Molecular Marker Confirmation for Member of Anopheles barbirostris Van Der Wulp 1884 in Different Localities

Vector and non-vector forms of Anopheles barbirostris have been recognized in Indonesia. However, because of their similarity in morphology, they were considered to be a single species. This information has led to the hypothesis that Anopheles barbirostris is a complex of species, which are morphologically indistinguishable from each other by ordinary methods. Objectives of the research was to identify the member of Anopheles barbirostris by PCR Assay. Samples were taken from two localities in Java, two in Sulawesi, two in Flores Indonesia, one from Thailand, one from China. The study was to develop a PCR-based technique of rDNA ITS2 region. Results showed that there are at least four species within the Anopheles barbirostris population studied, namely Anopheles barbirostris species DW, DX, DY and DZ. The length of the sequence amplified for species W, species X, species Y, and species Z were 339bps, 247bps, 165bps. and 157bps, respectively. Verification of the method was carried out with 270 mosquitoes from eight different field-collection sites using various sampling methods. Samples collected from Singaraja-Flores were identified as species W and X. All specimens collected from human bite outdoors were identified as species X; this species showed to be predominant among indoor light trap, indoor human bite and indoor resting collections Samples from Reo-Flores were identified as species W and X. All specimens from Manado and Palopo in Sulawesiwere identified as species Z. Similarly only species Y was found in samples from Thailand, while specimens from Salaman and Jambu in Java were identified as species W or species X. These species-specific molecular markers for the Anopheles barbirostris, complex appear to be reliable over a wide geographical area. However, larger number of samples is still needed from throughout the range of this species.Key words: Anopheles barbirostris, ITS2, PCR, Specific primer diagnosti

Molecular Marker Confirmation for Member of Anopheles barbirostris Van Der Wulp 1884 in Different Localities

Vector and non-vector forms of Anopheles barbirostris have been recognized in Indonesia. However, because of their similarity in morphology, they were considered to be a single species. This information has led to the hypothesis that Anopheles barbirostris is a complex of species, which are morphologically indistinguishable from each other by ordinary methods. Objectives of the research was to identify the member of Anopheles barbirostris by PCR Assay. Samples were taken from two localities in Java, two in Sulawesi, two in Flores Indonesia, one from Thailand, one from China. The study was to develop a PCR-based technique of rDNA ITS2 region. Results showed that there are at least four species within the Anopheles barbirostris population studied, namely Anopheles barbirostris species DW, DX, DY and DZ. The length of the sequence amplified for species W, species X, species Y, and species Z were 339bps, 247bps, 165bps. and 157bps, respectively. Verification of the method was carried out with 270 mosquitoes from eight different field-collection sites using various sampling methods. Samples collected from Singaraja-Flores were identified as species W and X. All specimens collected from human bite outdoors were identified as species X; this species showed to be predominant among indoor light trap, indoor human bite and indoor resting collections Samples from Reo-Flores were identified as species W and X. All specimens from Manado and Palopo in Sulawesiwere identified as species Z. Similarly only species Y was found in samples from Thailand, while specimens from Salaman and Jambu in Java were identified as species W or species X. These species-specific molecular markers for the Anopheles barbirostris, complex appear to be reliable over a wide geographical area. However, larger number of samples is still needed from throughout the range of this species.Key words: Anopheles barbirostris, ITS2, PCR, Specific primer diagnosti

Public knowledge, attitudes and practices towards COVID-19

In an effort to control the COVID-19 outbreak in Indonesia, the government implemented rules such as clean and healthy living behavior by all components of public, isolation, and early detection. Community knowledge, attitudes and practices (KAP) towards COVID-19 play an important role in determining the readiness of the community in accepting policies in the form of behavior change from the health authority. The aim of this study is to determine the KAP toward COVID-19 in Yogyakarta, Indonesia. A cross-sectional online survey of 155 householders was conducted between 04 May 2020 and 18 May 2020. Descriptive statistics, t-tests and one-way analysis of variance were conducted. Most public have understood about causes symptoms, mode transmission, high risk groups, isolation and quarantine, and proper use of disinfectants but people have not been able to distinguish between how to increase immunity and how to prevent COVID-19 transmission. The public has a positive belief that the government will succeed in controlling and managing a health crisis. Public behavior to prevent COVID-19 transmission is good, except the use of disinfectants and stop smoking or prohibiting family members from smoking. Differences in public knowledge, attitudes and practices towards COVID-19 occur in different gender and occupational groups

Risk factors related to malaria incidence at Santu’un village, Tabalong District, South Kalimantan Province

Santu’un Village is one of malaria endemic areas in Muara Uya Sub district, Tabalong District, South KalimantanProvince, Indonesia. Annual Parasite Incidences (APIs) from 2007 to 2009 were 11.4, 82.9, and 25.8‰ respectively.The majority of Muara Uya population are rubber tappers, gold miners and loggers in the forest. Moreover, most ofthe Muara Uya population do not use mosquito nets when sleeping at night. These occupations and communityhabits may contribute in high malaria incidence in Muara Uya. However, study concerning risk factor of malaria hasnot been performed yet. The study was conducted in order to evaluate risk factors that might correlate with malariaincidence in Santu’un village, Muara Uya subdistrict. Two hundred and ninety one subjects were enrolled in thisstudy. Those consisted of 70 people whom microscopically malaria positive and 221 people whom malaria negative.Household visits were conducted to each subject to perform interviews. Data collection of malaria risk factors wasobtained using questionaire. The results indicated that three main occupations, loggers, gold miners and rubbertapper, were significantly represent risk factors of malaria incidence. The community habits that also significantlycorrelated with malaria incidence were the not using of mosquito repellents and not using bed nets. Two clusteringcases were also observed during the year of 2010. The coordinate of the first cluster was 1.863500 S and 115.606700E with diameter of 0.65 km and the coordinate of the second cluster was 1.838667 S and 115.607200 E withdiameter of 0.67 km. In conclusion, in Santu’un village, the risk factor that correlated with the malaria incidencewere loggers, gold miners, rubber tappers. In addition the not using of mosquito repellents and not using bed netswere also risk factors of malaria incidence.Key words: malaria -risk factors-annual parasite incidence-Muara Uya-South Kalimanta

Pengaruh Lama Penyimpanan Telur terhadap Transovarial Infection Rate Virus DEN-3 pada Nyamuk Aedes aegypti

Transovarial transmission of dengue virus is important phenomenon causes dengue virus survival during inter-epidemic period. The purpose of this study was to analyze effect of egg storage duration on DEN-3 virus Transovarial Infection Rate (TIR) in Aedes aegypti mosquitoes. This was laboratory research with experimental design. Ae. aegypti was infected with the DEN-3 virus per-orally and allowed to undergo its gonotrophic cycle. Eggs stored at room temperature (25±3oC) and relative humidity (70±5%)  for 0, 1, 2 and 3 months. Hatched eggs then colonized until become adult mosquito. Sample was taken from colonies at 4, 8, 16 days old. Headsquash of mosquitoes was preparedwith Streptavidin Biotin Peroxidase Complex (SBPC) immunocytochemical method using DSSE 10 monoclonal antibody and detected the presence virus antigen. Transovarial infection rate was determined by counting percentage of positive samples. Data were analyzed by linear regression test. Result of this study showed that in 4, 8, 16 samples showed lowest TIR on 0 month storage (33,33%, 33,33%; 40,00%) and highest TIR observed on 2 month storage (76,67%; 66,67%, 76,67%). Statistical test showed significantly different result (p = 0,013) with α = 0,05 and R2 = 0,476. The duration of egg storage effect on DEN-3 virus TIR in Ae. aegypti mosquito with 47.6% contribution

A Study of Aedes aegypty Susceptibility Against Cypermetrin at Elementary Schools Yogyakarta

Introduction: Dengue Hemorrhagic Fever is transmitted by Aedes aegypti mosquitoes. Yogyakarta is an endemic area of DHF. One of the efforts of eradicating DHF in Yogyakarta was fogging with cypermethrin insecticide. Cypermethrin had been used since 2008 with a frequency of 1.469 times. However, repeated and prolonged application of chemical insecticide in an ecosystem unity may lead to vector resistance against the particular insecticide. Monitoring and early detection for the vector susceptibility status against insecticide was necessary in potential locations of DHF transmission, such as in elementary school. Detection of vector susceptibility status against insecticide could be conducted with susceptibility test using 0.05% cypermethrin impregnated paper.Objectives: To find out the status of Aedes aegypti susceptibility against cypermethrin insecticide and the relationship between the status of Aedes aegypti susceptibility with the frequency of cypermethrin application.Methods: An analytical observational research was conducted using cross-sectional design. Female Aedes aegypti from mosquito eggs collected at 72 state Elementary Schools in 45 villages in Yogyakarta were evaluated. Detection of Aedes aegypti susceptibility against cypermethrin was conducted using WHO standard method, namely, susceptibility test, which used 0.05% cypermethrin impregnated paper.Results: Aedes aegypti at elementary schools in Yogyakarta had been resistant against 0.05% cypermethrin, with an average mortality of 4.03%. No correlation between Aedes aegypti susceptibility in the elementary schools in Yogyakarta and the frequency of cypermethrin application. This might be attributable to the use of household insecticide and to the resistance history of the test animals.Conclusions: The population of Aedes aegyptiat the State Elementary Schools in Yogyakarta was found to be resistant against 0.05% cypermethrin. No relationship between Aedes aegypti susceptibility status and the frequency of cypermethrin application. Keywords: Aedes aegypti, Cypermethrin, impregnated paper, Susceptibility Test,  INTISARI Pendahuluan: Demam Berdarah Dengue adalah penyakit infeksi yang disebarkan oleh nyamuk Aedes aegypti. Kota Yogyakarta merupakan daerah endemis DBD. Salah satu cara penanggulangan DBD di kota Yogyakarta dengan pengasapan menggunakan insektisida sipermetrin. Penggunaan sipermetrin sudah dilakukan sejak tahun 2008 dengan frekuensi aplikasi sebanyak 1469 kali. Aplikasi insektisida kimia dalam waktu lama dan berulang di satuan ekosistem dapat menimbulkan resistensi vector terhadap insektisida yang digunakan.  Monitoring dan deteksi dini status kerentanan vector terhadap insektisida di kota Yogyakarta perlu dilakukan di lokasi yang berpotensi sebagai tempat penularan DBD diantaranya di sekolah dasar. Deteksi status kerentanan vector terhadap insektisida dapat dilakukan dengan susceptibility test menggunakan impregnated paper sipermetrin 0.05%.Tujuan: Mengetahui status kerentanan nyamuk Aedes aegypti di Lingkungan SDN di Kota Yogyakarta terhadap insektisida sipermetrin dan mengetahui hubungan status kerentanannya dengan frekuensi aplikasi insektisida sipermetrin.Metode: Penelitian ini merupakan penelitian observasional analitik dengan rancangan crosssectional. Subyek penelitian populasi nyamuk Aedes aegypti dari telur yang berasal dari 72 SDN di 45 kelurahan di Kota Yogyakarta. Deteksi status kerentanan nyamuk Aedes aegypti terhadap insektisida sipermetrin dilakukan dengan metode standar WHO susceptibility test menggunakan impregnated paper sipermetrin 0.05%.Hasil: Hasil uji kerentanan menunjukkan nyamuk Aedes aegypti di lingkungan sekolah dasar di kota Yogyakarta sudah resisten terhadap insektisida sipermetrin 0.05% dengan angka kematian nyamuk rata-rata 4.03%. Tidak ada korelasi antara status kerentanan nyamuk Aedes aegypti dengan frekuensi aplikasi insektisida sipermetrin. Hal ini bisa disebabkan karena penggunaan insektisida rumah tangga yang digunakan oleh masyarakat dan karena adanya riwayat resistensi dari nyamuk uji.Simpulan: Populasi nyamuk Aedes aegypti di lingkungan Sekolah Dasar Negeri di Kota Yogyakarta sudah resisten terhadap insektisida Sipermetrin 0.05%. Tidak ada hubungan status kerentanan nyamuk Aedes aegypti di lingkungan sekolah dasar negeri di Kota Yogyakarta dengan frekuensi aplikasi insektisida sipermetrin. Kata kunci: Aedes aegypti, Insektisida Sipermetrin, impregnated paper, Uji kerentanan, Sekolah Dasa

Spatial Analysis on Vulnerability to Dengue Hemorrhagic Fever in Kotabaru Subdistrict, Jambi Municipality, Jambi Province

Introduction: High incidence of DHF in Kotabaru Subdistrict was high even endemic in all villages. Utilization of Geographical Information System technology integrated with the wide of settlement land, the density of Aedes sp, maya index, resistance and transovarial of virDen, got a picture or map vulnerable areas prone event of DHF in Kotabaru Subdistrict.Objectives: This study to map areas vulnerable to dengue hemorrhagic fever is based on the highest incidence of DHF in Kotabaru Subdistrict, Jambi Municipality.Methods: This study is an observational survey with cross sectional design. The variable of the study was population density, the wide of settlement land, the density of Aedes sp. population, rainfall, vector vulnerability, and transovarial of virDen.Results: The result showed population density had no correlation to Dengue Hemorrhagic Fever (DHF) incidence, whereas the wide of settlement land (p=0.004) and population density of Aedes sp. (p=0.001) influenced DHF incidence. The number water containers controllable sites (89%) and percentage of Maya Index (p=<0.001) was high influenced DHF incidence. Rainfall not influenced DHF incidence. The vector resistance not influenced the high of DHF incidence and the high of DHF incidence was not influenced by transovarial of virDen. The high vulnerability to DHF in Kotabaru Subdistrict occurred in 2 villages and the intermediate vulnerability happens in 4 villages. The area with intermediate vulnerability generally have direct border to the village with high vulnerability category.Keywords: Spatial, Maya Index, Resistance, DH

Effect of temperature and storage duration of Aedes aegypti mosquito specimens artificially infected with dengue-3 virus on the results of immunohistochemical examination

To confirm the presence of any dengue viral in a mosquito, mosquito’s head can be squashed ona slide and stained with immunohistochemical staining. The remaining samples then can bestored in the cryo freezer at -80ºC to avoid specimen damage. However, for laboratories withlimited facilities, with only a refrigerator with a temperature range of -20ºC to 4ºC is available,examination to evaluate whether the dengue antigen can still be detected in specimens storedat these temperature is necessary. It was a quasi-experimental study. Three to five-day-old adultfemale Aedes aegypti mosquitoes were injected intrathoracically with dengue-3 (DENV-3) strainH-87 virus and then maintained for about 7 days. The dengue viral antigen on mosquitoes wasidentified using immunohistochemical method after stored at a temperature of 4oC, -20oC, -80oCfor 2, 4 and 8 weeks. Mosquito specimens that were not stored were used as a positive control.Kappa value was counted to analyze level of agreements between two observers. Two-wayAnova was used to analyze mean positive rates. Kappa value showed poor agreement (0.00-0.16) between two observers when the specimens were stored at 4oC for 2-8 weeks, and showedgood agreement (Kappa value of 0.77), when stored at -20oC for 4 weeks. The kappa valueshowed very good agreement (0.90-0.92) when the specimens were stored at -20oC for 2 weeks,and at -80oC for 2-8 weeks. Mean positive rates of the specimens stored at 4oC were significantlylower (p0.05). In conclusion, availability of thedengue viral antigen on mosquito specimens was influenced by temperature and storage durationof the specimens.Keywords: dengue virus - head squash - immunohistochemistry - kappa agreement - temperatur

Uji Efektifitas Atraktan pada Lethal Ovitrap terhadap Jumlah dan Daya Tetas Telur Nyamuk Aedes aegypti

AbstractControl of Aedes aegypti mosquito as dengue haemorrhagic fever (DHF) vector can be conducted by using the ovitrap modified into a lethal ovitrap. The addition of attractant substances to the ovitrap can attract more mosquitoes to come in to the trap, and prevent the mosquitoes to lay eggs in other places. The aim of this research was to compare the percentage of the number of eggs trapped, the number of eggs that hatched and the percentage of larval mortality in lethal ovitrap modified with the addition of two types of attractant. This research was an experiment research with a complete random design. The samples used were female bloodfed Ae. aegypti mosquito. The insecticide used was water extract of Annona squamosa seed, and the attractants used were hay infus at water with concentration of 20% and larval rearing water of the Ae. aegypti. Aquades used as control. The results showed that hay infusion was more effective than larval rearing water in attracting female Ae. aegypti mosquito to lay eggs. The highest mortality was found in the combination of lethal ovitrap and hay infusion. The combination could be an alternative controlling strategy for DHF management program in order to reduce the density of Ae. aegypti mosquito and minimize the dengue transmission in a region.Keywords : Lethal Ovitrap, attractant, Aedes aegyptiAbstrakPengendalian nyamuk Aedes aegypti sebagai vektor demam berdarah dengue (DBD) dapat dilakukan dengan menggunaan ovitrap yang dimodifikasi menjadi lethal ovitrap. Penambahan zat atraktanpada ovitrap dapat menarik lebih banyak nyamuk untuk datang ke perangkap yang dipasang dan mencegah nyamuk bertelur di tempat lain. Penelitian ini bertujuan untuk membandingkan persentase jumlah telur terperangkap, jumlah telur menetas dan mortalitas larva pada lethal ovitrap yang diberi tambahan dua jenis atraktan. Insektisida yang digunakan adalah ekstrak air biji srikaya (Annona squamosa), sedangkan atraktan yang digunakan adalah air rendaman jerami dengan konsentrasi 20% dan air bekas kolonisasi larva Ae. aegypti. Jenis penelitian ini adalah eksperimen dengan rancangan acak lengkap. Sampel yang digunakan adalah nyamuk Ae. aegypti betina yang kenyang darah. Hasil penelitian menunjukkan bahwa air rendaman jerami merupakan atraktan yang lebih berpengaruh dibandingkan air bekas kolonisasi Ae. aegypti dalam menarik nyamuk Ae. aegypti untuk bertelur. Mortalitas larva tertinggi terdapat pada kombinasi lethal ovitrap dengan air rendaman jerami.Kombinasi lethal ovitrap dengan atraktan air rendaman jerami dapat menjadi strategi pengendalian alternatif bagi pengelola program DBD untuk mengurangi kepadatan nyamuk Ae. aegypti dan meminimalisasi transmisi penyakit demam berdarah dengue di suatu wilayah.Kata kunci : Lethal Ovitrap, atraktan, Aedes aegypt

Analisis Spasial Karakteristik Habitat Perkembangbiakan Anopheles Spp di Desa Lifuleo Kecamatan Kupang Barat

Anopheles barbirostris and Anopheles subpictus are the primary vectors of malaria in East Nusa Tenggara. Anopheles mosquitoes generally breed at similar environment, including water streams, irrigation passages, water containers, paddy fields, impermanent ponds, water puddles, marsh, and brackish water. Modelling and spatial analysis play a role in identifying factors associated with potential breeding places for Anopheles mosquitoes, hence comprehension of breeding place characteristics and effective malaria control. The cross-sectional study used an observational-analytic approach. Study samples were identified larvae and breeding places of Anopheles spp. in Lifuleo village in West Kupang. We measured water salinity and pH, and we recorded coordinates of breeding places. Data analysis was performed by using Moran I index and spatial error model to identify factors associated with potential breeding places for Anopheles mosquitoes. Anopheles species found were An. barbirostris, An. subpictus, An. vagus, An. vagus var limosus, and An. indefinitus. All identified Anopheles larvae were found in brackish water, and in breeding places with high water salinity, with An. subpictus being able to survive the highest salinity (48‰). Univariate analysis demonstrated Io value of 0.00926, coefficient constant of 0.693868, and probability of 0.02252. The presence of Anopheles spp. was associated with breeding place habitat, daytime feeding, and presence of vegetations surrounding breeding places. Keywords: Anopheles spp, Lifuleo village, habitat characteristics, spatial. Abstrak Anopheles barbirostris dan Anopheles subpictus merupakan vektor primer malaria di Provinsi Nusa Tenggara Timur. Spesies Anopheles mempunyai habitat perkembangbiakan yang tidak sama yaitu aliran air, batas tangki, saluran irigiasi, sawah, kolam sementara, genangan air dekat pantai, genangan air di sungai, mata air, kolam ikan terlantar, rawa dan genangan air payau. Pemodelan dan analisis spasial dapat menjelaskan faktor-faktor yang memengaruhi habitat perkembangbiakan potensial bagi jentik Anopheles spp. Pengendalian malaria akan efektif apabila pengetahuan tentang karakteristik habitat perkembangbiakan dipelajari secara komprehensif. Metode penelitian menggunakan observasional analitik dengan pendekatan cross sectional. Sampel penelitian adalah seluruh habitat perkembangbiakan dan jentik Anopheles spp di Desa Lifuleo, Kecamatan Kupang Barat. Jentik diambil menggunakan pipet, kadar garam diukur menggunakan refractometer, pH diukur menggunakan pH meter dan koordinat diambil menggunakan aplikasi Avenza Maps yang terinstal pada Hand Phone android. Analisis data menggunakan Indeks Moran I dan Spatial Error Model untuk mencari faktor-faktor yang memengaruhi keberadaan jentik Anopheles spp. Spesies Anopheles yang ditemukan diantaranya; An. barbirostris, An. subpictus, An. vagus, An. vagus var limosus, dan An. indefinitus. Seluruh spesies Anopheles hidup pada habitat yang mengandung kadar garam tetapi An. subpictus mampu beradaptasi pada kadar garam tertinggi (48‰). Analisis univariat diperoleh Io sebesar 0.00926. Nilai Coefficient Constant sebesar 0,693868 dengan probability sebesar 0,02252. Spesies Anopheles yang hidup pada air payau yaitu An. barbirostris, An. subpictus, An. indefinitus, An. vagus, An. vagus varietas limosus. Keberadaan jentik Anopheles spp dipengaruhi oleh tipe habitat perkembangbiakan, Anopheles mengisap siang hari, dan adanya tanaman di habitat perkembangbiakan. Kata kunci: Anopheles spp, Desa Lifuleo, karakteristik habitat, spasia