Chemical Composition and Repellency Effect of Ferulago Angulate Plant Against Malaria Vector, Anopheles Stephensi

Background Malaria is a health problem globally. There are several vector control measures. Using repellent to protect human from biting of vectors is one of the best ways. The aim of this study is to evaluate the ecacy of plant against biting of malaria vector, An.stephensi under laboratory conditions. Methods Chemical constituents of Ferulago angulate plant were determined by using gas chromatography-mass spectrometry. Ecacy and the protection time of plant were evaluated on Anopheles stephensi . Results


Introduction
Malaria is the main vector borne diseases worldwide. According to the recent record of World Health Organization, 228 million cases have been reported in 2018 mainly in in African region [1]. According to the report of Ministry of Health of Iran, less than 89 locally-transmitted cases in 2017 have been reported.
The aim of country is to eliminate the disease by 2025 [2]. Malaria continues to be a main vector-borne public health problem in Iran. Using insecticide caused resistant in the vectors. There are several reports on resistant status of malaria vectors including An.stephensi [3][4][5] An.maculipenis [6]) , An.culicifacies ( [7][8][9] ) An. [10]). At the present, biopesticides are in the spotlight due to in icting of chemical pesticide hazards to humans, livestock, environment as well as the occurrence of the inducing resistance to different groups of insecticides among human and animal disease vectors. A lot of attention is being paid to natural products in vector control as they are environmentally safe, degradable and targetspeci c. Recent studies have demonstrated that use of repellents is one of the effective ways to control [11]. There are 9 species of plant in Iran including: F. contracta, F. macrocarpa , F. phialocarpa , F. stellate, F. angulate , F. Bernardi , F. carduchorum, F. subvelutina, F. tri da F. angulate is known as Chavil in the country .It is a perennial endemic aromatic herb from the nine Ferulago species growing particularly in Iran [12][13]. Its leaves have been traditionally used as antiseptic, pain reliever, in digestive disorders, to treat intestinal worms, snake bites, hemorrhoids, chronic ulcers, and ailments of the spleen [14] Furthermore, in Western Iran, this plant has been consumed as spice, and used as air fresher, decay preventer and avoring oil [15]. In a study to assess the in uence of geographical factors on essential oil composition, along with antiradical potential and phytochemical contents of Ferulago angulata (Apiaceae) extracts . Thirty-nine compounds were identi ed from the Eos of nine populations. α-Pinene was the predominant component. It can be considered as a perspective raw material in food and phytopharmaceutical industries [16] . Antioxidant and antibacterial activities of the essential oils of F. angulata collected from different natural habitats in the alpine regions of southwestern Iran showed positive results.. The essential oil of F. angulate could be serving as a potential source for use in the food, cosmetic and pharmaceutical industries [17]. Cytotoxic activities of Ferulago angulata extract on human leukemia and lymphoma cells by induction of apoptosis were determined [18] . This study was conducted to measure the chemical contents and to evaluate the repellent properties of plant essential oil against An. stephensi under laboratory condition on animal model. University. (Fig.2)

2.3.Mosquitoes rearing
Susceptible strain of An. stephensi were reared and maintained at 28±2 °C and 65±5% relative humidity (RH) under a 16:8 (L: D) photoperiod. Under insectary situation .Guinea pigs is used as blood feeding female mosquitoes for maturing the eggs.

2.4.Repellency test
Females of An.stephensi were used for the repellency tests. 12 hour before starting the experiments, the sucrose solution was picked up from the cage . Various repellency tests including protection time, failure time, effective dose and killing effects of EOs were carried out according ASTM E951-9 against 5-8 old female An.stephensi.

2.5.Extraction of essential oil of plant
All the extraction was carried out at Faculty of Pharmacology, Tehran University of Medical Sciences. Essential oils (EOs) of native medicinal plant of Ferulago angulate, were hydrodistilled in a Clevenger-type apparatus for 4-6 h and dried over anhyrdrous sodium sulfate. The EOs were stored in the dark sealed vials at 4 °C until starting the repellency tests maximum after 2 days past of EO preparation.

Plant essential oils analysis
Chemical composition of plant was analyzed using an Agilent 7890-5975 gas chromatography mass spectrometer. With a HP-5MS (5% Phenyl Methyl Silox) capillary column (30m×0.25mm, lm thickness 0.25μm), split ratio, 1: 1, and using a ame ionization detector. The GC was programmed at 50 °C for 0.5 min and then increased at 5 °C/min to 280 °C, and nally held with an isothermal for 3min. The injector temperature was 280 °C. The ow rate of the carrier gas was 1ml/min. The identi cation of compounds was performed by comparing their retention times and mass spectra with mass spectra from Wiley library [19] 2.7.Plants essential oils analysis Chemical composition of plant was analyzed using an Agilent 7890-5975 gas chromatography mass spectrometer. With a HP-5MS (5% Phenyl Methyl Silox) capillary column (30m×0.25mm, lm thickness 0.25μm), split ratio, 1: 1, and using a ame ionization detector. The GC was programmed at 50 °C for 0.5 min and then increased at 5 °C/min to 280 °C, and nally held with an isothermal for 3min. The injector temperature was 280 °C. The ow rate of the carrier gas was 1ml/min. The identi cation of compounds was performed by comparing their retention times and mass spectra with mass spectra from Wiley library.