Autogeny in Culiseta longiareolata (Culicidae: Diptera) mosquitoes in laboratory conditions in Iran
BMC Research Notes volume 13, Article number: 81 (2020)
Culiseta longiareolata is a cosmopolitan species and has implicated in the transmission of avian malaria, tularemia, and arboviruses. Despite the wide distribution of Cs. longiareolata in Iran, little is known about its biology and physiology. The current research was conducted to study the autogeny behavior in this potential vector. During 2018, larvae and pupae were collected from Nazloo region in Urmia City using standard methods. Mosquitoes were reared in cages and fed by 5% sugar in laboratory conditions and were then dissected in phosphate-buffered saline (PBS) under a stereo microscope.
In total, 230 adult female Cs. longiareolata mosquitoes were dissected. Egg rafts were observed in the ovary of only 10.86% unfed female mosquitoes. Autogeny behavior is a significant factor in the growth of population without a blood feeding. Therefore, it is necessary to study how autogenous reproduction affects mosquito-borne diseases.
Culicidae family is comprised of 113 genera and 3570 species  with a broad distribution throughout the world . In 1878, mosquitoes were the first arthropods officially introduced as the intermediate hosts of vertebrate parasites; however, they are now recognized as the most important arthropods affecting human health . Indeed, mosquitoes are the vector of various diseases such as Malaria, Filariasis, Yellow Fever, West Nile fever, Japanese Encephalitis, Rift Valley fever, etc. [4,5,6,7].
Typically, the development of eggs in blood-sucking insects is normally dependent on a blood meal ; nonetheless, there are some female mosquitos that can develop their eggs without a blood meal , a trait called autogeny [8, 10]. In a number of females of this group of insects, the first group of eggs and possibly the next can grow without blood feeding [11, 12]. For the growth, they often use the last larval stage food or feed on sugared salmon. This characteristic has been observed in many species of mosquitoes . Autogeny is necessary for some insect species such as Megarhinini because their semiflexible proboscis is specialized for nourishment from plants .
Culiseta longiareolata is a species of the family Culicidae, the Culicinae subfamily  and a vector of avian malaria , tularemia , and arboviruses such as West Nile fever [16,17,18]. This multivoltine, thermophilic, and ornithophilic species is distributed in Europe, Asia, and Africa, as well as in the Mediterranean Sea . It mainly develops in small water bodies, and the adults may enter houses and attack humans , though their primary hosts are birds . This mosquito species are readily distinguished from other Culiseta species , and its morphological characters include white stripes and points on legs, head, and thorax .
The purpose of this research was to investigate the autogenous reproduction in Cs. longiareolata. This result might be valuable in the survival of the vector species in the absence of a host and the vectorial capacity. Owing to the uncertainty about the influence of autogeny on the vector role of mosquito, it is essential to assess the basis of this autogeny.
Urmia, the largest city of West Azerbaijan Province, is located on a 70-mile-wide plain in a 18-km distance from Lake Urmia and situated in the latitudes of 44° 52″ 35″ S and 37° 39′ 13″ N. During the spring and summer in 2018, larvae and pupae were collected from Nazloo region (37°31′ 59.2″ N 45°02′ 54.5″ E) in Urmia by using standard dipping techniques . The collector’s name, global positioning system (GPS) coordinates of the area positioning, and date of capture were recorded. Following the sample collection of mosquitos with different larval stages from their habitats, they were transferred to the medical entomology laboratory in the Urmia University of Medical Sciences, School of Public Health (Urmia, Iran) for rearing. The laboratory temperature was set at 22–25 ℃, and room lighting was provided with a yellow light. The samples were kept for 10 h in the darkness and 14 h in the brightness. A plastic cuvette-like container with the dimensions of 45 × 25 × 5 cm was used to maintain immature stages (eggs, larvae, and pupes). More than half of the container was filled with the larval habitat water and kept in 22–25 ℃; approximately 200 larvae were placed in the container. After 2–3 days, the pupae were taken with a plastic sucker and placed in disposable glasses in cages (30 × 50 × 80 cm) until the emergence of adult mosquitoes (Fig. 1); the cages were covered with finely textured cloth. Adults were feed on a cotton impregnated with 5% sugar. The cotton was streaked with a glucose solution daily and was replaced with a new sugar solution every 3 days to prevent the growth of fungi. Pupae were identified to species level using the morphological key . The newly emerged males and females of Cs. longiareolata were then transferred to new cages for mating and dissecting in PBS, to obtain eggs under a stereo microscope. The test was repeated three times.
A total of 230 adult female Cs. longiareolata mosquitoes were used in the mating experiments in three replications (Fig. 2a). Within 15–21 days after emerging, the abdomen of mosquitoes enlarged. Each gravid mosquito was incubated in a separate cage for egg laying. Three days after incubation, eggs were laid on a cup of water (Fig. 2b) and transferred to new plastic containers. The color of eggs was grey immediately after laying, but 1–2 day(s) later, the color was changed to black or dark brown. The first instar of the larvae was emerged within 3–5 days (Fig. 2c, d). Of all the female adults, 10.86% produced eggs without a blood feeding. Interestingly, autogenous females of Cs. longiareolata had larger body size than anautogenous specimens.
There are numerous investigations related to Culicidae identification, distribution, insecticide resistance, and vector control in Iran [4, 7, 25,26,27,28,29,30]. Anopheles sacharovi experiences gonotrophic dissociation during autumn and winter in Northern Iran . Anautogeny has been reported in Culex pipiens in the central parts of the country , but there are no report on the autogeny in Cs. longiareolata in Iran. Therefore, the current study, for the first time, reports autogeny in Cs. longiareolata in the country.
Autogenous species have higher nutritional requirements than anautogenous species, in order to develop their egg batch . Temperature, environmental factors, geographical variation , seasonality , and food availability  help mosquitoes become autogenous. For instance, when larvae fed on an animal matter is more likely to be autogenous and to produce more eggs in Cs. longiareolata because of the high protein content of their food . Besides, very few populations of Cs. longiareolata are autogenous, and all the laboratory mosquito populations are anautogenous [37, 38]. In the present study, the autogeny rate was low (10.86%) in Cs. Longiareolata collected from Urmia, and larvae were gathered from a larval habitat near livestock and reared in water without any other nutrient additive. Our results regarding the larger body size of autogenous Cs. longiareolata mosquitoes confirmed that the growth of body size in Culicidae is a general phenomenon [39, 40]. However, relationship between the enhancement of the fertility and pupal mass and wing length has been documented in Aedes albopictus and Aedes geniculatus mosquitoes . Perhaps the larval nutrition is a significant factor in autogeny in Cs. longiareolata in Northwestern Iran, and the determination of physicochemical characters of larval habitat water may contribute to better understanding of Cs. longiareolata autogeny.
It is hypothesized that autogeny can affect different parameters of pathogen transmission, such as increasing the vector population and reducing the pathogen incubation period . Therefore, autogeny is an excellent survival mechanism for a particular mosquito species [9, 42, 43]. The present study showed a more extended survival period in females fed on a 5% sugar solution. The high survival rate enhances the ability of mosquitoes to transmit possible pathogens. However, autogenous species could transmit arboviruse pathogens more horizontally than anautogenous populations .
Cs. longiaerolata is one of the potential vectors of a number of vector-borne diseases such as avian malaria, bird flu, avian smallpox, and Malta fever. Urmia city is an excellent destination for many migratory birds because of its lakes and islands. The presence of migratory birds as the reservoir and Cs. longiaerolata as potential vector of infectious disease provide conditions for transmitting infectious diseases. However, their role of autogeny on transferring infectious diseases needs to be studied.
The major concern of this study is the examination of autogeny on specimens emerged from the field-collected larvae. In virtue of our limitations in insectary, we could not follow autogeny in the next generation. The current study was conducted only on specimens collected from the Nazloo region in Urmia district. The test of autogeny on samples from various regions and in large areas in the country is recommended.
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Global positioning system
Harbach R: Mosquito taxonomic inventory. 2013. http://mosquito-taxonomic-inventory.info/. Accessed 1 Feb 2020
Lane RP, Crosskey RW. Medical insects and arachnids; mosquitoes (Culicidae). London: Chapman & Hall; 2012. p. 120–221.
Mullen GR, Durden LA. Medical and veterinary entomology: Mosquitoes (Culicidae). San Diego: Academic Press; 2009. p. 207–60.
Azari-Hamidian S. Checklist of Iranian mosquitoes (Diptera: Culicidae). J Vector Ecol. 2007;32(2):235–42.
Onchuru TO, Ajamma YU, Burugu M, Kaltenpoth M, Masiga D, Villinger J. Chemical parameters and bacterial communities associated with larval habitats of Anopheles, Culex and Aedes mosquitoes (Diptera: Culicidae) in Western Kenya. Int J Trop Insect Sci. 2016;36(3):146–60.
Saidi S, Tesh R, Javadian E, Nadim A. The prevalence of human infection with West Nile virus in Iran. Iran J Public Health. 1976;5(1):8–13.
Gholizadeh S, Djadid ND, Nouroozi B, Bekmohammadi M. Molecular phylogenetic analysis of anopheles and cellia subgenus anophelines (Diptera: Culicidae) in temperate and tropical regions of Iran. Acta Trop. 2013;126(1):63–74.
Gaugler R. Medical Entomology for students: introduction to mosquitoes (Culicidae). Cambridge: Cambridge University Press; 2012. p. 1–33.
Laurence B. Autogeny in Aedes (Finlaya) togoi theobald (Diptera, Culicidae). J Insect Physiol. 1964;10(2):319–31.
Levin ML. Medical entomology for students. Emerg Infect Dis. 2014;20(8):1428.
O’Meara GF, Krasnick GJ. Dietary and genetic control of the expression of autogenous reproduction in Aedes atropalpus (Coq.) (Diptera: Culicidae). J Med Entomol. 1970;7(3):328–34.
Roubaud E. Cycle autogene d’attente et generations hivernales suractives inapparentes chez le moustique commun, culex pipiens L. C R Acad. 1929;188:735–8.
Marquardt WH. Biology of disease vectors: mosquitoes, the culicidae. San Diego: CA Academic Press; 2004. p. 95–112.
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.
Maslov AV, Ward RA, Rao P. Blood-sucking mosquitoes of the subtribe Culisetina (Diptera, Culicidae) in world fauna, vol. 81. Washington: Citeseer; 1989.
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. Parasites Vectors. 2011;4(1):230.
Hubálek Z, Halouzka J. West Nile fever–a reemerging mosquito-borne viral disease in Europe. J Emerg Infect Dis. 1999;5(5):643.
Romi R, Pontuale G, Ciufolini M, Fiorentini G, Marchi A, Nicoletti L, Cocchi M, Tamburro A. Potential vectors of West Nile virus following an equine disease outbreak in Italy. Med Vet Entomol. 2004;18(1):14–9.
Cranston P, Ramsdale C, Snow K, White G. Adults, larvae and pupae of British mosquitoes (Culicidae). Ambleside: Scientific Publication; 1987.
Maslov AV. Blood-sucking mosquitoes of the subtribe Culisetina (Diptera, Culicidae) in World Fauna. Washington D.C: Smithsonian Institution Libraries and the National Science Foundation; 1967.
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.
Becker N, Petrić D, Boase C, Lane J, Zgomba M, Dahl C, Kaiser A. Chemical control, vol. 2., Mosquitoes and their controlBerlin: Springer; 2003.
Focks DA. A review of entomological sampling methods and indicators for dengue vectors. Geneva: World Health Organization; 2004.
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):1–33.
Manouchehri A, Janbakhsh B, Rohani F. Studies on the resistance of Anopheles stephensi to malathion in Bandar Abbas, Iran. Mosq News. 1976;36(3):320–2.
Vatandoost H, Hanafi-Bojd AA. Indication of pyrethroid resistance in the main malaria vector, Anopheles stephensi from Iran. Asian Pac J Trop Dis. 2012;5(9):722–6.
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. J Arthropod-Borne Di. 2014;8(2):163.
Djadid ND, Gholizadeh S, Tafsiri E, Romi R, Gordeev M, Zakeri S. Molecular identification of Palearctic members of Anopheles maculipennis in northern Iran. Malar J. 2007;6(1):6.
Azari-Hamidian S, Norouzi B, Harbach RE. A detailed review of the mosquitoes (Diptera: Culicidae) of Iran and their medical and veterinary importance. Acta Trop. 2019;194:106–22.
Asadi Saatlou Z, Sedaghat MM, Taghilou B, Gholizadeh S. Identification of novel Glutathione S-Transferases epsilon 2 mutation in Anopheles maculipennis s.s. (Diptera: Culicidae). Heliyon. 2019;5(8):e02262.
Dehghan H, Moosa-Kazemi SH, Sadraei J, Soleimani H. The ecological aspects of Culex pipiens (Diptera: Culicidae) in central Iran. Iran J Arthropod-Borne DI. 2014;8(1):35.
Reisen WK, Milby MM. Studies on autogeny in Culex tarsalis: 3. Life table attributes of autogenous and anautogenous strains under laboratory conditions. J Am Mosquito Contr. 1987;3(4):619–25.
O’Meara GFI. Variable expressions of autogeny in three mosquito species. Invertebr Reprod. 1979;1(4):253–61.
Spadoni R, Nelson R, Reeves W. Seasonal occurrence, egg production, and blood-feeding activity of autogenous Culex tarsalis. Ann Entomol Soc Am. 1974;67(6):895–902.
Kay B, Edman J, Mottram P. Autogeny in Culex annulirostris from Australia. J Am Mosquito Contr. 1986;2(1):11–3.
Polis GA, Myers CA, Holt RD. The ecology and evolution of intraguild predation: potential competitors that eat each other. Annu Rev Ecol Evol S. 1989;20(1):297–330.
Al-Jaran TK, Katbeh-Bader AM. Laboratory studies on the biology of Culiseta longiareolata (Macquart) (Diptera: Culicidae). Aquat Insects. 2001;23(1):11–22.
Van Pletzen R, Van Der Linde TDK. Studies on the biology of Culiseta longiareolata (Macquart) (Diptera: Culicidae). Bull Entomol Res. 1981;71(1):71–9.
Clements AN. The biology of mosquitoes. Reprod Nutr Dev. 1992;1:509.
Hawley W. The effect of larval density on adult longevity of a mosquito, Aedes sierrensis: epidemiological consequences. J Anim Ecol. 1985;54:955–64.
Armbruster P, Hutchinson RA. Pupal mass and wing length as indicators of fecundity in Aedes albopictus and Aedes geniculatus (Diptera: Culicidae). J Med Entomol. 2002;39(4):699–704.
Hugo L, Kay B, Ryan P. Autogeny in Ochlerotatus vigilax (Diptera: Culicidae) from Southeast Queensland. Australia. J Med Entomol. 2003;40(6):897–902.
Tsuji N, Okazawa T, Yamamura N. Autogenous and anautogenous mosquitoes: a mathematical analysis of reproductive strategies. J Med Entomol. 1990;27(4):446–53.
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Khaligh, F.G., Naghian, A., Soltanbeiglou, S. et al. Autogeny in Culiseta longiareolata (Culicidae: Diptera) mosquitoes in laboratory conditions in Iran. BMC Res Notes 13, 81 (2020). https://doi.org/10.1186/s13104-020-04942-5
- Culiseta longiareolata