- Research note
- Open Access
Identification of symbiotic bacteria in the midgut of the medically important mosquito, Culiseta longiareolata (Diptera: Culicidae)
BMC Research Notes volume 13, Article number: 378 (2020)
The potential use of symbiotic bacteria for the control of mosquito-borne diseases has attracted the attention of scientists over the past few years. Culiseta longiareolata is among the medically important mosquitoes that transmit a wide range of vector-borne diseases worldwide. However, no extensive studies have been done on the identification of its symbiotic bacteria. Given the role of this species in the transmission of some important diseases and its widespread presence in different parts of the world, including northwestern parts and the West Azerbaijan Province in Iran, a knowledge about the symbiotic bacteria of this species may provide a valuable tool for the biological control of this mosquito. Accordingly, the present study was conducted to isolate and identify the cultivable isolates bacterial symbionts of Culiseta longiareolata using 16S rRNA fragment analysis.
The midguts of 42 specimens of Cs. longiareolata were dissected, and the bacteria were cultured on agar plates. After the purification of the bacterial colonies, 16srRNA region amplification and gene sequence analysis were performed, and the sequences were confirmed by biochemical methods. In the present study, 21 isolates belonging to the genera Acinetobacter, Aerococcus, Aeromonas, Bacillus, Carnobacterium, Klebsiella, Morganella, Pseudomonas, Shewanella and Staphylococcus were identified.
Acting as vectors of diseases, mosquitoes transmit a wide range of parasite and arbovirus pathogens which are of veterinary and medical importance [1, 2]. Some species of mosquitoes are widely distributed throughout the world and are involved in the transmission cycle of a notable number of mosquito-borne diseases.
Among the veterinary and medically important mosquito species is the multivoltine Culiseta longiareolata. This species is thermophilic and highly ornithophilic . It is widely distributed in Europe, Asia, Africa, and the Mediterranean Sea , and acts as the vector of some infectious diseases such as the avian malaria [5, 6], tularemia , and arboviruses like West Nile fever [7,8,9].
Since mosquito-borne diseases cause serious health problems in many parts of the world, identifying different aspects of the biology of mosquito is of great importance. Knowledge about the biological properties, environmental requirements, and food chains [10, 11] of mosquitoes can be utilized for biological control. The symbiotic microbiota associated with mosquitoes have been found to affect most of their biological activities [12,13,14].
The symbiotic microbiota associated with each mosquito and their role in the biological activities of the mosquitoes can provide a valuable tool for the biological control of disease vectors [15,16,17,18,19,20,21]. Symbiotic bacteria affect the development [22, 23], nutrition [24, 25], reproduction [26,27,28], defense mechanisms [29, 30], and immunity  of mosquitoes. To understand the effect of symbiotic bacteria in the biological control of mosquitoes or mosquito-borne diseases, accurate identification of the symbiotic bacteria associated with each vector is an important first step [32, 33].
Although symbiotic bacteria have been studied and identified in different mosquito species [33,34,35,36,37,38,39,40,41,42], so far, no study has been performed on the identification of bacterial symbionts of Cs. longiareolata.
Given the role of this species in the transmission of some important diseases and its widespread presence in different parts of the world, including the northwestern region [34, 43,44,45,46] and West Azerbaijan Province (which shares border with four countries) in Iran, the symbiotic bacteria of Culiseta longiareolata were investigated in this study. In the present study, the cultivable bacterial symbionts of Culiseta longiareolata were isolated, cultivated and identified using 16S rRNA fragment analysis.
Material and Methods
Field collection of Cs. longiareolata and isolation of midgut bacteria
Mosquitoes specimens were collected from three regions of Urmia County (1- Naz-Loo: 37.651213, 44.983285, 2- Ghahraman-Loo: 37.659869, 45.207550, and 3- Moallem 37.546660, 45.033280) in the West Azerbaijan Province in the Northwestern region of Iran (Additional file 1: Figure S1) during May–August 2018 using different previously described collection methods . The collection techniques used in this study included the standard dipping method for larvae collection, and hand catches, day and night landing catches on cows, total catch, and pit shelter collection for adult specimens. The specimens were transferred alive to the entomology laboratory of the Department of Medical Entomology in the School of Public Health, and species were identified using morphological characteristics-based keys .
Adult female specimens of Cs. Longiareolata were identified and used for gut bacteria isolation. These specimens were sterilized, and their midguts were dissected individually under sterile conditions, according to previously described methods [33, 39].
The dissected midguts were mashed and suspended in 500 μL of Brain Heart Infusion (BHI), and the suspension was incubated at 28 ± 2 °C and 200 rpm for 24 h. A 100 μL aliquot of the midgut contents was serially diluted up to 10−6 and plated onto Nutrient Agar (Merck, Germany) and incubated at 28 ± 2 °C for 24–48 h . Continuous sub-culture of each bacterial colony using the streaking method was done to isolate single purified colonies of the bacteria. The individual colonies of the bacteria were later used for DNA extraction and PCR, biochemical and phenotyping studies.
16S rRNA gene amplification and sequencing
All purified bacterial colonies were individually subjected to genomic DNA extraction using the FavorPrep™ Kit (Favorgen, Taiwan), according to the manufacturer’s instructions. The 16S rRNA universal primers and previously described PCR program were used to amplify the 16S rRNA fragment . The acquired PCR amplicons were sequenced by Microsynth (Swiss).
All acquired sequences were checked for the presence of probable chimeric sequences by the Mallard program (https://www.bioinformatics-toolkit.org). All suspicious sequences were removed from the data set, and the resulting sequences were analyzed. The sequences were compared to the databases of the Ribosomal Database Project (RDP II; Michigan State University: rdp.cme.msu.edu) and the GenBank (www.ncbi.nlm.nih.gov/BLAST). Isolates were identified at the Genus and Species level based on sequence comparison using the GenBank and RDPII entries.
Finally, sequencing results that were consistent with the results of the biochemical studies were considered as reliable and definitive sequence of the bacterial isolates.
The MEGA7  was used for phylogenetic analysis and tree construction. The Maximum Likelihood (ML) method was used for the phylogenetic tree construction based on the Tamura 3-parameter model  (1000 bootstrap replicates) analyses.
In the present study, five species belonging to three genera of mosquitoes were collected and identified (An. maculipennis, Culex modestus, Cx. pipiens, Cx. theileri and Cs. longiareolata) in three sites across the Urmia County.
After species identification, specimens of Cs. longiareolata were selected for the purpose of the study. The midguts of 42 specimens of Cs. longiareolata were dissected, and the bacteria were cultured on agar plates to obtain bacterial colonies. After the purification of the bacterial colonies, 16srRNA region amplification and gene sequence analysis were performed for the bacterial isolates, and the sequences were confirmed by biochemical methods. In the present study, 21 isolates belonging to ten genera of bacteria were identified. The bacteria genera identified in this study include, Acinetobacter, Aerococcus, Aeromonas, Bacillus, Carnobacterium, Klebsiella, Morganella, Pseudomonas, Shewanella, and Staphylococcus. All acquired sequences were deposited in GenBank. The accession nos. of the bacterial species have been presented in Table 1.
Among the ten identified bacteria Genera, six were Gram-negative (Acinetobacter, Aeromonas, Klebsiella, Morganella, Pseudomonas and Shewanella) and four Genera were Gram-positive (Aerococcus, Bacillus, Carnobacterium and Staphylococcus).
Among the 21 isolates from the midgut of adult Cs. longiareolata, Aeromonas was the most frequent symbiont with eight isolates. Two species belonging to each of the genera Aeromonas, Klebsiella, Morganella, and Pseudomonas were also isolated and identified from the midgut of adult Cs. longiareolata.
Interestingly, the phylogenetic analysis of the acquired sequences of the bacteria isolates showed distinct monophyletic clades based on gram staining properties of their cell wall (Gram-negative and Gram-positive bacteria) (Fig. 1).
Also, phylogenetic analysis of the sequences obtained from the present study and similar sequences retrieved from the GenBank revealed the placement of bacteria of the same species and Genera in common branch and clades (Fig. 2).
The present study is the first report on the bacterial symbionts associated with the midgut of Cs. longiareolata. This mosquito vector plays a notable role in the transmission and maintenance of the transmission cycle of important diseases such as avian malaria [5, 6], tularemia , and arboviruses like West Nile fever [7,8,9] as secondary a vector.
The results of the midgut symbiotic bacteria of this vector are consistent with the results of many studies conducted on other vectors. In previous studies, symbiotic bacteria isolated from the midgut of Aedes aegypti  and Cx. quinquefasciatus  were predominantly members of the genus Bacillus, Klebsiella, Pseudomonas and Staphylococcus, which is consistent with the results of the present study. In another study conducted in India, members of the Genus Aeromonas were isolated from Cx. quinquefasciatus , which is also in agreement with the present study.
Symbiotic bacteria belonging to the genera Morganella, Aeromonas, and Klebsiella have also been identified in Anopheles fluviatilis , which is similar to the findings of our study.
Concerning the result of the present study, which identified the predominant isolates in the midgut of Cs. longiareolata, this finding is in agreement with the results of the dominant bacteria in the midgut of An. stephensi and An. culicifacies [33, 39], Aedes aegypti .
The identification of suitable candidates for paratransgenesis in the use of symbionts for biological control of vectors is of major interest to researchers. Members of the Genus Pseudomonas have been suggested in some studies as suitable candidates for paratransgenesis [16, 32, 35, 57,58,59]. In the present study, members of the genus Pseudomonas were identified in Cs. longiareolata, which confirms the results of previous studies which have reported the wide range of presence of Pseudomonas bacteria in different mosquito species.
In the first part of the study, different mosquito species were collected and identified. We captured five species of mosquitoes (An. maculipennis, Culex modestus, Cx. pipiens, Cx. theileri and Cs. longiareolata) in the study area. Previous studies have also identified these mosquito species in the northwest of Iran [43,44,45,46].
The five species captured in this study are important vectors of human and animal diseases. The geographical location of the northwest region of Iran (shares border with four countries) and the climatic diversity, as well as the history of mosquito-borne diseases makes this region vulnerable to a wide variety of mosquitoes. The presence of these vectors in this region requires public health attention, and the design of appropriate control programs is necessary to prevent the occurrence of epidemics.
The present study identified bacterial symbionts of Cs. longiareolata. To the best of our knowledge, this is the first report of bacteria symbiont of Cs. longiareolata. Is is recommended that future research in this area focus more precisely on identifying the biological properties of the isolated symbiotic bacteria, their biodiversity, and the biological relationship with their hosts, with the aim of developing new symbiont-based control programs. Previous studies have suggested that members of the Genus Pseudomonas may be suitable candidates for paratransgenesis. The isolation of Pseudomonas spp. in the present study confirms the wide spread of this genus in mosquito species and may further support the use of this species as a candidate for paratransgenesis to control mosquito-borne diseases.
Only the symbionts of the adult stage of Cs. longiareolata were identified.
Data availability statement
The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.
- Cs. longiareolata :
- 16S rRNA:
16 s ribosomal `RNA
Clements AN. The biology of mosquitoes, volume 3 transmission of viruses and interactions with bacteria. Wallingford: CABI; 1992.
Mullen GR, Durden LA. Medical and veterinary entomology. Cambridge: Academic press; 2009.
Maslov AV, Ward RA, Rao P. Blood-sucking mosquitoes of the subtribe (Diptera, Culicidae) in world fauna, vol 81. Citeseer: Princeton; 1989.
Kampen H, Kronefeld M, Zielke D, Werner D. Three rarely encountered and one new Culiseta species (Diptera: Culicidae) in Germany. J Eur Mosq Control Assoc. 2013;31:36–9.
Huff CG. Susceptibility of mosquitoes to avian malaria. Exp Parasitol. 1965;16(1):107–32.
Schoener E, Uebleis SS, Butter J, Nawratil M, Cuk C, Flechl E, Kothmayer M, Obwaller AG, Zechmeister T, Rubel F. Avian plasmodium in Eastern Austrian mosquitoes. Malar J. 2017;16(1):389.
Bisanzio D, Giacobini M, Bertolotti L, Mosca A, Balbo L, Kitron U, Vazquez-Prokopec GM. Spatio-temporal patterns of distribution of West Nile virus vectors in eastern Piedmont Region Italy. Parasit Vectors. 2011;4(1):230.
Hubálek Z, Halouzka J. West Nile fever–a reemerging mosquito-borne viral disease in Europe. Emerg Infect Dis. 1999;5(5):643.
Medlock J. Possible ecology and epidemiology of medically important mosquito-borne arboviruses in Great Britain. Epidemiol Infect. 2007;135(3):466–82.KRSnowSLeach
Rueda LM, Debboun M. Taxonomy, identification, and biology of mosquitoes. In: Debboun M, Nava MR, Rueda L, editors. Mosquitoes, communities, and public health in Texas. Amsterdam: Elsevier; 2020. p. 3–7.
Goddard J. Mosquito-borne diseases. In: Infectious diseases and arthropods. Berlin: Springer; 2018. p. 39–89.
Strand MR. The gut microbiota of mosquitoes: diversity and function. In: Wikel S, Aksoy S, Dimopoulos G, editors. Arthropod vector: controller of disease transmission, vol 1. Amsterdam: Elsevier; 2017. p. 185–99.
Ricci I, Damiani C, Capone A, DeFreece C, Rossi P, Favia G. Mosquito/microbiota interactions: from complex relationships to biotechnological perspectives. Curr Opin Microbiol. 2012;15(3):278–84.
Engel P, Moran NA. The gut microbiota of insects–diversity in structure and function. FEMS Microbiol Rev. 2013;37(5):699–735.
Ricci I, Valzano M, Ulissi U, Epis S, Cappelli A, Favia G. Symbiotic control of mosquito borne disease. Pathog Glob Health. 2012;106(7):380–5.
Ricci I, Damiani C, Rossi P, Capone A, Scuppa P, Cappelli A, Ulissi U, Mosca M, Valzano M, Epis S. Mosquito symbioses: from basic research to the paratransgenic control of mosquito-borne diseases. J Appl Entomol. 2011;135(7):487–93.
Dodson BL, Hughes GL, Paul O, Matacchiero AC, Kramer LD, Rasgon JL. Wolbachia enhances West Nile virus (WNV) infection in the mosquito Culex tarsalis. PLoS Negl Trop Dis. 2014;8(7):e2965.
Soltani A, Vatandoost H, Oshaghi MA, Enayati AA, Chavshin AR. The role of midgut symbiotic bacteria in resistance of Anopheles stephensi (Diptera: Culicidae) to organophosphate insecticides. Pathog Global Health. 2017;111(6):289–96.
Coutinho-Abreu IV, Zhu KY, Ramalho-Ortigao M. Transgenesis and paratransgenesis to control insect-borne diseases: current status and future challenges. Parasitol Int. 2010;59(1):1–8.
Miller T. Paratransgenesis as a potential tool for pest control: review of applied arthropod symbiosis. J Appl Entomol. 2011;135(7):474–8.
Wilke ABB, Marrelli MT. Paratransgenesis: a promising new strategy for mosquito vector control. Parasites Vectors. 2015;8(1):342.
Braendle C, Miura T, Bickel R, Shingleton AW, Kambhampati S, Stern DL. Developmental origin and evolution of bacteriocytes in the aphid–Buchnera symbiosis. PLoS Biol. 2003;1(1):e21.
Koropatnick TA, Engle JT, Apicella MA, Stabb EV, Goldman WE, McFall-Ngai MJ. Microbial factor-mediated development in a host-bacterial mutualism. Science. 2004;306(5699):1186–8.
Baumann P. Biology of bacteriocyte-associated endosymbionts of plant sap-sucking insects. Annu Rev Microbiol. 2005;59:155–89.
Bäckhed F, Ley RE, Sonnenburg JL, Peterson DA. Host-bacterial mutualism in the human intestine. Science. 2005;307(5717):1915–20.JIGordon
Bandi C, Dunn AM. Inherited microorganisms, sex-specific virulence and reproductive parasitism. Trends Parasitol. 2001;17(2):88–94.
Hurst G, Jiggins FM. Male-killing bacteria in insects: mechanisms, incidence, and implications. Emerg Infect Dis. 2000;6(4):329.
Stouthamer R, Breeuwer JA. Wolbachia pipientis: microbial manipulator of arthropod reproduction. Annu Rev Microbiol. 1999;53(1):71–102.GDHurst
Piel J. A polyketide synthase-peptide synthetase gene cluster from an uncultured bacterial symbiont of Paederus beetles. Proc Natl Acad Sci USA. 2002;99(22):14002–7.
Oliver KM, Russell JA, Moran NA. Facultative bacterial symbionts in aphids confer resistance to parasitic wasps. Proc Natl Acad Sci USA. 2003;100(4):1803–7.MSHunter
MacDonald TT, Monteleone G. Immunity, inflammation, and allergy in the gut. Science. 2005;307(5717):1920–5.
Wang S, Jacobs-Lorena M. Paratransgenesis applications: fighting malaria with engineered mosquito symbiotic bacteria. In: Wikel S, Aksoy S, Dimopoulos G, editors. Arthropod vector: controller of disease transmission, vol 1. Amsterdam: Elsevier; 2017. p. 219–34.
Chavshin AR, Oshaghi MA, Vatandoost H, Pourmand MR, Raeisi A, Enayati AA, Mardani N, Ghoorchian S. Identification of bacterial microflora in the midgut of the larvae and adult of wild caught Anopheles stephensi: a step toward finding suitable paratransgenesis candidates. Acta Trop. 2012;121(2):129–34.
Bozorg-Omid F, Oshaghi MA, Vahedi M, Karimian F, Seyyed-Zadeh SJ, Chavshin AR. Wolbachia infection in West Nile Virus vectors of northwest Iran. Appl Entomol Zool. 2020;55(1):105–13.
Djadid ND, Jazayeri H, Raz A, Favia G, Ricci I, Zakeri S. Identification of the midgut microbiota of An. stephensi and An. maculipennis for their application as a paratransgenic tool against malaria. PLoS ONE. 2011;6(12):e28484.
Gusmão DS, Santos AV, Marini DC, Bacci M Jr, Berbert-Molina MA, Lemos FJA. Culture-dependent and culture-independent characterization of microorganisms associated with Aedes aegypti (Diptera: Culicidae)(L.) and dynamics of bacterial colonization in the midgut. Acta Trop. 2010;115(3):275–81.
Rami A, Raz A, Zakeri S, Djadid ND. Isolation and identification of Asaia sp. in Anopheles spp. mosquitoes collected from Iranian malaria settings: steps toward applying paratransgenic tools against malaria. Parasites Vectors. 2018;11(1):367.
Rani A, Sharma A, Rajagopal R, Adak T, Bhatnagar RK. Bacterial diversity analysis of larvae and adult midgut microflora using culture-dependent and culture-independent methods in lab-reared and field-collected Anopheles stephensi—an Asian malarial vector. BMC Microbiol. 2009;9(1):96.
Chavshin AR, Oshaghi MA, Vatandoost H, Pourmand MR, Raeisi A, Terenius O. Isolation and identification of culturable bacteria from wild Anopheles culicifacies, a first step in a paratransgenesis approach. Parasites Vectors. 2014;7(1):419.
Lindh JM, Terenius O, Faye I. 16S rRNA gene-based identification of midgut bacteria from field-caught Anopheles gambiae sensu lato and A. funestus mosquitoes reveals new species related to known insect symbionts. Appl Environ Microbiol. 2005;71(11):7217–23.
Terenius O, De Oliveira CD, Pinheiro WD, Tadei WP, James AA, Marinotti O. 16S rRNA gene sequences from bacteria associated with adult Anopheles darlingi (Diptera: Culicidae) mosquitoes. J Med Entomol. 2008;45(1):172–5.
Terenius O, Lindh JM, Eriksson-Gonzales K, Bussière L, Laugen AT, Bergquist H, Titanji K, Faye I. Midgut bacterial dynamics in Aedes aegypti. FEMS Microbiol Ecol. 2012;80(3):556–65.
Amini M, Hanafi-Bojd AA, Asghari S, Chavshin AR. The potential of West Nile virus transmission regarding the environmental factors using geographic information system (GIS), West Azerbaijan province, Iran. J Arthropod Borne Dis. 2019;13(1):27.
Bagheri M, Terenius O, Oshaghi MA, Motazakker M, Asgari S, Dabiri F, Vatandoost H, Mohammadi Bavani M, Chavshin AR. West Nile virus in mosquitoes of Iranian wetlands. Vector Borne Zoonotic Dis. 2015;15(12):750–4.
Khoshdel-Nezamiha F, Vatandoost H, Azari-Hamidian S, Bavani MM, Dabiri F, Entezar-Mahdi R, Chavshin AR. Fauna and larval habitats of mosquitoes (Diptera: Culicidae) of West Azerbaijan Province, northwestern Iran. Journal of arthropod-borne diseases. 2014;8(2):163.
Khoshdel-Nezamiha F, Vatandoost H, Oshaghi MA, Azari-Hamidian S, Mianroodi RA, Dabiri F, Bagheri M, Terenius O, Chavshin AR. Molecular characterization of mosquitoes (Diptera: Culicidae) in Northwestern Iran by using rDNA-ITS2. Jap J Infect Dis. 2016;69(4):319–22.
Silver JB. Mosquito ecology: field sampling methods. Berlin: Springer; 2008.
Azari-Hamidian S, Harbach RE. Keys to the adult females and fourth-instar larvae of the mosquitoes of Iran (Diptera: Culicidae). Zootaxa. 2009;2078:1–33.
Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991;173(2):697–703.
Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33(7):1870–4.
Tamura K. Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G+ C-content biases. Mol Biol Evol. 1992;9(4):678–87.
Moro CV, Tran FH, Raharimalala FN, Ravelonandro P, Mavingui P. Diversity of culturable bacteria including Pantoea in wild mosquito Aedes albopictus. BMC Microbiol. 2013;13(1):70.
Chandel K, Mendki MJ, Parikh RY, Kulkarni G, Tikar SN, Sukumaran D, Prakash S, Parashar BD, Shouche YS, Veer V. Midgut microbial community of Culex quinquefasciatus mosquito populations from India. PLoS ONE. 2013;8(11):e80453.
Pidiyar VJ, Jangid K, Patole MS, Shouche YS. Studies on cultured and uncultured microbiota of wild Culex quinquefasciatus mosquito midgut based on 16s ribosomal RNA gene analysis. Am J Trop Med Hyg. 2004;70(6):597–603.
Soleimani JM, Moosa-Kazemi SH, Vatandoost H, Shirazi MH, Hajikhani S, Bakhtiari R, Hydarzade S. Bacterial flora of the Anopheles fluviatilis SL, the vector of malaria in Southern Iran for proper candidate paratransgenesis. Int J Curr Microbiol App Sci. 2017;6(6):3275–85.
de O Gaio A, Gusmão DS, Santos AV, Berbert-Molina MA, Pimenta PF, Lemos FJ. Contribution of midgut bacteria to blood digestion and egg production in Aedes aegypti (Diptera: Culicidae)(L.). Parasites Vectors. 2011;4(1):105.
Van Arnam EB, Currie CR, Clardy J. Defense contracts: molecular protection in insect-microbe symbioses. Chem Soc Rev. 2018;47(5):1638–51.
Chavshin AR, Oshaghi MA, Vatandoost H, Yakhchali B, Zarenejad F, Terenius O. Malpighian tubules are important determinants of Pseudomonas transstadial transmission and longtime persistence in Anopheles stephensi. Parasites & vectors. 2015;8(1):36.
Raharimalala FN, Boukraa S, Bawin T, Boyer S, Francis F. Molecular detection of six (endo-) symbiotic bacteria in Belgian mosquitoes: first step towards the selection of appropriate paratransgenesis candidates. Parasitol Res. 2016;115(4):1391–9.
Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987;4(4):406–25.
Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985;39(4):783–91.
Tamura K, Nei M, Kumar S. Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci. 2004;101(30):11030–5.
This article is part of the results of the first author’s dissertation for fulfillment of MSc degree in Medical Entomology and Vector Control in the Department of Medical Entomology and Vector Control, School of Public Health, Urmia University of Medical Sciences, Urmia, Iran.
This study was financially supported by the Urmia University of Medical Sciences (UMSU), Urmia, Iran (Project no. 2704). The funder had no role in data collection, analysis, interpretation and presentation.
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This project was approved by the Ethics Committee of Urmia University of Medical Sciences (Ethic Committee Code: IR.UMSU.REC.1397.307).
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Additional file 1: Figure S1.
Location of West Azerbaijan Province and Urmia County and sampling localities, 1—Naz-Loo: 37.651213, 44.983285, 2—Ghahraman-Loo: 37.659869, 45.207550, and 3—Moallem: 37.546660, 45.033280 (Original basic map has been prepared from d-maps.com).
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Ghahvechi Khaligh, F., Vahedi, M. & Chavshin, A.R. Identification of symbiotic bacteria in the midgut of the medically important mosquito, Culiseta longiareolata (Diptera: Culicidae). BMC Res Notes 13, 378 (2020). https://doi.org/10.1186/s13104-020-05220-0
- 16S rRNA