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Background Endemic outbreaks of hantaviruses pose a critical public health threat worldwide. Hantaan orthohantavirus (HTNV) causes hemorrhagic fever with renal syndrome (HFRS) in humans. Using comparative genomic analyses of partial and nearly complete sequences of HTNV from humans and rodents, we were able to localize, with limitations, the putative infection locations for HFRS patients. Partial sequences might not reflect precise phylogenetic positions over the whole-genome sequences; finer granularity of rodent sampling reflects more precisely the circulation of strains. Methods Five HFRS specimens were collected. Epidemiological surveys were conducted with the patients during hospitalization. We conducted active surveillance at suspected HFRS outbreak areas. We performed multiplex polymerase chain reaction–based next-generation sequencing to obtain the genomic sequence of HTNV from patients and rodents. The phylogeny of human- and rodent-derived HTNV was generated using the maximum likelihood method. For phylogeographic analyses, the tracing of HTNV genomes from HFRS patients was defined on the bases of epidemiological interviews, phylogenetic patterns of the viruses, and geographic locations of HTNV-positive rodents. Results The phylogeographic analyses demonstrated genetic clusters of HTNV strains from clinical specimens, with HTNV circulating in rodents at suspected sites of patient infections. Conclusions This study demonstrates a major shift in molecular epidemiological surveillance of HTNV. Active targeted surveillance was performed at sites of suspected infections, allowing the high-resolution phylogeographic analysis to reveal the site of emergence of HTNV. We posit that this novel approach will make it possible to identify infectious sources, perform disease risk assessment, and implement preparedness against vector-borne viruses.

A total of 6,255 ticks belonging to three genera and six species (Haemaphysalis flava Neumann, Haemaphysalis longicornis Neumann, Haemaphysalis phasiana Saito, Ixodes nipponensis Kitaoka & Saito, Ixodes persulcatus Schulze, and Amblyomma testudinarium Koch) collected from May-August, 2013, at four southwestern provinces in the Republic of Korea (ROK) were submitted to the Armed Forces Research Institute of Medical Sciences and assayed for selected tick-borne pathogens. One pool each of H. flava and H. phasiana was positive by PCR and sequencing for a Francisella-like endosymbiont, while all pools were negative for Francisella tularensis, the causative agent of tularemia.

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 genus, Ornithoica Rondani, and two species, Ornithoica momiyamai Kishida, and O. unicolor Speiser, are reported for the first time from the Republic of Korea. A total of six species, including two new records, of louse flies were collected from 78 birds belonging to 35 species from Seoul and Incheon Metropolitan Areas and Gyeonggi, Gangwon, Gyeongsangbuk, Jeollabuk, and Jeollanam Provinces, 2005–2010. An updated checklist of Korean hippoboscids consisting of seven genera (Hippobosca L., Icosta Speiser, Lipoptena Nitzsch, Ornithoica Rondani, Ornithoctona Speiser, Ornithomya Lattreille, and Ornithophila Rondani) and 11 species is presented, including hosts, collection records, and repositories.

In total, 22,846 (17,793 culicines and 5,053 Anopheles spp.) female mosquitoes were captured by a Mosquito Magnet trap at Daeseongdong, a small village adjacent to the military demarcation line (center of the demilitarized zone) in northern Gyeonggi Province, Republic of Korea (ROK). Culicine mosquitoes were identified to species, placed in pools of up to 30 mosquitoes each, and screened for flavivirus using a SYBR Green I-based real-time polymerase chain reaction. In total, 51/660 pools positive for flaviviruses and confirmed by DNA sequencing of the NS5 region, were positive for Japanese encephalitis virus (family Flaviviridae, genus Flavivirus, JEV) (50 Culex tritaeniorhynchus Giles and one Culex bitaeniorhynchus Giles). The JEV maximum likelihood estimations (MLEs) (estimated number of viral RNA-positive mosquitoes per 1,000) for Cx. tritaeniorhynchus and Cx. bitaeniorhynchus were 9.7 and 0.9, respectively. This is the first report of a Cx. bitaeniorhynchus positive for JEV in the ROK. JEV is a local civilian and military health threat and a significant concern for nonimmune (unvaccinated) U.S. soldiers, civilians, and family members deployed to the ROK.

We used two mitochondrial loci (nicotinamide adenine dinucleotide dehydrogenase subunit 4 and cytochrome oxidase II) and a nuclear locus (28S-D2 spacer) for a total of 1337 bp to evaluate the relationships among the four subspecies of Aedes (Finlaya) japonicus Theobald. Ae. j. japonicus was recently introduced into the United States and has been expanding rapidly. We also included in our analysis a morphologically very closely related species, Aedes (Finlaya) koreicus Edwards, as well as three more distantly related species: Aedes (Finlaya) togoi Theobald, Aedes (Finlaya) hatorii Yamada, and Aedes (Aedimorphus) vexans Meigen. We found that the four subspecies in the Ae. japonicus complex are genetically quite distinct but seem to form a monophyletic group that surprisingly also includes Ae. koreicus, suggesting the need for a taxonomic reconsideration of the group. We also found that the two southern subspecies are more closely related to each other than to any of the remaining subspecies or to Ae. koreicus and may indicate an ancient north-south split of the lineage. Considering the overlap between Ae. j. japonicus and Ae. koreicus, but the stronger association between Ae. koreicus and humans, we are surprised it also has not expanded from its original range. As a proactive reaction to this possibility, we designed and tested a DNA-based rapid assay to differentiate Ae. koreicus from some of the species with which it may be confused in the United States. These Aedes are putative vectors of several important viral encephalitides.

Soldiers from the Republic of Korea and the United States conduct armistice military operations at Twin Bridges Training Area (TBTA) located near the demilitarized zone (DMZ) and are exposed to zoonotic disease pathogens that small mammals and their potentially disease-carrying ectoparasites transmit. TBTA is a 36 km2 rural training site with small villages and various forms of agriculture along its boundary. At TBTA, rodents, insectivores, and their ectoparasites are commonly found in association with unmanaged habitats of various densities of tall grasses, herbaceous plants, shrubs, briars, and crawling vegetation. Rodents and insectivores were collected during the winter (November-December 2005 and December 2006) and early spring (March 2007), and serologically tested for the presence of scrub typhus, murine typhus, and leptospirosis antibodies. Of the six species of small mammals collected, Apodemus agrarius, the common striped field mouse and known reservoir of scrub typhus, was the most frequently collected (96.1%), followed by Crocidura lasiura (2.5%), Micromys minutus (0.5%), Myodes regulus (0.5%), Mus musculus (0.3%), and Rattus rattus (0.1%). A. agrarius (56.1%), M. musculus (66.7%), M. minutus (25%), and R. rattus (100%) were positive for scrub typhus antibodies. Only A. agrarius (14.7%) and C. lasiura (4.5%) were positive for murine typhus antibodies, whereas only A. agrarius (1.5%) was seropositive for leptospirosis. Seroprevalence rates of scrub typhus and murine typhus based on weight and sex of A. agrarius are presented.

Malaria was eradicated and the Republic of Korea (ROK) declared "malaria free" in 1979. However, in 1993, a temperate strain of vivax malaria, expressing both latent and nonlatent disease populations, re-emerged near the demilitarized zone (DMZ), rapidly spread to civilian sectors near the DMZ, and increased exponentially in ROK military, veteran, and civilian populations through 1998. Malaria among all ROK populations decreased 5-fold from a high of 4142 cases in 2000 to a low of 826 cases in 2004, before increasing again to 2180 cases by 2007. Each malaria case in the ROK is reported in the metropolitan area/province where the diagnosis is made, which may be at some distance from the area where infection occurred. Therefore, it is difficult to ascertain transmission sites since approximately 60% of vivax malaria in Korea is latent with symptoms occurring1 month to 24 months after infection. A review of case diagnosis for civilian, veteran, and military populations shows that nearly all malaria south of Gyeonggi and Gangwon Provinces is the result of veterans exposed in malaria high-risk areas along the DMZ and returning to their hometowns where they later develop malaria. Thus, malaria currently remains localized near the DMZ with limited transmission in provinces south of Seoul and has not spread throughout Korea as previously hypothesized. This report describes the reemergence of vivax malaria cases in civilian and military ROK populations and U.S. military personnel and assesses variables related to its transmission and geographic distribution.

Environmental geospatial data and adult and larval mosquito collection data for up to 106 sites throughout the Republic of Korea (ROK) were used to develop ecological niche models (ENMs) of the potential geographic distribution for eight anopheline species known to occur there. The areas predicted suitable for the Hyrcanus Group species were the most extensive for Anopheles sinensis Wiedemann, An. kleini Rueda, An. belenrae Rueda, and An. pullus Yamada, intermediate for An. sineroides Yamada, and the most restricted for An. lesteri Baisas and Hu and the non-Hyrcanus Group species An. koreicus Yamada and Watanabe and An. lindesayi Yamada. The relative vectorial importance of these species is unknown, and all, except An. koreicus and An. lindesayi, are predicted to occur widely in the northwest of the ROK where malaria transmission has been sporadic since its resurgence in 1993. Our ENMs suggest that it is unlikely that An. koreicus and An. lindesayi are vectors, but we do not document consistent geographic differentiation that might incriminate any of the other species as vectors. Because all species are predicted to occur in North Korea, we also cannot reject the hypothesis that malaria infected mosquitoes from North Korea may have been the cause of the resurgence of malaria in the ROK. Ecological differentiation of the eight species is inferred from collection locations and 34 environmental layers based on remote sensing and global climatic averages. Interspecific differences were noted, and characterizing mosquito habitats by ground-based and remote sensing methods is proposed.

Malaria is a significant health threat to U.S. combat forces that are deployed to malaria-endemic regions. From 1979, when the Republic of Korea (ROK) was declared malaria free, malaria did not present a health threat to U.S. forces deployed to Korea until the early 1990s. In 1993, a temperate strain of vivax malaria expressing both latent (long prepatent incubation periods of usually 6-18 months after infection) and nonlatent (short prepatent incubation periods30 days after infection) disease reemerged near the demilitarized zone (DMZ) and once again presented a primary health threat to U.S. military populations in the ROK. Following its reemergence, malaria rates increased dramatically through 1998 and accounted for44% of all malaria cases among U.S. Army soldiers from 1997 to 2002. More than 60% of all Korean-acquired malaria among U.S. soldiers was identified as latent malaria. Nearly 80% of all latent malaria attributed to exposure in Korea was diagnosed in the U.S. or other countries where soldiers were deployed. These data illustrate the requirement for a comprehensive malaria education program, especially for those soldiers residing or training in malaria high-risk areas, to inform soldiers and providers of the risk of developing malaria after leaving Korea.

Anopheles sinensis Wiedemann (63.3%) was the most abundant Anopheles mosquito captured at cowshed resting collections in malaria high-risk areas (northern Gyeonggi Province) near the demilitarized zone (DMZ) in Korea during 2005, followed by Anopheles kleini Rueda (24.7%) and Anopheles pullus M. Yamada (8.7%). At cowshed resting collections in malaria low-risk areas (Jeonnam and Gyeongnam provinces), An. sinensis accounted for 96.8% of all Anopheles spp. collected, followed by An. kleini Rueda (2.7%), whereas no An. pullus were collected. Three species, An. kleini (50.9%), An. pullus (29.0%), and An. sinensis (13.8%), accounted for nearly all of the 224 Anopheles spp. captured by New Jersey light trap near the DMZ. In addition, An. pullus and An. kleini captured by New Jersey light trap near the DMZ and assayed by enzyme linked immunosorbent assay for Plasmodium vivax circumsporozoite antigen concentrations were higher than An. sinensis sensu stricto (s.s.), indicating higher levels of sporozoites. In laboratory studies of four concurrent artificial membrane feedings on malaria-infected blood from patients, F1 progeny of An. kleini and An. pullus had higher infection rates (8.8 and 7.5%, respectively) than An. sinensis s.s. (4.2%). These data suggest that An. kleini and An. pullus and An. sinensis are vectors of malaria in Korea. Further studies are required to determine the role of these species in the transmission of P. vivax in the Republic of Korea.

A seroepidemiological study of selected rodent-borne diseases (hantavirus [Seoul [SEO] virus], scrub typhus [Orientia tsutsugamushi], murine typhus [Rickettsia typhi], and leptospirosis [Leptospira interrogans]), as part of the U.S. military rodent surveillance and control program, was conducted from 2001 through 2005 at Yongsan Garrison, Seoul, Republic of Korea. Rodents were collected to determine the prevalence of rodent-borne diseases at a U.S. military installation in an urban environment. A total of 1,750 rodents representing three species was collected by using baited live traps (Tomahawk), glue boards, and poison baits (dead rodents observed but not assayed). The Norway rat, Rattus norvegicus (99.8%), accounted for nearly all of the rodents captured/observed. Only three roof rats, Rattus rattus (0.2%), and one house mouse, Mus musculus (0.1%), were collected. R. norvegicus rats were the only rodents that were serologically positive for SEO virus (9.6%), scrub typhus (2.8%), murine typhus (3.8%), and leptospirosis (4.6%). One of six rodents that were positive for SEO virus by immunofluorescent antibody test was positive for SEO virus antigen by reverse transcriptase-polymerase chain reaction. Infection rates for SEO virus, scrub typhus, murine typhus, and leptospirosis varied annually. Rodents were captured from 228 (20.7%) of 1,104 total buildings in Yongsan Garrison. The Yongsan commissary had the highest annual infestation rate (22 rodents per year), followed by Commisky's Club (18 rodents per year). Annual infestation rates were high for food service facilities, which often store perishable food products outdoors for short periods of time, attracting rodent populations; refuse from these facilities provides harborage and food for rodents. The effect of rodent populations outside the boundary of Yongsan Garrison was not determined.

As part of an operational effort to control the transmission of malaria after its reemergence in the Republic of Korea, a number of U.S. military and Korean civilian health workers assessed the situation at a large training site where two cases had been transmitted. The vector mosquitoes (Anopheles sinensis) probably had a limited flight range (< 1 km) in this situation based on low numbers in permanent light traps and biting collections at the billeting area compared with collections from light traps and larval surveys where the mosquito was abundant. The threat of malaria was proven by the presence of infected local Korean residents (13 of 105 sampled). However, only a small proportion of U.S. soldiers (39 of 78 surveyed) at the training site used personal protective measures. Further efforts should include improved techniques for informing and motivating soldiers to use personal protective measures and cooperation with Korean authorities to perform vector control in civilian areas bordering the training area.