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In total, 183,602 female culicine mosquitoes were captured by Mosquito Magnet, black light, and New Jersey light traps, and manual aspiration of resting blood-fed mosquitoes, in the Republic of Korea from 2008 to 2011. Culicine mosquitoes were identified to species, placed in pools of up to 30 mosquitoes each, and screened for flavivirus RNA by using an SYBR green I-based reverse transcription-polymerase chain reaction assay. Thirty-two of the 8,199 pools assayed were positive by quantitative polymerase chain reaction for Chaoyang virus (CHAOV), an insect-specific virus [26 Aedes vexans nipponii Theobald, 3 Culex pipiens L., 1 Aedes albopictus (Skuse), 1 Aedes bekkui Mogi, and 1 Armigeres subalbatus (Coquillett)]. The maximum likelihood estimations (estimated number of virus-positive mosquitoes/1,000 mosquitoes) for Ae. bekkui, Ae. albopictus, Ar. subalbatus, Ae. vexans nipponii, and Cx. pipiens positive for CHAOV were 5.37, 3.29, 0.77, 0.27, and 0.26, respectively. CHAOV is an insect-specific virus, and there is currently no evidence to suggest a role in animal or human disease.

One of the most significant modern day efforts to prevent and control an arthropod-borne disease during a military deployment occurred when a team of U.S. military entomologists led efforts to characterize, prevent, and control leishmaniasis at Tallil Air Base (TAB), Iraq, during Operation Iraqi Freedom. Soon after arriving at TAB on 22 March 2003, military entomologists determined that 1) high numbers of sand flies were present at TAB, 2) individual soldiers were receiving many sand fly bites in a single night, and 3) Leishmania parasites were present in 1.5% of the female sand flies as determined using a real-time (fluorogenic) Leishmania-generic polymerase chain reaction assay. The rapid determination that leishmaniasis was a specific threat in this area allowed for the establishment of a comprehensive Leishmaniasis Control Program (LCP) over 5 mo before the first case of leishmaniasis was confirmed in a U.S. soldier deployed to Iraq. The LCP had four components: 1) risk assessment, 2) enhancement of use of personal protective measures by all personnel at TAB, 3) vector and reservoir control, and 4) education of military personnel about sand flies and leishmaniasis. The establishment of the LCP at TAB before the onset of any human disease conclusively demonstrated that entomologists can play a critical role during military deployments.

Selected species of mosquitoes and Ornithodoros ticks were evaluated for their potential to transmit Karshi and Langat (tick-borne encephalitis virus complex) viruses in the laboratory. Although there was no evidence of replication of Karshi virus in either of the two mosquito species tested [Ochlerotatus taeniorhynchus (Wiedemann) or Culex pipiens (L.)], Karshi virus replicated in and was transmitted by all three species of Ornithodoros ticks tested (Ornithodoros parkeri Cooley, Ornithodoros sonrai SautetWitkowski, and Ornithodoros tartakovskyi Olenev). When inoculated with Karshi virus, 90% of Ornithodoros ticks (44/49) transmitted this virus by bite to suckling mice, and transmission continued to occur for at least 1 yr, the longest extrinsic incubation tested. After feeding on a suckling mouse with a viremia of approximately 10(5) suckling mouse subcutaneous lethal dose. units of Karshi virus per milliliter of blood, all three species of Ornithodoros tested became infected with and transmitted Karshi virus both trans-stadially and horizontally by bite to suckling mice. In addition, female O. tartakovskyi transmitted Karshi virus vertically to their progeny. In a continuation of a previous study, O. sonrai, orally exposed to Langat virus, were able to transmit this virus after3 yr, the longest interval tested. Therefore, Ornithodoros spp. should be considered as potential vectors and as possible long-term maintenance hosts for Karshi virus and other members of the tick-borne encephalitis virus complex.

Sand flies collected between April 2003 and November 2004 at Tallil Air Base, Iraq, were evaluated for the presence of Leishmania parasites using a combination of a real-time Leishmania-generic polymerase chain reaction (PCR) assay and sequencing of a 360-bp fragment of the glucose-6-phosphate-isomerase (GPI) gene. A total of 2,505 pools containing 26,574 sand flies were tested using the real-time PCR assay. Leishmania DNA was initially detected in 536 pools; however, after extensive retesting with the real-time PCR assay, a total of 456 pools were considered positive and 80 were considered indeterminate. A total of 532 samples were evaluated for Leishmania GPI by sequencing, to include 439 PCR-positive samples, 80 PCR-indeterminate samples, and 13 PCR-negative samples. Leishmania GPI was detected in 284 samples that were sequenced, to include 281 (64%) of the PCR-positive samples and 3 (4%) of the PCR-indeterminate samples. Of the 284 sequences identified as Leishmania, 261 (91.9%) were L. tarentolae, 18 (6.3%) were L. donovani-complex parasites, 3 (1.1%) were L. tropica, and 2 were similar to both L. major and L. tropica. Minimum field infection rates were 0.09% for L. donovani-complex parasites, 0.02% for L. tropica, and 0.01% for the L. major/tropica-like parasite. Subsequent sequencing of a 600-bp region of the "Hyper" gene of 12 of the L. donovani-complex parasites showed that all 12 parasites were L. infantum. These data suggest that L. infantum was the primary leishmanial threat to U.S. military personnel deployed to Tallil Air Base. The implications of these findings are discussed.

We evaluated the molestus form of Culex pipiens pipiens (L.) (hereafter referred to as "molestus") captured near Tashkent, Uzbekistan, for their ability to transmit Japanese encephalitis (family Flaviviridae, genus Flavivirus, JEV) and West Nile (family Flaviviridae, genus Flavivirus, WNV) viruses under laboratory conditions. These molestus were highly competent laboratory vectors of WNV, with infection and dissemination rates of 96 and 81%, respectively. Approximately 75% of female molestus that fed after development of a disseminated infection transmitted virus by bite. Therefore, approximately 60% of those molestus taking a second bloodmeal between 16 and 25 d after an infectious bloodmeal would be expected to transmit WNV by bite. In contrast, these molestus were less efficient vectors of JEV, with infection and dissemination rates of 51 and 25%, respectively. In addition, only 33% of individuals with a disseminated infection transmitted JEV by bite, indicating a significant salivary gland barrier. Therefore, only approximately 8% of orally exposed individuals would be expected to transmit JEV by bite if they took a second bloodmeal 16-25 d later. These data indicate that the molestus form of Cx. p. pipiens should be considered a potentially important vector of WNV in Uzbekistan and may become involved in the transmission of JEV, should this virus be introduced into Uzbekistan.