AIMSurv Aedes Invasive Mosquito species harmonized surveillance in Europe. AIM-COST Action

Study Extent

Temporal coverage: Sampling period was different for each team but covering the seasonal peak of the targeted AIM species was mandatory. Considering all participating teams, the first team started at 15/01/2020 and the last team ended sampling at 31/12/20. In general, sampling was conducted between May-June and October-November for most of the teams. Geographic coverage: 23 European countries and one African country: Albania, Austria, Croatia, Cyprus, France, Germany, Greece, Hungary, Italy, Kosovo, Luxembourg, Moldova, Montenegro, Morocco, North Macedonia, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Switzerland, The Netherlands and Turkey.

Sampling

The sampling protocol for Pan-European surveillance of AIM species (AIMSurv harmonized the sampling methods, frequency, minimum length of the sampling period and the form of reporting. There were minimum requirements (Minimum Requirements Protocol; MRP) about the type of samples (i.e., eggs in ovitraps), number of sampled sites, number of traps and frequency of collecting samples. Teams with more resources were suggested to follow a Recommended protocol (RP) to either increase number of samplings and/ or, additionally to eggs, sample other life stages such as adults.
The use of a common platform for data collection was also suggested, the VECMAP® App system was made freely available by Avia-GIS to all participants during AIMSurv activities.
For the MRP, all teams performed the survey in three sampling sites separated by 10 Km or more. Five oviposition traps (ovitraps) per site were placed and separated by 15 to 100 m. The type of ovitrap was selected by each team according to their availability in the region but usually consisted of 250 to 1000 ml capacity black containers filled with tap water. One scratched wooden tongue depressor (1.7x15 cm) per ovitrap, was used as a substrate for oviposition. Some teams used similar size pieces of Masonite board (when part of a pre-existing surveillance network).
The selected sampling sites shared a similar environment, when possible, in urban and/or peri-urban areas (e.g., a garden of single-family houses in residential urban/peri-urban areas, public parks near residential areas, recreational areas). The frequency of sample collection was biweekly over a minimum of three months that must include the population peak of the targeted AIM species (e.g., in Spain: from September to November).
The following parameters were recorded: latitude and longitude of the position of each trap; the name of municipality/county/district (according to the country) and locality; start and end date of each trapping event (e.g., a period of 14 days / 2 weeks for ovitraps); land use according to VECMAP® categories (urban, peri-urban and others); count of each life stage collected, including absences (0 values); finally, trap status (e.g., missing, broken, empty, etc.) per trapping event was also recorded. The more ambitious RP sampling included additional sampling sites sharing similar or different environments (e.g., urban areas, rural areas, high altitude areas, etc), again with five ovitraps per site, weekly sampling frequency and sampling length during the whole seasonality of the AIM species including start, peak and end of the mosquito season (e.g., May to November in Central Europe). In addition, sampling adults using one BG-Sentinel™ (Biogents, Germany) trap baited with BG-Lure™ (Biogents, Germany) and/or CO2 per site under a sampling frequency of one trap/night per week was also included. The use of VECMAP® to report the data was also suggested in the RP. Parameters to record were the same as for the MRP plus the daily or weekly record of meteorological parameters (maximum, minimum, average temperature) per site, collected using data loggers or local weather stations. For the processing of samples, collected eggs of AIM species were counted. For every location, a sub-sample (2 out of 5 ovitrap substrates per locality) of eggs was reared to confirm the species by larva/adult morphology, particularly in those areas where several AIM species are present (i.e., Ae. albopictus and Ae. japonicus). Alternatively, species were identified using MALDI-TOF MS or other molecular methods (e.g., DNA sequencing). Adult of AIM species collected in BG-Sentinel™ were identified by morphology, sexed and counted. Suggested identification keys were ECDC (2012) and MosKeyTool V2.1 (Gunay et al. 2018). Samples of adults were preserved in 96% ethanol and/or cold preserved at -20/-80 °C to confirm identification if needed (e.g., via molecular tools).

Quality Control

All participants in AIMSurv reported data using a harmonized template. All data reported has been curated and the terminology has been homogenized. Data has been validated using the validator available in GBIF.

Method steps

1. The sampling protocol for Pan-European surveillance of AIM species (AIMSurv harmonized the sampling methods, frequency, minimum length of the sampling period and the form of reporting. There were minimum requirements (Minimum Requirements Protocol; MRP) about the type of samples (i.e., eggs in ovitraps), number of sampled sites, number of traps and frequency of collecting samples. Teams with more resources were suggested to follow a Recommended protocol (RP) to either increase number of samplings and/ or, additionally to eggs, sample other life stages such as adults.
   The use of a common platform for data collection was also suggested, the VECMAP® App system was made freely available by Avia-GIS to all participants during AIMSurv activities.
   For the MRP, all teams performed the survey in three sampling sites separated by 10 Km or more. Five oviposition traps (ovitraps) per site were placed and separated by 15 to 100 m. The type of ovitrap was selected by each team according to their availability in the region but usually consisted of 250 to 1000 ml capacity black containers filled with tap water. One scratched wooden tongue depressor (1.7x15 cm) per ovitrap, was used as a substrate for oviposition. Some teams used similar size pieces of Masonite board (when part of a pre-existing surveillance network).
   The selected sampling sites shared a similar environment, when possible, in urban and/or peri-urban areas (e.g., a garden of single-family houses in residential urban/peri-urban areas, public parks near residential areas, recreational areas). The frequency of sample collection was biweekly over a minimum of three months that must include the population peak of the targeted AIM species (e.g., in Spain: from September to November).
   The following parameters were recorded: latitude and longitude of the position of each trap; the name of municipality/county/district (according to the country) and locality; start and end date of each trapping event (e.g., a period of 14 days / 2 weeks for ovitraps); land use according to VECMAP® categories (urban, peri-urban and others); count of each life stage collected, including absences (0 values); finally, trap status (e.g., missing, broken, empty, etc.) per trapping event was also recorded. The more ambitious RP sampling included additional sampling sites sharing similar or different environments (e.g., urban areas, rural areas, high altitude areas, etc), again with five ovitraps per site, weekly sampling frequency and sampling length during the whole seasonality of the AIM species including start, peak and end of the mosquito season (e.g., May to November in Central Europe). In addition, sampling adults using one BG-Sentinel™ (Biogents, Germany) trap baited with BG-Lure™ (Biogents, Germany) and/or CO2 per site under a sampling frequency of one trap/night per week was also included. The use of VECMAP® to report the data was also suggested in the RP. Parameters to record were the same as for the MRP plus the daily or weekly record of meteorological parameters (maximum, minimum, average temperature) per site, collected using data loggers or local weather stations. For the processing of samples, collected eggs of AIM species were counted. For every location, a sub-sample (2 out of 5 ovitrap substrates per locality) of eggs was reared to confirm the species by larva/adult morphology, particularly in those areas where several AIM species are present (i.e., Ae. albopictus and Ae. japonicus). Alternatively, species were identified using MALDI-TOF MS or other molecular methods (e.g., DNA sequencing). Adult of AIM species collected in BG-Sentinel™ were identified by morphology, sexed and counted. Suggested identification keys were ECDC (2012) and MosKeyTool V2.1 (Gunay et al. 2018). Samples of adults were preserved in 96% ethanol and/or cold preserved at -20/-80 °C to confirm identification if needed (e.g., via molecular tools).

Aedes Invasive Mosquito species are included, namely: Aedes albopictus, Aedes japonicus and Aedes koreicus. Native species of mosquitos are also included: Aedes caspius, Aedes detritus, Aedes geniculatus, Anopheles maculipennis, Anopheles plumbeus, Culex hortensis, Culex pipiens, Culex tritaeniorhynchus, Culiseta annulata and Culiseta longiareolata. Unidentified species of the genera Aedes and Culex are also included.

1. Aedes albopictus
   common name: Asian tiger mosquito rank: species
2. Aedes koreicus
   common name: Korean bush mosquito rank: species
3. Aedes japonicus
   common name: Rock pool mosquito rank: species
4. Aedes caspius
   common name: Salt marsh mosquito rank: species
5. Aedes detritus
   common name: Salt marsh mosquito rank: species
6. Aedes geniculatus
   common name: Tree hole mosquito rank: species
7. Anopheles maculipennis
   rank: species
8. Anopheles plumbeus
   rank: species
9. Culex hortensis
   rank: species
10. Culex pipiens
    common name: Northern house mosquito rank: species
11. Culex tritaeniorhynchus
    rank: species
12. Culiseta annulata
    common name: Banded house mosquito rank: species
13. Culiseta longiareolata
    rank: species

The AIMSurv harmonized surveillance of Aedes Invasive Mosquito species has been conducted in 23 European countries and one North African country: Albania, Austria, Croatia, Cyprus, France, Germany, Greece, Hungary, Italy, Kosovo, Luxembourg, Moldova, Montenegro, Morocco, North Macedonia, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Switzerland, The Netherlands and Turkey.