Species Composition and Geographic Distribution of Culicinae Mosquitoes and Their Possible Infection with West Nile Virus in Hormozgan Province, Southern Iran.

Background: West Nile virus (WNV) is a mosquito-borne virus that causes a febrile disease and may cause a fatal neurological illness in humans. We aimed to investigate the geographic distribution of Culicinae mosquitoes and their possible infection with WNV in Hormozgan Province, southern Iran. Methods: Field studies were carried out from June 2017 to May 2019. Different mosquito sampling methods were used monthly to collect mosquitoes from 22 sites. The Real-Time PCR technique was used to detect the virus infection in the mosquitoes. Results: Overall, 6165 mosquitoes were caught. The species were Culex theileri (33.25%), Cx. pipiens (20.45%), Cx. quinquefasciatus (10.51%), Aedes caspius (6.33%), Cx. tritaeniorhynchus (5.82%), Ae. vexans (4.10%), Cx. sinaiticus (3.62%), Cx. antennatus (3.29%), Culiseta longiareolata (2.81%), Cx. perexiguus (2.03%), Cs. subochrea (1.95%), Cx. mimeticus (1.49%), Cx. pusillus (1.38%), Cx. univittatus (1.27%), Cx. modestus (1.14%), and Cx. sitiens (0.57%). The molecular detection of virus infection in mosquitoes found to be negative for WNV. Conclusion: The presence of many species of mosquito vectors and high population traffic increase the risk of disease transmission is very high. Therefore, the way to restrict WNV infection factors is increasing the knowledge for personal protection measures to prevent mosquito bites.

Imported tires; a potential source for the entry of Aedes invasive mosquitoes to Iran.

INTRODUCTION: International trade of tires has been responsible for the introduction of invasive Aedes species into many countries. The present study aimed to determine the sources and volume of such trading in Iran and to establish and map points of entry for entomological surveillance. METHOD: A list of tire importers, type and number of tires and source of their shipment was compiled, and the map of the main location(s) of their depot in the country was produced from 2017 to 2018. Contamination of imported tires with mosquito eggs or larvae was also determined. The samples from all parts of the warehouse were taken randomly from 4 to 20 tires every 15 days. In tires with a trace of water inside, the existence of the egg and larva of Aedes mosquitoes was investigated by sticking the glue band to the hot spot of left water inside the unpacked tires. RESULTS: Approximately one-third of the annual tire requirements of Iran were imported from 15 countries, most of which were endemic to Aedes aegypti and or Ae. albopictus. It is obligatory to import only wrapped tires. About 10% of total country consumption is imported through informal markets and smuggled. However, the unofficially imported tires are usually not wrapped. The majority of tire imports were made through the southern ports of the country. The main sites for depots of imported tires were located in four Iranian provinces, namely Tehran, Isfahan, Fars, and Sistan and Baluchestan. The latter is extremely important given its border with Pakistan. Depot locations were mostly unstructured, and tires were kept in the open air. All these depot locations were at the edge of towns and residential areas. Priority areas for routine entomological surveillance were established. Such as the previous studies, surveillance of imported tires for the presence of Aedes eggs or larvae was negative in this study. DISCUSSION: Mapping the entry points of imported tires and their origin is crucial to determine and prioritize sites for entomological surveillance of invasive mosquito species. Strengthening collaboration with customs authorities and the association of tire importers is imperative in this effort. The development of national rules and regulations for tire import is necessary to minimize the danger of the introduction of invasive vector species into the country.

Repellent Efficacy of Eucalyptus globulus and Syzygium aromaticum Essential Oils against Malaria Vector, Anopheles stephensi (Diptera: Culicidae).

BACKGROUND: Mosquito species are highly considering as disease transmission as well as nuisance insects. One of the principal strategy to protect human from the mosquito bites is repellent agents. This study aimed to assess repellency of two organic essential oils, Eucalyptus globulus and Syzygium aromaticum from bites of malaria vector, Anopheles stephensi. METHODS: The study was conducted in 2019-2020. The components of essential oils of E. globulus and S. aromaticum was determined using gas chromatography/mass spectrometry. The unfed female mosquitoes aged 2-5 d old were used in all experiments. In vivo Klun and Debboun module bioassays were utilized on human-volunteer skin. The essential oils at serial concentrations were used to find repellent efficacy against Anopheles landings and bites. To find the synergistic effect, four combinations of the essential oils were tested. RESULTS: The main composition of E. globulus essential oil was 1,8-Cineol (78.20%), whereas that of S. aromaticum essential oil was 2-methoxy-3-(2-propenyl) (77.04%). Based on minimum effective dose (≤1% biting), 10% (v/v) of E. globulus showed high landing repellency (77.78%), whereas minimum effective dose of S. aromaticum at concentration of 1% had high landing repellency (88.89%). Among four combinations, the ratio of 1:1 of E. globulus (10%):S. aromaticum (1%) showed the most landing repellency (94.44%). CONCLUSION: The combinations of two essential oils had the most potential repellency effect against landing of mosquitoes. As essential oils are eco-friendly with less irritation for human skin, E. globulus and S. aromaticum essential oils are recommended as effective and safe mosquito repellents.

Investigation of LAMP Technique in Diagnosis Type of Plasmodium Species in Anopheles Mosquitoes :A Fast and Practical Technique to Detect Malaria Pathogens in the Field.

BACKGROUND: Malaria is one of the main parasitic diseases and a major health issue in some countries. This study aims to determine the rate and type of infections of Anopheles mosquitoes with malaria parasites using the molecular LAMP method in the Southeastern Iran. METHODS: In this study, 400 Anopheles mosquitoes were collected by the Zahedan Medical Insecticide Center in Nikshahr City, a high-risk area of malaria transmission in Sistan-Baluchestan Province. The mosquitoes were caught manually (by hand) in domestic (humans and animals), natural, and artificial outdoor places (Shelter pits). After DNA extraction, the LAMP method was used, which was compared with Multiplex Nested-PCR as a standard method. RESULTS: Out of 400 samples collected from Nikshahr City, 6 samples (1.5%) were infected with Plasmodium vivax. No Plasmodium falciparum or a mix (Plasmodium vivax and Plasmodium falciparum) was detected in this study. CONCLUSIONS: The results of this study indicate that in places with transmission of both species, i.e. Plasmodium vivax and Plasmodium falciparum, detection of malaria parasites by the LAMP method could be very useful in spotting infections in the field. Thus, molecular epidemiological studies could be conducted annually to monitor malaria in endemic regions. The results of this research show that contamination with mosquito malaria vectors is increasing in Nikshahr City, and it seems that more studies will be required to eliminate malaria until 2026.

Preparation of nanoemulsion of Cinnamomum zeylanicum oil and evaluation of its larvicidal activity against a main malaria vector Anopheles stephensi.

PURPOSE: There is a growing need to use green and efficient larvicidal as alternatives for conventional chemicals in vector control programs. Nanotechnology has provided a promising approach for research and development of new larvicides. Larvicidal potential of a nanoemulsion of Cinnamomum zeylanicum essential oil reports against Anopheles stephensi. METHODS: The nanoemulsion of was formulated in various ratios comprising of C. zeylanicum oil, tween 80, span 20 and water by stirrer. It was characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). All components of C. zeylanicum essential oil were identified by GC-MS analysis. The larvicidal potential of the oil and its nanoformulation were evaluated against larvae of An. stephensi. The stability and durability of nanoemulsion was observed over a period of time. RESULTS: Sixty one components in the oil were identified, cinnamaldehyde (56.803%) was the main component. The LC90 and LC50 values of C. zeylanicum essential oil were calculated as 49 ppm and 37 ppm, respectively. The nanoemulsion droplets were found spherical in shape. It was able to kill 100% of larvae in up to 3 days. It was stable after dilution and increased its larvicidal activity up to 32% compared with the essential oil. CONCLUSIONS: A novel larvicide based on nanotechnology introduced. This experiment clearly showed increasing larvicidal activity and residual effect of the nanoformulation in comparison with the bulk essential oil. It could be concluded that this nanoemulsion may be considered as safe larvicide and should be subject of more research in this field.

Level of Pyrethroid-Resistance Associated with Cytochrome P450 Expression in German Cockroach Blattella germanica (Blattodea: Ectobiidae) in the Field Collected Strains.

BACKGROUND: Cytochrome P450-dependent monooxygenases are a very important metabolic system involve in insecticide resistance. This study was conducted to find the association between the expression level of cytochrome P450 (CYP450) and permethrin-resistance level among four strains of the German cockroach Blattella germanica (L) (Blattodea: Ectobiidae). METHODS: Three field strains of German cockroach with different frequency of exposure to pesticides, and a laboratory susceptible strain were used in the present study. Insecticide susceptibility bioassays were carried out to detect resistance to permethrin. The concentration of CYP450 in each strain was determined using ion-exchange HPLC chromatography. Biochemical assays was performed to analyse CYP450 activities. RESULTS: The resistance ratios (RR) to permethrin among three field strains were 3.29, 4.10 and 6.17-fold comping with the susceptible stain. The CYP450 activity of three field strains was 1.6, 2.4 and 2.7 times higher than in the susceptible stain. The amount of CYP450 per mg of protein was significantly different between the susceptible and the three resistant strains. The resistant cockroaches showed a relatively high expression of CYP450 enzymes. A strong correlation was found between permethrin resistance level and total concentration of CYP450 enzymes. CONCLUSION: The results of current study show that more frequent usage of a pyrethroid insecticide cause the metabolic insecticide resistance to rise in German cockroach. Therefore, there is a ratio correlation between resistance level and monooxygenases activities in insect. Thus, the control program must be managed according to history of pesticide usage.

Preparation of a Nanoemulsion of Essential Oil of Acroptilon repens Plant and Evaluation of Its Larvicidal Activity agianst Malaria Vector, Anopheles stephensi.

Background: Extensive use of chemical larvicides to control larvae, has led to resistance in vectors. More efforts have been conducted the use of natural products such as plant essential oils and their new formulations against disease vectors. Nanoformulation techniques are expected to reduce volatility and increase larvicidal efficacy of essential oils. In this study for the first time, a larvicide nanoemulsion from the essential oil of Acroptilon repens was developed and evaluated against Anopheles stephensi larvae under laboratory conditions. Methods: Fresh samples of A. repens plant were collected from Urmia, West Azarbaijan Province, Iran. A clevenger type apparatus was used for extracting oil. Components of A. repens essential oil (AEO) were identified by gas chromatography-mass spectrometry (GC-MS). All larvicidal bioassay tests were performed according to the method recommended by the World Health Organization under laboratory condition. Particle size and the morphologies of all prepared nanoformulations determined by DLS and TEM analysis. Results: A total of 111 compounds were identified in plant. The LC50 and LC90 values of AEO calculated as 7 ppm and 35 ppm respectively. AEO was able to kill 100% of the larvae in 4 days. Conclusion: The nanoemulsion of AEO showed a weak effect on the larvar mortality. It may therefore be suggested that this kind of nanoemulsion is not appropriate for the formulation as a larvicide. It is important to screen native plant natural products, search for new materials and prepare new formulations to develop alternative interventions with a long-lasting impact.

Evaluation of Nanoemulsion of Eucalyptus globulus Oil as Potent Botanical Larvicide against Malaria Vector, Anopheles stephensi and West Nile Vector, Culex pipiens Under Laboratory and Semi-Field Conditions.

Background: Due to undesired environmental impact of insecticides as well as resistant of vectors to them, the development of organic and natural insecticides has been more considered. In the current study, we developed nanoemulsion of eucalyptus and investigated lavicidal activity of it against malaria vector, Anopheles stephensi and Culex pipiens under laboratory as well as semi-field conditions. Methods: An optimized nanoemulsion was prepared by mixing Eucalyptus oil, Tween 80 and ethanol at ratio of 1:2:1.5 in distilled water, then, stirred for 20 minutes at room temperature. The product was then used for bioassay tests against 3-4th instar larvae of Anopheles stephensi as well as Culex pipiens. Furthermore, a semi-field trial was carried out to evaluate larvicidal activity of nanoemulsion of eucalyptus. Results: Nanoemulsion of eucalyptus showed significantly high lavicidal activity comparing with bulk eucalyptus essential oil. The LC50 and LC90 value of nanoemulsion against An. stephensi were 111.0 and 180.8 ppm respectively and 29.5 and 73.7 ppm for Cx. pipiens, respectively. In the semi field condition, the Nanoemulsion of eucalyptus decreased 1-2nd instar larval density of Culicines and Anophelines to 90.1% and 85.2%, respectively. Conclusion: The nano formulation of eucalyptus oil showed high larvicidal activity. Therefore, nanoemulsion of eucalyptus oil can be used as an eco-friendly larvicide against mosquitoes.

Seroprevalence Study on West Nile Virus (WNV) Infection, a Hidden Viral Disease in Fars Province, Southern Iran.

BACKGROUND: West Nile Virus, a mosquito-borne flavivirus, causes a variety of symptoms in human, from asymptomatic infection to neuroinvasive disease. Several studies have been conducted on the seroprevalence of WNV infection in different areas from Iran. This study was performed to find the presence of antiviral antibodies in human serum among some high risk population and awareness of health care staff about symptom of the WNV infection. METHODS: Study performed in five geographical districts based on high population of immigrant and domestic birds and prevalence of the antiviral antibodies in horses which was reported previously. Totally 150 human blood samples were collected during 2018. The samples collected from patients referred to the clinics. The ELISA method used to detect IgG and IgM antibody against WNV. Logistic regression models used to analyze the effect of sex, age, keeping birds and urban/rural residence on the risk of infection. The awareness of health care staff about symptom of infection surveyed. RESULTS: From all blood donors, 41 samples (27.33%) showed positive to IgG antibody. From which 56.10% were males and remaining females. None of the mentioned factors had a significant relationship. Health care staff had less attention to the infection. CONCLUSION: Although the prevalence of antibodies was relatively high, due to the similarity to other viral diseases, health care staff had less attention to the disease. The study showed that people in these areas have been exposed to the virus. Further research activities are recommended for control of this arbovirus.

Biting and resting behaviour of malaria vectors in Bandar-Abbas County, Islamic Republic of Iran.

BACKGROUND: Blood feeding and resting behaviour of malaria vectors are the most influential factors in malaria transmission. AIMS: To measure blood feeding and resting behaviour, conventional mosquito sampling methods were performed in an area with potential for malaria transmission. METHODS: Adult mosquitoes were collected monthly from indoor/outdoor places by conventional sampling methods, and larval habitats were investigated. Enzyme-linked immunosorbent assay was used to measure the human blood index (HBI) of the mosquitoes. RESULTS: Anopheles stephensi, Anopheles culicifacies, Anopheles dthali and Anopheles fluviatilis, were collected. Overall, 1249 female Anopheles mosquitoes were captured on human and animal baits, but no human-vector contact occurred indoors. A. dthali, A. fluviatilis and A. culicifacies showed a greater tendency to outdoor resting places in contrast to A. stephensi, which had a propensity to indoor resting places. The seasonal biting activities of all species occurred at average temperatures between 23 and 27°C. HBI was measured as 27.2%, 20.7%, 19.1%, and 23.0% for A. fluviatilis, A. stephensi, A. culicifacies and A. dthali, respectively. CONCLUSION: Vector control strategy depends upon mosquito behaviour. Therefore, using appropriate sampling methods based on mosquito behaviour is critical for malaria control planning. Exophilic/exophagic habit of mosquito vectors leads to fewer human bites, resulting in biting protection. Exophilic behaviour also requires specific larvicidal operations in order to prevent and control malaria transmission.

Challenges and the Path Forward on Malaria Elimination Intervention: A Systematic Review.

BACKGROUND: This systematic review was conducted to highlights key challenges, and outlines important next steps to maximize the potential to contribute to the broader malaria elimination interventions. METHODS: This systematic review on malaria elimination intervention and challenges was undertaken searching six databases, between 1995 and 2018. Inclusion and exclusion criteria were set. The references were collated and categorized according to type of study, intervention, population, and health outcome. Articles selection based on title and abstract, retrieval of full text and additions of articles from reference lists and recommendations from experts. Disagreement in data extraction was solved by consultation of third reviewer. RESULTS: Overall, 4039 records were examined related to malaria elimination that initially identified by our designated electronic databases search. Overall, 35 studies contained 14 experimental studies (40%) and 21 analytic observational studies (60%) met the inclusion criteria for this review. Studies used a wide variety of malaria elimination interventions. Types of interventions either elimination-focused interventions or general interventions on educational, prevention and treatment of malaria are included. This review pointed out the variety of challenges for eliminate malaria among low and high endemic countries. CONCLUSION: Malaria elimination is facilitated by strong health systems, determined leadership, appropriate incentivization, an effective surveillance system, and regional collaborations. We have identified areas for elimination-specific interventions deserve more attention in the conduct and reporting.

Effect of Different Diets on Lifetime of Brown-Banded Cockroaches, Supella longipalpa (Blattodea: Blattellidae).

BACKGROUND: The brown-banded cockroach, Supella longipalpa, is not as common as the German cockroach in Iran. This species seeks out areas that are very warm most of the time, and prefer warmer area than what German cockroaches prefer. There is relationship between development of instars and diet of cockroaches. The aim of this study was to determine the effect of different diets on biology, life cycle on nymphal stages of S. longipalpa in laboratory condition prior to investigate the insecticide resistance status of this species in residence area in Iran. METHODS: The cockroaches were reared in the insectary of School of Public Health, Tehran University of Medical Sciences, condition and the population divided in four equal groups. The effect of four different diets on life cycle of S. longipalpa was studied to determine the effect of them on the lifetime of each nymphal stage. RESULTS: The diets significantly affected on growth and development of immature life stages of S. longipalpa. Based on introduced diets to the cockroache populations, total immature life cycle was 54, 58, 60 and 66 d for diets 2,4,1, and 3 relatively. However, the overall lifetime of S. longipalpa in average was about 225 days. CONCLUSION: As far as urban pest control is concerned, the result of this study will facilitate any operational programs for control of S. longipalpa. Among the different tested diets, diets 2 and 4 with less duration would be recommended for rearing of S. longipalpa in laboratory condition.

Isolation and Purification of an Antibacterial Protein from Immune Induced Haemolymph of American Cockroach, Periplaneta americana.

BACKGROUND: Antimicrobial peptides play a role as effectors substances in the immunity of vertebrate and invertebrate hosts. In the current study, antimicrobial peptide was isolated from the haemolymph of the American cockroach, Periplaneta americana. METHODS: Micrococcus luteus as Gram-positive bacteria and Escherichia coli as Gram-negative bacteria were candidate for injection. Induction was done by injecting both bacteria into the abdominal cavity of two groups of cockroaches separately. The haemolymphs were collected 24 hours after post injection and initially tested against both bacteria. Subsequently, the immune induced haemolymph was purified by high performance liquid chromatography (HPLC) to separate the proteins responsible for the antibacterial activity. RESULTS: The non-induced haemolymph did not show any activity against both bacteria whereas induced haemolymph exhibited high activity against M. luteus but did less against E. coli. Two fractions showed antibacterial activity against M. luteus. Finally the molecular weight of the isolated antibacterial proteins were determined as 72 kDa and 62 kDa using SDS-PAGE. CONCLUSION: Induced haemolymph of American cockroaches has the ability to produce peptides to combat against Gram-positive bacteria when an immune challenge is mounted. Further work has to be done to sequence of the protein, which it would be advantageous.

Health Workers Adjustment for Elimination of Malaria in a Low Endemic Area.

BACKGROUND: Malaria elimination efforts face with substantial challenges and the role of health workers in address this challenge, particularly advocates and mobilizes communities. The aim of the study was to explore perceptions of health workers in relation to eliminating malaria in order to better understand the level their involvement in malaria elimination efforts. A qualitative approach was adopted based on key informant interviews with 26 health workers who working at community-level in malaria low endemic areas, southern Iran. METHODS: Data were collected through key informant interviews. Data were analyzed using thematic content analysis. RESULTS: Findings reveal that the majority of participants concerned with the imported malaria cases, without to address an effective solution to the issue. Health workers had positive perceptions on their basic knowledge and opinions in relation to their field work with emphases to integrate methods. Participants expressed willingness to contribute to malaria elimination effort. They also emphasized on continuous training, resource mobilization, and support. In addition, their perceptions on malaria elimination policy such as sustained financial investment to achieve elimination and integrated management of vector control were rather negative. CONCLUSIONS: A mechanism should be considered that allow the health workers to feedback positively on their quality of their practice to health providers.

Community Eagerness and Participation for Supporting Eliminate Malaria.

BACKGROUND: This study was undertaken to identify key elements for obtaining community interest and motivate them to support eliminate malaria in the places that malaria is no longer a perceived threat. METHODS: Sequential explanatory mixed methods research design used to provide more comprehensive evidence research. A questionnaire was developed after reviewing the literature relating to community participation for malaria elimination and used to collect data from three native people, Iranian migrant and foreign immigrant groups. The variables included access to educational facilities, services, social support, social supporters, trust, and supportive norms were assessed. We also employed an ethnographic design involving: observation participant (PO), Key informant interviews (KIIs), and focus group discussions (FGDs). RESULTS: The results revealed that predisposing, enabling and reinforcing factors had association with community involvement. Less than 50% of total respondents reported have access to educational sessions, most of them were unaware of available and affordable services. Differences views regarding to social support influence have been found among respondents. Although patterns of social interactions were almost similar in all residents, frequencies of interactions were varied. Three thematic areas emerged including perceptions on involvement, potential barriers, and focus areas for the training community members. CONCLUSION: These findings indicate needs for greater understanding about how to strengthen community involvement with emphasize for community capacity building. This research has benefit to national and regional efforts to increase community involvement as a source for effective actions on malaria elimination.

Species Composition and Seasonal Activities of Malaria Vectors in an Area at Reintroduction Prevention Stage, Khuzestan, South-Western Iran.

BACKGROUND: The most part of Iran become malaria-free region and fall in prevention of re-introduction stage. These regions however are struggling with imported of malaria cases where malaria vectors exist. Therefore, understanding the situation of mosquito vectors is crucial. This study was carried out to find out the present situation of malaria vectors and malaria transmission potential in a malaria-free area. METHODS: The study was conducted in a malaria free area, Izeh County, Khuzestan Province during 12 months in 2011-2012. Five villages, including 2 in highlands and 3 in plain area, were selected randomly. The mosquito sampling methods were conducted using spray sheet and hand catch collection methods from indoor/outdoors, window trap and larvae collections. RESULTS: In total, 3352 female Anopheles were captured, 1826 mosquito from highland and 1526 from plain areas. Five species, An. stephensi, An. fluviatilis s.l., An. dthali, An. superpictus and An. pulcherrimus were identified. The seasonal activities were started from April to March. The abdominal conditions of collected mosquitoes from indoor/outdoor places pointed to exophilic propensity of An. fluviatilis.l. s.l. and endophilic behaviour for rest of the vectors. The results of window trap also confirmed these behaviors. The larval habitats of four species were widely dispersed and included spring, margin of rivers, irrigation channels, stagnant water and rice filed. CONCLUSION: Understanding the present situation of malaria vectors in free-malaria area is crucial particularly where is struggling with imported cases. The results of present study can be expanded to other area of northern Khuzestan for malaria vector control planning in reintroduction prevention stage.

Molecular characterization of calreticulin from Anopheles stephensi midgut cells and functional assay of the recombinant calreticulin with Plasmodium berghei ookinetes.

Transmission blocking vaccines (TBVs) that target the antigens on the midgut epithelium of Anopheles mosquitoes are among the promising tools for the elimination of the malaria parasite. Characterization and analysis of effective antigens is the first step to design TBVs. Calreticulin (CRT), a lectin-like protein, from Anopheles albimanus midgut, has shown antigenic features, suggesting a promising and novel TBV target. CRT is a highly conserved protein with similar features in vertebrates and invertebrates including anopheline. We cloned the full-length crt gene from malaria vector, Anopheles stephensi (AsCrt) and explored the interaction of recombinant AsCrt protein, expressed in a prokaryotic system (pGEX-6p-1), with surface proteins of Plasmodium berghei ookinetes by immunofluorescence assay. The cellular localization of AsCrt was determined using the baculovirus expression system. Sequence analysis of the whole cDNA of AsCrt revealed that AsCrt contains an ORF of 1221 bp. The amino acid sequence of AsCrt protein obtained in this study showed 64% homology with similar protein in human. The AsCrt shares the most common features of CRTs from other species. This gene encodes a 406 amino-acid protein with a molecular mass of 46 kDa, which contains a predicted 16 amino-acid signal peptides, conserved cysteine residues, a proline-rich region, and highly acidic C-terminal domain with endoplasmic reticulum retrieval sequence HDEL. The production of GST-AsCrt recombinant protein was confirmed by Western blot analysis using an antibody against the GST protein. The FITC-labeled GST-AsCrt exhibited a significant interaction with P. berghei ookinete surface proteins. Purified recombinant GST-AsCrt, labeled with FITC, displayed specific binding to the surface of P. berghei ookinetes in comparison with control. Moreover, the expression of AsCrt in baculovirus expression system indicated that AsCrt was localized on the surface of Sf9 cells. Our results suggest that AsCrt could be utilized as a potential target for future studies in TBV area for malaria control.