First Nationwide Monitoring Program for the Detection of Potentially Invasive Mosquito Species in Austria

Bakran-Lebl, Karin; Pree, Stefanie; Brenner, Thomas; Daroglou, Eleni; Eigner, Barbara; Griesbacher, Antonia; Gunczy, Johanna; Hufnagl, Peter; Jäger, Stefanie; Jerrentrup, Hans; Klocker, Lisa; Paill, Wolfgang; Petermann, Jana S.; Barogh, Bita Shahi; Schwerte, Thorsten; Suchentrunk, Carina; Wieser, Christian; Wortha, Licha N; Zechmeister, Thomas; Zezula, David; Zimmermann, Klaus F.; Zittra, Carina; Allerberger, Franz; Fuehrer, Hans‐Peter

doi:10.3390/insects13030276

Cited by 4 publications

(4 citation statements)

References 57 publications

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“…At each sampling event the water and the oviposition substrate were changed, and the inner surface of the container was wiped with a paper tissue. This study was part of a national monitoring programme for invasive Aedes species in Austria and the study design follows the AIMSurv protocol (Bakran-Lebl et al, 2022;Miranda et al, 2022).…”

Section: Trap Sites and Sampling Schemementioning

confidence: 99%

Wooden tongue depressors as oviposition substrate for the detection of container-breeding Aedes species in ovitraps – Do we miss something?

Bakran-Lebl,

Bartel,

Griesbacher

et al. 2024

J. Eur. Mosq. Control Assoc.

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Ovitraps are easy-to-use and inexpensive traps for detecting potentially invasive container-breeding Aedes mosquitoes, particularly the Asian tiger mosquito (Aedes albopictus). These traps consist only of a black container filled with water, and an oviposition substrate. Wooden tongue depressors are commonly used oviposition substrates, but they leave the inner surface of the container largely uncovered, which could be used for egg laying as well. The aim of this study is to investigate whether observations of container-breeding Aedes species can be missed if only eggs laid on the oviposition substrate are examined. In 2022, five ovitraps were set up at each of six locations in the province of Salzburg in Austria. Wooden tongue depressors, roughened with sandpaper were used as oviposition substrate. The water and the oviposition substrate were replaced with a fresh one at each of the weekly samplings during the observation period, and the inner surface of the container was wiped with a paper tissue. The eggs on the stick and tissue were counted and the species of the eggs determined (using morphological characteristics and multiplex PCR). We found eggs of three Aedes species, mainly from Ae. japonicus, but also from Ae. albopictus and Ae. geniculatus. All of those laid their eggs on the wooden tongue depressors as well as on the container wall. However, the tongue depressor was clearly preferred to the container wall. Only in 2.9% of the sampling events eggs would have been missed if only the tongue depressors had been investigated. However, this proportion decreased with an increasing number of eggs per sampling event. We can therefore conclude that, although the container wall is also regularly used by the mosquitoes to oviposit, the probability of overlooking the presence of container-breeding Aedes mosquitoes by examining only the eggs laid on the wooden tongue depressor is very low.

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Section: Trap Sites and Sampling Schemementioning

confidence: 99%

Wooden tongue depressors as oviposition substrate for the detection of container-breeding Aedes species in ovitraps – Do we miss something?

Bakran-Lebl,

Bartel,

Griesbacher

et al. 2024

J. Eur. Mosq. Control Assoc.

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“…[ 9 , 10 ]), proturans [ 11 , 12 ] to various insect taxa (e.g. Odonata:[ 13 ]; Orthoptera: [ 14 ]; Boreidae: [ 15 ]; Ceratopogonidae: [ 16 ]; mosquitos: [ 17 , 18 ]. With respect to Lepidoptera, DNA barcoding has been almost completed for Austrian butterfly species (superfamily Papilionoidea, [ 19 ]) and noctuid moths (superfamily Noctuoidea, [ 20 ]), and barcoding of the European species of microlepidopteran Gelechiidae and the leaf mining Gracillaridae included most of the species recorded from Austria [ 21 , 22 ].…”

Section: Introductionmentioning

confidence: 99%

A DNA barcode library of Austrian geometridae (Lepidoptera) reveals high potential for DNA-based species identification

Schattanek-Wiesmair,

Huemer,

Wieser

et al. 2024

PLoS ONE

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Situated in the Eastern section of the European Alps, Austria encompasses a great diversity of different habitat types, ranging from alpine to lowland Pannonian ecosystems, and a correspondingly high level of species diversity, some of which has been addressed in various DNA barcoding projects. Here, we report a DNA barcode library of all the 476 species of Geometridae (Lepidoptera) that have been recorded in Austria. As far as possible, species were sampled from different Austrian regions in order to capture intraspecific genetic variation. In total, 2500 DNA barcode sequences, representing 438 species, were generated in this study. For complete coverage of Austrian geometrid species in the subsequent analyses, the dataset was supplemented with DNA barcodes from specimens of non-Austrian origin. Species delimitations by ASAP, BIN and bPTP methods yielded 465, 510 and 948 molecular operational taxonomic units, respectively. Congruency of BIN and ASAP partitions with morphospecies assignments was reasonably high (85% of morphospecies in unique partitions), whereas bPTP appeared to overestimate the number of taxonomic units. The study furthermore identified taxonomically relevant cases of morphospecies splitting and sharing in the molecular partitions. We conclude that DNA barcoding and sequence analysis revealed a high potential for accurate DNA-based identification of the Austrian Geometridae species. Additionally, the study provides an updated checklist of the geometrid moths of Austria.

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“…japonicus in Styria in 2011 [ 43 ]. Aedes japonicus has been found in all provinces in Austria since its first report, has established stable populations, and can now be found in high numbers in all parts of Austria [ 44 , 45 ]. Aedes albopictus was the second exotic Aedes species to be reported in Austria.…”

Section: Introductionmentioning

confidence: 99%

“…While DNA barcoding using Sanger sequencing can additionally help with identifying cryptic species [ 56 , 57 ], it does not allow accurate identification of multiple species in one sample [ 58 ]. Collection of Aedes eggs from container breeding species is typically performed by using so-called ovitraps, which consist of a black container filled with water and a wooden spatula plunged inside for oviposition support [ 44 ]. Aedes mosquitoes of different species might lay single or multiple eggs on the spatula.…”

Section: Introductionmentioning

confidence: 99%

Comparison of a multiplex PCR with DNA barcoding for identification of container breeding mosquito species

Reichl,

Prossegger,

Petutschnig

et al. 2024

Parasites Vectors

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Background Identification of mosquitoes greatly relies on morphological specification. Since some species cannot be distinguished reliably by morphological methods, it is important to incorporate molecular techniques into the diagnostic pipeline. DNA barcoding using Sanger sequencing is currently widely used for identification of mosquito species. However, this method does not allow detection of multiple species in one sample, which would be important when analysing mosquito eggs. Detection of container breeding Aedes is typically performed by collecting eggs using ovitraps. These traps consist of a black container filled with water and a wooden spatula inserted for oviposition support. Aedes mosquitoes of different species might lay single or multiple eggs on the spatula. In contrast to Sanger sequencing of specific polymerase chain reaction (PCR) products, multiplex PCR protocols targeting specific species of interest can be of advantage for detection of multiple species in the same sample. Methods For this purpose, we adapted a previously published PCR protocol for simultaneous detection of four different Aedes species that are relevant for Austrian monitoring programmes, as they can be found in ovitraps: Aedes albopictus, Aedes japonicus, Aedes koreicus, and Aedes geniculatus. For evaluation of the multiplex PCR protocol, we analysed 2271 ovitrap mosquito samples from the years 2021 and 2022, which were collected within the scope of an Austrian nationwide monitoring programme. We compared the results of the multiplex PCR to the results of DNA barcoding. Results Of 2271 samples, the multiplex PCR could identify 1990 samples, while species determination using DNA barcoding of the mitochondrial cytochrome c oxidase subunit I gene was possible in 1722 samples. The multiplex PCR showed a mixture of different species in 47 samples, which could not be detected with DNA barcoding. Conclusions In conclusion, identification of Aedes species in ovitrap samples was more successful when using the multiplex PCR protocol as opposed to the DNA barcoding protocol. Additionally, the multiplex PCR allowed us to detect multiple species in the same sample, while those species might have been missed when using DNA barcoding with Sanger sequencing alone. Therefore, we propose that the multiplex PCR protocol is highly suitable and of great advantage when analysing mosquito eggs from ovitraps. Graphical Abstract

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First Nationwide Monitoring Program for the Detection of Potentially Invasive Mosquito Species in Austria

Cited by 4 publications

References 57 publications

Wooden tongue depressors as oviposition substrate for the detection of container-breeding Aedes species in ovitraps – Do we miss something?

Wooden tongue depressors as oviposition substrate for the detection of container-breeding Aedes species in ovitraps – Do we miss something?

A DNA barcode library of Austrian geometridae (Lepidoptera) reveals high potential for DNA-based species identification

Comparison of a multiplex PCR with DNA barcoding for identification of container breeding mosquito species

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