This study focused on the genomic analysis of Anopheles kleini and Anopheles pullus, both vectors of vivax malaria within the Anopheles Hyrcanus group. Using Illumina NovaSeq600 and Oxford Nanopore platforms, we identified 126 and 116 contigs, along with 40,420 and 32,749 genes from An. kleini and An. pullus, respectively. The assembled genome sizes were 282 Mb for An. kleini and 247 Mb for An. pullus, which are within a similar range to the sizes previously estimated by digital PCR (249 Mb and 226 Mb). We are currently also estimating the genome sizes of other Anopheles spp. and manually curating key genes determining vectorial capacity.

Malaria is notorious disease to transmitted by Anopheline mosquitoes. In Korea, Plasmodium vivax malaria was re-introduce in 1993 and exceed to more than 4,000 cases in early 2000s. Based upon our control efforts, it was sharply decreased and diagnosis 576 cases in last year. We made Action Plan in this year for certification of malaria elimination by WHO until year 2024. In this plan, we need to strengthen our capacity of vector surveillance and control. For vector surveillance, we will introduce daily mosquito population monitoring system. In 2020, every malaria high risk area install two mosquito counters and send information to main server. For vector control, we will try building capacity of malaria mosquito control people in the Public Health center and military section. To decrease no. of mosquitoes, we will concentrate our resource to cattle shed, which is main blood source of Anopheline mosquitoes. Based on the ecological characteristic of Anopheline, we will adopt Integrate Vector management (IVM) including physical, chemical and biological method. Based on the reconciliation with North Korea, we will support technical and material support for malaria elimination.

말라리아는 말라리아원충에 감염된 Anopheles속 모기에 의해 매개되는 질병이다. 한국의 Anopheles속 Hyrcanus group 모기 가운데 특히 Anopheles kleini, Anopheles lesteri, Anopheles pullus는 말라리아 주요 매개충으로 알려져 있다. 한국의 말라리아는 휴전선 인접지역을 중심으로 환자 발생이 지속되고 있으며 이와 더불어 한국의 기후는 점차 아열대 기후로 변화하고 있다. 이러한 기후변화가 한국의 말라리아 전파력에 미치는 영향을 예측하기 위하여 말라리아 위험지역 가운데 경기 서북부 지역인 파주, 김포, 강화 세 지역의 소 축사에서 2014년과 2015년 6월부터 9월까지 채집된 모기의 월별 종 분포를 분석하였으며, 이러한 종 분포와 기후조건과의 관계를 통계분석 하였다. 기후조건으로는 채집일 기준 14일 전까지의 평균기온, 풍속, 강수량, 습도를 이용하였으며 Hyrcanus group 모기 6종(Anopheles belenrae, An. kleini, An. lesteri, An. pullus, Anopheles sinensis, Anopheles sineroides)의 분포를 로지스틱 회귀분석을 통하여 분석하였고, 상관관계를 확인하기 위하여 정준상관분석(Canonical correlation analysis)을 실시하였다. 분석결과, 강수량과의 연관성은 찾지 못하였으나, An. kleini, An. lesteri, An. pullus는 기온이 상승했을 때 Hyrcanus group 내에서의 분포가 증가할 가능성이 있는 것으로 나타났다. 또한 An. sinensis는 풍속과 습도가 높을 경우 증가할 것으로 나타났으며 이를 제외한 나머지 종들은 풍속과 습도가 낮을 경우 증가하는 것으로 나타났다. 이러한 분석 결과를 바탕으로 현재 한반도의 기온이 점점 상승하는 환경은 말라리아 주요 매개충이 서식하기 좋은 환경으로, 기존의 북한과 인접한 DMZ 부근에서 발생하던 말라리아가 한반도 내 다른 지역으로 분포 지역이 확대될 가능성이 있음을 시사한다.

한국에서 말라리아는 1993년에 다시 등장했으며, 이후 증감을 반복하며 공간적 변화를 거듭하여 왔다. 지리 공간의 인문·자연·지리적 요인들은 복잡하게 상호작용하여 매개 모기의 생태와 말라리아 발생에 영향을 미친다. 본 연구는 말라리아의 발생 양상을 이해하고 이를 조절하는 데 기여하고자 그 공간적 패턴을 분석하였다. 본 연구는 Getis-Ord의 Gi* 와 평균 중심점을 이용하여 2001년부터 2014년 사이 말라리아 발생의 공간적 분포를 탐색적으로 분석하여, 그 시공간적 변화를 파악하였다. 또한 일반화 포아송 회귀모형을 이용하여 2001년, 2014년 말라리아에 영향을 미치는 인문·환경 요인을 비교·분석하고, 군사분계선으로부터의 거리 변수를 이용하여 그 공간적 자기상관을 설명하고자 하였다. 그 결과, 소 두수, 표준공시지가, 성비, 아파트 비율, 군사분계선 거리, 기후 요소, 논 비율이 말라리아 발생에 영향을 미치는 유의한 변수로 확인되었다.

Anopheles sinensis is an important vector for Plasmodium vivax and thus has been targeted with pyrethroids in Republic of Korea. Using the direct contact mortality bioassay, two field populations of An. sinensis from Ganghwa-gun and Goyang-si were characterized for their resistance to pyrethroids with RR ratio values (Resistant ratio to imidacloprid) of 125.6 to 203.8 folds and 80.0 to 120 folds, respectively. Genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the sodium channel genes to detect L1014S mutations. The homozygous susceptible Leu/Leu genotype in Ganghwa-gun and Goyang-si was 5.0% and 11.3% and the resistance genotypes were 95.0% and 88.7%, respectively. The homozygous Phe/Phe resistance genotype was the most prevalent as 35.4% in Ganghwa-gun and 44.0% in Goyang-si. Hence, this study suggests that malaria vector control programs should be prepared for the management of pyrethroid insecticide resistance.

Anopheles sinensis is an important vector for Plasmodium vivax and thus has been targeted with organophosphates in Republic of Korea. Using the direct contact mortality bioassay, two field populations of An. sinensis from Paju-si and Yeoncheon-gun were characterized for their resistance to organophosphats with RR ratio values (Resistant ratio to imidacloprid) of 446.9 to 756.3 folds and 502.1 to 861.7 folds, respectively. Genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the acethylcholine esterase to detect Gly119Ser mutations. The homozygous susceptible Ser/Ser genotype in Paju-si and Yeoncheon-gun was 20.0% and 20.9% and the resistance genotypes were 80.0% and 79.1%, respectively. The homozygous Gly/Ser resistance genotype was the most prevalent as 60.7% in Paju-si and 41.9% in Yeoncheon-gun. Hence, this study suggests that malaria vector control programs should be prepared for the management of organophosphate insecticide resistance.

Anopheles sinensis is an important vector for Plasmodium vivax and thus has been targeted with pyrethroids in Republic of Korea. Using the direct contact mortality bioassay, two field populations of An. sinensis from Paju-si and Yeoncheon-gun were characterized for their resistance to pyrethroids with RR ratio values (Resistant ratio to imidacloprid) of 96.8 to 167.2 folds and 34.2 to 98.4 folds, respectively. Genomic DNA was used for Allele-specific PCR (AS-PCR) genotyping of the sodium channel genes to detect L1014S mutations. The homozygous susceptible Leu/Leu genotype in Paju-si and Yeoncheon-gun was 3.3% and 37.1% and the resistance genotypes were 96.7% and 62.9%, respectively. The homozygous Phe/Phe resistance genotype was the most prevalent as 45.0% in Paju-si and 31.5% in Yeoncheon-gun. Hence, this study suggests that malaria vector control programs should be prepared for the management of pyrethroid insecticide resistance.

Control of the vector mosquitoes is critical for implementation of patient’s decrease. Vector mosquito control has been mainly carried out using pesticides in Republic of Korea (ROK), which has developed the resistance against selected insecticides. Voltage-gated sodium channel (VGSC) and Acethylcholine esterase (AchE) are the target of several classes of insecticides. Genetic mutations of VGSC and AchE have been documented to confer resistances to pyrethroids and organophosphates in mosquitoes, respectively. In our studies, the insecticidal resistable level of An. sinensis from four malaria risk areas near Demilitarized Zone (DMZ) was measured. The partial sequence of An. sinensis voltage gated sodium channel (VGSC) IIS6 domain for Kdr gene and ace1 gene for AchE was sequenced. An. sinensis from two areas demonstrated significantly higher resistance to pyrethroids and mutation rates of Kdr gene were comparatively higher. In all tested areas An. sinensis showed high insecticidal resistance to organophosphates and > 70% of mutation rates of ace1 gene ware higher. Understanding the mechanisms of insecticide resistance among vector mosquitoes populations is very important to efficient and effective vector control.

As the malaria surveillance agency, the Korean Center for Control and Prevention (KCDC) has been monitoring malaria vector mosquito density and Plasmodium vivax infection since 2009. Anopheline mosquitoes were collected at 35 sites in Incheon, northern Gyeonggi and northern Gangwon Province using black light trap daily from April to October 2015. P. vivax infection of malaria vector mosquitoes tested by polymerase chain reaction (PCR). In 2015, a total of 57,926 malaria vector mosquitoes were collected among 138,119 mosquitoes. Three P. vivax positive pools were detected among 1,556 pools (13,745 individuals) and its minimum infection rate was 0.22. According to the monitoring result of malaria vector mosquitoes, early summer (June-July) could be an appropriate time for a malaria elimination campaign until September.

Malaria is an infectious disease that cause chilling, fever and hepatosplenomegaly by protozoan in plasmodium invading red blood cell, carried by anopheles mosquitoes. Malaria is transmitting steadily in South Korea from 1993 when vivax malaria retransmission occurred to soldiers at the front line near North Korea. There are many attempts in academic fields to analyse the causes by carrying out correlation analysis considering weather elements and regional characteristics, also developing tools for analysis. However, existing analysis tools have many limitations relating multi-platform support, user associability, limitation of information and so on. This study is to improve these limitations by developing hybrid web app concepted malaria vulnerability map system. Thus, it is possible to check the real-time malaria risk information, even in mobile platforms, and maximize the support for ease of use by using the implements and game UI animation technology of mobile-specific functions. Further utilizing a frame animation technology is a game animation techniques visual representation, and visually provides a time zone of malaria risk information, it is attempted to increase the user access. The system is expected to utilized by experts in related fields and general users.

Melanization is a unique defence mechanism in arthropods involved in wound healing and pathogen encapsulation. Phenoloxidases (PPOs) are key enzymes of melanization, which mediate the enzymatic conversion of tyrosine to eumelanin. A serine-protease (SP) cascade, similar to the blood-clotting cascade of vertebrates, proteolytically activates prophenoloxidases to phenoloxidases. This proteolytic activation is tightly controlled by serpins and other melanization inhibitors. Melanization has been regarded as one of key immune responses against malaria parasites in mosquitoes. The ookinete melanization of both the simian malaria parasite, Plasmodium cynomolgi, and of the rodent parasite, Plasmodium berghei, prevent parasite development in the human malaria vector, An, gambiae. However, the recent studies revealed a melanization response regulated by Serpin-2 and two C-type lectins (CTL4 and CTLMA-2) was shown to result in ookinete melanization but did not affect the development of the natural human parasite P. falciparum in the mosquito. Instead of melanization, TEP1/APL1/LRIM1 complement-like pathway has been identified as major immune response that regulate parasite loads in the natural association of An. gambiae and P. falciparum. The studies by me and my colleagues revealed another melanization response independent on Serpin-2. Genome analyses of mosquitoes revealed a large expansion of the PPO, SP, and serpin genes potentially involved in the melanization pathway. This expansion was devoted to existence of at least two distinct SP-Serpin regulation modules in controlling separate melanization responses, tissue and hemolymph melanization, in the mosquito, Aedes aegypti. Tissue melanization regulated by Serpin-2 has role in melanotic tumor formation, but not in ookinete melanization. Hemolymph melanization regulated by Serpin-1 and a couple of SPs was activated by the infection of various pathogens and is involved in anti-malarial defense against the avian malaria parasite, P. gallinaceum. A new type of regulator, CLSP2, negatively modulate this hemolymph melanization. Cross-talk between hemolymph melanization and complement-like pathway will be discussed.

Density of malaria vector mosquitoes, Anopheles mosquitoes, and infection of Plamodium vivax from the vector mosquitoes were monitored at 9 surveillance points in Incheonsi, 12 in Gyeonggi province and 7 in Gangwon province in the Republic of Korea (ROK) from April to October of 2010. The seasonal numbers of Anopheles mosquitoes collected weekly showed a high degree of variability. A total of 4,435 mosquitoes were captured using black light traps. Members of the Anopheles mosquito group were the most abundant species and accounted for 71.8% of the mosquitoes collected. The majority of this species were collected from Incheonsi (86.2%), particularly from Ganghwagun (84.8%). During June, populations of this species steadily increased and peak at second week of July. Thereafter, populations of this species decreased by the second week of August and increased by the third week of August, again. Anopheles mosquitoes steadily decreased from the fourth week of August. First Anopheles mosquitoes were reported in Wolgotri (15th weeks/year), Ganghwagun, Incheonsi, followed by Tanhyundong (16th weeks/year), Pajusi, Gyeonggi Province and Daemari (17th weeks/year), Cheolwongun, Gangwon Province. In Ganghwagun in which the most abundant malaria patients (102 patients) were reported, Anopheles mosquitoes demonstrated the highest species portion (over 70%) among collected mosquito species from July to September. Infection rate of P. vivax from the Anopheles mosquitoes also was the highest in Ganghwagun. In Gangwon and Gyeonggi provinces, collection rate of Anopheles mosquitoes and infection rate of P. vivax were relatively lower than those of Ganghwagun. Reported malaria patient cases in Gangwon and Gyeonggi provinces also were relatively lower than those of Ganghwagun. However, detailed results showed that there were some sites in which any correlation between malaria patients and vector mosquitoes were not. This may be resulted from size of surveillance area. Smaller size of surveillance area was, higher accuracy of analysis on correlation between vector mosquitoes and patients was. Further studies on relationship between malaria vector and patients may need for more accurate analysis, such as increase of collection site and so on.

Density of malaria vector mosquito was monitored at 24 sites in Korea to estimate malaria epidemic from 2004 to 2009. Twenty four locations include six in Gwangwon province and nine in Kyonggi province and nine in Incheon metropolitan city. Mosquitoes collected by public health centers in 24 sites were sent to each RIPHE(Research Institute of Public Health & Environment) then the RIPHE identified the malaria vector mosquitoes. KCDC analysed the results. Number of malaria vector mosquito collected in 2009 shapely decreased over 33 % when compared with number of mosquito collected from 2004 to 2009. In 2009, malaria vector mosquitoes firstly were collected at Injegun on 2nd week of April (6-13, April) and the highest peak of collected mosquito was demonstrated on 3rd week of August. Number of collected mosquito sharply decreased below 10 at 3rd week of september and None of mosquito was collected 2nd week October. Decrease of number of malaria vector mosquito collected in 2009 seem to be related with lowest temperature around midnight and precipitation days per month.