaneously at each collec- Prior to extraction, the head, male genitalia and wings were
tion site. Collection times were not standardized as the aim was removed and later slide-mounted. Genomic DNA was extracted
to obtain maximum species diversity rather than relative abun- from the remainder of each specimen using Bioline’s Isolate II
dance data. Certain sites were visited at multiple time-points Genomic DNA Kit (Bioline, London, U.K.), with the standard

© 2017 The Royal Entomological Society, Medical and Veterinary Entomology, 32, 145–154
 Mosquitoes of Åland, Finland 147

Table 1. Collection sites, grouped into 1-km2 areas.

Site Latitude Longitude Elevation August

number Municipality (N) (E) (m) 2015 May 2016 July 2016 September 2016

1 Eckerö N 60∘ 12 E 019∘ 38 14 - FI785i, FI786i FI810i FI887i

2 N 60∘ 13 E 019∘ 32 6 - FI788i x FI888i
3 N 60∘ 13 E 019∘ 36 14 - FI787i - -
4 N 60∘ 14 E 019∘ 31 33 - - FI811i, FI812a -
5 Finström N 60∘ 13 E 019∘ 51 7 - FI771i, FI772i, FI819i, FI820a, FI897i, FI898i,
FI773i, FI774i FI827a, FI828i, FI899i, FI900a
FI829a
6 N 60∘ 14 E 019∘ 48 ? FI763a - - -
7 N 60∘ 14 E 019∘ 49 9 - FI775i, FI776i FI826i FI904i, FI905a
8 N 60∘ 14 E 019∘ 52 19 - FI770i - -
9 N 60∘ 15 E 019∘ 49 25 - FI782i x x
10 N 60∘ 16 E 019∘ 49 1 - - FI801a, FI808a, FI876a, FI884a,
FI809a, FI821a, FI885a, FI895a,
FI822a, FI823a, FI896a, FI906i,
FI830a FI907a, FI909a
11 N 60∘ 16 E 019∘ 50 0 - - FI824i, FI825i -
12 N 60∘ 18 E 019∘ 56 21 - FI777i FI805i FI883i
13 Geta N 60∘ 19 E 019∘ 51 22 - FI778i - -
14 N 60∘ 21 E 019∘ 46 5 - FI779i FI802a, FI803a FI877i, FI878a
15 N 60∘ 22 E 019∘ 55 2 - - - FI882i
16 N 60∘ 23 E 019∘ 48 5 - - FI804a -
17 N 60∘ 23 E 019∘ 55 5 - - x FI879i, FI880a,
FI901a, FI902i,
FI903i
18 N 60∘ 24 E 019∘ 54 9 - - - FI881i
19 Hammarland N 60∘ 12 E 019∘ 46 16 - FI783i FI806a, FI807i FI886i
20 N 60∘ 15 E 019∘ 42 29 - FI784i - -
21 Jomala N 60∘ 07 E 019∘ 59 33 - - - FI893i, FI894a
22 N 60∘ 10 E 019∘ 58 23 - FI769i - -
23 Lemland N 60∘ 00 E 019∘ 59 9 - FI789i - x
24 N 60∘ 03 E 020∘ 09 32 - - FI815i, FI816i -
25 N 60∘ 04 E 019∘ 57 11 - FI790i - x
26 Lumparland N 60∘ 04 E 020∘ 15 8 - - FI817i, FI818a FI889i, FI890a,
FI891i, FI892i
27 Mariehamn N 60∘ 06 E 019∘ 56 2 - FI768i FI813i, FI814i FI908i
28 Saltvik N 60∘ 17 E 020∘ 03 28 - FI780i x x
29 Sund N 60∘ 14 E 020∘ 04 4 - FI781i x x

When collections were attempted and how many/which successful adult or immature collections were made are indicated for each of the 29 areas.
Numbers commencing ‘FI’ are the original collection numbers, which correlate with the specimen numbers; a, adult collection; i, immature collection;
-, no collections attempted; x, collections attempted but no mosquitoes found. Only collections that were successful at least at one time-point are listed.

protocol. Polymerase chain reactions (PCRs) were set up in as alternatives. To sequence further along COI, primers UEA3
25-𝜇L reactions using Thermo Scientific™ DreamTaq™ DNA [adapted from UEA3 in Lunt et al. (1996) by Cook et al.
Polymerase (Thermo Fisher Scientific, Waltham, MA, U.S.A.) (2009)] (5′ -TATRGCWTTYCCWCGAATAAATAA) and
as follows: 2.5 𝜇L 10 × DreamTaq Buffer, 2.5 𝜇L dNTP mix Fly10 [adapted from UEA10 in Lunt et al. (1996) by Cook et al.
(2 mm each), 1.0 𝜇L × 10 𝜇m forward and 1.0 𝜇L × 10 𝜇m (2006)] (5′ -ASTGCACTAATCTGCCATATT) were also used.
reverse primers (see below), 0.125 𝜇L DreamTaq, 17.375 𝜇L Table 2 provides a summary of each primer pair that contributed
ddH2 O and 0.5 𝜇L DNA template (approx. 20 ng/𝜇L). to final sequences.
ITS2 was amplified using primers ITS2_5.8F (5′ -TGTGAA Thermocycler conditions were as follows: 94 ∘ C initial denat-
CTGCAGGACACATG) and ITS2_28R (5′ -ATGCTTAAATTTA uration for 5 min followed by 35 cycles of 94 ∘ C for 30 s, 48 ∘ C
GGGGGTA) (Collins & Paskewitz, 1996). COI was initially (ITS2 and UBC6F–UBC9R) or 50 ∘ C (LCO1490–HCO2198
amplified using the ‘universal’ barcoding primers LCO1490 and UEA3–Fly10) for 30 s and 72 ∘ C for 1 min, then a final
and HCO2198 (Folmer et al., 1994), but as they were not extension of 72 ∘ C for 7 min.
effective, UB6F (C1-J-1718F) (5′ -GGAGGATTTGGAAATT PCR products were visualized on a 2% agarose gel with
GATTAGTGCC) and UBC9R (C1-N-2191R) (5′ -CCCGGTAA GelRed™ (Biotium, Fremont, CA, U.S.A.) then purified using
AATTAAAATATAAACTTC) (Simon et al., 1994) were used the Thermo Scientific™ GeneJET Purification Kit. Sanger

© 2017 The Royal Entomological Society, Medical and Veterinary Entomology, 32, 145–154
148 C. L. Culverwell

Table 2. Molecular sequence data generated for Anopheles maculipennis sensu lato.

LCO1490- UBC6F- COI accession ITS2 accession

Specimen Le Pe Genus Species Sex HCO2198 UBC9R UEA3-Fly10 number number

FI 763-01 × × Anopheles messeae ♀ × ✓ ✓ KY971473 KY695113

FI 808-37 × × Anopheles messeae ♀ × ✓ ✓ KY971474 KY695114
FI 808-38 × × Anopheles messeae ♀ × ✓ ✓ KY971475 KY695115
FI 808-39 × × Anopheles messeae ♀ × ✓ × KY971476 KY695116
FI 808-40 × × Anopheles messeae ♀ × × ✓ KY971477 KY695117
FI 808-41 × × Anopheles messeae ♀ × ✓ ✓ KY971478 KY695118
FI 808-42 × × Anopheles messeae ♀ × ✓ ✓ KY971479 KY695119
FI 821-50 × × Anopheles messeae ♀ × ✓ ✓ KY971480 KY695120
FI 826-15 ✓ ✓ Anopheles maculipennis s.s. ♂ ✓ × ✓ KY971481 KY695121
FI 828-16 ✓ ✓ Anopheles maculipennis s.s. ♀ ✓ ✓ ✓ KY971482 KY695122
FI 906-10 ✓ ✓ Anopheles messeae ♂ × ✓ ✓ KY971483 KY695123
FI 906-11 ✓ ✓ Anopheles messeae ♂ × ✓ UEA3 KY971484 KY695124
FI 906-19 ✓ ✓ Anopheles messeae ♂ ✓ ✓ ✓ KY971485 KY695125
FI 906-35 ✓ ✓ Anopheles messeae ♀ × ✓ × KY971486 KY695126
FI 906-36 ✓ ✓ Anopheles messeae ♀ × ✓ ✓ KY971487 KY695127

Details of the primers used for successful sequences are given, along with their GenBank accession numbers. The COI sequence generated for each
specimen combined sequences from the three available primer pairs, indicated before the COI accession number. Successful reactions are marked with
‘✓’ and failed reactions with ‘x’. Specimen numbers correlate with the collection numbers given in Table 1 (e.g. FI828-16 is specimen 16 from collection
FI828). Specimens with associated larval and pupal exuviae are indicated in the columns marked ‘Le’ and ‘Pe’, respectively. ITS2, internal transcribed
spacer 2.

sequences were generated using the Big Dye™ Terminator Results

v3.1 Cycle Sequencing Kit (Applied Biosystems, Waltham, CA,
U.S.A.) with the same primers as used in the initial PCR round. In total, 3286 mosquitoes contributed to this study. Speci-
Sequences were edited using geneious version 8.1.8 (http:// mens belonged to at least 27 species representing eight genera
www.geneious.com; Kearse et al., 2012) then submitted to the (following Reinert et al., 2009). A summary of the results at
National Center for Biotechnology Information (NCBI) blast each time-point, compared with historical data, is presented in
website (Zhang et al., 2000) for comparison with highly similar Table 3.
nucleotide sequences (megablast).

New Finnish distribution

Mapping
Of the 95 specimens of Aedes that were collected, one larva,
Global Positioning System (GPS) coordinates were deter-
75 adult females and five adult males were not identified beyond
mined according to the WGS 84 system (dd∘ mm.mmm’) and
the possibility of being either Aedes cinereus Meigen or Aedes
collection locations were mapped using arcmap™ version
geminus Peus. Thirteen other adult males were identified to
10.2.1 (Environmental Sciences Research Institute, Los Ange-
les, CA, U.S.A.) (Fig. 1). As several locations were visited species based on critical examination of the gonostylus of their
multiple times, and some collections were made close together, genitalia. This revealed that both Ae. cinereus and Ae. geminus
the 88 collections were difficult to visualize in their raw format. were present in the collections.
Therefore, the collection data were divided into 29 1-km2 areas
based on the ‘dd∘ mm.’ part of the coordinate system, in order to
visualize them more effectively. Note that there are two points Molecular identifications
for collection areas 5 and 7, as the collections were made suffi-
ciently far from each other to warrant separate points, whereas Sequences with 95–100% BLASTN similarity were compared
in other cases the collections were made within approximately to the newly generated sequences for the 15 Anopheles spec-
100 m of one another. imens; of these, the sequences with 99–100% similarity were
then considered as potential identifications of the Anophelines.
Attention was paid to lower scoring BLASTN results, but lower
Specimen locations scoring results were not similar enough when sequences were
compared in Geneious, which is why they were discounted as
Specimens (pinned adults, slide-mounted exuviae, genitalia, potential identifications. A clear split was seen in the identi-
heads and wings) will be deposited in the Natural History fications, with 13 specimens coming out as potentially either
Museum, London and the Finnish Museum of Natural History, Anopheles messeae Falleroni or Anopheles daciae Linton, Nico-
Helsinki, following completion of ongoing research projects. lescu & Harbach, and two specimens as An. maculipennis. Fur-
Until then, they will remain with the author. ther comparison of the 13 ITS2 sequences potentially identified

© 2017 The Royal Entomological Society, Medical and Veterinary Entomology, 32, 145–154
 Mosquitoes of Åland, Finland 149

Fig. 1. Åland collection areas. Eighty-eight collections were made across the island (designated FIxxx in Table 1), which are herein represented by
29 separate 1-km2 collection areas, indicated by the numbered dots. Numbers correspond to the sites detailed in Table 1 and Tables S1–S3. Two points
are given for collection areas 5 and 7 as more than one location was sampled from within those 1-km2 areas.

as An. messeae or An. daciae, by aligning them in geneious with Sequences of the barcoding regions of COI and ITS2 for these
sequences of An. daciae deposited in GenBank by Nicolescu 15 specimens therefore revealed two species in the An. mac-
et al. (2004), revealed that the sequences generated in this study ulipennis complex: An. messeae (n = 13) and An. maculipennis
were An. messeae not An. daciae. None of the five mutations s.s. (n = 2). GenBank accession numbers for each sequence are
in ITS2 that Nicolescu et al. (2004) identified which determine listed in Table 2. Based on these sequences, An. maculipennis
An. daciae were present. Both new ITS2 sequences of An. mac- s.s. is reinstated on the species list for Finland.
ulipennis grouped 100% with An. maculipennis s.s. sequences in
GenBank, the next closest being Anopheles melanoon Hackett,
with 94% similarity. Other species records
The COI blastn results for the barcoding region of the
two potential An. maculipennis sequences returned both Anopheles: Anopheles claviger (Meigen) (n = 137) was col-
An. maculipennis and An. messeae (e.g. accession numbers lected across the season in larval collections (n = 104) and adult
KM258222.1 and KU877008.1) as potential identifications, collections (n = 29), but was found in the highest numbers in
while those of An. messeae grouped with others of the same July and September 2016.
species. To further investigate, the COI barcode regions were Aedimorphus Theobald: six female Aedimorphus vexans
aligned in geneious with the closest GenBank sequences (Meigen) individuals were collected in total from Finström and
identified as An. maculipennis and An. messeae. All sequences Geta at the end of the seasons in August 2015 (n = 1) and
of An. maculipennis grouped together, away from those of An. September 2016 (n = 5).
messeae, which also grouped together. When the sequences Coquillettidia Dyar: Coquillettidia richiardii (Ficalbi)
generated with primers UEA3 and Fly10 were later added to the was collected during July 2016 as adults (n = 131), par-
barcode regions, it proved more difficult to recover consistent ticularly host-seeking females, from locations in Fin-
identifications in the top hits, because the number of COI ström, Geta and Hammarland. Larval collections were not
fragments that extended beyond the barcode region was limited. successful.

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150 C. L. Culverwell

Table 3. Comparison of the current and historical mosquito collections from Åland.

September Found in Reported by

Genus Subgenus Species August 2015 May 2016 July 2016 2016 2015/2016 Utrio (1979)

AnopheIes AnopheIes claviger - ✓ ✓ ✓ ✓ ✓

maculipennis s.s. ? ? ✓ ? ✓ ?
maculipennis s.l. ✓ ✓ ✓ ✓ ✓ ✓
messeae ✓ ✓ ✓ ✓ ✓ ✓
Aedes - cinereus - ✓ ✓ ? ✓ ✓
geminus - ? ✓ ✓ ✓ -
Aedimorphus - vexans ✓ ✓ - ✓ ✓ -
Coquillettidia Coquillettidia richiardii - - ✓ - ✓ -
Culex Culex pipiens - - ✓ ✓ ✓ ✓
torrentium - - ✓ ✓ ✓ ✓
Neoculex territans - - ✓ ✓ ✓ -
Culiseta Culicella morsitans - ✓ ✓ ✓ ✓ ✓
ochroptera - - - - - ✓
Culiseta alaskaensis - - ✓ ✓ ✓ -
annulata - ✓ ✓ ✓ ✓ ✓
Dahliana - geniculata - - ✓ ✓ ✓ -
Ochlerotatus - annulipes - ✓ ✓ ✓ ✓ -
cantans - ✓ ✓ ✓ ✓ ✓
caspius - ✓ ✓ ✓ ✓ ✓
cataphyIIa - ✓ - - ✓ ✓
communis ✓ ✓ ✓ ✓ ✓ ✓
cyprius - - - - - ✓
dorsalis - - - - - ✓
excrucians - - - - - ✓
hexodontus - ✓ - - ✓ -
leucomelas - - - ✓ ✓ ✓
nigrinus - - - - - -
pionips ✓ - - - ✓ -
pullatus - ✓ - - ✓ -
punctodes* - - - - - -
punctor - ✓ ✓ ✓ ✓ ✓
sticticus ✓ - ✓ ✓ ✓ -
Woodius diantaeus - ✓ ✓ - ✓ ✓
intrudens - ✓ - - ✓ -
Total species reported ≥5 ≥15 ≥20 ≥ 18 ≥ 27 19

*Although no confirmed specimens of Oc. punctodes were collected, this species may yet be present in Åland since adult females of Oc. punctor and
Oc. punctodes are morphologically indistinguishable and often recovered from the same larval sites. Mosquitoes previously reported by Utrio (1979) are
compared to those collected in 2015 and 2016. Four species reported in 1979 were not collected during the 2015/2016 field trips; Culiseta ochroptera,
Ochlerotatus cyprius, Ochlerotatus dorsalis and Ochlerotatus excrucians. It was not possible to morphologically identify members of the Anopheles
maculipennis complex, nor to distinguish between females or immature stages of Aedes cinereus or Ae. geminus. Species marked ‘?’ therefore may or
may not appear in the unidentified material collected during the given time period.

Culex: three species of Culex were collected across the Dahliana Reinert, Harbach & Kitching: in July 2016, females
island: Culex (Culex) pipiens Linnaeus, Culex (Cux.) torrentium of Dahliana geniculata (n = 3) were collected from two munic-
Martini and Culex (Neoculex) territans Walker. Larval and ipalities, Finström and Lumparland. In September 2016, collec-
adult collections were successful in July and September, with tions were only successful in Finström (n = 7).
Cx. territans being restricted to larval collections. It was not Ochlerotatus: at least 12 species of Ochlerotatus were col-
possible to distinguish adult females of Cx. pipiens and Cx. lected from across the island: Ochlerotatus annulipes (Meigen),
torrentium, but the two species were identified by examination Ochlerotatus cantans (Meigen), Ochlerotatus caspius (Pallas),
of the genitalia of males reared from larvae. Ochlerotatus cataphylla (Dyar), Ochlerotatus communis (de
Culiseta: three species of Culiseta were collected from several Geer), Ochlerotatus hexodontus (Dyar), Ochlerotatus leucome-
locations: Culiseta (Culicella) morsitans (Theobald), Culiseta las (Meigen) (n = 1), Ochlerotatus pionips (Dyar), Ochlerotatus
(Culiseta) alaskaensis (Ludlow) and Culiseta (Culiseta) annu- pullatus (Coquillett), Ochlerotatus punctor (Kirby), Oc. sticti-
lata (Shrank). Culiseta (Culicella) ochroptera (Peus) was not cus, Ochlerotatus diantaeus (Howard, Dyar and Knab) and
encountered. Culiseta morsitans was consistently collected as Ochlerotatus intrudens (Dyar). Ochlerotatus punctodes (Dyar)
larvae from an open well in a wooded area at all three time-points was not found as larvae, but as available keys are not able to
in 2016. distinguish adult females of Oc. punctor and Oc. punctodes, it

© 2017 The Royal Entomological Society, Medical and Veterinary Entomology, 32, 145–154
 Mosquitoes of Åland, Finland 151

may still be present in Åland. Ochlerotatus sticticus (n = 181) previously published, but should not be viewed as definitive.
was collected as larvae (n = 32) and adults (n = 149) in July The diversity of species collected with comparison to published
and in September 2016. Although it was collected in July, only lists for the region highlights a need for further collections to
three specimens were collected compared to 178 collected in be made on the Finnish mainland, as well as on the other, scat-
September. tered, Åland islands. Sweden has a total of 50 recorded species
Four species that were previously found from Åland were not (Lundström et al., 2013; Statens Veterinärmedicinska Anstalt,
collected during this study: Cs. ochroptera (Peus), Ochlerotatus 2016) and with these results Finland now has 41, so it is very
cyprius (Ludlow), Ochlerotatus dorsalis (Haliday) and Ochlero- likely that other species are present. That said, a number of
tatus excrucians (Walker). the species recorded from Finland and Sweden have only been
reported from the extreme north or south of the countries, so the
Temporal distributions number of native European species from the islands would not
be expected to increase by a large number. Too few collections
A clear seasonal pattern was observed, as expected, in the were made to determine if the four previously reported species
mosquito species that were collected during this study. The that were not collected during this study (Cs. ochroptera, Oc.
single collection made in August 2015 is a brief snapshot of the cyprius, Oc. dorsalis and Oc. excrucians) are no longer present,
species potentially present on the island, with five confirmed or were just missed on these occasions.
species among 45 individuals. These included An. messeae, Eleven species collected during this study were not reported by
Am. vexans, Oc. caspius, Oc. pionips and Oc. punctor (Tables 3 Utrio (1979), but it is not possible to know for sure if they were
and S1). indeed present but not collected in the 1970s, or if they were later
In May 2016, despite attempts to locate adults, none were introduced to the island. Utrio (1979) included 20 640 specimens
encountered at any of the 23 collection sites, which resulted (including museum specimens) in his report as well as literature
exclusively in the collection of immature stages. Despite this, reports, which gave no detailed methods that could be used to
after a night of heavy rainfall there was evidence of adult infer where, when or how collections were made. It was clear
activity as fresh egg rafts of Cs. annulata (identified from during collections for the present study that certain species,
specimens subsequently reared from the eggs) appeared in including Cx. territans, were much more likely to be collected as
several locations. Excluding eggs, the species represented by immature life stages than as adults, or vice versa, leading to the
larvae present in May 2016 were restricted to An. claviger, Ae. question of whether Utrio biased sampling in favour of one life
cinereus/geminus, Cs. morsitans, Oc. annulipes, Oc. cantans, stage or another, or whether biases were introduced by sampling
Oc. cataphylla, Oc. communis, Oc. hexodontus, Oc. pullatus, at only one or two time-points during a given season. It seems
Oc. punctor, Oc. diantaeus and Oc. intrudens (Tables 1 and S1). likely, given his low specimen count and relatively low species
In July 2016, the species present changed slightly, whereby numbers, that collections were not comprehensive in the Åland
Oc. hexodontus, Oc. pullatus and Oc. intrudens were not archipelago in the 1970s and that more species were present at
collected, but in addition to the others collected in May, An. that time. While the climate has warmed marginally over the last
maculipennis s.s., An. messeae, Cq. richiardii, Cx. pipiens, Cx. 40 years (Pirinen et al., 2012), it is unlikely that this warming
torrentium, Cx. territans, Cs. alaskaensis, Cs. annulata, Da. would have contributed to the range shift in the species collected
geniculata and Oc. sticticus were collected (Tables 1 and S2). during this study, as they were also distributed across southern
In September 2016, the list of species collected was more Finland in the 1970s.
similar to the list of species encountered in July than in May,
with approximately 19 species collected (Tables 1 and S3),
including An. claviger, An. maculipennis s.l., An. messeae, Species of European interest and disease vectors
Ae cinereus/geminus, Am. vexans, Cx. pipiens, Cx. torrentium,
Cx. territans, Cs. morsitans, Cs. alaskaensis, Cs. annulata, A number of mosquito species within Europe are noted as
Da. geniculata, Oc. annulipes, Oc. cantans, Oc. caspius, Oc. being of interest for surveillance as they are either invasive
communis, Oc. leucomelas, Oc. punctor and Oc. sticticus. to Europe (European Centre for Disease Prevention and Con-
trol, 2012) or native but have a proven role in virus/parasite
Current Finnish mosquito fauna transmission (European Centre for Disease Prevention and Con-
trol, 2014). The European Centre for Disease Prevention and
Taking into account the current collections and historical Control (ECDC)’s list of the most significant native European
records/reviews (Utrio, 1979; Itämies, 1981; Huldén & Huldén, mosquito species includes 31 species [ECDC, 2014 (Table 2)].
2014), the list of Finnish species has increased from 38 to 41 Within Åland, 13 of the listed species are well established,
species with the inclusion of An. maculipennis s.s., Ae. geminus and are of greater interest from a public health perspective
and Oc. sticticus (Table 4). The list of species recorded from in view of their potential to vector arboviruses and parasites:
Åland rises from 19 to 31. An. maculipennis s.s., An. messeae, An. claviger, Ae. cinereus,
Ae. geminus, Cq. richiardii, Cs. morsitans, Cx. pipiens, Cx.
Discussion torrentium, Am. vexans, Oc. caspius, Oc. communis and Oc.
sticticus. Of these species, Cx. pipiens, Cx torrentium and Cs.
The present findings provide more detailed, up-to-date infor- morsitans are competent vectors of Sindbis virus in Sweden
mation regarding the species of mainland Åland than has been and in Finland (Sane et al., 2012; Hesson et al., 2015). Most
© 2017 The Royal Entomological Society, Medical and Veterinary Entomology, 32, 145–154
152 C. L. Culverwell

Table 4. List of Finnish mosquito species.

Genus Subgenus Species Species author

AnopheIes AnopheIes beklemishevi Stegnii & Kabanova, 1976

claviger (Meigen, 1804)
maculipennis* Meigen, 1818 s.l.
messeae Falleroni, 1926
Aedes - cinereus Meigen, 1818
geminus† Peus, 1970
Aedimorphus - vexans (Meigen, 1830)
Coquillettidia Coquillettidia richiardii (Ficalbi, 1889)
Culex Culex pipiens Linnaeus, 1758
torrentium Martini, 1925
Neoculex territans Walker, 1856
Culiseta Culicella morsitans (Theobald, 1901)
ochroptera (Peus, 1935)
Culiseta alaskaensis (Ludlow, 1906)
annulata (Shrank, 1776)
bergrothi (Edwards, 1921)
subochrea (Edwards, 1921)(in Wesenberg-Lund, 1921)
Dahliana - geniculata (Olivier, 1791)
Ochlerotatus - annulipes (Meigen, 1830)
cantans (Meigen, 1818)
caspius (Pallas, 1771)
cataphyIIa (Dyar, 1916)
communis (de Geer, 1776)
cyprius (Ludlow, 1919)
dorsalis (Meigen, 1830)
euedes (Howard, Dyar & Knab, 1913)
excrucians (Walker, 1856)
flavescens (Müller, 1764)
hexodontus (Dyar, 1916)
impiger (Walker, 1848)
leucomelas (Meigen, 1804)
nigrinus (Eckstein, 1918)
nigripes (Zetterstedt, 1838)
pionips (Dyar, 1919)
pullatus (Coquillett, 1904)
punctodes (Dyar, 1922)
punctor (Kirby, 1837)
riparius (Dyar & Knab, 1907)
sticticus‡ (Meigen, 1838)
Woodius diantaeus (Howard, Dyar & Knab, 1912)
intrudens (Dyar, 1919)

Based on collections made in 2015 and 2016, the number of species is increased from 38 to 41 with the inclusion of *Anopeheles maculipennis s.s.,
† Aedes geminus, and ‡ Ochlerotatus sticticus.

of the recent virus isolations from Finnish mosquitoes have for invasive species (European Centre for Disease Prevention
been performed on unidentified material, but historical isola- and Control, 2012). While no invasive species were found
tions have implicated a range of Ochlerotatus species as vec- during this trip, it does not negate the possibility for future
tors of Finnish arboviruses, including Oc. communis and Oc. introductions onto the islands if the environment becomes
punctor (Brummer-Korvenkontio et al., 1973), which are also favourable. Mainland Åland is at the centre of several regular
present in the Åland collections. There is therefore a potential passenger ferry routes that transport cars and commercial
for known mosquito-borne viruses to circulate in Åland in native vehicles between Sweden, Finland and Estonia through
species. no fewer than 16 ports around the three countries. It is
The climate is milder in Åland than on the Finnish mainland, not unreasonable to speculate that more southerly species
particularly during the winter months (Pirinen et al., 2012), and may enter the islands via this route, but the climate would
it is predicted that during the coming decades the climate will have to be suitable for them to become established in the
warm by several degrees, thereby increasing the potential range region.

© 2017 The Royal Entomological Society, Medical and Veterinary Entomology, 32, 145–154
 Mosquitoes of Åland, Finland 153

Anopheles maculipennis s.s. as the specimens were lost and unverifiable. Sweden has a
well-established population of Oc. sticticus that occurs across
There are many reports of ‘An. maculipennis’ from Fin- the south of the country (Lundström et al., 2013) and is predicted
land, but they do not, in most cases, distinguish between to spread much further northwards over the coming decades
An. maculipennis s.s. and Anopheles beklemishevi (Stegnii with climate warming (Schäfer & Lundström, 2009). Åland
& Kabanova). Lokki et al. (1979) noted this problem, and in falls within the known north–south species range that has been
their publication on Finnish An. maculipennis s.l., categorically reported in Sweden, so it is not surprising that Oc. sticticus is
stated that ‘all references to the occurrences of Anopheles mac- well established on the island given the proximity to Sweden.
ulipennis maculipennis in Finland are incorrect … ’. Huldén Results from Åland and Sweden combined challenge the status
& Huldén (2014) did not cite any references but stated that of mainland Finland as being free from this species.
this species was ‘not recorded in Finland’, claiming that all
prior references referred to An. beklemishevi or An. messeae.
Although few specimens of this species were collected during Supporting Information
the present study, the species is demonstrated to be present in
Åland and also has a distribution in southern Finland (C. L. Additional Supporting Information may be found in the
Culverwell, unpublished data). online version of this article under the DOI reference: DOI:
10.1111/mve.12272
Table S1. Specimen details for collections made on 30 August
Culex pipiens
2015 and 14–17 May 2016. Species confirmed as present are
indicated by ‘✓’, and absent by ‘-’. Ochlerotatus punctodes is
Recent and historical references have been made to Cx. pipiens
indicated as ‘?’ as adults females are indistinguishable from
biotype molestus being present in Finland, and in northern
those of Oc. punctor and no males or larvae were collected.
Europe as a whole (Utrio, 1976; Hesson et al., 2016; Lindström,
Collections are indicated as ‘A’ = adult and ‘I’ = immature
2017); however, this biotype was not considered within the
stages.
scope of this study. Although there is a possibility that the
species inhabits Åland, the molecular work for this species will Table S2. Specimen details for collections made on 6–9 July
be conducted in another study will examine Cx. pipiens from 2016. Species confirmed as present are indicated by ‘✓’, and
mainland Finland and Åland together. absent by ‘-’. Members of the Anopheles maculipennis complex
requiring molecular identification are marked as ‘?’ as at least
one or other of An. maculipennis s.s. or An. messeae was
Dahliana geniculata
collected at the given site. Culex pipiens and Cx. torrentium were
identified from characters of the male genitalia; thus, collections
A single biting female of Da. geniculata was collected at
with females only have a ‘?’, but it is possible that one or
Rauma, Finland for the first time in 1979 (Itämies, 1981), but
both species are present. Ochlerotatus punctodes is indicated
was not reported again thereafter. Itämies’s report, written in
as ‘?’ as adult females are indistinguishable from those of Oc.
Finnish, was overlooked in the distribution chart for European
punctor, and in most collections Oc. punctor was confirmed
mosquitoes compiled in 1999 (Snow & Ramsdale), so the
from larvae, which were not available for potential specimens
species was widely considered to be absent from Finland.
of Oc. punctodes. Collections are indicated as ‘A’ = adult and
Thus, when Dahl & Blackmore (2001) compiled the European
‘I’ = immature.
distribution of Da. geniculata, they also stated that it was
absent from the country, citing Snow & Ramsdale (1999). Table S3. Specimen details for collections made on 6–9 Septem-
This statement was an accidental error, however, as Dahl had ber 2016. Species confirmed as present are indicated by ‘✓’, and
confirmed the identification of the specimen collected by Itämies absent by ‘-’. Members of the Anopheles maculipennis complex
and therefore its presence in Finland (C. Dahl, pers. comm.). requiring molecular identification are marked as ‘?’ as at least
In the literature review of Finnish species, Huldén & Huldén one or other of An. maculipennis s.s. or An. messeae was col-
(2014) stated that this species was possibly an accidental visitor lected at the given site. Culex pipiens and Cx. torrentium were
to Finland, but it appears to be established on Åland. This is identified from characters on the male genitalia; thus, collec-
likely because Åland has many large, old, deciduous trees, which tions with females only have a ‘?’, but it is possible that one
provide optimal larval habitats. While larvae were not collected or both species are present. Ochlerotatus punctodes is indicated
during this study, the reason is more likely to be lack of time as ‘?’ as adult females are indistinguishable from those of Oc.
and lack of ladders to collect from higher tree holes, given that punctor, and in most collections Oc. punctor was confirmed
females were found from two locations on the island in both July from larvae, which were not available for potential specimens
and September 2016. of Oc. punctodes. Collections are indicated as ‘A’ = adult and
‘I’ = immature.
Ochlerotatus sticticus
Acknowledgements
Despite historical reports that Oc. sticticus has been collected
in Finland (Frey, 1932 in Syrjämäki, 1960; Utrio, 1979), Huldén This project was primarily funded by Societas Pro Fauna et Flora
& Huldén (2014) removed this species from the national list Fennica, but also in part by the Sigrid Jusélius Foundation and

© 2017 The Royal Entomological Society, Medical and Veterinary Entomology, 32, 145–154
154 C. L. Culverwell

the University of Helsinki Integrative Life Sciences Graduate Lokki, J., Saura, A., Korvenkontio, P. & Ulmanen, I. (1979) Diagnos-
Program. Many thanks to Joni Uusitalo for providing the ing adult Anopheles mosquitoes. Aquilo Seriologica Zoologica, 20,
specimens collected in 2015, and to Dr Ralph Harbach, Prof. 5–12.
Olli Vapalahti and Dr Eili Huhtamo for commenting on the Lundström, J.O., Schäfer, M.L., Hesson, J.C. et al. (2013) The geo-
manuscript. Thanks also to Martin Snickars for providing the graphic distribution of mosquito species in Sweden. Journal of the
shape file for Åland used to construct Fig. 1, and to Ruut European Mosquito Control Association, 31, 21–35.
Uusitalo for assisting with mapping software. Lunt, D.H., Zhang, D.-X., Szymura, J.M. & Hewitt, G.M. (1996)
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