* New Zealand (NZ) Endemic (E); NZ Introduced (I); non-NZ control (C) mosquito species. † Developmental stage at time of collection. ‡ Not applicable. GPS = global positioning system.
184KRAMER AND OTHERS and adults maintained as described previously. Females were fed weekly on defibrinated rabbit blood (Hema-Resources) sweetened with 10% sucrose. Eggs were collected weekly, allowed to embryonate, and stored in a small plastic bag in a closed container containing a moist sponge for humidity. When needed, the eggs were hatched under vacuum and larvae reared in low densities.Aedes albopictus. Eggs were kindly provided in April 2007 by D. Seversen, Notre Dame University. Mosquitoes were reared and maintained in a colony, and eggs stored as described for Ae. aegypti .Culex pipiens ( USA ) . A colony was established from mosquitoes collected by M. Hutchinson in 2002 in Pennsylvania. Mosquitoes were reared and adults maintained as described previously except mosquitoes were maintained in 46 cm 3 cages. Adult females were fed weekly on defibrinated goose blood (Rockland Immunochemical) sweetened with 10% sucrose.Culex quinquefasciatus ( USA ) . Mosquito larvae were provided by D. Fonseca, Rutgers University in 2008. The colony originated from Benzon Research, Inc. (Carlisle, PA) who obtained the mosquitoes from Virginia Polytechnic Institute and State University. The adults were maintained at 26°C as described previously.Mosquito feeding preferences. Blood preference and optimum feeding time were determined before the vector competence assays for each mosquito species ( Table 2 ). A variety of defibrinated bloods were offered including guinea pig, sheep, rabbit, bovine, and goose using pledgets, sausage casing (porcine casing rinsed in the brine solution, knotted at one end, filled with the virus suspension, knotted, and warmed in a 30°C water bath), and membranes. The results ( Table 2 ) guided the subsequent feedings.Cells and viruses. Aedes albopictus cells (C6/36; ATCC CRL-1660) were grown in minimal essential medium (Invitrogen, Carlsbad, CA) supplemented with 10% fetal calf serum (Hyclone, Logan, UT), 2 mM L-glutamine, 0.3% sodium bicarbonate, 100 U of penicillin/mL, and 100 μg of streptomycin/ mL and 1 U of non-essential amino acids/mL. African Green monkey kidney cells (Vero; ATCC CC1-81) were grown in the same medium but without non-essential amino acids.The history of the 12 virus strains used in this study are listed in Table 3 . The viruses included five flaviviruses ( Flaviviridae , Flavivirus ): WNV, MVEV, DENV serotype 2 (DENV2), Yellow fever virus (YFV), and JEV; and four alphaviruses ( Togaviridae , Alphavirus ) BFV, RRV, CHIKV, and SINV. Viruses used in the vector competence assays were generally prepared on the day of use from a fresh harvest of infected C6/36 cells; however, several studies used virus thawed from a frozen stock with a known concentration of virus, as noted in Tables 4 and 5 .Vector competence assays. Peroral infection. All work with infectious blood meals was conducted in the BSL3...