Heung-Chul Kim

Characterization of Imjin Virus, a Newly Isolated Hantavirus from the Ussuri White-Toothed Shrew (Crocidura lasiura)

ABSTRACT Until recently, the single known exception to the rodent-hantavirus association was Thottapalayam virus (TPMV), a long-unclassified virus isolated from the Asian house shrew (Suncus murinus). Robust gene amplification techniques have now uncovered several genetically distinct hantaviruses from shrews in widely separated geographic regions. Here, we report the characterization of a newly identified hantavirus, designated Imjin virus (MJNV), isolated from the lung tissues of Ussuri white-toothed shrews of the species Crocidura lasiura (order Soricomorpha, family Soricidae, subfamily Crocidurinae) captured near the demilitarized zone in the Republic of Korea during 2004 and 2005. Seasonal trapping revealed the highest prevalence of MJNV infection during the autumn, with evidence of infected shrews clustering in distinct foci. Also, marked male predominance among anti-MJNV immunoglobulin G antibody-positive Ussuri shrews was found, whereas the male-to-female ratio among seronegative Ussuri shrews was near 1. Plaque reduction neutralization tests showed no cross neutralization for MJNV and rodent-borne hantaviruses but one-way cross neutralization for MJNV and TPMV. The nucleotide and deduced amino acid sequences for the different MJNV genomic segments revealed nearly the same calculated distances from hantaviruses harbored by rodents in the subfamilies Murinae, Arvicolinae, Neotominae, and Sigmodontinae. Phylogenetic analyses of full-length S, M, and L segment sequences demonstrated that MJNV shared a common ancestry with TPMV and remained in a distinct out-group, suggesting early evolutionary divergence. Studies are in progress to determine if MJNV is pathogenic for humans.

Anopheles kleini, Anopheles pullus, and Anopheles sinensis: Potential Vectors of Plasmodium vivax in the Republic of Korea

Abstract 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.

Laboratory Transmission of Japanese Encephalitis, West Nile, and Getah Viruses by Mosquitoes (Diptera: Culicidae) Collected near Camp Greaves, Gyeonggi Province, Republic of Korea, 2003

Abstract We conducted experimental studies to evaluate mosquitoes captured in Paju County, Gyeonggi Province, Republic of Korea, for their ability to transmit West Nile virus (family Flaviviridae, genus Flavivirus, WNV), Japanese encephalitis virus (family Flaviviridae, genus Flavivirus, JEV), and Getah virus (family Togaviridae, genus Alphavirus, GETV) under laboratory conditions. Both Culex pipiens pallens Coquillett and Culex tritaeniorhynchus Giles were highly susceptible to infection with WNV, with infection rates >65% when allowed to feed on chickens with viremias of ≈107 plaque-forming units (PFU) of virus/ml blood. In contrast, Cx. tritaeniorhynchus were significantly more susceptible to JEV or GETV (infection rate 100%) than were the Cx. p. pallens (infection rate 3% for JEV and 0% for GETV) captured in the same area when allowed to feed on chickens with viremias of ≈105 PFU of virus/ml blood. The detection of JEV in field-collected Cx. tritaeniorhynchus in Gyeonggi Province in 2000 and the demonstrated ability of this species to transmit this virus support the importance of the continued vaccination of Koreans against JEV and indicate a risk of infection for nonvaccinated individuals.

Malaria in the Republic of Korea, 1993-2007. Variables Related to Re-emergence and Persistence of Plasmodium vivax Among Korean Populations and U.S. Forces in Korea

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 occurring >1 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.

Plasmodium vivax malaria among U.S. forces Korea in the Republic of Korea, 1993-2007.

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 periods < 30 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 for > 44% 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.

Isolation and genomic characterization of Chaoyang virus strain ROK144 from Aedes vexans nipponii from the Republic of Korea.

During June 2003, mosquito surveillance was conducted at a US Army installation and a US Military training site 2 km south of the demilitarized zone, Republic of Korea. Mosquitoes were collected using Mosquito Magnets™, sorted to species, and assayed for the presence of arboviruses. From the 3,149 mosquitoes that were sorted into 126 pools, one Aedes vexans nipponii pool (out of 73 pools) tested positive for flavivirus RNA by reverse transcription-PCR. After isolation from C6/36 cell culture supernatant, the viral genome was sequenced and found to be 98.9% related to Chaoyang virus, a potential arthropod-specific flavivirus. This report details the first identification of Chaoyang virus in the Republic of Korea and highlights its relationship to other flaviviruses.

Seroepidemiological survey of rodents collected at a U.S. military installation, yongsan garrison, Seoul, 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 Commiskys 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.

Overwintering of Anopheles lindesayi japonicus Larvae in the Republic of Korea

Abstract Mosquito larval surveillance for environmental monitoring and pest-control purposes was conducted monthly at dredged soil-dumping areas during the construction of a new harbor in Yongcheon Bay, approximately 5 km SE of Jinhae, on the SW side of Namsan (Mt. Nam) and across the bay from Su-do (Su Island) in Gyeongsangnam Province, Republic of Korea (ROK) from November 2007 through April 2008. During this study, mosquitoes collected as overwintering larvae were Aedes togoi in brackish rock pools along the seashore and Anopheles lindesayi japonicus along the vegetated margins of a slow-flowing drainage ditch and associated freshwater ground pools containing green algae. Overwintering An. lindesayi larvae also were collected along stream margins and stream pools of moderate- to fast-flowing mountain streams near Chungju (Chungcheongbuk Province) (October 2007 and March 2008) and Munsan (Gyeonggi Province) (September 2007 and April 2008). First and second instars were collected and identified in late September 2007 through February 2008. During March and April, collections were primarily 3rd and 4th instars, and by the end of April, pupae were collected. This is the first report of An. lindesayi japonicus overwintering as larvae in the ROK.

Surveillance of vivax malaria vectors and civilian patients for malaria high-risk areas in northern Gyeonggi and Gangwon Provinces near the demilitarized zone, Republic of Korea, 2003–2006

After re‐emergence of malaria in 1993, a continued increase in Plasmodium vivax cases was observed from 1993 to 2006 in northern Gyeonggi and Gangwon Provinces adjacent to the demilitarized zone separating North from South Korea. Annual parasite incidence per 1000 people ranged from 0.33 in 2004 to 0.89 in 2006. While malaria case rates declined (22.6%) in 2004, they increased 75.1% in 2005 and 51.7% in 2006 from the previous years. An initial incorrect diagnosis of 46.8% of malaria cases as common cold resulted in a mean delay of 1.3 days for the detection malarial parasites. Of the total cases, 10.2% from December to May were due to latent intrinsic incubation infections acquired the previous malaria season and the rest of the cases from June to November were either latent or short incubation infections. Overall, the peak anopheline population occurred from July to September, resulting in a similar peak in malaria cases. While malaria cases increased during 2005–2006, anopheline populations, based on trap indices, were not significantly different during 4 years of surveillance. To decrease the malaria patient infective period to mosquitoes, public health centers in Paju and Cheorwon in 2006 prescribed chloroquine + primaquine at days 0–3 after initial malaria diagnosis followed by an additional 11 days of primaquine (early primaquine treatment), rather than chloroquine on days 0–3 and primaquine on days 4–17 (delayed primaquine treatment). A reduction in the malaria parasite incidence during 2007 was recorded for the two locations offering the early primaquine treatment relative to other locations using the delayed primaquine treatment.

Association of Temperature and Historical Dynamics of Malaria in the Republic of Korea, Including Reemergence in 1993

Plasmodium vivax malaria reemerged in the Republic of Korea in 1993 after it had been declared malaria free in 1979. Malaria rapidly increased and peaked in 2000 with 4,142 cases with lower but variable numbers of cases reported through 2011. We examined the association of regional climate trends over the Korean Peninsula relative to malaria cases in U.S. military and Republic of Korea soldiers, veterans, and civilians from 1950 to 2011. Temperatures and anomaly trends in air temperature associated with satellite remotely sensed outgoing long-wave radiation were used to observe temporal changes. These changes, particularly increasing air temperatures, in combination with moderate rains throughout the malaria season, and distribution of malaria vectors, likely supported the 1993 reemergence and peaks in malaria incidence that occurred through 2011 by accelerating the rate of parasite development in mosquitoes and increased numbers as a result of an expansion of larval habitat, thereby increasing the vectorial capacity of Anopheles vectors. High malaria rates associated with a favorable climate were similarly observed during the Korean War. These findings support the need for increased investigations into malaria predictive models using climate-related variables.