Establishing a Culex Colony from Field-Collected Eggs.

Culex larvae are well adapted to growing and developing in containers, and therefore collecting and rearing field-collected Culex to adulthood in the laboratory is relatively straightforward. What is substantially more challenging is simulating natural conditions that encourage Culex adults to mate, blood feed, and reproduce in laboratory settings. In our experience, this is the most difficult hurdle to overcome when establishing new laboratory colonies. Here, we detail how to collect Culex eggs from the field and establish a colony in the laboratory. Successfully establishing a new colony of Culex mosquitoes in the laboratory will allow researchers to evaluate physiologically, behaviorally, and ecologically relevant aspects of their biology and better understand and manage these important disease vectors.

Rearing and Maintaining a Culex Colony in the Laboratory.

After overcoming the significant obstacle of getting adult Culex mosquitoes to reproduce and blood feed in the laboratory, maintaining a laboratory colony is much more achievable. However, great care and attention to detail are still required to ensure that the larvae have adequate food without being overwhelmed by bacterial growth. Additionally, achieving the appropriate densities of larvae and pupae is essential, as overcrowding delays development, prevents pupae from successfully emerging as adults, and/or reduces adult fecundity and alters sex ratios. Finally, adult mosquitoes should have constant access to H2O and nearly constant access to sugar sources to ensure that both males and females have adequate nutrition and can produce the maximum number of offspring. Here, we describe our methods for maintaining the Buckeye strain of Culex pipiens and how other researchers might modify them to suit their specific needs.

Points to Consider When Establishing and Rearing Culex Mosquitoes in the Laboratory.

Culex mosquitoes transmit several pathogens to humans and animals, including viruses that cause West Nile fever and St. Louis encephalitis and filarial nematodes that cause canine heartworm and elephantiasis. Additionally, these mosquitoes have a cosmopolitan distribution and provide interesting models for understanding population genetics, overwintering dormancy, disease transmission, and other important and ecological questions. However, unlike Aedes mosquitoes that produce eggs that can be stored for weeks at a time, no obvious "stopping" point exists in the development of Culex mosquitoes. Therefore, these mosquitoes require nearly continuous care and attention. Here, we describe some general considerations when rearing laboratory colonies of Culex mosquitoes. We highlight different methods so that readers may choose what works best for their experimental needs and laboratory infrastructure. We hope that this information will enable additional scientists to conduct laboratory research on these important disease vectors.

Light Pollution Disrupts Seasonal Differences in the Daily Activity and Metabolic Profiles of the Northern House Mosquito, Culex pipiens.

The Northern House mosquito, Culex pipiens, is an important disease vector, and females are capable of surviving the winter in a state of overwintering diapause. This species' diapause response has been extensively studied, and recent evidence suggests that the circadian clock is involved in measuring seasonal changes in daylength to initiate the diapause response. However, differences in the circadian activity of diapausing and non-diapausing Cx. pipiens have not been thoroughly investigated. Additionally, recent findings indicate that artificial light at night (ALAN) can disrupt mosquito diapause, potentially prolonging the mosquito biting season. We compared the circadian locomotor activity of mosquitoes reared in diapause-averting, long-day conditions and diapause-inducing, short-day conditions with and without ALAN to elucidate the interplay between circadian activity, diapause, and light pollution. We also uncovered metabolic differences between mosquitoes reared under diapausing and non-diapausing photoperiods with and without ALAN by measuring the concentration of protein, fructose, glycogen, water-soluble carbohydrates, and lipids. We found that ALAN exposure altered several diapause-associated phenotypes including slightly, but not significantly, increasing activity levels in short day-reared mosquitoes; and preventing some short day-reared mosquitoes from accumulating lipids. ALAN also significantly reduced glycogen and water-soluble carbohydrate levels in long day-reared mosquitoes. Based on our findings, light pollution may decrease insect fitness by perturbing metabolism, and may also impact several phenotypes associated with insect diapause, potentially extending the mosquito biting season and preventing insects in urban environments from overwintering successfully.

New records, distribution, and updated checklists of old world Phlebotomine sand flies, with emphasis on Africa, southwest Asia, and central Asia.

This article includes new records, distribution, and updated checklist of Phlebotomine sand flies (Psychodidae, Diptera) in the Old World (Africa including West Indian Ocean Islands, Southwest Asia, and Central Asia) based on specimen collections housed in different repositories worldwide. About 124 species have primary types housed in 5 repositories including holotypes (45 species, 4 subspecies), syntypes (28 species, 3 subspecies), "types" (14 species), allotypes (10 species), paratypes (36 species, 3 subspecies), lectotypes (13 species), and cotype (5 species), mounted on 671 slides. New abbreviations were proposed for 2 subgenera in the genus Phlebotomus and 6 subgenera in the genus Sergentomyia. New country records were noted in Phlebotomus (4 species in 4 subgenera in 7 countries) and Sergentomyia (10 species in 4 subgenera in 8 countries). For species diversity in the Old World, Phlebotomus includes 92 species and 7 subspecies in 9 subgenera, while Sergentomyia includes 166 species and 16 subspecies in 12 subgenera. A total of 95 species and 7 subspecies of 2 genera (Phlebotomus and Sergentomyia) were recorded in Africa while about 26 species and 16 subspecies in Southwest Asia and Central Asia.

Records and Distribution of New World Phlebotomine Sand Flies (Psychodidae, Diptera), With Special Emphasis on Primary Types and Species Diversity.

This article includes the records and distribution of Phlebotomine sand flies (Psychodidae, Diptera) in the New World based on the specimen collections housed in 2 repositories, the US National Museum of Natural History and the Museum of Entomology, Florida State Collection of Arthropods. Approximately 128 species have primary types housed in the 2 repositories, including holotypes (47 species, 3 subspecies), "types" (7 species), allotypes (52 species, 6 subspecies), lectotypes (4 species), paratypes (93 species, 10 subspecies), and neoallotype (1 species), mounted on slides, with a total of 1,107 type slides. For species diversity, collection data from 24 countries in the sand fly database were analyzed according to the number of species present, specimen records, decade of collections, and countries where collections were conducted.