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. 2022 Mar 15;17(3):e0264300. doi: 10.1371/journal.pone.0264300

The Asian tiger mosquito Aedes albopictus (Skuse) in Kosovo: First record

Nesade Muja-Bajraktari 1, Përparim Kadriaj 2, Ferdije Zhushi-Etemi 1, Kurtesh Sherifi 3, Bulent Alten 4, Dusan Petrić 5, Enkelejda Velo 2,*, Francis Schaffner 6
Editor: Carina Zittra7
PMCID: PMC8923454  PMID: 35290988

Abstract

The Asian tiger mosquito, Aedes albopictus, is an invasive mosquito species that is considered a potential vector of about 22 arboviruses, among which dengue, chikungunya and Zika. Here we report the first record of Aedes albopictus in the territory of the Republic of Kosovo. The first finding, in July 2020, was driven by a photo of an adult mosquito published by a citizen in social media. The subsequent field investigation in July 2020 confirmed the presence of adult mosquitoes by human landing catch and collection of eggs in ovitraps at the village Zhur. Monitoring was performed for 7 weeks with ovitraps and BG-Sentinel adult traps at 36 sampling stations from 7 localities, in the Southern part of Kosovo, on the border with Albania. Fifty-two out of 81 ovitrap samples resulted positive for the presence of Ae. albopictus. A total of 2,711 eggs are collected in 22 out 36 stations and a total of 78 adults from 4 out 4 stations with BG Sentinel trap and 15 adults with handled electric aspirator. Our finding shows that the tiger mosquito is expanding its geographical range in the Balkans, southeastern Europe.

Introduction

Aedes (Stegomyia) albopictus (Skuse, 1894) (Diptera: Culicidae), commonly called the ’Asian tiger mosquito’, is widespread throughout the tropical and subtropical regions of the world. During the last 30 years, it has invaded many countries worldwide, including in the Mediterranean Basin. The species is currently considered one of the top 100 invasive species globally and the most widespread invasive mosquito species in Europe [1, 2]. It represents a severe threat to public health due to its aggressive daytime human-biting behavior and its vectorial competence for numerous arboviruses of the family Flaviviridae (e.g., dengue, West Nile, and Japanese encephalitis viruses), Bunyaviridae (e.g., Rift Valley fever, Potosi, Cache Valley, and La Crosse viruses), Togaviridae (e.g., chikungunya and Ross River viruses) [3–7]. After Ae. aegypti, Ae. albopictus is the secondary vector of dengue and dengue hemorrhagic fever [8]. In Europe, the species was incriminated as the vector in outbreak of, among others, chikungunya in 2007 northeastern Italy [9], dengue in 2010 in Croatia [10], dengue and Zika cases between 2010 and 2020 in France [11–13].

The first report of the tiger mosquito in the European continent dates back to 1979 in Albania [14]. It is thought to have been imported in shipments and containers from China in the mid-1970s. Later it has been detected in Italy in 1990 and has spread throughout the European continent via various routes [1]. In the surroundings of Kosovo, this species was recorded in Montenegro [15], in North Macedonia [16], and Serbia [17]. In Serbia, Ae. albopictus was intercepted in two districts in the western and southwestern part of the country. It has been present for the past nine years on the Croatian border (Batrovci, northwest of Serbia) [18], and on the Montenegro border since 2014 [19].

Distribution models predict that Ae. albopictus will continue to expand, depending on transport, environmental, and climatic changes [20–22].

This worldwide expansion is mostly based on transport and dissemination of dormant egg via the international trade of used tires [23] and shipments of the Asian plant ‘‘lucky bamboo” (Dracaena spp.) [24, 25] and by public and private ground transport from heavily infested areas [22]. In 2017, a research for tiger mosquito in Kosovo was conducted at the borders with Macedonia and Albania within the VectorNet project framework. It resulted negative, although Kosovo showed favorable conditions for the development of this mosquito species [26].

The primary objective of our study was, following a citizen report, to confirm the presence of the tiger mosquito in the territory of Kosovo and to determine its distribution.

Materials and methods

Study area

The present study was conducted in the municipality of Prizren (42.2166 N, 20.7333 E) and in the city of Suhareka (42.2248 N, 20.2248 E) in July, August and September 2020. The Municipality of Prizren, occupies the southern position in the Dukagjini Plain and southwestern Kosovo. The average altitude is about 450 m above sea level and includes nearly 640 km2 of the Kosovo’s entire surface. Sharri mountain, Prizren plain and downstream area of the Drini I Bardhë define the relief. Sharri mountain is also an essential geographical element since it constitutes a watershed between the Adriatic Sea and the Aegean Sea basin. The climate of this part of Kosovo is classified as Mediterranean. The city of Suhareka is located in the southern part of Kosovo. The average altitude is about 455 m. It is characterized by a rich hydrography.

Collection and identification

We used ovitraps which are commonly used for detecting the females’ presence via egg laying [27], and entomological aspirator (GeniccoSrl, Italy, Model: JF0825S1H—R) and BG-Sentinel™ traps (Biogents, Germany) to collect the adult mosquitoes. The ovitraps (500 ml black plastic cups) were filled with tap water and equipped with a masonite strip (12.5 × 2.5 cm) for egg deposition. The plastic cups were modified by two holes, punching 3 cm from the top of the cup to prevent water overfilling. A total of 36 ovitraps were randomly distributed in 7 localities [Vërmicë (5), Zhur (11), Vlashnje (5), Prizren (8), Atmaxhë (3), Landovicë (3) and Suharekë (1)]. The distance between the traps was 100 m at minimum. At four localities (Vërmicë, Zhur st.1, Zhur st.2, Prizren) we also used BG-Sentinel traps baited with BG-Lure and CO2. The ovitraps were placed on the ground, in shaded and accessible places, under vegetation, with free space above at least 1 m (Fig 1A–1D).

Fig 1. Examples of traps operating at several mosquito collection sites.

Fig 1

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Ovitraps in: a Vlashnje (Tyrecentres); b Zhur st.1 (Privat residence garden); c Prizren (Privat residence garden); d Vërmicë (Restaurant veranda); e Odour-baited adult traps (BG-Sentinel) Vermice (Restaurant garden); f Zhur st.1 (Resident garden); g Prizren (Resident garden). Catching with aspirator: h Vlashnje (Tyrecentres). Adults resting in: i Prizren (on the human body); j Zhur (Plastic bottle); k Vlashnje (Inside the tire, resting on the surface water); l Zhur st.1 (first specimen of Aedes albopictus caught while landing on human body).

The ovitraps were left at the same place during a sampling period of 10 successive days. After 10 days, the masonite strips were collected from the traps and transferred to the laboratory at the university of Prishtina for identification based on their color, size, shape and surface sculpting [28]. The adult mosquitoes were identified using the MosKeyTool identification key [29]. Another technique used in this research was aspiration from human bait. This technique was used in three localities (Zhur st.1, Vlashnje, Prizren, (Table 1) for 30 minutes (Fig 1H, 1I and 1L).

Table 1. Results of Aedes albopictus adult trapping with BG-Sentinel traps (BG) or handheld electric aspirators (A) in the Prizreni municipality (July -September 2020).

Locality Latitude (N) Longitude (E) Method of capture Sampling area Sampling period# No. of adults No. of adults
            Ae. albopictus Cx.pipiens s.l
Vërmicë 42.166918  20.572473  BG Restaurant garden with vegetation 25-07-20 0 7
09-08-20 15 3
25-08-20 12 3
Zhur st.1 42.16604  20.61539  BG Private residence garden 25-07-20 27 3
09-08-20 10 0
25-08-20 5 0
Zhur st.2 42.161245  20.623351  BG  Private residence garden 25-07-20 0 2
09-08-20 4 5
25-08-20 0 0
Prizren 42.223997  20.734394  BG  Private residence garden  25-07-20 0 2
09-08-20 2 5
25-08-20 3 7
Zhur st.1 42.168164 20.615606 A House yard 01-08-20 1 0
10-08-20 3 0
Vlashnje 42.198731 20.667758 A Tire storage 03-09-20 9 0
Prizren 42.223997 20.734394 A House with vegetable garden  30-08-20 1 0
08-09-20 1 0
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#Entomological survey period: I: 25–27 July 2020; II: 08–10 August 2020; III: 25–27 August 2020.

One BG-Sentinel trap was run for two consecutive nights every two weeks and for three periods (Table 1). The traps were set in the private houses’ backyard (Fig 1E–1G). In total we did perform 12 BG-Sentinel trap-nights.

Results

The presence of Ae. albopictus was registered in Kosovo for the first time at the end of July 2020 in Zhur village. The annoyance and bites that the mosquito caused to citizens during this period encouraged citizens to post a mosquito photo on a social media. This information suggested the tiger mosquito to be present in this area and to perform a field survey. Our first catch, a male mosquito (Fig 2), was caught by hand on 23.07.2020 in a private residence garden at Zhur st.1 (Table 1). It was identified in the laboratory of the Institute of Public Health, Tirana, Albania, as belonging to Ae. albopictus.

Fig 2. First specimen of Aedes albopictus caught in Zhur st.1, date 23.07.2020 (photo: N. Muja-Bajraktari).

Fig 2

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In total, 52 ovitrap samples revealed positive, yielding 2,711 eggs collected at 36 sampling stations from 7 localities (Fig 3).

Fig 3. Distribution map of the ovitraps in Prizreni region showing positive and negative site for the presence of Aedes albopictus.

Fig 3

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The general administrative maps were extracted from the Natural Earth (https://www.naturalearthdata.com/) and then modified subsequently according to the information presented in the map, using Q-GIS version 3.18.

Using ovitraps, we collected eggs of Ae. albopictus but also of the native species Aedes (Dahliana) geniculatus (Olivier, 1791) (Table 2) at the Prizreni municipality. Eggs of Ae. albopictus were found at 22 out of the 36 sampling stations, while 14 remained negative. We did collect 440 eggs of Ae. albopictus at Vërmicë, 1,187 at Zhur, 139 at Vlashnje and 119 at Atmaxha, respectively. In the city of Prizren we collected 786 eggs from three stations, while 40 eggs were counted from ovitraps in 3 stations at a Truck Terminal, on the periphery of the city of Prizren. We didn’t found any egg in the Landovica and Suhareka city (Table 2).

Table 2. Location of ovitraps in the Prizreni municipality and numbers of eggs collected for Aedes albopictus and Aedes geniculatus.

Localities ID Sampling period# Latitude (N) Longitude (E) Sampling area No. of eggs
            Ae. albopictus Ae. geniculatus
Vërmicë 01/1 I, II, III 42.167278 20.577672 Near the road 68 63
01/2 I, II, III 42.168801 20.582681 Near the road 54 0
01/3 I, II, III 42.166918 20.572473 Restaurant garden 202 0
01/4 I, II, III 42.164574 20.567957 Restaurant 112 0
01/5 I, II, III 42.163792 20.563543 House veranda 4 0
Total           440 63
Zhur 02/1 I, II, III 42.168164 20.615606 Private residence garden 346 0
02/2 I, II, III 42.166543 20.607746 Private residence garden with pets 196 0
02/3 I, II, III 42.161062 20.622807 Private residence garden 168 48
02/4 I, II, III 42.15736 20.618477 Private residence garden 62 0
03/1 I, II, III 42.165116 20.623798 Private residence garden 175 0
03/2 I, II, III 42.161867 20.611347 Private residence garden 240 0
03/3 I, II, III 42.223801 20.734074 Private residence garden 0 0
03/4 I, II, III 42.15956 20.629809 Private residence garden 0 0
04/1 IV 42.163807 20.632446 Private residence garden with chicken 0 0
04/2 IV 42.166677 20.627916 Private residence garden 0 0
04/3 IV 42.15726 20.624888 Near forest 0 0
Total           1,187 48
Vlashnje 05/1 II, III 42.200968 20.660929 At the gas station near the road 76 0
05/2 II, III 42.204696 20.659614 Inside the tires 30 0
05/3 II, III 42.201094 20.664804 Inside the tires 33 0
05/4 II, III 42.203438 20.668572 Tire center garden 0 0
05/5 II, III 42.200968 20.660929 Tire center garden 0 0
Total            139 0
Prizren 06/1 I, II, III, IV 42.223288 20.741278 Private residence garden 438 4
06/2 I, II, III 42.228297 20.739493 Private residence garden 176 0
06/3 I, II, III 42.226967 20.729043 Private residence garden 172 0
06/4 I, II, III 42.223801 20.734074 Private residence garden 0 0
Total           786 4
Truck terminal 07/1 IV 42.249954 20.729769 Truck terminal garden 3 0
07/2 IV 42.256972 20.733839 Restaurant garden 16 0
07/3 IV 42.254285 20.72666 Near the road 0 0
07/4 IV 42.250268 20.721548 Truck terminal garden 21 0
Total           40 0
Atmaxhë Total 08/1 III 42.245329 20.699713 At the gas station near the road 73 0
08/2 III 42.24801 20.696709 Near the car wash 46 0
08/3 III 42.243715 20.702604 Hotel garden 0 0
          119 0
Landovicë 09/1 IV 42.259853 20.688001 Supermarket forecourt 0 0
09/2 IV 42.264082 20.6842 Near the road 0 0
09/3 IV 42.255862 20.684525 Private residence garden 0 0
Suharekë 10/1 I, II, III, IV 42.363973 20.832803 Bus station 0 0
Overall total           2,711 115
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#Entomological survey period: I: 23–02 July 2020; II: 02–12 August 2020; III: 12–22 August 2020; IV: 02–12 September 2020.

Eggs of Ae. geniculatus were found only at 3 sites from 3 locations, i.e. Vërmicë, Zhur and Prizren (Table 2).

Four adult females were collected with an aspirator at a house yard, nine adult mosquitoes at a tire center with numerous tires holding rainwater, (Fig 1h and 1K) and two males at a house vegetable garden. Using BG-Sentinel traps, we collected 78 adult mosquitoes (38 females and 39 males) at four sampling stations in gardens of residential houses or restaurant within three entomological survey periods (Table 1).

Forty-eight eggs were successfully hatched and were reared to adults (18 males / 30 females). All adult mosquitoes were morphologically identified and classified as Ae. albopictus. During the research period, we also identified 115 eggs of Ae. geniculatus caught with ovitraps (Table 2) and 34 adult mosquitoes of Culex pipiens s.l. Linnaeus, 1758 caught with BG-Sentinel trap. (Table 1).

Discussion

The Asian tiger mosquito Ae. albopictus, originating from Southeast Asia, has undergone a significant expansion of its range in the last few decades [30].

In Europe, the tiger mosquito was first reported in Albania in 1979 [14], than ten years later in Italy [31], France [32, 33], Spain [34], Belgium [35], Switzerland [36], Greece [37], Montenegro [17, 38], Croatia [15], Bosnia and Herzegovina [15], Slovenia [15, 39] and North Macedonia [16]. The movement of cars has helped a lot in distributing Ae. albopictus species [40]. The first identification of the tiger mosquito was made near the border with Albania. It is thought that the way of its introduction was done through land routes strictly through the movement of vehicles.

Our study reports an established population of the Asian tiger mosquito in the municipality of Prizren in the southern part of Kosovo.

The ovitraps were selected as a research method due to their high sensitivity to low mosquito density, low price and practical use in the field [41]. A female tiger mosquito can lay eggs in several ovitraps placed in different areas; however, it depends on the sites’ attractiveness [42]. Ovitraps can help control the mosquito population by eliminating the eggs, which results in a lower number of mosquitoes [16]. The first Ae. albopictus specimen is registered in Zhur, a village close to the border with Albania. This specimen recorded in a garden with many artificial breeding places (for container-breeding species). The ability of the tiger mosquito to use both natural and artificial containers for larval habitats facilitates the widespread occupation of urban and peri urban environments [30], ensuring a close connection between the species and the human population and increasing the risk of vector-borne disease pathogen transmission in these areas [43].

The other sampling station, Vërmicë, is the closest to Albania’s border, which argues the significant presence of eggs in the ovitraps and the large number of adults caught with BG-Sentinel traps. The border zones are considered highly vulnerable for introduction of potential invasive species. The number of cars and other vehicles coming from Albania stop for a considerable time at the border crossings, respectively in Vërmicë, so introducing the tiger mosquito through this road is indisputable. After that, from the border with Albania continues the highway, Morinë-Prizren-Prishtinë, therefore it is expected that mosquitoes will spread in other parts of Kosovo, precisely through the land route. Also, the third sampling station in Vlashnje is characterized by the significant presence of used tires, which served as breeding sites. At the monitoring station inside the city of Prizren, the number of eggs in ovitraps was high, suggesting presence of a high number of adults. The recording of Ae. albopictus in this part of Kosovo is an important finding, demarcating new boundaries of the distribution range of the species in the Balkans and suggesting an increasing mosquito-borne disease threat to public health in that region. Based on our results, Ae. albopictus may be, by 2020, absent from the Landovica locality, in the periphery of Prizren, and from Suhareka, located about 41 km away from the Albanian border.

As a potential invasive species, it is expected that Ae. albopictus will expand its distribution to other parts of the country. This already happened with other invasive insect species recorded in Kosovo like the western conifer seed bug Leptoglossus occidentalis. This insect was reported from two localities in the east of the country, in the area of Batllava Lake, as well as in two localities from the foothills of Bjeshkët e Nemuna Mountains, in the western part of Kosovo [44].

Conclusions

Since the presence of the tiger mosquito in the Republic of Kosovo is confirmed, it is expected that the Health authorities will develop a monitoring system for both mosquito and pathogen surveillance to survey and prevent the risk of pathogen transmission. Community involvement has been very successful in other countries, therefore we suggest to further encourage community involvement (citizen scientists) for an early detection of this invasive mosquito species in other parts of the country.

Supporting information

S1 Table. Results of the investigation for the presence of Ae. albopictus eggs at 36 stations in Kosovo and Ae. albopictus adult trapping with BG Sentinel traps (BG) or handheld electric aspirator (A).

(XLSX)

Click here for additional data file. (21.5KB, xlsx)

Acknowledgments

The work was done within the framework of AIM-COST Action CA17108.

We would like to express our gratitude to Prof. Dr Silvia Bino, Head of the Department of Epidemiology and Control of Infectious Diseases, Institute of Public Health in Tirana, Albania, for her continuous support of our research and Mihaela Kavran that provided papers for research conducted on Serbia.

We are grateful to Mr. Kreshnik Morina from Prishtina, Kosovo, who assisted us to prepare the new map of the Fig 3 of this manuscript. We also want to thank in particular Ms. Hatixhe Çollaku from Zhur who posted in social media the photo of the first specimen of Aedes albopictus and Mr. Artan Muja and all citizens for their cooperation to provide information on mosquito’s presence and for giving us permission to investigate in their properties during the fieldwork.

Data Availability

All relevant data are within the paper and its Supporting Information file.

Funding Statement

Ovitraps used in this study were provided by ECDC VectorNet project to KS, NMB, and FS (framework contract OC/EFSA/AHAW/2013/02-FWC1 funded by the European Food Safety Authority (EFSA) and the European Centre for Disease prevention and Control (ECDC)).

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Decision Letter 0

Carina Zittra

29 Nov 2021

PONE-D-21-33477The Asian tiger mosquito Aedes albopictus (Skuse) in Kosovo: first recordPLOS ONE

Dear Dr. Velo,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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Carina Zittra, Ph.D., Mag.rer.nat.

Academic Editor

PLOS ONE

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Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

********** 

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #2: N/A

********** 

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

********** 

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: Yes

********** 

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The authors describe the first records of tiger mosquitoes in Kosovo. The manuscript is of high relevance in the field and provides novel information.

Major comment: English language editing is needed.

Minor comments:

entire document – e.g. Line 31: Change Aedes to Ae. when needed (not at first mentioning and at beginning of sentences)

The definition “invasive” should be used with care. It has not proven to outcompete native mosquitoes in Kosovo. It is recommended to change to “potential invasive”

Line 64: Mosquitoes transmit pathogens and not diseases.

Line 82: Dracaena in italics

Line 109: The link to BG is not necessary. It is better to mention BG-sentinel generation 2. Moreover it is not mentioned if CO2 and lure were used. Info is give in line 139 – recommended to move to line 109

Line 182: s.l. not in italics

Line 180-190: Many countries are missing – if all European countries where Ae. albopictus was reported are planned to be listed

Mosquito alert is mentioned but no data shown. Maybe discuss that this app might increase the number of reports (the reviewer has seen tiger mosquitoes from the Kosovo when specifying for Mosquito Alert)

Reviewer #2: The manuscript is well written and provides interesting findings from Kosovo. There are some issues however to be addressed and most of them are commented directly in the manuscript pdf document. The abstract is somehow too long, and it would be nice to be shortened. In Introduction or somewhere in the Discussion section it would be advisable to give an overview of other invasive insects known from Kosovo, so that the paper is put also in the context of broader invasion challenge. English must be improved as well, although most of the linguistic corrections are done inside the document. I suggest to publish the paper after these revisions.

********** 

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Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: Yes: Halil Ibrahimi

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

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Attachment

Submitted filename: PONE-D-21-33477_reviewer 3.pdf

Click here for additional data file. (1.2MB, pdf)
PLoS One. 2022 Mar 15;17(3):e0264300. doi: 10.1371/journal.pone.0264300.r002

Author response to Decision Letter 0

5 Feb 2022

Response to Reviewers

Dear reviewers,

Thank you very much for the given comments. We find them very useful and tried to answer to each of them.

Answer to Journal requirements

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdfand

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

Answer: Manuscript is corrected according to the Journal requirements

2. In your Methods section, please provide additional information regarding the permits you obtained for the work. Please ensure you have included the full name of the authority that approved the field site access and, if no permits were required, a brief statement explaining why.

Answer: Since the village Zhur where the first specimen of Aedes albopictus was recorded does not belong to a protected area, there was no need to ask for the permit to conduct our research. There was no need for permit to carry out the study also for the other localities. Citizens were very collaborative and allowed us to collect in their properties.

3. We note that Figure 3 in your submission contain [map/satellite] images which may be copyrighted. All PLOS content is published under the Creative Commons Attribution License (CC BY 4.0), which means that the manuscript, images, and Supporting Information files will be freely available online, and any third party is permitted to access, download, copy, distribute, and use these materials in any way, even commercially, with proper attribution. For these reasons, we cannot publish previously copyrighted maps or satellite images created using proprietary data, such as Google software (Google Maps, Street View, and Earth). For more information, see our copyright guidelines: http://journals.plos.org/plosone/s/licenses-and-copyright.

Answer: A new map has been designed by another author for the purpose to be used in our Manuscript according to the PLOS ONE policy.

The author of the new map, Mr. Kreshnik Morina (Geodesic) submitted a confirmation about the authorship and granted the permission to the authors of the Manuscript to use the content of the map for publication. He also explained that the general administrative maps were extracted from the Natural Earth (a public domain map dataset) https://www.naturalearthdata.com/ which is free to use in any type of project and then modified subsequently according to the information presented in the map, using Q-GIS version 3.18 (an open source GIS program).

4. We note that Figure 1 includes an image of a [patient / participant / in the study].

As per the PLOS ONE policy (http://journals.plos.org/plosone/s/submission-guidelines#loc-human-subjects-research) on papers that include identifying, or potentially identifying, information, the individual(s) or parent(s)/guardian(s) must be informed of the terms of the PLOS open-access (CC-BY) license and provide specific permission for publication of these details under the terms of this license. Please download the Consent Form for Publication in a PLOS Journal (http://journals.plos.org/plosone/s/file?id=8ce6/plos-consent-form-english.pdf). The signed consent form should not be submitted with the manuscript, but should be securely filed in the individual's case notes. Please amend the methods section and ethics statement of the manuscript to explicitly state that the patient/participant has provided consent for publication: “The individual in this manuscript has given written informed consent (as outlined in PLOS consent form) to publish these case details”.

If you are unable to obtain consent from the subject of the photograph, you will need to remove the figure and any other textual identifying information or case descriptions for this individual.

Answer: In the image of the figure is the first author of the manuscript during the research in the field. She included her photo in the Manuscript.

5. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Answer: The references are checked carefully.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

________________________________________

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #2: N/A

________________________________________

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

________________________________________

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: Yes

________________________________________

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The authors describe the first records of tiger mosquitoes in Kosovo. The manuscript is of high relevance in the field and provides novel information.

Major comment: English language editing is needed.

Answer: The English editing is done as suggested.

Minor comments:

entire document – e.g. Line 31: Change Aedes to Ae. when needed (not at first mentioning and at beginning of sentences)

Answer: It is changed as suggested

The definition “invasive” should be used with care. It has not proven to outcompete native mosquitoes in Kosovo. It is recommended to change to “potential invasive”

Answer: It is corrected. In the revised text is used potential invasive.

Line 64: Mosquitoes transmit pathogens and not diseases.

Answer: In the line 61 it is stated that Ae. albopictushas potential to transmit 22 arboviruses-pathogens, of the family Flaviviridae

Line 82: Dracaena in italics

Answer: it is corrected

Line 109: The link to BG is not necessary. It is better to mention BG-sentinel generation 2. Moreover it is not mentioned if CO2 and lure were used. Info is give in line 139 – recommended to move to line 109

Answer: It is corrected as suggested by reviewer.

Line 182: s.l. not in italics.

Answer: Corrected.

Line 180-190: Many countries are missing – if all European countries where Ae. albopictus was reported are planned to be listed

Answer: Other European countries where Ae.albopictus is reported are added.

Mosquito alert is mentioned but no data shown. Maybe discuss that this app might increase the number of reports (the reviewer has seen tiger mosquitoes from the Kosovo when specifying for Mosquito Alert).

Answer: The first author of the Manuscript used Mosquito Alert to report first record of Ae. albipictus in her country.

Reviewer #2: The manuscript is well written and provides interesting findings from Kosovo. There are some issues however to be addressed and most of them are commented directly in the manuscript pdf document.

The abstract is somehow too long, and it would be nice to be shortened.

Answer: Abstract is shortened as suggested.

In Introduction or somewhere in the Discussion section it would be advisable to give an overview of other invasive insects known from Kosovo, so that the paper is put also in the context of broader invasion challenge.

Answer: As suggested by the reviewer, an overview of other insect invasive species reported from Kosovo is given in the Discussion.

English must be improved as well, although most of the linguistic corrections are done inside the document. I suggest publishing the paper after these revisions.

Answer: The language corrections are done.

Beside the correction based on the reviewers’ comments and suggestions, some other minor correction are done in the Manuscript aiming to improve its quality.

________________________________________

6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: Yes: HalilIbrahimi

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Attachment

Submitted filename: Response to Reviewers.docx

Click here for additional data file. (25.4KB, docx)

Decision Letter 1

Carina Zittra

9 Feb 2022

The Asian tiger mosquito Aedes albopictus (Skuse) in Kosovo: first record

PONE-D-21-33477R1

Dear Dr. Velo,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Carina Zittra, Ph.D., Mag.rer.nat.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Acceptance letter

Carina Zittra

1 Mar 2022

PONE-D-21-33477R1

The Asian tiger mosquito Aedes albopictus (Skuse) in Kosovo: first record

Dear Dr. Velo:

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org.

If we can help with anything else, please email us at plosone@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Carina Zittra

Academic Editor

PLOS ONE

Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    S1 Table. Results of the investigation for the presence of Ae. albopictus eggs at 36 stations in Kosovo and Ae. albopictus adult trapping with BG Sentinel traps (BG) or handheld electric aspirator (A).

    (XLSX)

    Click here for additional data file. (21.5KB, xlsx)
    Attachment

    Submitted filename: PONE-D-21-33477_reviewer 3.pdf

    Click here for additional data file. (1.2MB, pdf)
    Attachment

    Submitted filename: Response to Reviewers.docx

    Click here for additional data file. (25.4KB, docx)

    Data Availability Statement

    All relevant data are within the paper and its Supporting Information file.

    Articles from PLoS ONE are provided here courtesy of PLOS

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