Anopheles Aconitus Research Articles

Anopheles diversity, biting behaviour and transmission potential in forest and farm environments of Gia Lai province, Vietnam

BackgroundDespite recent reductions in Vietnam, malaria transmission persists in some areas in forests and farmlands where a high density of Anopheles mosquitoes relative to other environments occurs. To inform effective malaria control measures, it is important to understand vector bionomics and the malaria transmission role of Anopheles spp. in the highland regions of Vietnam. This study was conducted to quantify the abundance, composition and biting behaviour of the Anopheles mosquito population, and the proportion of Plasmodium spp. infected mosquitoes collected from forest and agricultural farm sites in Gia Lai province, Vietnam.MethodsForest and agricultural farm sites in Gia Lai province were selected for mosquito collections (total eight sites). Mosquito collection was performed by Human-baited Double Net Trap (HDNT), animal-baited traps (ABT) using cattle, and CDC light traps. Captured mosquitoes were identified morphologically, and salivary glands of Anopheles mosquitoes were examined for sporozoites using microscopy. Plasmodium infection was determined by Polymerase Chain Reaction (PCR), and identification of blood meal type was determined by PCR and diffuse serum agglutination assay.ResultsA total of 1815 Anopheles mosquitoes belonging to 19 species were collected by ABT (n = 1169), HDNT (n = 471) and CDC light trap (n = 175). Anopheles abundance and diversity varied by district and environment. Capture by HDNT of Anopheles of vectorial concern was observed between early evening and early morning. Plasmodium vivax infection was determined by PCR in two Anopheles dirus specimens captured by HDNT in forest sites. Blood from a range of hosts could, including human blood, could be detected in species considered primary and secondary vectors An. dirus, and Anopheles aconitus, and Anopheles maculatus, respectively.ConclusionsA low number of Anopheles spp. considered primary vectors of concern and very low numbers of Plasmodium spp. infected Anopheles mosquitoes were captured at the end of the rainy season in the Central Highlands of Vietnam. However, capture species of vectorial concern by HDNT throughout the early to late evening demonstrates that use of additional personal protective measures could supplement current preventative measures, such as bed nets to prevent exposure to vectors of concern in this region.

Myzomyia and Pyretophorus series of Anopheles mosquitoes acting as probable vectors of the goat malaria parasite Plasmodium caprae in Thailand

Unlike malaria parasites in humans, non-human primates, rodents, and birds, ungulate malaria parasites and their vectors have received little attention. As a result, understanding of the hosts, vectors, and biology of ungulate malaria parasites has remained limited. In this study, we aimed to identify the vectors of the goat malaria parasite Plasmodium caprae. A total of 1019 anopheline and 133 non-anopheline mosquitoes were collected from goat farms in Thailand, where P. caprae-infected goats were discovered. Anopheline mosquitoes were identified using molecular biological methods that target the cytochrome c oxidase subunit 1 (cox1), the cytochrome c oxidase subunit 2 (cox2) genes, and the internal transcribed spacer 2 (ITS2) region. Pool and individual mosquitoes were tested for P. caprae using the head-thorax parts that contain the salivary glands, with primers targeting three genetic markers including cytochrome b, cytochrome c oxidase subunit 1, and 18S small subunit ribosomal RNA genes. Additionally, goat blood samples were collected concurrently with mosquito surveys and screened to determine the status of malaria infection. This study revealed nine mosquito species belonging to six groups on goat farms, including Hyrcanus, Barbirostris, Subpictus, Funestus, Tessellatus, and Annularis. The DNA of P. caprae was detected in Anopheles subpictus and Anopheles aconitus. This is the first time An. subpictus and An. aconitus have been implicated as probable vectors of P. caprae.

Molecular characterization of anopheline mosquitoes from the goat malaria-endemic areas of Thailand.

Despite the fact that over a 100 anopheline mosquito species have been identified as human malaria vectors, little is known about ungulate malaria vectors. Consequently, we focused on investigating the bionomics and genetic characterizations of anopheline mosquitoes in goat malaria-endemic regions. We also attempted to screen for ungulate malaria potential vectors. A total of 1019 female anopheline mosquitoes were collected from six goat farms in four provinces of Thailand from 2020 to 2021. Mosquitoes were morphologically identified and subsequently confirmed using the mitochondrial DNA barcoding region-cytochrome oxidase c subunit I (MtDNA-COI), mitochondrial DNA-cytochrome c oxidase subunit II (MtDNA-COII), and ribosomal DNA internal transcribed spacer 2 (rDNA-ITS2) sequences. The current study reveals the genetic characteristics and distribution of nine mosquito species within the Anopheles and Cellia subgenera. Four dominant species, including Anopheles peditaeniatus, Anopheles subpictus, Anopheles vagus, and Anopheles aconitus exhibited significant intraspecific gene flow within their corresponding species. Although malaria parasites were not found in 126 mosquito pools, meaning more investigation is necessary, the current study adds to the existing DNA barcoding data collection and improves the current understanding of the genetic structure and distribution of anopheline mosquito species, which could be useful for effective control of mosquito-borne diseases.

UPDATED KNOWLEDGE ON MALARIA VECTORS IN MALAYSIA: A SCOPING REVIEW

Malaria is transmitted by the Anopheles mosquito. The list of human and zoonotic malaria vectors in Malaysia is being updated in this scoping review. Search were conducted at four electronic bases. Title, abstract, and full-text reviews were used to determine which studies were eligible for further assessment. A total of 631 papers were identified until June 2021, yielded to 30 studies were reviewed. Human malaria vectors in Malaysia are identified as Anopheles maculatus, Anopheles sundaicus, Anopheles balabacensis, Anopheles donaldi, Anopheles campestris and Anopheles flavirostris. Vector for zoonotic malaria are Anopheles balabacensis, Anopheles introlatus, Anopheles cracens, Anopheles letifer, Anopheles donaldi, Anopheles hackeri, Anopheles latens, Anopheles collesi and Anopheles roperi. On the other hand, Anopheles aconitus, Anopheles kochi and Anopheles pujutensis are recognized as the potential malaria vectors. In order to implement effective mosquito control measures, it is crucial to comprehend and be knowledgeable about malaria vector profile. This will help stakeholder to develop and establish holistic and efficient strategy framework for integrated vector management (IVM) and develop new strategies for the elimination of malaria based on available evidence.

The Absence of Sporozoites in Anopheles Mosquitoes in the Malaria Pre-elimination Area

Introduction: The existence of sporozoites in the salivary gland of Anopheles is a prominent factor in the transmission and spread of malaria. Malaria control programs must be comprehensive including the treatment of patients, management of the vector population, and efforts to prevent human-vector contact. The present study aimed to track the presence of sporozoite in the Anopheles mosquito population in a malaria pre-elimination area and evaluate the implementation of a malaria elimination program. Methods: This cross-sectional study was conducted in a malaria pre-elimination area over a period of one year. Mosquitos were caught using human bait and in resting sites close to cattle pens. Sporozoites were detected microscopically and circumsporozoite protein (CSP) was tested using polymerase chain reaction technique. Results: A total of 212 Anopheles mosquitoes were caught. Anopheles maculatus, Anopheles aconitus, Anopheles subpictus, Anopheles barbirostris, Anopheles vagus, and Anopheles kochi species were identified. Salivary gland of 111 mosquitoes were dissected and CSP was detected in 101 specimens. Neither type of test found any sporozoites in the captured Anopheles. Conclusion: The absence of sporozoites in Anopheles in the pre-elimination area is a good sign of the ongoing malaria control program. Breaking the life cycle of the parasite that causes it by minimizing contact with vectors is an effective method that should be continuously improved.

Diversity of mosquito species and potential arbovirus transmission in long-tailed macaque (Macaca fascicularis) breeding facilities

Background and Aim:Mosquito-borne viral infections are diseases that reduce human and animal health levels. Their transmission involves wildlife animals as reservoirs and amplifying hosts, including long-tailed macaques (Macaca fascicularis), and potentially transmits to humans and vice versa. This study aimed to determine the species diversity, richness, and biting activity of mosquitoes in a long-tailed macaque breeding area facility and discover the presence of Flavivirus and Alphavirus as the two main arboviruses reported to infect macaques.Materials and Methods:Human landing catch, light trap, and sweep net methods were used for mosquito collection around long-tailed macaques cages at parallel times for 12 h (18:00–06:00) for 12 nights. Mosquito species were identified to the species level based on the morphological identification key for Indonesian mosquitoes. Mosquito diversity was analyzed by several diversity indices. Mosquitoes caught using the human landing catch method were pooled based on mosquito species for viral ribonucleic acid extraction. Reverse transcription-polymerase chain reaction (RT-PCR) detected the non-structural protein 5 of the Flavivirus region and the non-structural protein 4 of the Alphavirus region. This study used the man-hour density and man-biting rate formulas for mosquito density.Results:Ten mosquito species were collected, namely, Aedes albopictus, Anopheles aconitus, Anopheles minimus, Anopheles vagus, Armigeres foliatus, Armigeres subalbatus, Culex gelidus, Culex hutchinsoni, Culex tritaeniorhynchus, and Culex quinquefasciatus. The number of mosquitoes caught using the light trap method had the highest abundance. In contrast, the number of mosquito species caught using the sweep net method had lower diversity than the other two methods. Seven mosquito species were obtained using the human landing catch method. The mosquito species with the highest density was Cx. quinquefasciatus within the observed densest period from 20:00 to 21:00. Negative results were obtained from RT-PCR testing on five species detected using universal Flavivirus and Alphavirus primers.Conclusion:The occurrence of mosquitoes in long-tailed macaque breeding facilities can be a source of transmission of zoonotic vector-borne diseases between animals and humans and vice versa.

Morphological characteristic of malaria vector Anopheles aconitus (Family: Culicidae) revealed by advanced light and scanning electron microscope

Abstract. Supriyomo, Soviana S, Novianto D, Musyaffa MF, Tan S, Hadi UK. 2022. Morphological characteristic of malaria vector Anopheles aconitus (Family: Culicidae) revealed by advanced light and scanning electron microscope. Biodiversitas 23: 3546-3552. Anopheles aconitus (An. aconitus) is one of the primary vectors for malaria in several areas in Indonesia, especially in Java Island. Scanning electron microscope (SEM) is a powerful tool that can differentiate Anopheles spp. Morphology that is difficult to identify by using a light microscope only. The adult of An. aconitus used in this study have been reared in the laboratory. This research is conducted to better understand the Anopheles aconitus' external morphology, such as the head, proboscis, wings, thorax and legs, using advanced light and SEM. Results from an advanced light microscope showed that An. aconitus has palpus with the same length as proboscis with one narrow basal end and two broad distal ends; meanwhile, the apical half of proboscis is pale. Anopheles aconitus has wings with four pale spots on the costa side. Legs are uniformly dark with only very narrow, rather faint pale bands at the joining of segments. Using SEM, we found several types of sensilla trichodea: short-sharp tipped, short-blunt tipped, and long-shaped tipped in antenna, apical proboscis, maxillary palps, abdomen, and legs. Antenna is also covered with microtrichia and sensilla. This research is important for basic necessity in fully understanding and determining of mosquito bionomic due to the development of effective and efficient mosquito vector control strategies.

Impact of a spatial repellent product on Anopheles and non-Anopheles mosquitoes in Sumba, Indonesia

BackgroundThe East Nusa Tenggara province, Indonesia, contributed to 5% of malaria cases nationally in 2020, with other mosquito-borne diseases, such as dengue and filariasis also being endemic. Monitoring of spatial and temporal vector species compositions and bionomic traits is an efficient method for generating evidence towards intervention strategy optimization and meeting disease elimination goals.MethodsThe impact of a spatial repellent (SR) on human biting mosquitoes was evaluated as part of a parent cluster-randomized, double-blinded, placebo-controlled trial, in Sumba, East Nusa Tenggara. A 10-month (June 2015–March 2016) baseline study was followed by a 24-month intervention period (April 2016 to April 2018)—where half the clusters were randomly assigned either a passive transfluthrin emanator or a placebo control.ResultsHuman-landing mosquito catches documented a reduction in landing rates related to the SR. Overall, there was a 16.4% reduction (21% indoors, and 11.3% outdoors) in human biting rates (HBR) for Anopheles. For Aedes, there was a 44.3% HBR reduction indoors and a 35.6% reduction outdoors. This reduction was 38.3% indoors and 39.1% outdoors for Armigeres, and 36.0% indoors and 32.3% outdoors for Culex species. Intervention impacts on the HBRs were not significant and are attributed to large inter-household and inter cluster variation. Anopheles flavirostris, Anopheles balabacensis and Anopheles maculatus individually impacted the overall malaria infections hazard rate with statistically significance. Though there was SR-based protection against malaria for all Anopheles species (except Anopheles sundaicus), only five (Anopheles aconitus, Anopheles kochi, Anopheles tessellatus, An. maculatus and An. sundaicus) demonstrated statistical significance. The SR numerically reduced Anopheles parity rates indoors and outdoors when compared to the placebo.ConclusionEvidence demonstrating that Anopheles vectors bite both indoors and outdoors indicates that currently implemented indoor-based vector control tools may not be sufficient to eliminate malaria. The documented impact of the SR intervention on Aedes, Armigeres and Culex species points to its importance in combatting other vector borne diseases. Studies to determine the impact of spatial repellents on other mosquito-borne diseases is recommended.

An inventory of human night-biting mosquitoes and their bionomics in Sumba, Indonesia.

Mosquitoes are important vectors that transmit pathogens to human and other vertebrates. Each mosquito species has specific ecological requirements and bionomic traits that impact human exposure to mosquito bites, and hence disease transmission and vector control. A study of human biting mosquitoes and their bionomic characteristics was conducted in West Sumba and Southwest Sumba Districts, Nusa Tenggara Timur Province, Indonesia from May 2015 to April 2018. Biweekly human landing catches (HLC) of night biting mosquitoes both indoors and outdoors caught a total of 73,507 mosquito specimens (59.7% non-Anopheles, 40.3% Anopheles). A minimum of 22 Culicinae species belonging to four genera (Aedes, Armigeres, Culex, Mansonia), and 13 Anophelinae species were identified. Culex quinquefasciatus was the dominant Culicinae species, Anopheles aconitus was the principal Anopheles species inland, while An. sundaicus was dominant closer to the coast. The overall human biting rate (HBR) was 10.548 bites per person per night (bpn) indoors and 10.551 bpn outdoors. Mosquitoes biting rates were slightly higher indoors for all genera with the exception of Anopheles, where biting rates were slightly higher outdoors. Diurnal and crepuscular Aedes and Armigeres demonstrated declining biting rates throughout the night while Culex and Anopheles biting rates peaked before midnight and then declined. Both anopheline and non-anopheline populations did not have a significant association with temperature (p = 0.3 and 0.88 respectively), or rainfall (p = 0.13 and 0.57 respectively). The point distribution of HBR and seasonal variables did not have a linear correlation. Data demonstrated similar mosquito-human interactions occurring outdoors and indoors and during early parts of the night implying both indoor and outdoor disease transmission potential in the area-pointing to the need for interventions in both spaces. Integrated vector analysis frameworks may enable better surveillance, monitoring and evaluation strategies for multiple diseases.

Durio zibethinus Murr Extracts as Potential Larvacide Against Anopheles aconitus Donitz and Anopheles maculatus Theobald (Diptera: Culicidae)

Malaria is endemic in most parts of Indonesia. Anopheles aconitus and Anopheles maculatus are examples of dominant malaria vectors in Indonesia. Plant secondary metabolites can be used as malaria vector control that is safe for the ecosystem, one of which is Durio zibethinus leaves which are thought to have active compounds so that they can be used as botanical larvicides. This study aimed to determine the potency of D. zibethinus leaf extract against An. aconitus and An. maculatus and to determine the concentration, type of extract and content of secondary metabolites of D. zibethinus leaves. Determination of LC50 and LC90 using probit analysis. Maceration and remaceration were used to produce the extract and phytochemical screening to determine the active compound of D. zibethinus. The WHO insecticide bioassay testing procedure was used for the larvicide test. Ethanol extract of D. zibethinus leaf as a larvicide for An. aconitus second and third instar and larvae of An. maculatus second and third instar had LC50 ​​respectively: 480; 520; 510; and 540 ppm and LC90 respectively: 750; 760; 760; and 810 ppm. D. zibethinus leaf aqueous extract as larvicides against larvae of An. aconitus second and third instar and larvae of An. maculatus second and third instar had LC50 values ​​respectively 14,500; 16,400; 22,100; and 23,300 ppm and LC90 respectively, 26,100; 27,200; 30,600; and 33,700 ppm. Ethanol extract of D. zibethinus leaves was more effective as a larvicide. Alkaloid, saponins and tannins were found in the leaves of D. zibethinus through phytochemical screening.

Human-Plasmodium Like in Domestic-goat Blood in Malaria Endemic Areas in Purworejo Indonesia

Background: Etawa crossbreed goats are cultivated livestock by Jatirejo Kaligesing Purworejo villagers. This village is the area with the highest malaria incidence in the Province of Central Java in 2017. The Annual Parasitic Incidence (API) is 0.32%. Verified malaria vectors are Anopheles aconitus and Anopheles maculatus. Studies show that vectors prefer goat blood over humans. The research aims to trace the presence of human-plasmodium in Etawa-goat blood. Method: A cross-sectional study was conducted to explore the presence of h-Plasmodium in domestic goats’ blood in pre-elimination malaria areas. 97 Etawa-goats’ samples were taken by purposive sampling. Microscopic detection of human-plasmodium in Etawa-goat’s blood was carried out on thick blood preparations by Giemsa staining. Results: Human-plasmodium like was detected on four slides, with the findings of three slides resembling P. vivax trophozoites and one slide resembling P. falciparum trophozoites. Conclusion: The presence of parasites similar to P. vivax and P. falciparumin goats’ blood is important initial information to continue studies on the transmission of h-Plasmodium in domestic livestock. Replication studies in high case incidence areas are needed to complete the microscopic visualisation and are supported by molecular testing.

Species diversity and breeding site of mosquito larvae (Diptera: Culicidae) in Macaca fascicularis breeding area

Mosquito larvae play an essential role in the ecological, and many of them can spread human and animal diseases, including in Macaca fascicularis. Information on mosquito species and their habitats can provide an overview of the role of mosquitoes in the spread of vector-borne diseases in M. fascicularis captivity area. This study aimed to identify species diversity of mosquito larvae, species affinity and association, and the larval breeding sites around M. fascicularis captivity area in Bogor, West Java. Mosquito larvae were collected from 102 sites using a 350 ml dipper. Mosquito larvae that were successfully collected consisted of 11 species; Aedes albopictus, Ae. aegypti, Armigeres subalbatus, Anopheles aconitus, An. kochi, An. vagus, Culex fuscocephala, Cx. pseudovishnui, Cx. tritaeniorhyncus, Cx. quinquefasciatus, and Cx. vishnui. Co-occurrence in mosquito larvae as many as 13 compositions, with the highest co-occurrence in Ae. albopictus and Cx. quinquefasciatus that was 11 times. There were seven types breeding sites for the larval mosquitoes, i.e containers, ditches, creeks, ponds, artificial ponds, groundwater puddles, and rice fields. We conclude the existence of mosquito larvae and the availability of their breeding site in M. fascicularis captivity area can be a potential transmission of pathogens between mosquitoes and hosts

Effect of Mindi Leaf Powder (Melia azedarach L) on Mortality of Anopheles aconitus Instar III Larvae

Background: Malaria is a disease caused by Plasmodium parasite infection wwhich is transmitted through bite of an Anopheles mosquito. Efforts to control malaria can be carried out in various ways, starting from treatment, prevention, to breaking the transmission chain through vectors, both mosquitoes and larvae. Mindi tree contains Azadiractin compounds which are larvicides. Study aims to determine effect of mindi leaf powder (Melia azedarach L) on mortality of Anopheles aconitus instar III larvae.Methods: A true experimental study with a post test only control group design, the object of research was Anopheles aconitus Instar III larvae. The research was conducted at Laboratory of Parasitology, Faculty of Medicine, Universitas Islam Sultan Agung, Semarang. 20 larvae / 100 ml were used. Air temperature and water pH were controlled during observation. Assessment by counting number of mortality larvae and the number of live larvae from each treatment was compared to the control. The data obtained were analyzed by the Kruskal-Wallis statistical test. The LD50 and LD90 were analyzed using Probit analysis.Results: The dose of 0.2 g/100 ml was the minimum dose that killed 100% of larvae of Anopheles aconitus (p = 0.01). In the probit test, LD50 was 0.01 g/100 ml of aquades, while LD90 was 0.06 gram/ 100 ml of aquades.Conclusion: Dose can served as larvicidal (LD90 )was 0.06 g/100 ml of aquades., while LD50 was 0.01 g/100 ml of aquades.

Anopheles Mosquito Species in Betung Bedarah, Tebo Regency, Jambi Province

Background: Indonesia is a tropical country which is a good breeding ground for mosquitoes. Mosquitoes are still one of arthropods that contribute to occurrence of infectious diseases, including malaria, which is a disease that is a health problem in Jambi Province.Objective: To determine types of Anopheles mosquito spesies in Betung Bedarah Tebo, Jambi.Methods: A discriptif study to identifed distribution of mosquitoes in Betung Bedarah, Tebo Regency used human landing collection and resting collection methods. Collection of mosquito samples used an aspirator and light trap. Analysis used tabulation.Results: Of the 64 mosquitoes collected, 25% were Anopheles mosquitoes which consisted of 43.7% Anopheles barbirostris, 25% Anopheles vagus, 18.7% Anopheles aconitus, and 12.6% Anopheles kochi.Conclusion: Most of mosquito species collected at Betung Berdarah Tebo were Anopheles barbirostris.

New insights into malaria vector bionomics in Lao PDR: a nationwide entomology survey

BackgroundIn Laos, the malaria burden remains high despite a significant reduction of cases during the last decade. In the context of the disease elimination by 2030, a nationwide entomological survey was conducted to better understand the distribution, abundance and behaviour of major malaria vectors (Anopheles spp.) in the country.MethodsMosquito collections were implemented in ten villages from ten provinces during the rainy and dry seasons of 2014 and 2015 by using human landing catch (HLC) and cow bait collection (CBC) methods. After morphological identification in the field, molecular identification of the sibling species of Anopheles mosquitoes from the Funestus, Leucosphyrus, and Maculatus groups were determined using PCR specific alleles. A screening of Plasmodium falciparum and Plasmodium vivax infections in the vectors was carried out by quantitative PCR assays.ResultsA total of 14,146 adult mosquitoes representing 25 different Anopheles species were collected and morphologically identified. Molecular identification revealed the presence of 12 sibling species within the main primary vector groups, including Anopheles maculatus, Anopheles rampae, Anopheles sawadwongporni, Anopheles pseudowillmori, Anopheles dravidicus, Anopheles minimus, Anopheles aconitus, Anopheles pampanai, Anopheles harrisoni, Anopheles dirus, Anopheles baimaii, Anopheles nemophilous. Anopheles maculatus and An. minimus were predominant during both the dry and rainy seasons, but showed highly zoophilic preferences (Zoophilic index of 98% and 95%, respectively). Overall, 22% of the total malaria vectors were collected between 10:00 PM and 5:00 AM indoors when people are sleeping. Twenty-seven percent of primary and secondary vectors were collected outdoors before 10:00 PM or after 5:00 AM, times when people are usually awake and outdoors. Only two specimens were positive for P. falciparum, one An. aconitus from Phongsaly and one An. minimus from Vientiane ProvinceConclusionsThe results indicate that people living in rural areas in Laos are constantly exposed to malaria vectors throughout the year and specifically outdoors. The use of LLINs/IRS remains important but innovative tools and new strategies are needed to address locally, the early and outdoor malaria transmission. Lack of expertise in general entomological methods may further exacerbate the situation.

Model pengendalian nyamuk Anopheles Aconitus dengan asap “lampu Templek” dari Malathione yang dibandingkan dengan alfa-cypermetrine

Malaria, one of vector borne disease has been still a major Public Health problem in Indonesia. The disease is spread by Anopheles biting, for example Anopheles aconitus. So far, efforts which have been conducted by the government to control the disease is house spraying program with thermal fogging and also Ultra Low Volume (ULV) with Malathione 96 EC. A commercially available insecticide Fendona 30 EC is commonly used equal to Malathione 96 EC in its application. The application of insecticides through “Templek lamp” would be predicted more applicable due to its simple use by the community. The purpose of this study was to learn the efficacy of trial using “Templek lamp” of Malathione 96 EC and Fendona 30 EC to find the Lethal Concentration 90% (LC90), Lethal Time 50% (LT50) and 90% (LT90) of insecticides to Anopheles aconitus mortality. An experimental research with posttest only control group design was used in this study. Female An. aconitus, age 2-5 days were purposely selected. They were fed with 10% sugar and tested by Glass chamber method with double control groups. Groups of 20 An. aconitus each were tested with various concentrations of the insecticides, i.e. 0.1%, 0.5%, 1.0%, 2.5% and 5.0% in triplicate. The result showed that based on probit analysis, Fendona 30 EC LT50s at the aforementioned concentrations were 7.25, 5.81, 3.48, 1.16 and 0.54 hours respectively. On the other hand Malathione 96 EC LT50s at the same concentrations were 8.13, 5.48, 3.96, 2.69 and 0.57 hours. At the same concentrations, Fendona 30 EC LT90s were 61.76, 54.96, 20.56, 5.95 and 1.44 hours respectively, while Malathione 96 EC LT90s were 86.37, 43.62, 26.60, 18.86 and 1.48 hours respectively. It was also observed that LC90 for Fendona 30 EC and Malathione 96 EC was 0.688% and 1.048% respectively. Analysis of variance showed significant difference (p<0.05) between concentrations of each insecticides. Further studies are still required to confirm this observation.

Complete mitochondrial genomes of Anopheles aconitus and Anopheles splendidus and phylogenetics analysis of known mtgenomes in the subgenus Cellia (Diptera: Culicidae: Anophelinae)

The complete mitochondrial genome (mtgenome) sequences of Anopheles aconitus and A. splendidus were sequenced and analyzed in this study. They are 15,359 bp and 15,362 bp long, respectively and contains 13 protein-coding genes (PCGs), 22 tRNA genes (tRNAs), 2 rRNA genes (rRNAs), and 1 AT-rich control region (CR). The codon UUA (Leu) are prodominarily used in all 13 PCGs. ATN is mainly used as the initiation codon in theses PCGs except for COI and ND5 genes, which use TCG and GTG as the initiation codon, respectively and TAA as termination codon except for COI, COII, COIII, and ND4 that use the incomplete termination codon T. All of the tRNAs have the typical clover-leaf structure except for tRNASer(AGN), which lost the dihydrouridine (DHU) arm. The CRs have the highest A + T content of 92.97 and 93.18% in these two species, respectively. The phylogenetic relationships of 20 species in the subgenus Cellia were constructed using Maximum Likelihood based on concatenated nucleotide sequences of 13 PCGs with the selected best-fit GTR + I+G model. The 20 species are clearly divided into four monophyletic series: Pyretophorus, Neomyzomyia, Neocellia, and Myzomyia, and the Neomyzomyia is basal to the other three series. The Neocellia and Myzomyia are suggested to be sister groups and the Pyretophorus is proposed to sister with Neocellia + Myzomyia. This study provides a basis for further study on mtgenome and phylogenetics in the subgenus Cellia.

Diversity and biting patterns of Anopheles species in a malaria endemic area, Umphang Valley, Tak Province, western Thailand

Malaria is highly endemic in Umphang Valley, a district in the western edge of Tak Province, along the boundary with Kayin State of Myanmar. Although there are high indigenous malaria cases in this area every year, nothing about malaria vectors and their transmission role have been investigated before this study. The objective of this work is to characterize the Anopheles species diversity and trophic behavior of malaria vectors in the transmission area of Umphang Valley. Females of Anopheles mosquitoes were collected every two months during a two-year period. Mosquito collections were using standard collection technique, indoor and outdoor human landing collections and outdoor cattle bait collection. Anopheles mosquitoes were identified using morphological characters and multiplex AS-PCR assay for the identification of sibling species within groups and complexes present. From a total of 16,468 Anopheles females, 2723 specimens (16.54%) were collected from humans and 13,745 specimens (83.46%) were captured from cattle. From human landing collections, 2447 specimens (89.86%) of Anopheles minimus were obtained, followed by 119 Anopheles peditaeniatus (4.37%), 62 Anopheles maculatus (2.28%), 17 Anopheles dirus (0.6%), 15 Anopheles aconitus (0.5%) and 6 Anopheles sawadwongporni (0.2%) respectively. Seven putative malaria vectors, including An. minimus, An. dirus, An. baimaii, An. sawadwongporni, An. maculatus, An. pseudowillmori and An. aconitus were documented from this study and trophic behavior of each respective species were observed. Such information is definitely crucial for defining the vector capacity of each single species and for designing appropriate vector prevention and control strategies against target vector species.

BIONOMIK Anopheles spp SEBAGAI DASAR PENGENDALIAN VEKTOR MALARIA DI KABUPATEN KEBUMEN JAWA TENGAH

Malaria is one of the serious diseases in Indonesia and is the target of Central Java Provincial Government in Malaria Elimination Program. One attempt to eliminated malaria is by knowing the vector bionomics, Anopheles spp as the basis of the policy control. The research was conducted in the location indicated by malaria case in Wagirpandan Village, Rowokele District Kebumen Regency. The study was conducted in two sampling sites, taking samples of adult mosquitoes and larvae. Environmental parameters observed include pH, temperature, rainfall and humidity and vegetation. The results of this study found seven species which were Anopheles aconitus, An. Annularis, Anopheles barbirostris, An. balabacencis, An. kochi, An. maculatus. An. vagus. The peak activity of the blood sucking of Anopheles spp around 08.00-09.00; 10.00-11.00 pm and 04.00 – 05.00 am outside the house and cattle pens. The proportions of parous mosquitoes are caught 42,8% in Cuntelan and 69,49% in Borang. All mosquito except An. Annularis and An. Kochi found were confirmed as malaria vectors.

Molecular characterization of Anopheline (Diptera: Culicidae) mosquitoes from eight geographical locations of Sri Lanka

BackgroundGenus Anopheles is a major mosquito group of interest in Sri Lanka as it includes vectors of malaria and its members exist as species complexes. Taxonomy of the group is mainly based on morphological features, which are not conclusive and can be easily erased while handling the specimens. A combined effort, using morphology and DNA barcoding (using the markers cytochrome c oxidase subunit I (COI) gene and internal transcribed spacer 2 (ITS2) region, was made during the present study to recognize anophelines collected from eight districts of Sri Lanka for the first time.MethodsCytochrome c oxidase subunit I and ITS2 regions of morphologically identified anopheline mosquitoes from Sri Lanka were sequenced. These sequences together with GenBank sequences were used in phylogenetic tree construction and molecular characterization of mosquitoes.ResultsAccording to morphological identification, the field-collected adult mosquitoes belonged to 15 species, i.e., Anopheles aconitus, Anopheles annularis, Anopheles barbirostris, Anopheles culicifacies, Anopheles jamesii, Anopheles karwari, Anopheles maculatus, Anopheles nigerrimus, Anopheles pallidus, Anopheles peditaeniatus, Anopheles pseudojamesi, Anopheles subpictus, Anopheles tessellatus, Anopheles vagus, and Anopheles varuna. However, analysis of 123 COI sequences (445 bp) (16 clades supported by strong bootstrap value in the neighbour joining tree and inter-specific distances of >3%) showed that there are 16 distinct species. Identity of the morphologically identified species, except An. subpictus, was comparable with the DNA barcoding results. COI sequence analysis showed that morphologically identified An. subpictus is composed of two genetic entities: An. subpictus species A and species B (inter-specific K2P distance 0.128). All the four haplotypes of An. culicifacies discovered during the present study belonged to a single species. ITS2 sequences (542 bp) were obtained for all the species except for An. barbirostris, An. subpictus species B, An. tessellatus, and An. varuna. Each of these sequences was represented by a single species-specific haplotype.ConclusionsThe present study reflects the importance and feasibility of COI and ITS2 genetic markers in identifying anophelines and their sibling species, and the significance of integrated systematic approach in mosquito taxonomy. Wide distribution of malaria vectors in the country perhaps indicates the potential for re-emergence of malaria in the country.