William J. Sames

Prevalence of tick-borne encephalitis virus in ticks from southern Korea

The prevalence of tick-borne encephalitis virus (TBEV) in southern Korea was determined by collecting ticks using tick drags. A total of 4,077 of 6,788 ticks collected were pooled (649 pools) according to collection site, species, and developmental stage and assayed for TBEV. The TBEV protein E and NS5 gene fragments were detected using RT-nested PCR in six pools of nymphs collected from Jeju Island (2,491 ticks). The minimum field detection rates for TBEV were 0.17% and 0.14% for Haemaphysalis longicornis and Haemayphysalis flava nymphs, respectively. The 252 bp NS5 and 477 bp protein E gene amplicons were sequenced. Phylogenetic analysis showed that the NS5 and protein E genes of the Jeju strain were clustered with Western subtype (98.0% and 99.4% identity, respectively). The Western subtype of TBEV is endemic in Korea, including Jeju Island. The study of vector and zoonotic host susceptibility to TBEV is required to better understand its potential impact on public health.

Geographic distribution and ecology of potential malaria vectors in the Republic of Korea.

ABSTRACT 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.

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.

Ecological surveillance of small mammals at Firing Points 10 and 60, Gyeonggi Province, Republic of Korea, 2001–2005

Abstract Throughout Korea, small mammals are hosts to a number of disease-causing agents that pose a health threat to U.S. and Korean military forces while they conduct field-training exercises. A seasonal rodent-borne disease surveillance program was established at two firing points (FP), FP-10, and FP-60, and conducted over five years from 2001 through 2005 in response to hantavirus cases among U.S. soldiers. The ecology of these sites consisted primarily of tall grasses associated with semi-permanent and temporary water sources (drainage ditches and a small stream) and dry-land agriculture farming. Eight species of rodents and one species of insectivore were collected, including Apodemus agrarius, Micromys minutus, Mus musculus, Rattus norvegicus, Tscherskia triton, Microtus fortis, Myodes regulus, and Crocidura lasiura. The striped field mouse, A. agrarius, (primary reservoir for Hantaan virus, the causative agent of Korean hemorrhagic fever), was the most frequently collected, representing 90.6% of the 1,288 small mammals captured at both sites. Reported herein are the ecological parameters, seasonal population densities, and seasonal population characteristics associated with small mammals collected at two military training sites in the Republic of Korea.

Mosquito species distribution and larval breeding habitats with taxonomic identification of Anopheline mosquitoes in Korea

In 2005, adult and larval mosquito surveillance was conducted at selected sites in Korea to associate larval habitats with species distribution of mosquitoes of the Anopheles Hyrcanus Group (An. sinensis, An. lesteri, An. pullus, An. belenrae and An. kleini) and other mosquito species. Anopheles specimens belonging to the Anopheles Hyrcanus Group were identified to species level by molecular confirmation using the internal transcribed spacer (ITS)‐2 within nuclear ribosomal (r)DNA. A total of 6644 mosquitoes from resting and light trap collections (4451; 67.0%) and larval collections (2193; 33.0%) comprising 32 species and nine genera (Culex [11], Anopheles [8], Ochlerotatus [5], Aedes [3], Armigeres [1], Coquillettidia [1], Mansonia [1], Tripteroides [1] and Lutzia [1]) were collected. Larval habitats were characterized into 14 categories. Of a total of 4534 Anopheles spp. collected (3766 resting and light trap collections and 768 larval collections), Anopheles sinensis (3194; 70.4%) was the most frequently captured, followed by An. kleini (813; 17.9%), An. pullus (299; 6.6%) and An. belenrae (129; 2.8%). Four species of Anopheles (An. lesteri, An. sineroides, An. koreicus and An. lindesayi) were infrequently collected (<3.0%) at all sites surveyed by all methods of collection. Anopheles kleini, An. pullus and An. belenrae were collected in greater proportions in malaria high‐risk areas north of Seoul, and were infrequently collected in other parts of Korea, where An. sinensis was the predominant Anopheles spp. captured. A total of 2110 culicine mosquitoes (685 adult collections and 1425 larval collections) comprising 24 species and eight genera were collected.

Seasonal Distribution of Ticks in Four Habitats near the Demilitarized Zone, Gyeonggi-do (Province), Republic of Korea

This study describes the seasonal distribution of larvae, nymph, and adult life stages for 3 species of ixodid ticks collected by tick drag and sweep methods from various habitats in the Republic of Korea (ROK). Grasses less than 0.5 m in height, including herbaceous and crawling vegetation, and deciduous, conifer, and mixed forests with abundant leaf/needle litter were surveyed at United States (US) and ROK operated military training sites and privately owned lands near the demilitarized zone from April-October, 2004 and 2005. Haemaphysalis longicornis Neumann adults and nymphs were more frequently collected from April-August, while those of Haemaphysalis flava Neumann and Ixodes nipponensis Kitaoka and Saito were collected more frequently from April-July and again during October. H. longicornis was the most frequently collected tick in grass habitats (98.9%), while H. flava was more frequently collected in deciduous (60.2%) and conifer (57.4%) forest habitats. While more H. flava (54.1%) were collected in mixed forest habitats than H. longicornis (35.2%), the differences were not significant. I. nipponensis was more frequently collected from conifer (mean 8.8) compared to deciduous (3.2) and mixed (2.4) forests.

Tick surveillance of small mammals captured in Gyeonggi and Gangwon Provinces, Republic of Korea, 2004–2008

Abstract A total of 4,575 ticks were collected from 5,953 small mammals captured from March 2004 to December 2008 at 19 military training areas and 6 US military installations, Gyeonggi and Gangwon Provinces, Republic of Korea. Ixodes nipponensis (98.9%; 753 nymphs, 3,771 larvae) was the most frequently collected tick, found on six of the 11 small mammal species captured, followed by Ixodes pomerantzevi (1.1%; 17 females, 9 nymphs, 24 larvae) found on two species. While only one T. sibiricus was collected, it was infested with 44 ticks, with an infestation rate of 100.0%, followed by Rattus norvegicus (16.7%, n=18), Apodemus agrarius (14.6%, n=5,397), Crocidura lasiura (7.5%, n=265), Microtus fortis (7.3%, n=82), Myodes regulus (5.7%, n=53), and Micromys minutus (4.8%, n=63). No ticks were collected from Apodemus peninsulae (n=3), Mus musculus (n=58), Tscherskia triton (n=12), and Mogera wogura (n=1). Ixodes nipponensis nymphs were most frequently collected from small mammals from March to April, while larvae were more frequently collected during September. Ixodes pomerantzevi was collected only during February–April 2008, and a single Haemaphysalis flava nymph was collected from A. agrarius in August 2004.

Comparison of Dragging and Sweeping Methods for Collecting Ticks and Determining Their Seasonal Distributions for Various Habitats, Gyeonggi Province, Republic of Korea

ABSTRACT As part of the 65th Medical Brigade tick-borne disease surveillance program to determine the abundance, geographical and seasonal distributions, and tick-borne pathogens present in the Republic of Korea, dragging and sweeping methods were compared to determine their efficiency for collecting ticks in grass and deciduous, conifer, and mixed forest habitats at military training sites and privately owned lands in northern Gyeonggi Province near the demilitarized zone from April-October, 2004–2005. Three species of Ixodid ticks, Haemaphysalis longicornis, Haemaphysalis flava, and Ixodes nipponensis, were collected. Overall, H. longicornis adults and nymphs were most frequently collected from grass and deciduous forest habitats, accounting for 98.2 and 66.2%, respectively, of all ticks collected. H. flava adults and nymphs were most frequently collected from conifer and mixed forests, accounting for 81.6, and 77.8%, respectively, of all ticks collected. I. nipponensis adults and nymphs accounted for 9.3% of all ticks collected from mixed forests, were less commonly collected from deciduous (4.1%) and conifer (4.1%) forests, and infrequently collected from grass habitats (0.9%). Overall, there were no significant differences between dragging and sweeping methods for the three species when the areas sampled were similar (sweeping = 2 × the area over the same transect). Adults and nymphs of H. longicornis were most commonly collected from April-August, while those of H. flava and I. nipponensis were most commonly collected during April-July and again during October. Larvae of all three species were most frequently observed from July-September.

Detection of Rickettsia felis and Rickettsia typhi and Seasonal Prevalence of Fleas Collected from Small Mammals at Gyeonggi Province in the Republic of Korea

Fleas were collected from live-captured small mammals to identify flea-borne pathogens, host associations, and seasonal prevalence of flea species, as part of the 65th Medical Brigade rodent-borne disease surveillance program at 20 military installations and training sites, Gyeonggi Province, Republic of Korea, 2005-2007. A total of 1251 fleas were recovered from 2833 small mammals. Apodemus agrarius, the striped field mouse, accounted for 93.1% (2,637/2,833) of all small mammals captured, followed by Crocidura lasiura (3.1%), Mus musculus (1.3%), Microtus fortis (0.7%), Myodes regulus (0.7%), Micromys minutus (0.5%), Rattus norvegicus (0.4%), Tscherskia triton (0.1%), Apodemus peninsulae (< 0.1%), Rattus rattus (< 0.1%), and Mogera robusta (< 0.1%). A total of 6/11 species of mammals captured were infested with fleas with infestation rates ranging from a high of 26.3% (A. agrarius and M. regulus) to a low of 5.3% (M. fortis). Flea indices among infested mammals were highest for R. norvegicus (2.50), followed by C. lasiura (2.20), A. agrarius (1.71), M. regulus (1.20), M. musculus (1.0), and M. fortis (1.0). The predominant flea species collected were Stenoponia sidimi (56.5%), followed by Ctenophthalmus congeneroides (38.3%) and Rhadinopsylla insolita (3.9%). The minimum field infection rates [number of positive pools/total number of fleas (600)] for Rickettsia typhi and for Rickettsia felis were 1.7% and 1.0%, respectively.

Ecology of Hantaan virus at Twin Bridges Training Area, Gyeonggi Province, Republic of Korea, 2005–2007

ABSTRACT: The Twin Bridges Training Area (TBTA) in the Republic of Korea consists of dirt roads, barren training areas, and forested hillsides adjacent to linear and broad expanses of tall grasses, herbaceous, and scrub vegetation. Of the six species of small mammals, the striped field mouse, Apodemus agrarius, was the most frequently captured (96.1%). Apodemus agrarius capture rates varied from 17.7 to 33.2% during three trapping periods. Gravid females were observed during November–December 2006 (8.4%) and March 2007 (5.1%). In 2005, the overall seroprevalence of Hantaan virus (HTNV) was high (34.4%) and lower during surveys in 2006 (14.2%) and 2007 (13.8%). Seroprevalence was directly correlated with weight increase of A. agrarius.