The Comparative Field Evaluation of Four Different Traps for Mosquito Surveillance in the Republic of Korea.

Monitoring mosquito populations is essential for controlling mosquito-borne diseases, and the selection of mosquito traps should be tailored to specific surveillance objectives. Here, we tested four mosquito traps for their efficiency and applicability: the Nozawa-style black light trap (BLT), BG-sentinel trap II (BGT), UV-LED Blackhole Plus Mosquito Buster trap (LED), and digital mosquito monitoring system (DMS). The traps were rotated weekly for a 24 h cycle at the same location for 13 weeks. Overall, 1649 female mosquitoes belonging to seven genera and sixteen species were collected by the traps. The traps exhibited differences in both the number of collected individuals and species composition. The BLT showed superior collection efficiency in terms of the number of collected individuals and species evenness, whereas the BGT showed the highest species diversity among all the traps. Thus, the BLT and BGT are the best choices for effective mosquito surveillance based on trap performance. Additionally, despite the relatively low efficiency of the LED and DMS observed in this study, the LED is known to be efficient when used for indoor conditions such as cowsheds, while the DMS has an advanced function that can automatically count the number of mosquitoes. Thus, our findings provide significant guidelines for planning new mosquito surveillance projects in the ROK.

Molecular Identification of Spotted Fever Group Rickettsiae in Ticks in the Republic of Korea.

The Rickettsia species transmitted by ticks are mostly classified within the spotted fever group rickettsiae (SFGR), which causes tick-borne rickettsiosis. Although efforts have been made to investigate their prevalence in the Republic of Korea (ROK), research has been limited to certain areas. Furthermore, the pooling method for ticks does not fully reflect the exact infection rate. Therefore, we aimed to perform molecular identification of SFGR in ticks to elucidate the current prevalence of tick-borne rickettsiosis in the ROK. The SFGR of ticks was identified using polymerase chain reaction targeting the 17 kDa antigen, ompA, and gltA, followed by sequencing for species identification and phylogenetic analysis. In total, 302 ticks belonging to four species (Haemaphysalis flava, H. longicornis, Ixodes nipponensis, and Amblyomma testudinarium) were collected between April and November 2022. The overall SFGR infection rate was 26.8% (81/302 patients). Both adult and nymphal ticks and the SFGR infection rate increased during April-May, reaching their peaks in June, followed by a marked decline in August and July, respectively. Phylogenetic analysis revealed three species (R. monacensis, R. heilongjiangensis, and Candidatus R. jingxinensis) of SFGR. Thus, our results emphasize the importance of tick surveys for the prevention and management of tick-borne rickettsiosis.

Molecular Analysis of Tick-Borne Bacterial Pathogens from Ticks Infesting Animal Hosts in Kyrgyzstan, 2021.

This study investigated the prevalence of Anaplasma and Ehrlichia in 494 engorged ticks collected from various animal hosts, including cattle, horses, sheep, chickens, dogs, and cats, in six regions of northern Kyrgyzstan. Ten tick species, belonging to two families and six genera, were identified based on CO1, 16S rRNA, and ITS2 genes: Argas persicus (26.5%), Haemaphysalis punctata (18.0%), Dermacentor spp. (16.0%), Rhipicephalus annulatus (11.8%), R. turanicus (10.9%), D. marginatus (7.7%), Hyalomma scupense (4.5%), Hy. marginatum (3.8%), R. sangineus complex (0.6%), and Ornithodoros lahorensis (0.2%). PCR analysis revealed a 15.0% (74/494) overall infection rate of Anaplasma and Ehrlichia. Anaplasma species were found in six tick species and were identified as A. bovis (n = 44), Anaplasma spp. (n = 20), A. ovis (n = 5), and A. capra (n = 2). Ehrlichia species were found only in H. punctata (n = 5) and identified as E. chaffeensis (n = 1) and Ehrlichia spp. (n = 4). Additionally, two H. punctata were co-infected with Anaplasma and Ehrlichia. This is the first study to investigate tick-borne bacterial pathogens in ticks collected from animal hosts in Kyrgyzstan. Our findings contribute to a better understanding of the epidemiology and emergence of tick-borne infections in Kyrgyzstan.

Characterization of genotype V Japanese encephalitis virus isolates from Republic of Korea.

Japanese encephalitis (JE), caused by the Japanese encephalitis virus (JEV) infection, continues to pose significant public health challenges worldwide despite efficient vaccines. The virus is classified into five genotypes, among which genotype V (GV) was not detected for a long period after its initial isolation in 1952, until reports emerged from China and the Republic of Korea (ROK) since 2009. The characteristics of the virus are crucial in estimating its potential epidemiological impact. However, characterization of GV JEVs has so far been limited to two strains: Muar, the original isolate, and XZ0934, isolated in China. Two additional ROK GV JEV isolates, NCCP 43279 and NCCP 43413, are currently available, but their characteristics have not been explored. Our phylogenetic analysis revealed that GV virus sequences from the ROK segregate into two clades. NCCP 43279 and NCCP 43413 belong to different clades and exhibit distinct in vitro phenotypes. NCCP 43279 forms larger plaques but demonstrates inefficient propagation in cell culture compared to NCCP 43413. In vivo, NCCP 43279 induces higher morbidity and mortality in mice than NCCP 43413. Notably, NCCP 43279 shows more severe blood-brain barrier damage, suggesting superior brain invasion capabilities. Consistent with its higher virulence, NCCP 43279 displays more pronounced histopathological and immunopathological outcomes. In conclusion, our study confirms that the two ROK isolates are not only classified into different clades but also exhibit distinct in vitro and in vivo characteristics.

Current Status of Trypanosoma grosi and Babesia microti in Small Mammals in the Republic of Korea.

Small mammals, such as rodents and shrews, are natural reservoir hosts of zoonotic diseases, including parasitic protozoa. To assess the risk of rodent-borne parasitic protozoa in the Republic of Korea (ROK), this study investigated the status of parasitic protozoa, namely Trypanosoma, Babesia, and Theileria, in small mammals. In total, 331 blood samples from small mammals were analyzed for parasites using PCR and sequenced. Samples were positive for Trypanosoma grosi (23.9%; n = 79) and Babesia microti (10%; n = 33) but not Theileria. Small mammals from Seogwipo-si showed the highest infection rate of T. grosi (48.4%), while the highest B. microti infection rate was observed in those from Gangneung-si (25.6%). Sequence data revealed T. grosi to be of the AKHA strain. Phylogenetic analysis of B. microti revealed the US and Kobe genotypes. B. microti US-type-infected small mammals were detected throughout the country, but the Kobe type was only detected in Seogwipo-si. To our knowledge, this is the first nationwide survey that confirmed T. grosi and B. microti infections at the species level in small mammals in the ROK and identified the Kobe type of B. microti. These results provide valuable information for further molecular epidemiological studies on these parasites.

Geographical Characteristics of Culex tritaeniorhynchus and Culex orientalis Microbiomes in Korea.

Mosquitoes, the primary vectors of arboviruses, harbor a diverse microbiome that plays a crucial role in their development, immunity, and vector competence. The composition of the mosquito microbiome is heavily influenced by the environment and habitats. Therefore, identifying the relationship between the habitat and the mosquito's microbial community can improve the overall understanding of mosquito biology. However, The microbiome profiles of Culex tritaeniorhynchus and Culex orientalis, known as transmission vectors of the Japanese encephalitis virus, are poorly understood. Using 16S rRNA Illumina sequencing, we hereby investigated the microbial profiles in these two mosquito species collected in several areas in the Republic of Korea. Thirty-six prevalent bacterial families were identified from these mosquito species. The microbial composition variations were primarily influenced by the mosquito collecting sites. Moreover, species biomarkers were identified by utilizing the regional specificity of the mosquito microbiome. Based on the microbiome profiles representing high similarity, Culex orientalis may share an ecological niche with Culex tritaeniorhynchus.

Molecular Prevalence of Blastocystis sp. from Patients with Diarrhea in the Republic of Korea.

Blastocystis sp. is the most common intestinal protozoan affecting human health worldwide. Several studies have reported the prevalence of Blastocystis sp. in various regions of the Republic of Korea. However, limited data are available on the prevalence and subtype (ST) distribution of this parasite among regions. Therefore, we investigated the prevalence and ST distributions of this parasite in the Republic of Korea. For this purpose, 894 stool specimens were collected from patients with diarrhea and tested for the presence of Blastocystis sp. using PCR analysis. The isolates were subsequently subtyped. The overall prevalence was 11.6%. Of the 104 isolates, ST3 was the most prevalent, followed by ST1. Additionally, a single case of the rare subtype ST8 was identified, representing the first reported case in the Republic of Korea. The results suggested that the predominance of ST3 observed in this study reflects human-to-human transmission with low genetic diversity within the ST, while ST1 transmission is likely correlated with animals. In the future, to better understand Blastocystis sp. transmission dynamics, human, animal, and environmental factors should be studied from a "One Health" perspective.

Molecular Detection and Phylogenetic Analysis of Tick-Borne Encephalitis Virus from Ticks Collected from Cattle in Kyrgyzstan, 2023.

Ticks are important vectors of the tick-borne encephalitis virus (TBEV). In Kyrgyzstan, the livestock farming trade and nomadic lifestyle enable tick-borne diseases to be imported from neighboring countries, but there are few relevant studies. In this study, we collected 40 ticks from cattle in Kyrgyzstan. Molecular marker analysis identified the ticks as Ixodes persulcatus (97.5%; n = 39) and Haemaphysalis punctata (2.5%; n = 1). Real-time PCR screening revealed two ticks to be positive for TBEV, but only one tick was amplified using nested PCR targeting the TBEV envelope (E) and non-structure 5 (NS5) gene. The obtained sequences belonged to the TBEV Siberian subtype and phylogenetic tree analysis results confirmed that the virus was related to the Bosnia strain. We also performed next-generation sequencing, which confirmed the TBEV Siberian subtype. Continuous research and surveillance of TBEV in Kyrgyzstan are required to provide further information on tick-borne diseases.

Molecular Detection of Anaplasma, Ehrlichia and Rickettsia Pathogens in Ticks Collected from Humans in the Republic of Korea, 2021.

Tick-borne pathogens (TBPs), transmitted by the bites of ticks, are of great medical and veterinary importance. They include bacteria, viruses, and protozoan parasites. To provide fundamental data on the risk of tick contact and public health strategies, we aimed to perform a molecular investigation on four tick-borne bacterial pathogens in ticks collected from humans across the Republic of Korea (ROK) in 2021. In total, 117 ticks were collected, including Haemaphysalis longicornis (56.4%), Amblyomma testudinarium (26.5%), Ixodes nipponensis (8.5%), H. flava (5.1%), and I. persulcatus (0.9%). Among the ticks, 20.5% (24/117) contained tick-borne bacterial pathogens, with infection rates of 17.9% for Rickettsia (Candidatus Rickettsia jingxinensis, R. tamurae, R. monacensis, and Candidatus Rickettsia tarasevichiae), 2.5% for Anaplasma (A. phagocytophilum, A. capra, and A. bovis), and 0.9% for Ehrlichia (Ehrlichia sp.). Additionally, the co-detection rate for R. monacensis and A. phagocytophilum was 0.9%. To our knowledge, this is the first report of A. capra and A. bovis detection in ticks collected from humans in the ROK. This study contributes to the understanding of the potential risk of tick contact and provides fundamental data for establishing a public health strategy for tick-borne disease management in the ROK.

The relationships between microbiome diversity and epidemiology in domestic species of malaria-mediated mosquitoes of Korea.

Microbiota in the mosquito plays an important role in their behavior and vector competence. The composition of their microbiome is strongly influenced by the environment, especially their habitat. The microbiome profiles of adult female Anopheles sinensis mosquitoes from malaria hyperendemic and hypoendemic areas in Republic of Korea were compared using 16S rRNA Illumina sequencing. In different epidemiology groups, the alpha and beta diversity analyses were significant. The major bacterial phylum was Proteobacteria. The most abundant species in the microbiome of hyperendemic mosquitoes were the genera Staphylococcus, Erwinia, Serratia, and Pantoea. Notably, a distinct microbiome profile characterized by the dominance of Pseudomonas synxantha was identified in the hypoendemic area, suggesting a potential correlation between the microbiome profiles and the incidence of malaria cases.

Kudoa septempunctata Spores Cause Acute Gastroenteric Symptoms in Mouse and Musk Shrew Models as Evidenced In Vitro in Human Colon Cells.

Kudoa septempunctata is a myxosporean parasite that infects the trunk muscles of olive flounder (Paralichthys olivaceus) and has been reported to cause foodborne illnesses in humans. However, the molecular mechanisms underlying K. septempunctata spore toxicity remain largely unknown. In this study, the gastroenteropathy of K. septempunctata was examined in human colon adenocarcinoma cells as well as experimental mice inoculated with spores. We found that K. septempunctata decreased transepithelial resistance and disrupted epithelial tight junctions by deleting ZO-1 in Caco-2 monolayers. Additionally, serotonin (5-HT), an emetic neurotransmitter, was increased in K. septempunctata-inoculated cells. In vivo, K. septempunctata spores induced diarrhea in suckling mice (80% in ddY and 70% in ICR mice), with a minimum provocative dose of 2 × 105 K. septempunctata spores. In house musk shrews, K. septempunctata induced emesis within 1 h and induced serotonin secretion in the intestinal epithelium. In conclusion, K. septempunctata may induce diarrhea and emesis by increasing intestinal permeability and serotonin secretion.

Imported human babesiosis in the Republic of Korea, 2019: two case reports.

Human babesiosis is a tick-borne disease induced by the genus Babesia and has been significantly reported in the Republic of Korea. This report shows the cases of 2 patients with human babesiosis who traveled to the USA in 2019. The 2 patients experienced fever and had travel histories to babesiosis-endemic regions. The diagnoses of both cases were verified by the identification of Babesia-infected red blood cells on blood smears. One patient was found to be infected with Babesia microti using polymerase chain reaction (PCR) for 18S rRNA, which discovered the phylogenetic link to the B. microti strain endemic in the USA. The 2 patients recovered from fever with subsequent hemoparasite clearance. Babesiosis could be diagnosed in anyone with histories of travel to babesiosis-endemic countries and tick bites. Furthermore, Babesia-specific PCR is required for determining geno-and phenotypic characteristics.

Positive rates for Enterobius vermicularis eggs among preschool children in Yeosu-si, Jeollanam-do, Korea (2017-2021).

This study aimed to evaluate the positive rates for Enterobius vermicularis eggs among preschool children in Yeosu-si, Jeollanam-do, the Republic of Korea (Korea) over a period of 5 years (2017-2021). Perianal swab samples, obtained using cellotape, from 10,392 preschool children in 26 districts were examined microscopically for E. vermicularis eggs. The test results were notified through the local health center, and the families of children who tested positive were advised to provide them anthelmintics treatment. The annual positive rates were 5.0%, 5.2%, 4.4%, 2.2%, and 1.0% in 2017, 2018, 2019, 2020, and 2021, respectively. The overall positive rate was higher in boys than in girls (P< 0.05), and children aged 5-7 years were at a higher risk of being infected than those aged 0-4 years (P< 0.05). Although the rates of infection by E. vermicularis in the survey area, Yeosu-si, were still in the 1% range , the results of this study suggest that they can be significantly reduced through continuous intervention centered around the test-treatment strategy.

A Rare Case of Imported Cutaneous Leishmaniasis Caused by Leishmania infantum in the Republic of Korea, 2021.

Leishmaniasis is a neglected tropical disease and an infectious disease transmitted by sandflies that occurs worldwide. In the absence of physicians seeking to identify the causes of disease in non-endemic areas, appropriate diagnoses cannot be made, thereby hampering effective treatment. In this report, we examined a nodular lesion on a patient's chin by performing a biopsy and molecular analysis. The biopsy finding led to the identification of a Leishmania amastigote. On the basis of PCR analysis of the internal transcribed spacer 1 gene and 5.8 S ribosomal RNA with a subsequent BLAST search, we identified the causal organism as Leishmania infantum. The patient, who had visited Spain from 1 July to 31 August 2018, was accordingly diagnosed with cutaneous leishmaniasis and was administered liposomal amphotericin B, which successfully treated the skin lesion. Travel history plays an important role in the diagnosis of leishmaniasis, and physicians should bear in mind that travelers can also introduce diseases and pathogens to non-endemic areas. Identification of Leishmania at the species level will increase the efficacy of treatment.

Severe fever with thrombocytopenia syndrome virus from ticks: a molecular epidemiological study of a patient in the Republic of Korea.

Severe fever with thrombocytopenia syndrome (SFTS) is a tick-borne infectious disease caused by Dabie bandavirus, commonly called SFTS virus (SFTSV). In the Republic of Korea (ROK), 1,504 cases of SFTS have been reported since the first human case was identified in 2013 until 2021. However, no case exists to provide molecular evidence between questing tick and patients with confirmed SFTS in the same living environment. In this study, we investigated the presence of ticks near the area of a patient infected with SFTSV. Ticks were collected by flagging and dry ice-baited traps at three spots in the vegetation around the patients' residence in Chuncheon City, Gangwon Province (ROK). Among the tick samples collected, the presence of SFTSV was genetically determined using reverse transcription PCR, followed by the phylogenetic analysis of the tick virus sequences and SFTSV found in the patient. In total 1,212 Haemaphysalis longicornis ticks were collected, and SFTSV was detected at a minimum infection rate of 5.3% (33 pools/618 tested ticks). The sequences of SFTSV in ticks were 99.6-100% identical with the patient's SFTSV in the M segment. To the best of our knowledge, this study is the first case to provide a molecular correlation between SFTSV in questing ticks collected from residence and patient with SFTS in the ROK. The present results provide useful information for the epidemiological investigation of patients with SFTS using ticks as vectors of SFTSV.

Treatment with Extracellular Vesicles from Giardia lamblia Alleviates Dextran Sulfate Sodium-Induced Colitis in C57BL/6 Mice.

Inflammatory bowel disease (IBD) is a chronic and recurrent illness of the gastrointestinal tract. Treatment of IBD traditionally involves the use of aminosalicylic acid and steroids, while these drugs has been associated with untoward effects and refractoriness. The absence of effective treatment regimen against IBD has led to the exploration of new targets. Parasites are promising as an alternative therapy for IBD. Recent studies have highlighted the use of parasite-derived substances, such as excretory secretory products, extracellular vesicles (EVs), and exosomes, for the treatment of IBD. In this report, we examined whether EVs secreted by Giardia lamblia could prevent colitis in a mouse model. G. lamblia EVs (GlEVs) were prepared from in vitro cultures of Giardia trophozoites. Clinical signs, microscopic colon tissue inflammation, and cytokine expression levels were detected to assess the effect of GlEV treatment on dextran sulfate sodium (DSS)-induced experimental murine colitis. The administration of GlEVs prior to DSS challenge reduced the expression levels of pro-inflammatory cytokines, including tumor necrosis factor alpha, interleukin 1 beta, and interferon gamma. Our results indicate that GlEV can exert preventive effects and possess therapeutic properties against DSS-induced colitis.

Molecular Detection of Anaplasma phagocytophilum and Ehrlichia Species in Ticks Removed from Humans in the Republic of Korea.

Human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME) are zoonotic tick-borne diseases transmitted via tick bites. To determine the state of human Anaplasma and Ehrlichia infections caused by tick bites in the Republic of Korea (ROK), we conducted a nationwide investigation of human cases of tick bites in 2020. A total of 180 ticks were obtained, comprising Haemaphysalis longicornis (70.0%), Amblyomma testudinarium (17.8%), Ixodes nipponensis (6.1%), H. flava (4.4%), and I. persulcatus (1.7%). In three cases (1.7%; 95% CI: 0.3-4.9), A. phagocytophilum was detected in Ixodes ticks using primers for Anaplasma-specific genes (16s rRNA, ankA, and msp4). Conversely, Ehrlichia sp. was only detected in H. longicornis, in two cases (1.1%; 95% CI: 0.1-4.0). To the best of our knowledge, this is the first record of Ehrlichia sp. in ticks parasitizing humans in the ROK. As concerns remain about the possibility of HGA and HME transmission, continuous monitoring and management of the pathogens and vectors are necessary.

Status of Helminthic Infections in Residents around River Basins in the Republic of Korea for 10 Years (2011-2020).

The positive rate of Clonorchis sinensis is the highest among intestinal parasites in the Republic of Korea (Korea). More than 1.2 million people were at risk of C. sinensis infection in Korea in 2012. An intensive control program is being implemented for residents of the 5 major river basins to reduce helminthic infections, including C. sinensis infection. This study evaluated the continuous intensive control program for parasitic diseases including clonorchiasis in areas near the 5 major river basins in Korea over the past 10 years (2011-2020). A total of 335,020 fecal samples (one sample per resident) prepared by the modified sedimentation technic were microscopically examined. Those who expelled helminth eggs were treated with anthelmintics through local health centers and re-examined 3 months later. The overall positive rate of helminths egg was 7.1%. The annual positive rates were dramatically decreased from 14.4% (2011) to 5.9% (2020). The egg positive rate was highest in C. sinensis (5.3%), followed by heterophyid flukes (1.5%) and Trichuris trichiura (0.2%). The prevalence of C. sinensis was significantly higher in males (7.6%) than in females (3.7%), and the highest in the 50-59 years (7.0%) age group. Our results are beneficial to establish prevention and control policies against helminthiases including clonorchiasis in endemic areas in this country.

Endemicity of Zoonotic Trematode Metacercariae in Fish from Deokcheon-gang (River) in Sancheong-gun, Gyeongsangnam-do, Republic of Korea.

The endemicity of zoonotic trematode metacercariae (ZTM) was investigated with total 871 freshwater fishes (19 species) from Deokcheon-gang (a branch stream of Gyeongho-gang) in Sancheong-gun, Gyeongsangnam-do, Korea for 3 years (2018-2020). All fishes were examined with the artificial digestion method. The metacercariae of Clonorchis sinensis (CsMc) were detected in 233 (36.3%) out of 642 fish in 11 positive fish species (PFS), and their infection intensity was 27 per fish infected (PFI). Especially, in index fish, Puntungia herzi, of CsMc infection, prevalence was 64.2% and infection intensity was 37 PFI. Metagonimus spp. metacercariae (MsMc) were found in 760 (87.5%) out of 869 fish in 18 PFS and their infection intensity was 228 PFI. In sweet smelt, Plecoglossus altivelis, the prevalence of MsMc was 97.6% and their infection intensity was 3,570 PFI. Centrocestus armatus metacercariae were detected in 209 (29.4%) out of 710 fish in 8 PFS and their infection intensity was 1,361 PFI. Echinostoma spp. metacercariae were found in 293 (42.6%) out of 688 fish in 15 PFS and their infection intensity was 5 PFI. Metacercariae of Clinostomum complanatum and Metorchis orientalis were also detected in 2.7% and 21.2% fish in 4 PFS and their infection intensities were 3.1 and 3.4 PFI respectively. By the present study, it was confirmed that some species of ZTM including CsMc and MsMc are more or less prevalent in fishes from Deokcheon-gang in Sancheong-gun, Gyeongsangnam-do, Korea.

First Identification and Genotyping of Enterocytozoon bieneusi and Prevalence of Encephalitozoon intestinalis in Patients with Acute Diarrhea in the Republic of Korea.

Encephalitozoon intestinalis and Enterocytozoon bieneusi can cause diarrhea in humans, especially severe diarrhea in immunocompromised patients. However, there have been few studies on Enc. intestinalis and Ent. bieneusi in patients with acute diarrhea in the Republic of Korea (ROK). In this study, fecal samples were collected from 1241 patients with acute diarrhea in 2020. Among these, 24 cases of Enc. intestinalis and one case of Ent. bieneusi were detected via PCR amplification of small subunit ribosomal RNA. Genotyping of the internal transcribed spacer region sequence revealed that the detected Ent. bieneusi genotype was in Group 1. This study provides the first evidence that Ent. bieneusi exists in humans in addition to animals in the ROK. To identify the causative agent, continuous monitoring of Enc. intestinalis and Ent. bieneusi is necessary for patients with acute diarrhea in the ROK.