Development of Plant-Based Larvicide and Herbal Mosquito Repellent Fast Card with Reference to Identification of the Functional Bioactive Compounds Effective Against Culex Mosquito
Abstract:Warm and humid climate creates ideal conditions for mosquito breeding. The ability of these vectors to spread a number of diseases to humans causes millions of deaths every year. Indiscriminate use of synthetic insecticides leads to the development of resistance in vector mosquitoes and along with this, these pesticides cause biological magni cation of toxic components and affects adversely the non target organisms including human being. Commercially available, chemically manufactured mosquito repellent fast c… Show more
“…Several species of mosquitoes have a strong invasive capacity [ 1 ]. At present, mosquito-borne diseases are a great global public health challenge [ 2 , 3 ]. Due to the complexity of mosquito-borne pathogens, eradication of vectors has become one of the more effective strategies for controlling infectious diseases.…”
Background
Mosquitoes are an important vector of viral transmission, and due to the complexity of the pathogens they transmit, vector control may be the most effective strategy to control mosquito-borne diseases. Chitin is required for insect growth and development and is absent in higher animals and plants, so regulating the chitin synthesis pathway can serve as a potentially effective means to control vector insects. Most of the current research on the chitin synthase (CHS) gene is focused on chitin synthase-1 (CHS-1), while relatively little is known about chitin synthase-2 (CHS-2).
Results
The CHS-2 gene of Ae. albopictus is highly conserved and closely related to that of Aedes aegypti. The expression of CHS-2 in the third-instar larvae and pupal stage of Ae. albopictus was relatively high, and CHS-2 expression in adult mosquitoes reached the highest value 24 h after blood-feeding. In the fourth-instar larvae of Ae. albopictus, CHS-2 expression was significantly higher in the midgut than in the epidermis. Silencing CHS-2 in Ae. albopictus larvae had no effect on larval survival and emergence. The expression of four genes related to chitin synthesis enzymes was significantly upregulated, the expression level of three genes was unchanged, and only the expression level of GFAT was significantly downregulated. The expression of chitin metabolism-related genes was also upregulated after silencing. The level of chitin in the midgut of Ae. albopictus larvae was significantly decreased, while the chitinase activity was unchanged. The epithelium of the midgut showed vacuolization, cell invagination and partial cell rupture, and the structure of the peritrophic membrane was destroyed or even absent.
Methods
The expression of CHS-2 in different developmental stages and tissues of Aedes albopictus was detected by real-time fluorescence quantitative PCR (qPCR). After silencing CHS-2 of the fourth-instar larvae of Ae. albopictus by RNA interference (RNAi), the expression levels of genes related to chitin metabolism, chitin content and chitinase
activity in the larvae were detected. The structure of peritrophic membrane in the midgut of the fourth-instar larvae
after silencing was observed by paraffin section and hematoxylin–eosin (HE) staining.
Conclusion
CHS-2 can affect midgut chitin synthesis and breakdown by regulating chitin metabolic pathway-related genes and is involved in the formation of the midgut peritrophic membrane in Ae. albopictus, playing an important role in growth and development. It may be a potential target for enhancing other control methods.
Graphical Abstract
“…Several species of mosquitoes have a strong invasive capacity [ 1 ]. At present, mosquito-borne diseases are a great global public health challenge [ 2 , 3 ]. Due to the complexity of mosquito-borne pathogens, eradication of vectors has become one of the more effective strategies for controlling infectious diseases.…”
Background
Mosquitoes are an important vector of viral transmission, and due to the complexity of the pathogens they transmit, vector control may be the most effective strategy to control mosquito-borne diseases. Chitin is required for insect growth and development and is absent in higher animals and plants, so regulating the chitin synthesis pathway can serve as a potentially effective means to control vector insects. Most of the current research on the chitin synthase (CHS) gene is focused on chitin synthase-1 (CHS-1), while relatively little is known about chitin synthase-2 (CHS-2).
Results
The CHS-2 gene of Ae. albopictus is highly conserved and closely related to that of Aedes aegypti. The expression of CHS-2 in the third-instar larvae and pupal stage of Ae. albopictus was relatively high, and CHS-2 expression in adult mosquitoes reached the highest value 24 h after blood-feeding. In the fourth-instar larvae of Ae. albopictus, CHS-2 expression was significantly higher in the midgut than in the epidermis. Silencing CHS-2 in Ae. albopictus larvae had no effect on larval survival and emergence. The expression of four genes related to chitin synthesis enzymes was significantly upregulated, the expression level of three genes was unchanged, and only the expression level of GFAT was significantly downregulated. The expression of chitin metabolism-related genes was also upregulated after silencing. The level of chitin in the midgut of Ae. albopictus larvae was significantly decreased, while the chitinase activity was unchanged. The epithelium of the midgut showed vacuolization, cell invagination and partial cell rupture, and the structure of the peritrophic membrane was destroyed or even absent.
Methods
The expression of CHS-2 in different developmental stages and tissues of Aedes albopictus was detected by real-time fluorescence quantitative PCR (qPCR). After silencing CHS-2 of the fourth-instar larvae of Ae. albopictus by RNA interference (RNAi), the expression levels of genes related to chitin metabolism, chitin content and chitinase
activity in the larvae were detected. The structure of peritrophic membrane in the midgut of the fourth-instar larvae
after silencing was observed by paraffin section and hematoxylin–eosin (HE) staining.
Conclusion
CHS-2 can affect midgut chitin synthesis and breakdown by regulating chitin metabolic pathway-related genes and is involved in the formation of the midgut peritrophic membrane in Ae. albopictus, playing an important role in growth and development. It may be a potential target for enhancing other control methods.
Graphical Abstract
“…Tropical and subtropical regions are particularly vulnerable because the warm and humid climate creates ideal conditions for mosquito breeding [1][2][3][4][5]. For example, 120 million individuals have lymphatic filariasis transmitted by Culex spiculosus, and about 80% of them reside in tropical nations including Bangladesh, India, Indonesia, Malaysia, Nigeria, and the Philippines [6]. According to the World Health Organization, the parasitic infection of malaria transmitted by Anopheline mosquitoes causes an estimated 219 million cases worldwide and gives rise to more than 400,000 deaths annually.…”
Two series of novel sophoridine derivatives were designed, synthesized, and evaluated for their anti-mosquito activity. SOP-2g, SOP-2q, and SOP-2r exhibited potential larvicidal activity against Aedes albopictus larva with LC50 values of 330.98, 430.53, and 411.09 ppm, respectively. Analysis of structure–activity relationships indicated that the oxime ester group was beneficial for improving the larvicidal biological activity, whereas the long-chain aliphatic group and fused-ring group were introduced. Furthermore, the larvicidal mechanism was also investigated based on the inhibition assay of acetylcholinesterase (AChE) and the morphological observation of dead larva treated with derivatives. Results indicated that the AChE inhibitory activity of the preferred three derivatives were 63.16%, 46.67%, and 35.11%, respectively, at 250 ppm concentration. Additionally, morphological evidence demonstrated that SOP-2q and SOP-2r induced changes in the larva’s intestinal cavity, caudal gill, and tail, thereby displaying larvicidal action against Ae. albopictus together with AChE inhibition. Therefore, this study implied that sophoridine and its novel derivatives could be used to control the population of mosquito larva, which may also be effective alkaloids to reduce the mosquito population density.
“…The developed card is found to cause less gas emission than that of commercial one which is better for the human health. Also, it is revealed by the High Performance Liquid Chromatography (HPLC) analysis that propionic acid in Duranta is responsible for the mosquitocidal properties [11].…”
There are many life-threatening insects in nature and the most dangerous ones are mosquitoes. People have taken herbal precautions against them since ancient times. Although several chemical and synthetic insect repellents exist in the market, the plant-based protection methods have never been given up such that there is a trend towards producing and commercializing plant-based insect repellents in recent years. In the light of ethnobotanical studies, new insect repellent plants and formulations are being explored, developed and tested. In literature, there are a lot of research on plant-based repellents some of which are reviewed in this article. Information about the most frequently used plant-based repellents and related studies are presented. Different application methods of the plant-based repellents on the textiles are explained with their a few examples in literature. Also, common test methods used for determining the effectiveness of insect repellents and repellent treated textiles are stated and the importance of the standardization of these test methods is emphasized. It is understood that usage of plant-based insect repellents in practice can be beneficial and useful if stayed within their safe concentration limits.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.